University of Tennessee, Knoxville

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Masters Theses Graduate School

6-1937

Quality control of market silk

William Coley Patton

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Recommended Citation
Patton, William Coley, "Quality control of market silk. " Master's Thesis, University of Tennessee, 1937.
https://trace.tennessee.edu/utk_gradthes/9153

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To the Graduate Council:

I am submitting herewith a thesis written by William Coley Patton entitled "Quality control of
market silk." I have examined the final electronic copy of this thesis for form and content and
recommend that it be accepted in partial fulfillment of the requirements for the degree of
Master of Science, with a major in Animal Husbandry.

C. E. Wylie, Major Professor

We have read this thesis and recommend its acceptance:

T. B. Harrison, Paul W. Allen

Accepted for the Council:

Carolyn R. Hodges

Vice Provost and Dean of the Graduate School

(Original signatures are on file with official student records.)

 my 18, 1937

To the Coimaittee on G-raduate Study:

I submit herewith a thesis written by Mr. W. 0. Patton and entitled

"Quality Control of Market Milk", and recommend that it be accepted for

nine quarter hours credit in partial fulfillment of the requirements for

the degree of Master of Science in Agriculture, with a major in Dairying*

jor Professor

At the request of the

Committee on Graduate Study,
we have read this thesis,
and recorafiiended its acceptance.

Cc) CdLtju^
2£BZZZZZ
Accepted by the Committee

Dean

A."
 QmLlTY OOHTROL OJf MARKBT MILK

A SHSSIS

9al3raitt«d to the Graduato (knuitteo

of
Tko UhlTorslty of TaonoBsee
in
Partial TUlfilliaont of the HequireBMnjl^.
for the degree of
Master of Scienoe
in
Agrioulture

W. 0. PATTON

June 1937

•". A

Jim
 PBSFAOS

Tho laaterlal in. this thaals waa aacured from a study of aome of tha

faetora affecting the quality of loarket milk* The problem was planned so

that the information could be of pr^tioal value to dairymen, milk

dealers, health officials, and oonsumere*

A survey of the literature on this aubjeot and related subjects is

given in Part I* Many helpful suggestions on methods of procedure and

presentation of results were found in this survey* Due credit has been

given those earlier investigators whose work was included. The data in

Part II were secured from an investigation xoade by the writer during two

years of graduate study at the Dhiversity of Tennessee and with the

cooperation of the !3aoxville Bureau of Health*

The writer wishes to sKpress his sincere appreciation for the

assistance given by tiie mambers of the IMiversity Dairy Staff and

students. He is espeoially indebted to Professors 0* S. %lie and

Thos. B* Harrison of the Dhiversity of Tennessee Dairy Department for

^ their able guidance during the investigation and writing of this thesis*

W* 0* Patton
V)

K)

ii 14051''^^
 TABLI5 OF CONTENTS

Pages

BTTHODOCTICaT «. *. •. *. .. ». • «. •. Til

PAST I, SORVir OP LITERATDHE;

Flafors of Market Milk • • • • • « •.. «• • • • • • * « 1

Feed and Weed Flavors * • • • • • • • * «• * • • • « 8
Abnormal Conditions of the Cow • • • • • # * • • • • « 9
Handling of mik 10
Unolean Flavors ••• •• ••• • ••« • • • • « 10
Pasteorination • • • • • • • « * •••• •• • • 18
Oxidized Flavors •• • * ••• • •• • • • • • • 18

Creaming of ^larket Milk •.• • • « « • 14

Formation of Cream Layer • * • • • • • i « 18
Cooling end Holding Tan^sratures •• • • * • • • • • • 16
Pasteurization TcBaperatures « • • • • • • • • ••• • 18
Holding Time ... ...• SO
Agitation and Punning 81
Homogenization 83

Baoteriology of Market Milk 28

Methods of Analysis . • • « • • • • • « • • • • • • • 86
Merosoopio Ivtethods of Counting • « • 87
Agar Plate Counting Lfethod 27
Methylene Blue Heduotase Test • •.• • • • • • • 89
Effect of Cooling . •.. « • • • • • • • • • • • » • 87
Effect of Pasteurization 89
Sterilization of Utensils and Equipment • « • • •.• 40
Storage Temperatures * 87
Aoidity SB
Keeping Quality ••••••.• 89

Summary of Survey of Literature 60

part II. ORIGINAL INVESTIOATIONl

Objects of Investigation 61
Plan of Investigation 62
Methods of Proeedure 63

ill
 TABLE OP OOOTffllTS (Ooixtlau«d)

fltftors of Macrktt mik 65

Mothods of Seorloi; * • • • • • • • • • • • • • • • • « 66
Inflaenee of deration • • • • . * • • • • • • • « • •• 66
Pastourization Tonparatures * • • • • • • • • •• • • • 74

Oreaming of Merkat Milk 78

liethoda of Measuring • • • • • • • • * • • •• • • • • 78
In the Bottle • • • • * •• • • • • • • • • • « 79
Oz^duated Cylinders • • • • • •• • • **•••• 79
Pasteurization Teo^eraturea • « •• • * • • ••«•• * 00
Influenoe of Storage • • • • • • •«• • # •••• « • 8£

Baateriology of I^arkat Milk 84

Methods of Analysis • « . « • •••• • • • • • • •• 84
Agar Plate Counting • • • •• • • •• • • *• * • 86
Methylene Blue Reduotase • * • • • « • • • • •« 88
Influenoe of Cooling •. • • • • • • • • • • • • • • • 96
lilffioienoy of Pasteurization • • • • • • • • • • • •• 99
Sterilization of Cans 101

106

Blbliogragphy #• • • • • • • •• • * . loe

if
 TABLES Ur TEXT

Tabltt

I* SuBBary of Initial Trials (koqmring Sterilization of Cans

by Steam and by Hot Mr 60

II» Geriaicidal Potency of Various Types of Ohlorins

Preparations Against B. Coll •««« .• • * • • • • 54

III« Plata Counts on B* Coll Inooulnra After Treatments ^?ith

Various Chlorine Cermicidea Containing Approximately
SCO p*p.ia. of Available Chlorine in Cold Solution « • 54

IV* Plate Count on J^ilk Can Rinse Inoculum After TSxposure to

Germicidal Solutions Containing 800 p«p*m. of
Available Chlorine in Cold Solution • 55

V. Growth of Bacteria in '/Ilk Shea the Milk is Held at SO® F»

and 68° F... * *, 67

VI* Scores for CSood Flavored Milk Cooled by Different Methods

Average of 3 Trials • • •.. * * 70

VII* Scores for Poor Flavored Milk Cooled by Different litethods

Average of 6 Trials 71

VIII. Scores for Medium Flavored f<fllk Cooled by Different Methods 79

IX* Flavor Scores for Mlk Sonnies Pasteurized at Different

Tos^eratures • •. « • • * • • • •• * .• • • * • • 76

X* Creaming of Ifllk Pasteurized at Dlfferwait Tei^aratures • • 8S

XI* Creaming of Milk Pasteurized at Different Temperatures • • 83

XII. Correlation Between Bacteria Counts in Thousands par ce*

and Reduction Time in Hours •• 62

XIII* Influence of Cooling on Bacterial Growth •*« • • • • • M

XIV* Effioienty of Pasteurization • • * • • * ... • • • * • • 96

XV* Number of Bacteria per ec* of Rinse Water Before and

After Sterilization of Cans bj- steam • • . • * .• • 102

XVI* Number of Bacteria per oc* of Rinse Water Before and

After Sterilization of Cans by Dry Heat • • * • * • 103

XVII* Number of Bacteria per co. of 500 oc. Sterile Rinse Water
Before and After Sterilizing with Chlorine
Solutions Containing 50 Parts per Mllion of
Available Chlorine • . • .. * . * • • •* 104
 FIOURSS

1* Student Score Oard 67

2* Surfaee Tul>ttlar Cooler » • • «* • ••• • • • « • 69

3. VSXlln Preheator 76

4* "Oie Cherry Burrell Spray-type Peeteurlaer 81

5. riatorial and Kquipmwit Used '' or jMaJdng The Feductase Teat • • » 89

6* Correlation Between Bacteria Counts in Thousands per oc. and

Peduotion Tine in Hours • • • • • • * • • « • • • • • • • • 94

7» Correlation Between Bacteria Counts in Thousands per oa« and

Reduction Tine in Hours 85

•y .:, -

.'^ v'J; .. J:

rf''.

'^1 ..
' ' ' . ^ . .-'i. ' "*

«■

Ti
 II'^TRODTTCTIOH

Dairy products oonstitut© appixuxiiaately one fifth of the diet of the

sTorage family in the United States* They appear daily on alnKtst every

family table. This requires over ten billion pounds of milk eaoh year.

Aln»>st one half of this moount is eonsumad as fluid or market milk*

Unusual care must be ezeirelaed throughout the production and handling of

this milk in order for it to be of high quality when it reaches the

oonsumer.

Quality in milk ia a general tena beoauso it is governed by many

factors, and yet, it may ba applied as a specifie torn to mean a very

highly standardiaad product* In this investigation, quality refers to

the relative merits to be placed upon market milk as governed by the

factors used in its production and handling. The factors included in

this study are associated with the flavor, creaming, and bacteriology of

milk* They determine to a great extent the standard of any market milk

supply, ^hroper control matlMds are usually associated with high quality

milk* Improper methods ara usually associated with a corresponding

sacrifice in quality*

Host of the available information on the quality of market milk has

been secured throu^-h studies of the various factors affecting separate

indioations of quality. In problems which include only one point, a

detailed series of trials can be made* In this problem, however, fectora

affecting three of tha major indications pertaining to the quality of

oommeroial market milk were studied* For tha most part these studies were

made for the purpose of obtaining some valuable iriformation from souroea

vii
oomparatiTa to a eonn^rclal e^ale, rather than from a laboratory aoale,

whleh is the eoiamon procedure* In Tiew of the fact that toethode of

control were very difficult for experiments which were conducted on a -

coimneroial scale, several trials waare made for each series and the

averages taken for reporting the results*

Several different viewpoints may be considered In making a study of

the quality control of market milk* The producer and the distributor are

interested in the economic control of the factors whleh affect the quality

of their milk. There are those features which are primarily of intoarest

to the oonsUiTtar, and those features required by law or the legal health

authorities. The eonsuraer is prinarlly interested in the flavor of hie

milk, and the cream layer appearing on it* He depends on the health

authorities for the xuitritioua and sanitary qualities* The health

officials are governed largely by the city ordinances and the state dairy

laws which define milk as to the minimum requirements of fat, serum solids,

and the maximum bacteria counts* Sinca palatability does not permit legal

definition, the cons'naer must employ his senses of taste and amell for

deteimiinlng this quality in milk*

vili
 PiHT I. SURVEY OF UTERATIJRE

Flavors of Market Milt.

Flavor Is tho iMst lta|)ortant quality in narknt nilk» The oonsunlng

publio expects and domands that this food be palatable* r^llk which has

an unpleasant flavor and aroioa will be used only in lisiited quantities;

whereas* it should be used in increasing amounts because of its hi^ food

value* When it is off-flavor* children oftentimes refuse to drink it*

No other food can take the place of milk in the diet of growing children.

Uany adults are fond of milk and drink it because it is refreshing

and nutritious. The producer and the distributor should exercise the

utmost care in the production and processing of market milk ao that there

can be no developments of off-flavors in the milk.

To most people good milk has a pleasant taste which is probably due

to a combination of various ingredients found in milk; namely, fat, milk

sugar* casein* albumin, and the minerals.

Normal milk is sweet because about of it is milk sugar. When it

is kept at worm tenqjeratures, it turns sour because of the activity of

certain bacteria* Milk readily absorbs odors that surround It and thereby

may develop an off-flavor* The food the cow eats may greatly affeot the

flavor of the milk.

Cows Vary somewhat in the quality of the milk they produce* A cow

far alon-r in her lactation period is apt to give milk with a somewhat salty

flavor* Individual differences* however* are overcmne to a oertain extent

when the milk from a number of cows is mixed.

 Anoag the teimui used to desorlbe the off-flavors lu milk, feed le

probably the most eooncm* Neoct to feed may come bitter, garllo or onloa,

and unolean* There are at present sixtemi flavor oritielams on the score

eard reooonended by the tMted states Department of Agriculture*

Feed and Weed Flavors*

Milk plants and eonsamera are objecting to milk with imdesirable

flavors and odors. This Is a loss to the producer and the industry as a

whole* The producer at one time mis primarily interested in prev^xting

the milk from souring before it reached the plant, giving very little

attention to the objectionable flavors caused by toproper feeding* The

producer of today is face l with the problem of minimizing bacteria counts

and preventing objectionable flavors caused by iii^>roper feeding* This may

best be eooomplished by a tborou^ study of the differwa.t feeds which

cause undesirable flavors in milk, and the proper time for feeding to pre-
53
vent these off-flavors*
22
Gan^le and Sally, after a systematio study of the effect of feeding

silage upon the flavor and odor of milk, reported a wide variation between

individual cows under normal condition which received the sane feeds*

When silage was fed one hour before milking, it was absorbed so quickly

that it was noticeable in Idxe milk from most of the animals in the ezpaoN*

Iments* In order to avoid undesirable flavors, it was reoonsnsnded that

silage be fed soon after milking, and that not more than 15 to 25 pounds

of com or legume silage be fed after eaoh milking* These investigators

also found that prong)t aeration of warm milk roaoved slight silage flavors

and reduoed the more pronounced ones*

 Aoong the other inTdstigators mho confirmed the couoluslonB of Gemhle
3
and Kelly, as a whole or in part, was C» J« Baboook, who niade an exten-

slTe study of feed flawore in milk during the period of 1923-19S5 InolusiTe*

These inreetigatore reported on the use of turnips, green alfalfa, eahbage,

potatoes, garlio, green rye, and green peas* The above feeds wex^ found to

produce objectionable flavors and odors if fed within one hour before

milking* Where fed in moderate quantities, four to five hours before

milking, only eli^t off-flavors and odors resulted from most of the above

feeds*

50
Boadhouee and Henderson found that garlio could be detected in

sauries of milk takan one minute after feeding and through the seventh hour

following* de little as 10 pounds of green alfalfa or 5 pounds of alfalfa

hay, fed two hours before milldng, oaused a distinot feed flavor and odor

which was undesirable in market milk* Larger quantities caused a more

pronounced off-flavor. When oat hay was substituted for about one half of

the amoxint of alfalfa, the flavor was less pronounced. Ibderate quo&tities

of com silage, 5 to 12 pounds, fed one to two hours before milking,

produoed an undesirable flavor; and when cows were fed 10 pounds, one hour

before milking, a deoided f<Md flavor end odor were present, whioh eould

be detected by the oonsomer of the milk* Cows that were fed Imy in whioh

a muety odor was observed gave milk with a muaty flavor and odtur, which were

not present idien this feed was discontinued* When the Juice extracted from

SS poimda of green alfalfa was given to oowa by force feeding just before

milking, the following results were obtained: during the first twenty

minutes, no feed flavor was present; a sli^t feed flavor at twenty to

twenty-five minutes; and a noticeable feed flavor and odor at the end of
thirty minutes* The most distinot feed flaror was present troa forty-fiti
to sixty minutes after drsuohing.
51
Boadhouse, Kegau, and found that there was a marloed differenoe,

as to the off-flavors present in indiwidual oow*s milk, vdien fed feeds of

the sane kind and naount* A pronoimoed flavor was found in the milk when

alfalfa hay and ^reen alfalfa were fed to a group of eows one to four hours

before milking, and a less pronouneed flavor was found when fed four hours

or more before milking*

Most of Ihe soluble feeds give the intensity of their flavor begin

ning within thirty minutes after feeding and reaohlng a maxiinm at the

two or three-hoia: period* Practically all of this flatvor has gone after the

fourth hour* Ihider this division may be found pasture grasses and most of

the hays and com silage, all of which impart distinot feed flavors to milk*

These flavors are usually not objectionable to consumere, but in some oases

the milk is rojootea* Hay and silaga should, whore conditions will warrant

it, be fed after milking* These flavors may also be reduced by immediate
S3
cooling at the barn over surfaeo ooolers and by pasteurization of the milk*

Suoeulent feed flavors are more eaasoonly fotmd in milk than l^ose

flavors caused by conoentratee or hay* Silage made from com, alfalfa, soy

beans, cabbage, ttimipe, n^e, and kale materially affect the flavor of

milk; rye, oowpeas, potatoes, dried beet pulp, and carrots affect the flavor

only sli^tly; and com, oats, peas, pmBS>kins, and sugar beets have y&sy

little effect on the flavor*

Where proper feeding methods are employed, the intensity of the flavor
 5

ot moat feeds will not be notioed by the sTsrage eonsaoiT* ^weyey, ehen

very strong feeds are eonennsd Is wmsaal lare^ (xuantities, oabbage for

exainple, the flawor of the fflllk will be affeeted In soiss oases for twelve
53
hours after feeding.

Feeding the oows at thoes when least objeotionable flavors will result

very often oonfliots with other ohores around the dairy. This gives rise

to 8<»se of the improper feeding practices which are conanonly followed.

50
Roadhouse and Henderson found that very few of the conoentrate feeds

ooimBonly fed to dairy oows produced objeotionable flavors* They report a

pleasing rather than an objeotionable flavor in most instanoes. With idieat

bran fed in qtuintities of from 4|- to 7 pounds one hour before milkingt they

reported a pleasing flavor. There was more flavor pj>odaoed in the milk

sAien the bran was fed than in the control 8aiiQ>lee where no bran was fed

idien other feed had been withheld for five hours previous to milking. The

flavori however, was an inprovement over the control sasples. Where

pounds of dried beet pulp were fed one hour before milking, there was

apparently no change in flavor, but when this amount was increased to Sj^
pounds one hour before milking, a slii^t off*flavor was present, thou£^

not objectionable. Seven pounds of dried beet pulp produced milk with a

slight off-flavor. It was not as desirabls as the control whan fad both

one and two hours before milking, but this flavor would not ordinarily be

noticeable to the oonsumer. Besides, 7 pounds is mors than most feeders

reoomoesd.

Where a grain mixture oonsistlng of 100 parts barley, 50 parts wheat

 6

braa» SO parts ooeonat insal, 2 parts salt, and 3 parts boas moal vas tsd

In 8-pomid quantities two hours before milldng, a flavor was produced which

was a little less pleasing than the control ssi^les* ?rith 7^ pounds of

this mixture fed two hours before milldLng, the off-flavor was more

pronomoed. When 1 pound of ootton seed meal was added to the preceding

ration, all samples gave a satisfactory flavor but loft an unpleasant

50
after-flavor*

Besides the oconmn feeds for cows, there are many weeds which produce

xmdesirable flavors and odors in milk* Wxva. such weeds are present on

pastures, the cows should be kept off, where conditions will permit, until

the weeds are eradicated* Sfhen the cows are permitted to graze irtxere these

weeds are present, they should be removed several hours before milking*

Oarlic flavof may be ecHaswhat reduced If cows are removed from seven to ten
3
hours before milking, while bitter weeds may take a longer time*

One of the most common objectionable flavors in milk, cream, and

butter is that caused by garlic or wild onion* When oows are first turned

on pasture in the spring, great care should be taken to prevent their

eating onions* In many oases it causes great losses to the producer by hie

milk being returned* Whmx the flavor has entered the milk, it is very
4
difficult to neutralize or ranove* Babcock made a very extensive stuf^ of

this to determine: (1) in what way the flavor entered the zoilk, (2) the

length of time required after consumption for the flavor and odor to be

noticeable, and (3) the length of time after oonsusption that the flavor

would remain in the milk* He concluded that garlic flavor and odor wexre

present in the milk one minute after the feed was consumed* The intensity
of this flaTor InoxNsasod with the time to twi minutoa* A very strong

garlio flavor was present where ooi??s wore ailowod to eat one pound of garllo

four hours before milking* The intensity of the flavor was Inveansely

proportional to the time interval between oonsomption of the feed and

milktog* Allowing the oows to inhale garlic odors aleo produoed unde8i3>«

able flavors* ^he:re blood teets were takm tvoa. cows which had been fed

garlio thirty minutes beforehand, positive reealte were reported. Stronger

odors were found where fortyfive minutes had elapsed*

MaoDonald and Crawford found that the onion flavor in milk was

largely associated with the fat* This produces ths greatest injury to

oream - the most valuable part of the milk* Boiling, steaming, or blowing

air through the milk will take out part of this flavor, but these methods

injure the milk* The publio health servioe forbids the addition of any**

thing to milk > even clean water* This makss the uae of ohsmicals out of

the question*

Mnoral oil with suoh a iB>oolfio gravity to insure a good separation

is suitable for removing onion or garlio flavor from milk. It is an exsel-

lent solvent for the material which is responsible for the onion flavor in

milk* This oil esparates from the milk and leaves no foreign material la

the milk*

Their reooHHtmded proosdure is vdry simple* All that la neesMary

is to diluts the off*flavorod milk with a good gxrade of mineral oil, four

parts milk to oxui part oil, mix the two together well, allow the mineral
oil to separate, and remove it from the milk* Ono troatment will umially
 8

reaato-ftt a 8ll(^t onion flavor, wMlo a sMOnA traataoant la raaoiaiiaiidad for

milk containing a stroog onion flaTor. This double treatment process

xisoessitates the use of nmr oil for the second trea'teisnt if best results

are desired* The mineral oils may be washed and used sewMral times, thus
35
reducing the cost*

MaoDonald and Crawford found that the bitter flavor oaused ly the

eomaon dog fennel was associated with the milk serum and not fat, as

was the case with onions* Tbis makes the problem ewen more eonylex* It

oan be washed from oream by using skim milk whleh is free from the bittw

flavor, but this would not be reeommended for removing it frcm milk,

because ^is would require separation* Talbut says that a combination

of mowing, fertilizing, and careful graning will help to check the growth

of bitt«r weeds*

Wylle^' of Tennessee conducted an InveBtigation to determine the

practical value of the i''teeT)onald procees for the removal of onion flavor

from milk and to determine the relation of the process to ocnamereial

possibilities. Briefly, the procedure was the addition of 5 pints of

mineral oil to 7 gallons of milk and the mixture heated to 115° F. in a

40-gallon pasteurizer vat. Ths mixture waa ailoft^d to set undisturbed for

three minutes for the oil to rise. The milk was then drawn throuid^ a

"cotton pad Btraincr" and exaained for onion flavor* "Tharo was little if

any evidence of onion flavor or odor in the milk." The milk after a second

treala&ent showed no onion flavor. The oil was prepared for use again by

washing with cold water, a 10!^ solution of washing soda, then heating with

live ateam until it was sterilized. There was less than one p^ cent of the

milk loet and lees than three per cent of the oil lost during the treatment.
Abnoimal Coaditloas of the Cow*

Jkatm^ th« undesirablQ flavors vl^ieh asgr ba attributed to abnoxml

oonditions of tba oan are thoee reaultixtg from udder dleturbauoes and those

vhieh magr be found in mlllc draom from eovs ehieh are advanced la laetatlon,

particularly those which have been milking more than a y^r« "Hiese flavors

are not so easily recognized by the consumer, but may be noticeable to the

extent that he eould know that eomsthing was wrong with the milk*

Cows affected with udder disturbances resulting fTcna inflsraoation of

the udder tissues xsay produce milk with an abnormal Cfflsposition which would

influence the taste somewhat* When these chronic inflflnBatlons of the

udder are present, the milk from the Infected quarters may taste salty for
SO
the remainder of the lactation period*
50
According to experiments by Iioadhouse and Henderson to oon^are the

chloride percentages in the loilk from the quarters which ware ioown to be

affected with mastitis with that from quairters which had not been affected

witii mastitis, it was found that the chloride content of the infected

quarters was hl^er in every case, and that in most oases It was over 50^

higher* The lactose per cent in each ease was found to be lower, and

consequently, a hi^ ohlorida->laet08e number in each case* The milk

secured from the quarters of the udders which were infected in chronie

rather than acute oonditions appeared to be normal whea the samples were

tested for composition* When oospared with milk from the other quarters

it had a salty taste. The flavor was not as pleasing as that from the

control gzoup of cows*

 10

A Similar salty tasto is tha most otmaon dsfeot foand la milk from

oovs afliraneod in laotatlon period. Whan this taste aeooB3>aaie8 adranood

lactation, the milk from all quarters is affeoted. The reec«Bisad«A proos*

dure is to oease milking the eovs that are glTlng these results. When they
50
are again fresh the milk will. In most oases, he normal.

Handling of Milk.

The flavor of milk after secretion by the sow usually oannot be

iB^roved. On the other hand, there Is a natural tendency for It to dete->

riorate. It Is the problem of the producer, the distributor, and the

oonsumer to preserve the natural flavor of milk, and to prevent the develop«

ment of undesirable flavors which are so closely associated with In^roper

eooling. Inefficient sterilization of equij^sent, esQjosure of milk to mm*

50
light or hot metals, and imsaxiltary surrovmdings in general*

naalean Flavors. The dairy farmers of the tJhltsd States suffer great

losses each year because too large a proportion of the milk th«^ produce

Is Inferior In quality. The estimated loss Is many millions of dollars.

Scmm of the primary causes for this great loss are sour and off*flavoxed

milk and cream. These Inferior products ere not readily marketed, and whem
30
the dairyman does find a market, he usually gets a oc]iBq>aratively low price.

Milk of hl^ quality not only brings better prices but It also helps to

Increase oon8U]q;)tloB. Milk of hig^ quality may be defined as that milk

coning from healthy cows, Is of good flavor and free from dirt, and contains

very few bacteria, none of t^lch are harmful. This does not mean that milk

is ''clean* In the strict sense of the word because that would exclude milk
 11

irhloh eontai&ad may Wetorla or forai^i aatariftl nSmterar* All milk,

ozoopt that prodoaad under Tory exoeptlozial oonditlozus, has aoma basterla
^ 30
present*

Oiseaae produoing baoteria are not so oonsaon in loillc. When thsy azti

present they may bo attributed to diseased oows, unhealthy attendants

handling the milk, or oontaminated surroundings oeoised from water, flies,

or filth* Helatiwely large numbers of baoteria get into the milk from

neglect of proper cleaning of the oows, and lack of cleaning and ster*

ilizing the milk utensils. The use of small top milking pails helps to

reduce the number of bacteria and enount of dirt getting into the milk*

Bacteria find ideal conditions and food material in milk for rapid

growth, unless the milk is held at low teaqparaturea* The minute organisms

are 8ingle«>celled plants which eaimot be seen with the naked eye* Some of

thma divide to form two bacteria at maturity, and, under favorable condi

tions, each of these repeat the proceaa of division in thirty minutes,

thereby resulting in two additional grown individuals. The optimum tcm^

perature range for the ecnnon types of ozganisms to develop in milk is

from 80^ to 100^ F* Considerable multiplioation takes place at 70° ?*,

while at from 40® to 50® F. the growth is retarded.®*'
Many of the ooamon types of organisms found in milk do not cause a

change in the flavor. Other types mi^t change the flavor with no

apparent change in the appearance of the milk. Some of the most coionon

types of baoteria change both the flavor and appearance of milk. As a

result of these organisms producing acid from the milk sugar, many dif

ferent flavors may ocour* ffiiara proper control aaasures are eaployed.
 12

the riaT0378 produoed from bacterial growth should be reasonably olean*

However, where this growth is assoeiated with filth and dirty surroundingB
30
the flavors are usually very unoleaa.

Pasteurization vniile no definite figures are available to adiow

what pero^tage of market milk sold in the Halted States is paet«Htrizedy

so one will diq;>ttte its rapid iserease. This inerease alone represents

the endoreement of the general public* The larger eities are laoxe inolined

toward pasteurization than the smaller ones, probably dhis to the relatively

long period between production and oonsin^tion, and the faot that epidemie
29
diseases are more ecsznon in the larger cities*

Pasteurization is desirable for bacteria rodtietion; however, the

cream line and the change of the flavor have led to some objection on the

paxt of the consumer* If pasteurization is to be generally adopted, it

most be eaecn^lished at temperatures low enough to avoid cooked flavors*

This can be done only with the most accurate scuipment and under the siqter-

vision of szperienced operators* The solution to the problem of avoiding

cooked flavors is the adoption of ecme method which does not depend on

uncertainties for heat conduction*^

Oxidized yiavora* l^ere are several terms in coamon use to describe

the flavor sooMitimea found in laarket milk due to oxidation of the milk fat*

ikaong them may be Ibond oxidized, metallic, eappy, cardboard, oily,

tallowy, and papery* TSiis flavor may be caused in a number of different

ways, or by a oombination of factors, and yet the flavors produced ere

TOmewhat similar* This flavor varies in its intensity wi^ the time of

eoiposare to adverse conditions and ite stage of development* It can

 13

aaoally lid tx>ac«d to the oontaet of milk vlth oertain raetala. So»o of

thofio metole aie acted upon 1:^ the lalllc in sueh a tiey that a tsaali amount

of the metal is disQclved. This In tuim foiae metallic salts which con-

50
stituto the undesirable f],avor»

Since pasteurization has been so generally adopted, this flaror has

beo<nae more noticeable* This may be partly explained by eajrlng that the

mlllc ecsnes in contact with more metal than before pasteurization, and the

metals are mom soluble In hot milk* Refrigeration, which permits longer

holding of the milk botii by the dietributor and the eomemosr, thus allow

ing more tlrna for this oxidized flavor to develop, has been anothmr faetor
50
to oauee milk to derelop thoao objeotlonable flairors*
S4
Guthrie foxuid that certain cowe have i^rodused milk which developed

an oxidized flavor after one or two days in storage* This apparently was

true when there was no oontact with metals or no exposure to sonli^t*

so
Roadhottse and Headerson reooesasnd the use of ehrosHaiokel-iron and

chrom-niekel-alloy in adlk Vats and other eqnipmsnt with which hot milk

ocHsas in contact* Oopper, brass, bronze, aonel metal, and nickel silver

caused oxidized milk flavors in their experiments, and a lose in weight

when brou^t in contact with hot milk*

:7here milk is exposed to direct suulie^t in clear glass bottles, it

develops a definite oxidized, or tallowy, flavor* "Ihie flavor le notice

able to tiie eonstcoer, though he may not deteet the cause* ?aien ejg;>osed

for only a short time, ten minutes during saamer months, this flavor amy

affect palatability* This is unllka oxidized flavors due to metals which

develop In from one to lairee days,^

 u

Crea3Bl3ag cf ?aLlte.

MarlQBt milk Is Tsry often Judged by the eonsumer eusoording to the

anoiat of oreem appearing on the bottle. The amount of oream. forming on

milk is of special cimittsrcial Importance. The distributor etreeeee the

oream layer ae a sales adyantage* It glyes the oonsuner some Idea of the

richness of milk in regard to Its butterfat content. Ifoweyer, thla Is

sometimes a poor Indication of the per cent of butterfat In milk. The

consumer very often rmaoves part of the upper layer for table oream. This

graylty separation was once depended upon for obtaining oream for ehumlng,
10
but the oentrlfugal separator has about eliminated that practloe.

Another yery important eonsideration for creaming properties of milk

lies in the fact that the range of tensperatures la yery narrow betwosn

idiere pathogenlo bacteria are destroyed and where the cream layer is
, o o,
shortened. Ordinary paeteurlzatlcoi temperatures (142 • 145 } do not

materially affeot the oreemlng ability of maikst zallk whon the holding

time is not longer than thirty minutes* T«gB^>srature8 aboye 145° F. for
thirty mlnutss, or above 165° F. momentarily, prodoee a permanent preclp*
Itatlon of the calcium salts present in milk which do not diseoXve whon

the temperature Is lowered. Th9 preolpltatlon of these ealolum l<m8

10
partly destroy the ereaming properties of milk.

Dahlberg and Marquardt^^ say that the yolums of ereem rleing on milk
depends upont (1) the per oent of fat in the milk befoira creaming begins,

(2) the per cent of fat left in the eklm after separation, and (3) the per
cent of fat in the layer aboye the ereem line.
 15

roTngLtiotL of Crsgg I^yer*

Kie fat la allk ezista ao ymvy emit rotmfi elot>ulea» which riae to th«

top heoauso they are less than nins-tenths as heayy as skimllk* Hbm fat

glohulea, liiim separated as indirldualsy rlsa only a azoall fx-aetlon of an

inch per hour* The tijne required for an Indlrldual glohule to rise the

entire depth of a xollh bottle (approximately 7 inohea) would be around

826 hours, or ^ days* Since cream usually forms a ^od layer in 1«o hours,
10
It Is self evident that the fat does not rise as individual globules*

There Is no oonslusive evidence to prove the eause of the aggregation

of fat into clusters» yet It le eesrtaln that these clusters are forxaed*

1%ey are very Irregular In tdiape and size, and are loosely held together by

some unltnovn force* Mlk with excellent creaming piop<n>tles has aeny

oosparatlvely large clusters whleh rise faster than the emaller ones* It

le the rate of rising of the smaller clusters which dstexmlnea the rate of

eremsing, beoause they must rise before a distinct line is formed, indicat-
10
ing occplete creaming*
11
Sahlberg eaplains the foimation of these e lusters of fat globalsa as

being the neutralization and combination of the minute particles of fat as a

result of certain physical and chemical changes produced idisn the negative

anions attract the positive oations* This causes the fat glo)mles to come

together in clusters*

The calcium salts in milk are present in such great amounts that they

are not all dissolved, teit some remain in auepwsion* In wazm milk there is

a maximum number of caloit&a lone and thus a maximum number of positive

chargea, beoause the solubility increases with temperature, tharely reducing

 Id

tha formation of fat olusters* Wlian the warm milk is quickly oooled to

40^ F* or helov, ths ealoium ion moTooMmt is raduaad and olunQ>iug is

spaadsd iQ>» In that way the theory of best oreamiisg at low taa^eratuze

has been adranoed*^^

Cooling and Holdiim Tamperatures*

12
Dahlberg and Marquardt found that cooling teoperaturee ware

the greatest influences on Ihe ereaming ability of aarkst milk. On raw

milk with good creaming properties, the xoaximum cream roltime may be

reached in two to four hours. This volume may be gradually decreased with

ege for the next twenty-four hours. The cooling and holding teaapsraturBS

affset the rate at which the cream rises, the marirmtm length of the layer,
and the shrinkage which occurs after the creaming process ime been

completed.

For ideal creaming, milk should be held at temperatures of from 35^

to 40^ F* Where milk was allowed to cream at an awerage temperature of
38*^ F. for 24 hours, a cream volume of 4.2 per oent for eaoh 1 per cent
of fat in the milk was found, idiile at F., milk from the Sane smoples
gave cream volumes of 2.8 per oent for eaoh 1 per o«it of fat.^
26
WsBsaer found that a greater oream WJluraa appeared en milk when

held at tatiq>erature8 somewhat nser that of ios water, than an milk at

higher temporatures. When milk is oreamed at room tesqieTatures, there is

a tendency for the fat to pack closer together. He conclndes that the

original temperatures, at which milk is held before cremaing begins, have

little affect on the oream layer. Separation and remixing causes a sli^t

decrease. Milk creamed at low tenqperatures, then allowed to stand at xocaa

 17

toDoperatures, sbowad a noticeable decrease in oresni rolume, approxiiaately

8^ in raw milk* A marked increase of cream Tolume is noted on milk creamed

at room temperature for B4 hours, then held at ice water taaperatures for

creaming* This increase, 50'^ in eome oasea, is possibly due to age*

57
Trout found that a greater cream volume appeared on bottles creamed
a O
in air at 40 F* than vh^ creamed in ice water at SB F* at the S4-houtr

period. However, the creaming efficiency, as shown by ^e moount of fat

remaining In the milk serum, was in favor of the ice water taaQ)erature.

He explained this by saying that there was a greater concentration of fat

o
in the cream layer creamed in ice water than when creamed at 40 F.

Likewise, the fat test was lower in the milk serum creamed in ice water

than in lailk serum oreamsd at 40^ F.

Creaming at 70^ F. ie a relatively slow process. A distinct line doea

not appear until after aiz hours. Cream rising at 7(fi F., when examined
under a microscope, reveals a greater percentage of large fat globules than

cream which rises at 32° to 40° F. The percentage of fat found in the
cream layer where milk was allowed to set at 70° F. was higher than where
o 57
creamed at 32 F.

o ^
Haw milk creamed at 32 F. showed that 92^ of the total cream had

risen after two hours, 93^ after four hours, end the maziimaa, or lOO;^,

after six hours. At the end of 24 hours the volume had shrunk to approx

imately 90;t. When creamed at 40° F., raw milk reached a volume
after 24 hours. In most eases where large volumes of cream appeared in a

short time, the cream seamed to be bulky or coarse, and a decreaee in

volume resulted between the sixth hour and 24-hour periods. A shrinkage

in cream volume of approximately 10^ was found betwem the sixth hour and

24-hour period when rear milk was oreamsd at 32° F.; whereas, that creamed
 18

at 40 F* shoaad an inerearM» of omr 3^ In eroara Toluno during the mem

periods.®^

"Paatearizatlon Tetaperatures.

The eonnon praotloe for oarket loillc planta of today ia to paeteurine

the oilllc at teB^ecrattu'es of fiKHS 140^ to 14^ F« and hold it at theee

teB^esraturee for 30 xBlnuteet after vhleh it is oooled to 40^ to SO* F*
This procedure often affeote the eream lajrer* My proeedure vhieh will

retard the natural rising of the eream should he of espeeial interest to

57
the plant operator*

Trout57 found that milk pasteurized at 14^

-A
F* for 30 minutes showed

an arerage depth of vaevm layer of approximataly 14^ when ereamed at ioe

water teo^eraturef ae eomparsd to 11^ at 40* F#* and 6% at 70* F* The

per cent of fat remaining in the serum of pasteurized oilfc irtten ereamed

at 33* to 40* F. was more than twice as great as that found in the sesnna
of row milk ereamed under eimilar eonditione* 7exy Httle differenee in

the fat peroratage rwsaaining in Mlk serum was found in pasteurized and

raw milk idien creamed at 70* F«

Botis and Baboook^* say that heating milk to 140* F* for 30 minutes
will kill the pathogenio haoteria present, prowided aU of the milk is

heated to that tetq>orature and held throu^iout the 30 minutes* lliere is

a tendeney for milk plant operators to pasteurize as near the minimum

required lew as possible to avoid oooked flavors and so as to not

injure the cream line*

The United States UepartiaBnt of i^grlculture reooononds that all of

the milk he heated to a teiqperature not lower than 143* F* and held at
 IS

this toonporatura for xu>t less than thirty minutes* When this is properly

done, an ample margin of safety is not only insured, but a greater destruc

tion of bacteria will result than when railit is pasteurized at 140° F* and
held for the same period of thirty minutes* Only about of the bacteria

in millc r^aain alire when milk is pasteurized at 142^ F. for thiirty

29
minutes*

45
Parker eonfinaod the statement of Professor ^etor, of the Swiss

Dairy School in Berne, that the quickness of creaming as well as the

volume of oream increased up to a pasteurization temperature of 141*8° F*

Beyond this point these properties decreased, but even at 145*4° F* they
were greater than in raw milk*

Dahlborg and Marquardt^® state that there is no known way of restoring

the creaming properties of milk when pasteurization temperatures were over
o o
145 F. for thirty minutes, or 165 F, momentarily*
37
Marcuesen points out that the temperature of pasteurization exerte

an influence on the quantity of eraam rising on pasteurized milk* He eaya

that the volume of cream rising on milk which has been heated to 145° F*
for thirty minutea is alwa3ra loss than the volume of cream rising on the

same milk when pastetarizod at 142° F. for thirty minutea* The average
decrease in volume when pasteurized at 145° F. was 13^ as coiapared to that
pasteurized at 142° F*
Trout^"^ states that pasteurizing milk at 145° F* for thirty minutea
decreases the oream volume from 9 to 16^, the decrease depending on the

temperature at which the milk was held for oreaming*

 80

ss
tiarqaardt and Da&lbere nads a study of the oreanLng of milk

pasteurized at hi^ temperatures and found that the impaired oroaming

properties vars more noticeable and the differences were greater at

ireadings taken at the end of the second and fourth hours* Hovever, a

permanent reduction of the oreen layer was noted in most cases where tra*

peratures of ISO'^ - 165'' F* were usedy their aoplanation being that the

common methods of control were insufficient to hold all of the milk at

uniform time exposures*

Holding Time*

Hither method of pasteurization, whether It is the high teBq;)erature

short holding or low temperature long holding process, is satisfactory for

oareamlng of milk, provided the time of exposure to beat is in the proper

proportion to the temperature* By this is mesnt that milk heated to 142*^

F. for thirty minutes retains very good creaming properties. Twj^peratures

higher than 1^'* f» for thirty minutes sema to retard the natural rising of
the cream* Holding periods for more than thirty minutes at 142*^ F* also
seem to retard the natural rising of tiia cream. Furtheruiore, it is

possible to heat milk as high as 160^ F* for a period of twenty seconds

without affecting the cream voluaMj whereas, the neoct twenty seconds of
38
holding at 165^ F* greatly reduces the cream Toltaae*^ Marquardt and
38
Hahlberg foxmd that the holding time after the milk had reached the
o
145 F* required more attention when pasteurizing at high t«Bq>erature8,
13
than up to that tei:q)eratux>e* Dahlbarg and Marquardt , after a very

extensive study of the influ^ioe on the time that milk was held at pae«*
 21

teurizlug tei^peraturds, ooneluded that the reduetlon in oream toIum wbb a

"tlme-tai^aratura relationship". They found that a longer time eas

neoeasary for coBS)loto oreaming where milk was held longer than thirty

aittutsa at paateurization taEQierattU'ea. For eaCaBqple* milk heated to 148^ 7«

and held at that teogjerature for 150 to 160 minutee to Z hours) showed

a greater oream Tolume at the 24~hour pcnriod than at the S;^hour periods.
39
liarquardt says that frequently the oream line of milk la Impaired by

pasteurization tenQ>sratures whioh are too hig^, but numerous other things

Influenoe the oreaming of milk. He also says that the faetora which

influence cream rising in milk are rery often beyond the control of the

distributor.

Agitation and Pumping.

Considerable disagreement eziste among recent workers as to the affect

of agitation of milk on its creaming properties. Most authorities agree on

the critical temperatures at ahich pujifl^ing has the most harmful effect u®on

the cream rolume. Uhittaksr and others^® reported that their results would
indicate that the tendency appeared to be slightly in fawor of holdizig milk

during pasteurization without agitation. HowsTer* their results wore some-'

what inconsistent. Their oonolusiona were that a reasonable aaiunt of

agitation during tbe thixty->mlnuta holding period seemed to haTe no appro*

eiable effeot on the oream layer •.• • Holding and agitating at pas

teurization temperatures had considerably leas effect on the creaming

pjropertioa than agitation at 60® F. to 100° F.
 St

32
KiXboume roportod that ono of the moat inaportant feetore Influeoolng

the oreaalxig ebillty of milk was the amount of agitation to which the milk
57 10
wae subjeoted, eapeeially while hot* Trout and Dahlberg found that

agitation at paeteurlzation temperatures had -rsrjr little effeet on the

oreaiaing of milk*
57
Trout foimd from an ayerage of 92 trials that milk pasteurized at

145^ F. for thirty mlsutee ^To a oreaa Tolume eguii^tlsnt to 85^ of the

original raw milk whan sao^les w«n> taken from the eat* When the milk was

panped at 145^ F*, a slight deorsase of approzimatsly 8^ was eyident* The

samplee taken from milk cooled heek to 135^ F* in the yat« than pu^ed at
this temparature* gaye a oraam yoluma squivalent to 90^ of the original

sample* Other series of samples taksn at 125^* 115^* and 105^ F* gaye
0
similar cretmi yolumes, but where the milk was puim;>ed at 100 F* or below,

the depth of the eream layer was reduced materially*

17
Erh made a study of the effects of agitation on the cream layer of

milk, extending over a one»year period. He found that my agitation between

the temperatures of 40** and 105** F« semaed to Impair the oreamlng properties
of milk, the degree depending on the amount and kind of agitation and the

distance from the mean* He also stated that the d amage caused by agitation

was greater in pasteurized milk than in raw milk*

There is a correlation between partial chuming of the fat globules and

17
the reduction of cresa line as shown by examination under the microscope*
O 0
dgltation of milk at temperaturee of from 105 to 144 F* had no effeet on

the eream line, neither did mioroseopio examination show any partial
 17
ohumlniS of tlio fat globuloa* Srb euggoata that idtan the orean layer

has been lapaired by agitation, reheating to 140° f« Bemeataniy will

restore ita rielng* TiM id«atlty of the fat glcibales is partly lost when

the nillc is re~heated to the laelting point of fat*

]tox)fienigation>

A ooiBparatiyely now item in the bottle milh trade has been introdaoed

in SOBS seetions of the I^ted States; priioarily in the North and East*

Sinoe this is a new produet, information is limited to a few artieles in

the dairy journals* A oosg^lete history was not found, but apparently it

is just getting into eaaBasroial trade* Scnne of the distributors are

calling it '*Bono** milk • The dairy at Ohio State Uhiwersity has been

eelling homogenized milk for more than two years with considerable growth

in dBiand*

An intereating editorial recently appeared in one of the poptaar

53a
dairy journals wi-^ roferenee to the oonstimer'e reaction toward "Homo**

milk. Tho author aays that **irolumeB have baen written about irtiy pMple

should drink more milk* Little, howerer, has been aaid about making milk

eo good that eweryone would drink moro of it***

63a
Tho proeesB, as deserlbed by the writer ,Involwes heating the milk

to 142° F*, then passing it through a machine that puts a pressure of

8,CK)0 to 3,000 pounds par equare inoh upon the milk* The fat glohulea are

thereby brokon into many esaaller partioles* In this pulverized

condition the fat globules do not elump together to foxm a oream layer*

The milk le then pasteuriSMid and bottled in the usual aaaner*

 M

Diatieians hare loiie oontanded that the more finely dirided food eaa *

the easier It would be digested* Theoretioally, ''Hcaao'* milk should he

more readily digested than regular milk*

^Is type of milk la ideal for infant feeding and is gaining in

53a
popularity for oooklng as well as for general home oonsus^tlon#

^ ^ 17'

"♦ •-

\ ■'
 25

Bacteriology of Klarket Milk*

Otte of the lojat importjsuit faotora to consider in the handling of

market milk, intended for huioan conouinption, is that of bacterial control#

A great deal of attention has been focused on this problmi recently,

especially where a system of strict grading has beooias effeotlTO*

Additional information on this subject should j>royre to be intaresting and

beneficial.

Bacteria are minute organisms so small that they cannot bo seen

without the use of a microscope* It was once believed that they belonged

to the animal kingdom, but It is now agreed that they are amall one«cell,

colorless plants. They are nonaally found in milk, air, water, and

practically everytdiere. Kormal milk, howovor, before any contamination,

contains relatively few bacteria when drawn.

The problem of bacterial growth is of especial interert to the dairy

industry and ahould be understood and properly controlled skilled

operators. While some bacteria grow at temperaturea commonly used for

pasteuriBation of milk, and othora at the freezing point of water, a

temperature range of from 70° to 100° F. with an optimum of around 95° F.

is favorable for the development of most bacteria. The common oloments
31
necessary for bacterial growth are moisture, food, and oxygen.
18
Evans found that the bacterial flora of milk from apparently normal

udders ranged from 0 to 300,000 per oublo oentimater. There wore several

different types of organisms present. Jfest of them were of the streptococcus

groups* About IS^ of tho maploe oontalnod so fe« baetsria that their

signlfloaiioe was negligible*

After milk is drawn there are many souroes of eontamination. Sc»ae of

the more eonmoa ones are the body of the oowf the air, the attendant, and

the utensils and equipment* Speoifio disease produoing organisms may be

3X
introduoed from the attendant, flies, oontaninated water supply, or maxure*

In studying the aotion of bacteria on milk, awTeral influwaoea may be

oonsidered*

Hethods of Analysis*

It would not only bo impractloable» but physically impossible to

aotxially eount the number of baoteria in a quart of milk or eren in a drop

of milk* In analyzing milk for baoteria, recourse must be had to the

O
making of estimatee of numbers, not actual counts.

The methods ocnsaonly einployed for estimating the number of baoteria

in milk are: (1) direct microscopic, (3) Agar plate counting, (3) Frost
little plate, and (4) Methylene Blue Reductaee teat* All of the abowe

mentioned methods haws their adwantagee and dieadwantages* rtowewer, this

should not iiiQ>ly that nona of them are satisfactory, beeause each of them

has a place in the control of bacteria in milk.

Mioroacopto Methods of Oeuntimt Bacteria* One of the easiest ways of

coxmting objects is by direct observation* This is almost in^ossible with

the case of baoteria because they are too small to be seen with the naked

aye* However, sinoe the discovery of the miorosoope, more accurate

estimatee of bacteria may be made by direct observation* This method of

estimation is ocw^aratlvely new and has proved very valuable. There are
 27

oertatn optloal limitations when eiBploying the mloroscope, possibly due to

the (1) difficulty of Inaccurate measurements in such minute quantities

required for observation, (S) presence of dead cells, and (3) growth mA
8
diYision during preparatory steps for exmaination* On the other hand,

some idea of the types of organionus present may be had and results are
87
qttioldy obtainable*

Agar Plate Methods of CountInia! Bacteria* IMe method employs the

uae of inoculating some nutrient Jelly, which is transparent, with

meaeured quantities of the milk to be analyzed* This mixture of the

nutrimit ie then incubated until tbe original bacteria hare produced suffl^

dent growth ao that they may be counted with a low«powar magnifying lens*

This method is an Inqsrowwoent orer that introduced by Eboh in 1881,

which was conducted with the use of gelatin as a means of isolating pore

cultures of bacteria. Nutrient agar has supplammited gslatln for analysis

of milk largsly through the work of the laboratory aeotion of the American

Public Health Association* It is now reeogdzed by most authorities as a

6
very eatisfaetory growing media*
27
A modification of this method wae In^duced by ?rost about 1915,

who employed the ssme media end preparations as are used in the large plate

method* "^nie greatest distinction being that in the "Little Plate" method,

the colonies which develop on the media are counted under a compound

mioroseope before they eould be seen with the unaided eye* Its advantages
8
and dieadvantagos are about the Same es for the regular agar plate method.
 S9

Soaie of the limitations for the eulture media methods of estimating

the haeterla in milk are)

a* All baeteria do not grow under the preraillng eonditlons*

btt Seme colonies may grow ao rapidly that other colonies are

depressed, therefore en inaeoarate estimation would result*

e* Growth from Indiwiduals cannot be distinguished from growth

produced by olmops or clusters*

d* The possibility of oontenlnation from technique is always preswxt*

e« Meaysurements for dilutions eanaot be abeolutely aoourate*

Of the above, faulty technique may have one of the greatest influences

on inaccuracy* On the other hand, eklll end care nay reduce the dif*

fioulties to a aiainHn, but these handioapa have not yet been oompletely
Q
eliminated*

8
Brew oonoluded that aetual eounta in numbers of baeteria in milk were

impossible as well as unxieeeesary* The plate method or the direct method

may serve as an index to the quality of milk at hand, but neither of them

will indicate the aetual nmaber of baotexla pzesasit* Laboratories are

fortunate in getting the eon^parison of real value that they do find with

the inaocuraoiee that eate koown to exist*

There is no necessity for knowing the actual number of bacteria in a

given sample of milk. It matters not whether a eaaple of milk contains

5,000 baeteria per cubic centimeter or 6,000 bacteria par cubic centimeter*

Howwer, it is desirable in many instances to Imow the bacterial index of

a milk supply* That is, If it would fall in a ralativsly hi|^, madium, or

 29

low eXass« Sl^er Vxtt plate matiiod or the dlreet eoo&t m8Qr be relied
7
to give this lafonnation*

CoaBDarlaon betiroea the inlcroscopie and agar plate mettoda of

s
oottntlng« Breed eajns that with the knowledge of oonditlons md technique

under which both teete are eonduotad, the oonoa assumption that the agar

plate method gives more eeourate results thim Idie dlreet method has no

basic walue* He points out that for the plate count to be ecouzsite it

BBtst not contain more then 300 colonies. This maziaBa& must be adjusted

b7 dilutions and measures which introduce error. Consequently, the plate

method would be accurate only when low count milk was examined. In

contrast to this, ths direct microscopic method requires only a «Bw>n but

constant amotuxt of milk regardless of the esqpected count. Therefore, it

a
ie leas subjected to error in milk with high counts. Breed, in sumnerls-

ing the above, says that a failure of eome investigators to appreciate

these differences has caused unjustified orltloism for bacterial analysis,

but both methods are satisfactory.

The Methylene Blue Beductase Test. The reductaee teat is based on

the fact that the color Inparted to milk by the addition of a small omount

of methylene bine solution disappears more or less quickly, depending on

yKhethBT the bacterial content of the Sfflaplo of luilk is hi?h or low. By

the use of solutions of this dye made up to a definite strength and added

in known dilutions, consistent results may bs eiqpeoted. However, it la

very difficult to catimate the number of bacteria present In the eample of

milk.
 30

"OiOM baoterla 9aam>nly foaad In nllk but idiioh do not doTdlop very

rapidly are of no oeaBeroial inportaaoe. Therefore» a laethod whioh

detenaines the aetual nunlMr that will grow in ndllc is of aost Ixoportanoe

to the fluid lailk indusftry*

A brief oonsideration of all the mothoda suggested for detexmining the

baoterial estimation would seem to indieate that the reduotase method

should come nearer to aeaeuring the bacteria growing In the milk than any

of the other methods* the principle being that the color disappears in

proportion to the growth of the baoterla* This test* however* Is made at

tmaperatures at whloh milk should not be held* The gix>vth and development

of some types of organisms may result whloh* idien held at comparatively

S8
low temperatuxea, would not function*

One logioal way to analyze the basic sotmdness of a given test would

be to do as the chemist does; that Is* to make samples of solutioixs of

known strength and oonduet the test on these to determine If the reaxats

are oonslstent* l%e plate count makes use of thle to stms extent by

making the different dilutions Into sterile water blanks before plating.
It has be«n shown by Barthel, 1917, that sterile milk free flrtHa

oxygen* upon heating or by passing hydrogen* nitrogen* or carbon dioxide

through it, will reduoe methylene blue solutions in about two hoxnrs when

proteeted from atmospheric orygsn* This indicates that mom constituent

of the milk has a reduoing power* If the milk alone did the reducing,

about two hours would be a wlnlwam tlnui, while if the baoterla form a

reductase (an ensyme), as is the ease* the reduction tins would mueh
shorter*^
 31

The outstandlJis differenose botweea tb» roduetase test and ths other

nsthods for approxlniatiiii; the baeterlal content are:

1* The plate and little plate methods giye us counts, ehlle

the reduetase test glTes us aa estimation or an index to quality*

3* The oethylene hlue test is less toxie in dilutions ussd, thus

glTlng a point in its favor*

for the analysis of milk belof dslivered to distributing end manufao*

turing plants, the reduetase test is gaining in favor* A supplement to

this test has also bsen proved worthy of oonslderation• that of the

JsBtts Oreen* It has been said to have given a more aecurats test thsn the

rsduotass method* The prooedure is the same, except one part of the dye
S8
is used to 200,000 parts of milk*

All who have made a careful study of the comparison of the plats and

reduetase teat have found that the agreement was not very close* Either

of them gives a fair indloation of the quality of milk and the oars used

in its production* There is a dlreet relation between the rsduetlon tlnm

and the leucoooytes present, which would indicate that the reduetase method
49
aids in ploking out abnoraal milk*

A suBBoary of this may be illustrated aa followsi

Beduotion time in hours •> 1, 2, etc*

Leuooocyte In thousands ** T
Millions » M

1 £ 3 4 S « 7 8 9 10 10-f
im m m aoor soot soot isot 45t 40t ist 13T

Ordinarily, tiis methyleoe blue reduotase test for therDoophylio baaterla

in milk is the same as for raw milk, except that the tubes are iooubated at
o
pasteurizing temperatures (1A2*14S) iastsad of the ueuel 93 f, A soaU
 . ■X.

38

amount of glyraol or light oil, e.g* mineral oil» ohouW be added to form a

layer on top of the milk to prevent the absorption of oxygen from the air

which may interfere with the tost. Tlio results should be recorded in

30 minutes, 45 minutes, 2 hours, and 5 hours. These organiama appear to be

associated with milkstona. Possibly, erewlcea in vats protect them*

Pasteurizing milk, holding it for 24 hours, then remixing it with fresh

milk, and repasteurizing causes this* At the above incubation temperatures

all sauries which retain the blue color over 8 hours contain vory few
52
thaiwjphylio bacteria.

A combination of the loethylene blue and fermentation teat serves aa

an index to help to determine the kinde of bacteria present.

After reduction Incubate 18 hours.

£• If the eolor is reduced in less than

5;V hours, there is an i,ndioation of
improper cooling.

a. A smooth, firm aux>d indicates

growth of lactic acid bacteria.

Channels showing goe are the

result of;

(1) Wet hand rallking

(2) Dirty oows
(3} Improper oooling

3. If curd has appearance of being chewed up,

this may be caused by proteolytic bacteria.
Thesa organisms are usually associated with
utensils which have not been properly
washed and sterilized. Unfortunately, all
milk does not oomo in these elasees; the
classes overlap, thus requiring some
interpretation of results.
 m

The relations betVNMa plate oouat aaiA reduetion tiiixi as glten. by tbe

Cr. S« Pablie Health Service in the code which accar£5)anies the staniarA

miUc erdiaaBoe are;

l>eB8 than 50,OCX) per ol* 8 hours

50,0(X> * 200,000 G-B hours
210,000-1,000,000 Si-6 houM
Over 1,000,000 less than 3|^ hours

Tbm above oorrelatlens tore for raw milk*

Deooloriaation will evwitually set up beeause of an enzprne temed

aldellxyde reduotaae* Where a siaall eaniount of fOimalin is added, milk

reduces in 10-24 hours*

Orgsoaiaaui foomd in tiie afldev do not deoolorise iMthylene blue at Idie

sane rate as thoi» found on utensils* !76ithsr do they grow as fast as

the utensil flora* The organion S* laotis eauranee osqrgen re^ldly, making
44
methylene blue teet praotieal*
IP
Szperimente were eonduoted by Fay to detexinine the reduetion and

plate count of milk to which formalin had been added* Dilutions were made

where the final would have 1:100 end up to 1:1,000,000 parts of pure

(BOBO) formalin* The dilutions were made in the milk and held at 21^ C*
(70*' F*) for 24 hours* The standard plate oount on the milk was 18,000*
After 24 hours the milk oontaining eoneentratione of 1:2,500 or more shosed
•••

no eoloniee in the 1/10 dilution plates* There was a very marked depre^e-
in oount with a 1:20,000 dilution, slight growth was evident in 1:25,OOO

dilution, and appreeiable growth was evident in hii^er dilutions. It may

be oeen fron these restats that dilutions weaker thm 1:29,000 failed to

retard bacterial growth sufficiently to keep milk within legal standards*

 34

10
fl9y*s results indleated that the a:ax>unt of fozraaldehyde naeessary

to inhibit growth varied with different samples of milk and those seiq;>le8

carrying hii^ counts required relatively large amounts of (ECHO) formalin*

The reduction time for the original sample of milk was 660 minutes

(11 hours)* Addition of fozmalint IU.00,000 parts concentration, shortened

the reduotien time to S59 minutes or 4 1/3 hours; similarly, the lower

dilutions redueed quicker, the 1:1,000 requirixg 21 minutes* However,

with a dilution of ItSSO the reduction time was 440 minutes or 7 X/Z hours,
and the 1:100 dilation inhibited reduotion*

Uelng dilutions of 1:100 to 1:10,000 (ECHO) the reduotion time

deoreased to a few minutea, apparently due to oheoleal changee rather than

10
Mologioal factors**'

The methylenc blue reduotion method helps to pick out suspicious

saa^Iee of added foimaalin; however, it anet be followed hy more aecurate

methods to establish proof. Dilutions as hi^ as 1:15,000• 1:25,000 must

he xised if growth is retarded without being noticeable to the taste* The

addition of formalin would require accuracy to .0125 oo., or 5 drops per

gallon* This would neoessltate more aoouraoy idum most producers oould

dspend on. Any samples with a low plate count that reduce in less thsm
19
one hour may be said to be suspicious of ECHD added*
19
Ihy , in giving the nomal limits of variation of the methylene blue

reduotase test, aaye that the resulta seera to indloate that duplloate

samples oheok closer than duplicate plate counts* He points out that

where 1774 tests were made on 19 sanples of milk with epecific controX,! VhO
 38

x«dtietai8« teat gave aa 68 per era.t agreaoMBit to that of the dlreet ooont

and aoaafirhat the aasw reaolta for aedioent*

19
The advantagea aa auggeated Fay are that it ia atuplet easy to

Interpret, and requirea very little expense for laateriala* It ia a good

test for ntall planta and eitiea. Many larger eitlea use it for a general

aixrvey, then aupplaomit it with more elaborate teats*

14a
In en abatraet givm by Bevareau, the results indieated lOiat the

OMiparison of the different nethoda may be summarized as follows: The

reduotase test was able to grade the milk into four olasses equally as well

as any of the others. The Brom thymol blue was also a good indication of

quality*

A dipper is just aa satisfaetory as is a pipette provided it is

thoroughly rinsed after each saQq>le, preferably with oold water, then hot

water, and then a ohlorine solution* After a thorotig^ washing, the t«d>e'a
should be either steaiaad, subjeotsd to boiling water, or filled with

ehlorine water at 200 P*P*M* before using again* The burette ia quieker

and juat as aocurate as a pipette for adding the nMthylene blue solution*

The saB^les any be plaeed directly in the test tubes and should be started

as soon as is practical; if they are to be held over 2 hours, they should

bo kept at frcm 50° to 32° F. The aeiqplea may be mixed by placing the
thumb or a piece of rubber over the mouth of the tube* Vhen milk reduces
15
qtilckly, use the microscopic tost to verify results*

The reduotase teat apparently is a better indication of quality of

19a
milk due to delayed eooling than is the plate count*
 Apparently, |;«»uitrio rnaam is mora dosirable than arithEiatio

naaii tor reoording eounta of baeterla bawd on the time sveet miUc will

renaln aweet* Th« 6varm baaad on arithmetlo linear gra^a m»«ois worthleas.

When used aa an index the logarithmatie raluea of the plate count tium

Xeaa Tariation on milk with a low baeterial content, and the reduetioa

method showe leaa variability with milk of hi^ booterial content. One

method eeaeis to be about as accurate aa the othffir whan the wntire xtmtga of

leeeping qualities is oonsidMWd. Both are more accurate than the actual
19
baeterial plate eotmt becauee they conform more to a ourra relation.

Hileman eogi^sted using a eingla solution ataln that waa stand*

erdisad hy B. W. Heaman. "Wa are not actively using the methylene blue

reduction method for gpradlng milk thoni^ we are satisfied it could be uaad

as well ae the direct method.** Profeaaor Troy of Gomell Thiivarsity ^^t

the entire eunner of 198(1 in a large plant comparine the dirMt and

reduetaaa mathoda, and found vary good agraement on tha idiole. Neithar of

the two methods ara entirely satisfactory on low count milk. Tha plate

count le probably more satisfactory than either of the above for milk

containing aiaall noahere of bacteria.

 87

Effect of GooXlng»

Bacterial growth aod naltiplieation is sstoh more ii^portant thaa the

original couftt* counts are not oanraonly found where growth la retarded

aeon €ifter silIkliMft* Tehle 7 iidiows how rapidly hactsrla nultlply in « waxn

medluca*

Isgproper cooling, or the failure to cool, is hy far the greatest

influence on hig^ bacterial ocnints in milk which is reeeiTsd on the ererage

market* Milk held at tsagieratures near those at whieh it is drawn p«nait

bacteria to reproduce euid double in number once every half hour, ivt this

rate, one single orgaalflm would multiply to over 1,000 in 5 hours* ITesh

milk eontaining 10,000 bacteria per cubic centizaster might contain at ths

mid of S hours, 10,000,000* Fortunately, the growth does not taks plaee at

such a trmaendous rate* Sosm of the organisms do not deTslop daring the

first few hours, lAlle others die without producing si^ growth* Sowever,
31
there remains a rapid growth wh«a milk is not properly cooled*
3!L
Felly and Oleiasnt say that milk abould be cooled as soon as it is

drawn from ths cow* ihera cooling is delayed, the multiplication of

bacteria takes place, resulting in a high count* Ooollng milk in ocas bgr

placing it in cold water does not chill the milk as qhiokly as hf allowing

the milk to flow over a surface cooler* The (quickest method for cooling

is the moat desirable from a quality viewpoint* In addition to the

bacterial control by quick cooling, the surface cooler pexnits the milk to

be shipped more quickly, especially the moming*s milk, and saTes time in

cooling*
 m

^ aaya that watar at 80® to 80® f* is atailabXs for ooolisog

fflllk oa raost dairy faras# Witto watar at this tso«>€(raturB, a Tsry rsyld,

offioienti and sooaimLoal oooler aay ba Inatalled* % sslaotlng a durabXs

set of ooils, one that he easily eleana^* the oost of sooling OT»r a

surfaeo oooler should he about the sasDS as for eaa ooolins# ^eh eoir*s

joilk should ha cooled just as soon as loilllBf is eoi^lete without vaittag

for the reraalndar of the cows to be finished. Great oaro ^uld bo given

to the op«rati(m of the oooler in order to regulate it so that a oontinuous,

thin streea of ailk will flxm throxigbout the entire nilking period. Wum

it is not nsoessary to cool the milk to bs i^ppsd immediately, an e^para.

tus asy be set up to allow the milk to be ooolsd corer a surfaes eoolsr part

of the way, then finished in a cooling tank. Where this method ia practiced,

cold water flowing through the tubes eoola the milk to within a tmi degrewi

of the tsiiqperature of the water#

14
Downs and Lewis found that oooling elsan milk with a low oount to

50® « 60® 7. was a satisfactory procedure, end that oooling clean and
medium olaan milk to 80® - 60® F. was a fairly satisfactory proeedurs*
When dirty or high count ia not cooled below 60® 7., the baeterial growth
ourve undesirable praaP^Mss#
 m

Sffsot Pasteurigatloa»
3i
and OlAsent S€&* that about 09 par eant of tlio l^toria found

in milk sra uaually idllod by pastemrlssatlon* poroantaea klllad will

insry eomewbat vitb tba original count and tha typaa of orgaiaiaaa praaont*

Tba peiroantage of deoraaso may not be a satisfactory Indication of the

quality of milk, because the efficiency of pasteurization may be as big^ m

and the final count still be as high as 100,000 bacteria per cubic

eentimeter*

The types of organisms present may influsnoe the efficiency of

pastaurization due to the ability of some bacteria to surrive ordinary

55
paateurization taiaparaturos* lyers and Johnson found that four eeparata

groups of bacteria surrxTed pasteurization. Samely, acid formers, inert,

alkali formers, and peptonizing orgonlmse* Shey found furthcnr that *^10

paroontage of acid formers was iiu;roaG»d by pasteurization, imd the other

groups were decreased* fhe average xiusiber of lactic acid forming bacteria

that survive pasteurization is about S per amt of the number in the raw

milk*

After a good grade of milk hae been paeteurlzed, the typee of oxfsm^

laas develop which may cause the milk to appear peptlonlzed without tht

development of any acid, Ifhen the grade of milk is only fair, the aoid

group may develop and outgrow all of the other groupe, and produce a

normal curd* A very poor quality of raw milk may, after pastaurization,

first beeeaae curdled as with a normal curd, then chnoge to a broken-Dq^ curdl
l^uitwurlzetion of milk at to 148^ ?• for 30 minutes destroys
any pathogsnic bacteria that may be present. It reduces ^e ntsaber of
Si
lactic acid orgnxiinHi and l^ral^ adds to the keeping quality of the milk.
 40

Starlllgatlon of tTtenalXg anfl Bquliaaant*

91i» sterilization of utttoalXa and equiiatiinat is one of the essential

factors in the production of Biilk with low bacteria counts. The minute

organisBBS which wery often cause off-flavors^ souring, and sometimes huBMUi

illness, multiply very rabidly on moist surfaces of improper sterilized

milk utensils and equipment* JUst the mere washing of milk containers is

not satisfactory* The surfaces with which the milk cones in contaet ehould

be thoroiighly washed, treated in some way to destroy the baotearia, and kept

dry tmtil ready for use a^Ltn* They should then be sterilized to prevmt

eonteoBination of the milk*

Some of the comaon methods of sterilizing dairy equipment are;

(1) hot water, (s) eteam, and (3) chemical* All of iihm have been used

very extensively, plant practices being one of the major influences on Idle

selection of the type*

9
Hot Water* Oaulfiold, Riddel, and Fay of Kiansas sey^ that boiling

water is effective as a sterilizing agent only when the uteneil may be

sulmierged for several minutes in water that is boiling vigorouiBly* The

c«aaoa practice of scalding utensils by pouring water over the anrfeoe is

very inadequate as a metbod of sterilization; however, this is the method

most commonly used in Idtiio as shown by a survey made by Theophllua and

54
Atkason • A survey of 204 membwre of a dairy hez^ izopsrovement association

showed that 830 or 89*4^ used hot water* Of the 830 using hot water for
sterilization puzposes, 210 heated the water en kitchen stoves, and the

remainder used various other means* The survey showed that about 4 gallons

of hot boiling water was required to properly sterilize the dairy utensils
 41

as«d by thfl ordlsaxy amalX produowr vhioli ustially eonsisted of 3 oens» a

strainor, a eooler, and th© separator parts*

Praoha» m99t«e, and CQxa^»ers^^ of Illinois mad© a very extenslt©

study of sterilisation ©itli hot water from the aicamlnatlon of 170 freshly

washed sans (unsteasMid) which showed the presence of large anabMni of

baeterla* Had these freshly wajihed eans been filled with sterile lallky

the germ oontent of the mlllc would hawe varied from 197 to 2,S57»000

baeteria per ee* with an average of 128*592 baeterla per oo* Their results

from the obseirratlon on 170 eans suggest that milk cans* wairibed In the

ordinary masnar, contain wxffiolent gexm life to heavily inoculate the mlllc

later placed In thsau The results of successive rinsings with sterile

water chow that while the gexm life removsd by the first rinsings wilh

sterile water amounts to a considerable portion of germ life In the ean» It

Is by no meeoMi the entire germ life present* Accordingly, the gexm content,

as determined in this manner, Is imoh lower than the actual number of

baeterla present in the can under Investigation*

In ths preceding srperlmsnt It was ehoim that freshly washed cans

Invariably harbored large numbsre of baeterla* Dairy utensils, however,

are not eoniaonly used for milk Immediately after they are washed* This Is

eepeelally true of cans In which milk Is shipped from the ttaem to Ihe

plant* Such cans are usually washed and steamed at the plant, them covered

with the lids and returned to the farm where they are frequently used for

milk without any further treatment* Very oftmct, one or two days will elapse

between the time the oans are washed and when they are used*
 42

47
Pruoha, fheetdr, and OhaiabaM alao atudlad th« g«xm Ufa in 160

Swgallon oans at the time they would ordinarily he used* These eana were

examined after being washed at a particular dairy* 100 of then were steaaiei

while 60 were left uneteamed*

The steaming eonsisted of holding eaoh ean ower a Jet of eteam for

25 eeoonds at 15«>potmd8 pressure* The pressure of the steam was measured

by a guage placed between the Talve and the Jet opening* Other experiments

on steaming oans in this manner showed that if cans so treated were filled

with milk immediately afterward, they rarely erer added more than two

bacteria per oe* to tho milk* 50 of ths steamed cans and SO of those x>ot

staamsd were inwerted on a x^ok with tiie lids off* The other SO steamed

oans and 10 not ateamsd were closed iomdiately after washing* All the

cans were then kept SO hours in a vwm hawing a humidity of 40 and a tmo-

parature of 60^ to 70^ 7* The number of baoteria found in each ean was wery
47
interesting*

The 50 oans that were washed, steamed, and held 30 hours uncowered

and inverted on a rack were dry and free from objectionable odors with only

a few baoteria present. Only 3 of the SO sans had mors than ons million

baoteria and 36 of thsa had loss than 100,000* 2f the baoteria in the SO

oans had been added to 400 gallons of lailk, the gexm content of the milk

would hawe been inoreaaed by only 6 bacteria per oo* Whether any bacterial

growth took place in the oans during the 30 hours was not known, but tho

results show that ths mans so treated hawe a negligible effect t^n the
47
germ content of milk#
 m

50 oans tliat vecro vaslxed, stoamod, aad held 30 hours with Urn lids

oa eere still doap end laost of thsu had a pxtmotmoed odor* These eaas had a

soioh larger number of baeteria than those vhieh ware steasMidt Dmootered, and

Inyerted on the rsah* Only 3 of the 50 eons had less than 100,000 baoteria

per ee* If the baoteria found in these SO ei#t-gallon easis vere added to

400 gallons of xailh, its oontsnt would be Infficeaaed by 1,315 baeteria psif

00*

The 90 eans whloh were washed but sot etesmed, end held 30 hours

unoovered cmd inserted on a raok were di^, and none of them had a disagree-

able odor* Tho nuaabdra of baoteria in them were mooh larger than in the

eans steamed and interted<» Only one of the 50 eans had lees than one

xoillion baoteria, in £4 of than the numbers of baoteria were between one

million and ten million, and 4 eans eontained over a billion baeteria eaoh*

If the baoteria found in all of these cans were added to 400 gallona of

adUc, its gsxra content would be Inereaeed 97,164 baoteria p«r eo* the

effeot of drying the oans upon the germ life parosent is evident frcm a

ooQ^arison of the atewe xesults with those obtained from the freidUy welded
47
oans*

iSaoh of the 10 oans whioh were washed but not steaiaed, ooweiwd, and

held 30 hours had a disagreeable odor, and tdiey elso eontained a large

number of baeteria* Hine of this number eontained over a billion bacteria

eaoh* If the total ntmdier of baoteria ibund in these 10 ei^t-gallon eans

were added to 80 gallojus of milk, its gsxm oontont would have bem. inoreased

by 198,730 baeteria per oe*'*''

It Is evident frena the above results that rapid bacterial growth took

place In the oans whioh were oovered end allowed to stend 30 hours*
 44

Baoteriea growth la gwaei^al id hyt (1) tco^wraturo, (£) faad,

aad (3) oolsture* All of the oaas in the above experimeate wero held at the

same t^eratare and were washed in the aaoe dairy by the sane ^eratoTy

0O tiiat the prlnctpai dlffereaee between the oovered and unoovered esne was

the presenoe of moisture in the oovered eans* These reeolts point to the

oonolusioa that it is very diffioult to wash oans so that no baoterlal food

is left in tlMBit and if the oens are oovered without drying end are allowed
47
to stand for several hours* the bacterial growth inoreases rabidly*
4?
Baoterlal studies were made ly Fruoha and others using oans whieh

were being washed end roturoed to the farm ready for In order to

maintain the usual eondltions in this dairy, no interfermxoe was made in

any of the r^sular operations and the men doing the worh ware not aware of

the ejq^eriment* £lo record oould be obtained of the exact treatBKmt of each

oan, Imt in general, eaoh oan was washed* rinsed, stewied over a Jet, and

covered with a lid* Steaodng of the cans Tarled team 5 to 20 seconds*

The treatment of the oans at the farms was not uniform* Scsse of them

were inverted on raclcs with the lids off, and others were not opened until

they were used* The tistB intervening between the washing of the cans and
47
their use varied from 6 to 40 hoxirs*

Just before the cans were filled with milk, they ware rinsed with ane

liter of eterlla water, and the bacterial content of this water was

determined*

Of the 91 cans examined, 3 ^wed lesa than one million bacteria; 67

Hallowed between «u million and one hundred million; and 31 showed over one

hundred million baeteria per can* If all the bacteria found in the 91 cans
 45

wided to 728 gallons of mlUc (the total eapacity of the eaas)» the
4n
haoterlal oontent of this stilk would be inoreaeed 23tS^ bacteria per e««

^e aboTo cans were inspected prior to the bacteriological examination

and were found to be free from any dirt* and were dry in most eases* From

extensiwe study of cans on a large nuatber of fanai and in dairies, the

inrestlgators were of the opinion that the 91 eans ware oleaner and in a
4?
better oondltion than the areirage oan used for milk*

SteasB^ ^nie most effectiwe method of sterilizing dairy equipment,

40
aoooz^ng to Csulfield, Hiddell* and ?ay » is by exposing utensil to

lire steam In a closed cabinet for 10 to IS minutes* Stosm jets eomnonly

mi:s>loyed in dairy planta for sterilizing cans, pails, and other equipment

are frequently not effective beeauee of the short period of exposure* Whan

exposed to a jet of steam for less than one minute, as is very frequently

the case, little destruction of bacteria might be expsoted* fhe principal

advantage of the use of a steam jet is that the utensils are heated so that

subsequently they will become thoroughly dry*

54
Theophilus and Atioeson made a study using slectrleally-heated sliwia

sterilizers. They used three sterilizers made by different eosipaalM* 1!lui

sterilizers studied were of the same size (4~oaa size), similar in construc

tion, end all were of the oabinet type*

The sterilizers were studied from two viewpoints: first, as a msans of

sterilizing; and second, as a means of heating water for washing puj^osea

around the dairy* In the first part of the study, 4 eans (two 10-gallon

and two 5-gallon) used in the Ihiiverslty of Idaho Oreemary wara washed but

not sterilized* One of tdie lO-gallon oans was rinsed with 800 oc* of
 49

sterile vater end the haoterial content of this wat«r was detexaslned hjr the

standard plate method. The sterilizer was then operated aocordlng to the

directions of the nanafaetarer* After sterlllzetlon, the oan prerioosly

checked for bacterial oontaalnatlon was again oheoked Iry the same method.

Sterilizing effioieney was expressed by the percentage of bacteria

54
destroyed.

In ? of 13 trials with mitk electrically-heated steam 8teriliaer« the

tttansils were held in the sterilizer 20 minutes after the electric heat was

automatically cut off \fy the thermostatic control. In three trials the

utensils we3re held IS minutes and in three other trials they were held only

0 minutes. The electric energy used in all trials was measured in kil>watt

hours. The tenperature of the cabinet was measured by an accurate long

stem themcmeter. the bulb of which was at the uppermost portion of ths
54
can.

nie average of 7 trials, rdien ths utensils were held in the sterilizer

20 minutes after the maxiiagn temperature was reached, showed each of the

sterilizers to be 99.9 per cent efficient. Also an average of 99.9 per cant

was obtained in 3 trials when ths utensils were hold in the eterilizw

19 minutes. The sterilizers showed a sterilizing efficiency of 99.5^, 99.1^^

and 99*9^ respectively in three trials when the utensils were hold in the

sterilizer S minutes. Ik>t only do the averages of those different trials

show a high degree of offieieney. but in no inetanoo in any of tbeeo trials

with any of the sterilizsre did the sterilizing effioieney diwp below 99.(

The bacteria contamination of the original eans. measured as previously

outlined, varied greatly, eoctending to as high as 14 million per oo.

 47

After BteriUzatioai hoverer, tha hlghaet Daoterial oomt obtaiaed la 39

54
trials was 31 bacteria per oc«

TbeM results iadieate that the oabiaet-type eleetrloally-heated eteam

sterilizer bas a degree of sterilizing effici(moy> that of the atari*

lizers studied thesre was praotioally no difference la efficienoy, and that

leaTing the utensils in the cabinet longer than 5 minutes after the

tsE^perature is reached is not necessary* The third point is of particular

iaportanoe when time is a factor in the sterilizing processi especially

when more than one run of the sterilizer is necessary to handle all the

utensils*

Blevea trials with each of the eterilizers averaged £2*7 minutes*

85 minutes» and 31*1 minutes respeotively from the time ^e electricity was

turned on until the thexmostatio control cut off the heat* These sterl*

lizers required 8, 16*5, and 18*0 pounds of water respectively for ateri*

lization, and the thermostatio control operated at 190°, 193°, and 197^ F*
Thus, the time of operation Is related to the amount of water heated for

steam and the oat*off teoo^erature* Therefore* when the utensils are held

in the eabinet for 5 minutes, the ocs^lete sterilizing prooess varies from
54
88 to 3d minutes with the three sterilizers*
AA
Fasson and Hotis made some studies on the care of milk utenells on

the farm* They say that if the steam is evenly distributed in the oabinet

sterilizer* and a reliable thermometer in the top of the eabinet showe that

a temperature of 800° F* or more has been maintained for 5 minutes or xoore*

it is certain that the number of bacteria on the utensils will be greatly

reduoed; a period of time longer than 5 minutes gives a margin of safety*

 48

Utensils aaoh as eaas, pails* and bottles should always be plaeed in the

oablnet in an inverted position* If they are plaeed open end up it will

tahs Innger to heat the utensils, and the condensed stem cannot drain from

thenu

Results show that the bact^ia contained la 10»gallon cans after they

had been washed and rinsed were practically all killed by steaming the cans

In a Oalvanlzed-iron Box Steamer* lach of these eaiui contained on the

avereee at least 80,000,000 bacteria before being treated, as compared to

46
S,100 after the t3*e8taent»
46
Paason and Hotia say that the effectiveness of the Jet depends upon

the steam pressure used, the size of the opming throu^ vhieh the steam is

ejeoted, and the length of the tium the utensils are ateamed* They aay that

it usually requires about half a minuta to steam a lO-gallon oan thorou^ily

if the staem pressure is BO to 85 pounds* Where a atosm Jet la uead, the

utensils should be steamed until they are too hot to handle with the bare

hands* After trea-^nt in this mamer tfaay wiU baecaoe dry from their own

heat if placed right«sida~cq> and left tuxcovered for a few minutes before

they are inverted on the reek*

'Sto influence of an unsteamed bottle filler upon germ content of milkf

47
eooording to Pxnieha, Whoeter, and Chambers , idxen the bottle filler ime

carefully washed and steamed, exerted no appreciable effect upon the germ

content of the milk paasing through it. When it was similarly washed hut

not ateamed, the gexm oonteut of the milk in the first bottles was inereased

on the average by 96,900 bacteria per cc. The continued use of the bottle

filler gradually washad the larger part of the gexm life txom. the machine,

thereby reducing the contamination*

 49

I>ry Heat* A <KHB!parati'V9l7 reosnt deraXopanat Is that of 8o*oallftA

**dr7 heat" sterilization of milk utensils* The method ooisaonly so^loyed is

to transfer the utensils direotly from the wash vat into a speoially

designed cabinet which is heated with electricity or scnes efficient type of

burner* This method* when properly used* is very efficient* It has the

additional adtantage that the utensils ere absolutely dry at the eonpletion

of the sterilizing process*

43
Nicholas, Sperry* end Tonney made the following report on Humidified

Hot Air sterilizers: Two pails were used as a oheok on the bacterial

efficiency of this sterilizer* Saaaples for bacteriological analysis were

taken from each pail by rinsing with one liter of sterile saline solution

before and after smehanioal washing and hot air sterilization* These pails

were hand washed in water* using an accepted dairy cleanser* rinsed in clear

warm water* drained and plaoed in the sterilizer for treatment* The results

obtained from a direct plate count* using one cubic centimeter of the rinse

water as a unit of eomparison* show the efficiency to be over 99^

14
Cahlberg and Marquardt conducted ezpe rimsnte with an ^sco Sterilizer

idiioh had an electric heating unit and a mereoid control* Cans were plaoed

in the unit to determine sterilization efficiency at tenperatuzms ranging

from 160^ to 200^ F* fbr various periods of holding* In this work the cans
were washed and then rinsed with SOO eo* of contaminating water containing

a miscellaneous mili^grown infection and a high contamination of colon*

aerogenese bacteria as in(*ioatod by gaseous fermentation* One contaminated

sen* after draining* was washed with 500 co* of sterile water* From this

sterile water rinse, 0*1 cc* and 0*2 eo*, and 1 eo* were plated in duplicate

to obtain a knowledge of the bacterial condition of the cans*

 50

In later wxk., 1 to 100 and 1 to ItOOO dllutlone were also mde* %e

results were all giwen as the haeterlal oontemlnation per ean, this figure

haTing t>e6n oaloulated from the count on 1 oo« from the 500 oo. (ahout

1 pint) of sterile rinse water used in each can* After the heat treatment

the oans were rinsed with 500 oc* of sterile water* and 0*1 and S eo*

portions were plated* Also a plate eount of 1die contaminating material was
14
made In duplicate in a dilation of 1 to 100*000»

llie contaminating material was prepared as foUowst Into a solution

oontaining 1% of raw milk and of dehydrated broth, 1 ga. of dried

pulverized cow manure was added per 100 ec* of solution. This solution was

inoubated for 34 hours at F. This contaminating material was mixed

flora representing many types of organisms eneountered in the dairy industry.

The data presented in Table I gives the preliminary results In which

the tei^eraturea were secured by dry heat and by steam in the sterilizer

heated ly gae. They show that either steam or hot air can be succsssfuUy

ussd to sterilize utensils on the farm. Exposure to a temperature of 21g^ F.

for 15 minutes was sufficient for either steam or hot air. The results show

further that a teunperature of 230° 7. for 25 miimtes gave only alightly

o 14
better results than 212 F. for 15 minutes.

TABLE I

samuxf 07 IOTTIAL trials OCaiPARIKO sttoilizatiok of cars bt steam and by

HOT AIR

No. of Type of Temperature Time in Bacterial content of sterilized cans

Trials P.eat Degrees F. Minutes Minimum No. Average Maximum

5 Steam 212 15 500 4,500 10,000

22 Hot air 212 IS 2,500 6,500 13,500
5 Hot air 230 25 0 2,500 10,000
 61

TIm oema irere washed before heat treataeat with 500 oo» of water

containing in excess of 100»000«000 organisiQa per oc« After the heat treat»

swnt the cans were cooled and rinsed with 500 co. of sterile water. The

bacterial counts represent the total nuoher present in the water after the

rinsing.

Chemical sterilization. Many studies amploying the use of chlorine

sterilizers in the dairy industry have been made during the past few years.

Their introduction was first opposed on the ground that chlorine may be

intentionally placed in the milk as a praservative or gain access by accident,

OK

thus reducing its quality. BSle and Heecher state that active ohlnrine,

in hypochlorites, acts as a germicide idien put directly into milk and that

bacteria counts are redticed in proportion to its concentration in the milk.

59
Zoller studied the rate of decomposition of hypochlorite and showed that

sterile milk reduced the active chlorine content as rapidly as milk contain*.

lag large numbers of bacteria, from which he concluded that chlorine in the

form of hypochlorite whem placed in milk exerted no selective action upon the

chemical con^lezes of the bacterial cell. He further ixi^lies, differing from

Halo and Bleecher^, that the gexmicidal action of chlorine in milk was
negligible in concentrations idiich did not affect its flavor and odor. The

studies of loohoad and Johns33 and of Puxoha47 tend to support the eonolusions

that the germicidal action of chlorine la relatively low in the presence of

milk. The former subjected it to treatment with various eoaounts of hypo

chlorite for 18 hoxirs without materially reducing the bacteria count or

changing the oxirdling time of treated and untreated milks. The latter states

that 5^ of milk in a chlorine rinse solution practically destroys its

germicidal actions.
 58

5
Baksr , studying sewage disposal, states that aotlTe ohlorlne showed

greatttP geroioldaX powers thra. did other oxidation agmts, suoh as pertBsn*

gttoate whloh possess a higher oxidation potential* He concluded that its

ability to kill bi^teria was probably the result of reactions other than
S 33 4i2
oxidation* Ayyar , Loohead and Johns , and liyeTB and Johzison found that

chlorine coiqpouQds decreased in germloidal effeetiweness with increased

alkalinity*
40
Madge and LawXer found that the extent of the gexmioidal action of

alkaline solutions wm detezioined largely by the neasiire of tioe, t«EQ>era-

ture, and pH values* Mjers^ found that increasing the pH of alkali

washing eoBQ)ound solutions augoanted; whereas, a sisdlar inorwient in

chlorine solutions reduced their gexmioidal effectiveness* He also

ooB^ared the effectiveness of neutral sodium hypochlorite solutions contain

ing 100 parts per million of active chlorine in water to a solution of 0*5

normal sodium hydroxide and ftnmd chlorine to be considerably more effective

than the alkaline solution* A neutral sodium hypochlorite solution proved

to be much more active than an alkaline solution, destroying 99^ of suspended

spores in less than a minute; whereas, it took 22 minutes to aoeoBQ>li8h the

seme results when alkaline hypochlorite solutions were used*

Chloramine products have always shown less efficiency than hypochlorites*

47
Prucha reports that 200 parts per million of chlorine from chloramine-T

ocm^toundB gave good results in practical tests, but equally good results

were secured when only 50 parts per million of hypochlorite solutions were
S3
used* Lochead and Johns tested represantative chlorine oompounds by

observing their action in reducing the smabers of viable bacteria derived

trom iHtro eulturvs of tha types oonBaonly preseat in milk and found ehloramine
4S
products mieh slower in action tlun kypoehlorites* ^ers and Jolmson and
41
%er8 seeurad similar results* Thay reooismend higher conoantration and

longer oontast periods idiare chloramiBS products are used instead of hype**

ehlorites*

■^parently* the available ohlorine content of a solution* vdimi

detemnined by chemical tests* does not giye a true pieture of its potency
42
idien used as a gezmioide* i^yers and Johnson studied the geznicidal

effectiyenass of twalye eotanaroial ohlorine compounds relatiye to their

ohlozlne concentration* Their obaeryations seean to agree with those of

33
Loohead and Johns * as well as preyious studies made by ISyeaen* That ie*

their results indicate that hypoohlorites i^ch approach neutrality axe laore

effective than tiiue alkali types* althouj^ the ohlorine concentration of the

hypoohlorites may be only half as great* The organic o<Hq;K>«nde studied*

although IStiey were acid in reaction* required double the strength of chlorine

to give results as effectlye as those afforded by hypoohlorites of low

alkalinity*

Tonney* Qreer* and Leibig®® found that extremely low conoentratione of

ohlorine would destroy bacteria when foreign organic matt^ was absent*

B* coll* the most ireslstant of 503 strains of the 48 species studied* suoeuntb

in from 15 to 90 seconds whan 250 parts per million were used*

34
The following ooB^touzids were used in emperiments by Loveless and the

results designated by letters assigned to Idheeu sodium hypoohlorite (A)*

caloium hypoohlorite (B), ohlonaiUfT <D), calcium hypochlorite with soda

ash (B)*
 54

Table II shows the results seoured from tests made with three types of

hypoohlorlte eompotmds to detexmlae their oomparative effeotlTeness in

killing the sasoe test organism* The solutions contained approximately 100

parts per million of actire chlorine* Triplicate tests were made and the

aTarage percentages of these surviving determined*

TABLB II

GOTIOIDAL POTOTOY OF VARIOUS TYFSS OF CHLORINS PREPABATIOKS AOADTST B. COLI

Percentages of bacteria BurriTing

1 Solutions 100 p*p*m* 106 p*p*m*
Seconds Initial
Exposed Count A B E d2
15 165,000 0 2*0454 3.7621 17*57
30 440,000 0 0.8181 1*0122 3*63
45 573,000 0 0*0000 0*0000 0.311
60 578,000 0 0.0000 0.0000 0.0346

^ Initial counts of the diluted broth culture used*

^ Also contained 0*1^ ^2^4*
The results seem to indicate that the hypoehlorltes act wary jrapidly,
34
and that so far as this organism is concemed they are very effective*

TABLB III

PLATS COURTS ON B* OOLI INDOULIW AFT® TRSATMSRTS WITH VARIOUS CHLORINB

aSRfUCIDES COmiNDTO APPROXBSATBLT 200 p*p.m* OF AVAILABLE CHLORINS IN COLD
SOLOTION. INITIAL COUNT 15,400,000.

(Six trials for each ccaapound)

Colony plate count after exposure t|0 chlorine
Compound 15 sec. 30 sec* 45 sec* 60 sec*

E (Average count) 6,133 1,533 67 33

B ♦» " 0 67 33 133
A » « 0 33 0 0
D W W 939,000 89,000 18,233 533
 m

Cociparison of the results obtained in these later trials with those

in which a solution containing approximately 100 p»p#m« was used show a

similar trend but not to the semie degree* l%.e increased concentration of

awailable chlorine gawe inoreased genoioidal efficiency, eren in cold

solutions* Aoidifisd and hot solutions were wary efficient in Mib destrue*

tion of B. Coli* It would aeem that from the resiilts obtaiiuid ueing

sterile water at 180^ F*, that some of the increased affioieiMsy in the hot
solution is probably due to its tesiperature as well as to its aotiwe

chlorine concentration or content*

Tests were also made on four gexmleidal products used in praotioal

farm ezperlmsnts. Cold water solutions were made up in 2CX> eo* of sterile

water blanks to contain approximately SCO p*p*ia* of ayailable ohlorine*

The same inoculum was used*

TABLE IV

PLATE COTJOT ON MILK CAN RINSE INOCULUM AFTI® EXPOSURE TO OERMICIDAL SOLU
TIONS CONTAININO 200 p*p.m* OF AVAILABLE CHLORINE IN COLD SOLUTION.
INITIAL COUNT OF IHOCUICll 3,110,000 per ce.

Colony Plate Count after e23>OBure to ohlorine

Compound 15 sec* 30 sec. 45 sec* 60 see*

5 (Average Count) 10,513 4,567 2,133 433

B u n 133 67 0 0
A * * 0 0 0
D !♦ « 46,200 35,500 15,000 6,033

The results obtained in these trials further indicate a more rapid

action of inorganic chloride ccm^otmds as compared to organic forms. Liquid

«}dium hypoohlorite apparently has the greatest speed and efficiency of the
 56

eampoonds tested. The amalX eurrlTal In the ease of all hypoohlorltee

tested had a tendonoy to make this type of germicide preferable to othera

for rapid rinsing with cold solutions when qpaed of action and effioienoy

were considered.
34 . »
The above results seoured by LoTeless indicate that (1) when

solutions containing 200 p.p.m. of available chlorine were used, the higher

concentrations inereaaed the germicidal efficiency in cold solutions;

(s) acidified and hot solutions were even more efficient; and (3) idiere

sterile water solutions at 160° F. were used containing 200 p.p.au. the
efficiency was apparently greater than in any of the other trials.
Stor&ji^ Temperature*
Tha nuober of baotoria found in milk at deXiToxy tim dapanda raxy

largely upon tba tanparature at vhioh the milk haa been kept during storage*

The bacteria oonmonly found in milk have an optimum growth tans>eratuze of

around 95® F* At 50® to 70® F* the rate of growth is much slower, and at
o
4rO F#. and below the rate of growth is wary slow* However, a few types of
30
organisas produce some growth at fraesing tesDQ)«ratures.
30
Kially and Baboock Illustrate the rate of bacterial growth as shown
in Table Y.

TAILF 7

GROOTH OF BAOTSRU IN MHJC WM TH8 MILK IS TTTCTJI AT 30® and 60** F.

Tamperatuxas Number of badberla per cubic centimeter

of milk
at begin at end of at end of at end of at end of
ning 6 hrs. 12 hrs. 24 hrs. 40 hrs.

50® F,.• , 10 12 15 41 62

68® P. ♦ , , 10 17 242 61,280 3,574,990

At the rate of growth as shown above, milk with a oount of 1,000 would

inerease to 4,000 in 24 hours idien held at 50® F.; whereas, the sans milk
held at 6a!® ?• would have a oount of 6,000,000 bacteria per cubic centimeter.
27
Hnmer found that the storage teaperaturas not only influenced the

rate of growth, lut also had an effect on the types of organisms that wotild

develop. At tnaperaturee around 70® F, the develoim»nt of the S. Lactis

organism is vary rapid. These organisms tend to utilise the bacterial food

at 70 F,, thereby checldng the growth of the other types present.

 88

56
Iborston and Olaoa say that pasteurization of low ooimt milk sometlmss

produoea a renin-Xito eaaanas idiieh oauaes a alow deTeloi^Mnt of acid, and

coagulation of the nilk at low acidltieB» possibly due to the change In

56
baeteirial flora, fhey found no tendmiey for pcq^tonizing bacteria to out

grow other types at storage temperature of around 40** F* Hbwerer, there was
a decided tendency for the pastetirized-bottled milk to dewelop a "cappy**

flawor after two days storage at temperatures of around 40^ F*

All milk should be held at taopax'atures not higher than 40^ 7. until
delivery. There is only a slij^t increase in bacteria in pasteurized Biilk
29
d\iring the first 84 hours wh«a held at this temperature.

Acidity.

Hi^ acidity In market milk is an indication that the milk is being

soured by idie action of bacteria on the milk sugars. This is, of course,

a very undesirable characteristic. This acid flavor and odor, when present

in milk, is noticeable to the average consumer at an acidity of around

•3 per cent, while e3Q)arlenoed judges begin to notice the imdesirable flavor

at around .2 per cent, because it does not have the desired keeping
3X
qualities.

Acidity in milk may be attributed to two causes, (1) the popssenee of

acid phosphates, and (2) the formation of lactic acid as decomposition

products of milk sugar which has been acted upon by the bacteria commonly

found in milk. High acidity may be best prevented by quick cooling to a low
31
temperature, and holding in this state until delivery.
 59

Bheplag Qoallty*

Ono of the prliaary requisites of nertet mllic is that of gooA teeplag

qttalities. The consumar desires zollk vhioh will keep from one laeal to

another without souring* llridentlyt the foregoing pages hare oorered oany

of the faotors wfaioh influenoe the keeping gtuadity of milk* A oonhinatlon

of oooXing* pasteurization, and beoterial eontrol largely determine the

leng'ttx of tisis that milk will keep*

Conn found that a speeimen of milk whioh had been held in a cold

room for four days had a final bacterial count of ten million bacteria per

oubio cffioitiBetw; whereas, a sample of the same milk after standing in a

wazm room for sewan hours Inereased a hundredfold*

In 03Cder for milk to keep well, the producer, distributor, and

eonsumar must oooperate to prevent exposure to warm te^eratures, sunlight,

and unsanitary surroundings*

 60

Staaaary of Surrey of Literature*

!• There are Basy feeds which haye a tendency to t^tart a eXl^t oft-

flayor to BdUu

&• Feeding silage soon after loilking produces a better flayoxed milk than

feeding it before milking*

3. Garlic and wild onion usually produce undesirable flavors when cows are

allowed to sat them anytime within several hours before milking*

4* Relatively few of the concentrate feeds cossnonly fed to dairy cows

pzoduce objectionable flavors in milk*

5* The flavor of milk after secretion cannot usually be improved; there

is a tendency for it to deteriorate*

6* Ordinary pasteurization tenQ)eraturea have a tendwucy to injure the

flavor and creaming of milk*

7* Raw milk forms a maxliaaa cream layer more quickly then pasteurized milk*

8* The direct microscopic» agar plate counting, and msthylene blue reduc

tion test are satisfactory methods for making bacterial estimates in

milk.

9* Duplicate samples of the reduction method check closer than duplicates

for the other methods in determining the bacteria in milk*

10* In general* the quickest method for cooling milk is the most desirable

from a quality viewpoint*

11* Approximately 99^ of the baoteria coBmoxUy found in milk Is killed by

pasteurization*

12* ISffeotive sterilization of milk utensils and equipment may be aooom*

plished by the use of steam, hot water* dry heat* or ohsmioals*

IS* Chemical sterilization is appsrontly the n^st effeotive method for

use on milk utensils and equlpsmat*

14* Relatively low storage temperatures are desirable for holding market
milk*
 PARf II. ORIOIHAL INTSariOATIOH

Objoote of Investieatloa.

Xm To stuidy tho influe&eo of tlM» followiag faoiors on tho

flatov of BtarlsBt mllki

Aeration

B« Pasteurization

S* To study the oreaming of market milk as affeoted hyt

A* Pasteurization taoqieirature

B. Storage tizos

3« To study the baoterial oontent of mariost milk eoxd Its

relation toi

A. Cooling

B. Pasteurization

C« Sterilization of oaiis

ftl
 6S

PXan of lavaatlnatlon.

This ixi-rosti^tioii wea aaaentlBlly dividad into three aeotiono,

so that a detailed atiidy of several of the faotors vhieh Influenee the

quality of narket millc ooald be niade* All of the faotors lAleh determine

quality do not enter eaoh milk distributor's probl(Ha, even though the

fundamental principles are olosely relateda

A raw milk dealer is not confronted with the partial destruction of

the oreaming ability of milk due to heating; vhereasi this ia a eoioBoa

problem in the pasteurization of milk* On the other hand, milk ehioh has

been properly pasteurized and given careful attention after pasteurization

ia said to be free traa any danger of cexrying disease^produoing bacteria*

Pasteurized milk very often has a slight cooked flavor. This flavor

is not objectionable to n»>st people, especially after they have become

accustomed to pasteurized milk* However, the pasteurized flavor has led to

oonsideirable objeotlon on the part of some oonsun^rs*

The plan of this investigation included a study of seme of the different

flavors c<xanonly found in market milk, the oauaea for the preaenoe of theae

flavors, and an effort to deteaanina the factors neoasaary for preventing or

reducing the intansity of aome of the undesirable flavors by methoda

praotioad by oomnarcial milk diatributoira*

It was farther planned to study the effect of pasteurization tempera*

tores and storage time on the creaming ability of milk by determining

variations whioh would normally be noticeable to the consumer.

An attffispt was made to deteimine the effeote of cooling milk on the

bacterial growth, the efficiency of bacterial destruction by pasteurization.

 es

and tho effaet of aterlllzation of eans as a aoorca of reducing oontaaisa^

tlon eoaanonly encountered in tlie handling of laartot milk. Sterilization,
as applied in this problem, shall refer to the partial destruotion of the

bacteria preecmt.

The plan of the inrestigation was outlined so that most of the trials

could be made on a eoraaercial scale, and not on a laboratoxy scale which

would not be practicable for modem dairies and milk pasteurizing plants.

Methods of Proeedure.

The major part of this investigation was limited to problems directly

related to the market milk industry# Most of the problems are very ocammm
in creameries# The data presented were secured from eoperiments oon5>arative
to methods ocmmenly employed in the commercial routine of hondUng aiHc,
The equipment ueed for the experimental work was designed for modem dairiea
and milk plants, and for technical work in quality control laboratories#

The milk used in this study was obtained frma the University of
Tennessee dalzy herd and the I&ioxvllle Mlk Producers* Association# The
Uhivereity herd at the tim of the eapeariiaent consisted of Holstelns and
Jerseys, while the herds supplying the Association were of mixed breeding
with the Jersey breed predominating#

Duplicate samples of each lot of milk were taken for each trial for the
flevor tests# The sanq>les were held in quart milk bottles except as other

wise indicated. These samples were held in the storage room at 40® F#
When the milk was ready to bo scored for flavor, it was waxmed to a
 64

t««jporature of fre»n 80** to 90® F. and aoorad at that temperature.

ISight students, with considerable experience in the judging of milk,

were aTailabls for scoring the seaaplee for flavor. Three students were

used for eaoh series of samples; however, the same three were not always

present throughout the Investigation. These three students were selected

from two different judging teams. >ach of the two teams made excellent

records at the Students Hational Dairy Products Tudging Contest, 1935 and

1936.

A detailed procedure of the scoring swthods may be found in the

section devoted to flavors of milk.

The creaming tests wore made by two different laethods. When the

experiment was firet begun, the cream volume was measured as it appeared

on the bottle. The chief object was that of representing the comparative

cream volume of the milk as it could be seen by the oonsumier. This method,

however, v^as later replaced by the use of 100 ec. graduate cylinders* The

preparation and handling of the samples will be discussed under the section

devoted to the methods of meaaurinf^ the creaming of milk.

The bacteria analyses in this investigation were based on the technique

as given in the standard methods of milk analysis and reoommended by the

American Publie Health Aesociatlon. The reductase tests were conducted in

the dairy laboratory at the University of Tennessee. The agnr plate counting

was conduotsd, in part, in the dairy laboratory of the University of

Tsnneasee, and, in part, by the milk sanitation division of the Knoxville

Bureau of Health. The methods used for securing the data on bacteriology

studies will bo given in the section devoted to the bacteriology of market

milk.
 «9

glaTors of Magtet

S^lk is eithc»r pleaaiog, fXat» or dlsagnwabla ia flator to the

ordiaary ooaawuHr* gortunatoly, tho fluid milk ludustry has sstabllshed a

reputation for good-flarored odlk* As a result of the oonfidenee vhioh the

eonsumsr has for the palatabillty of milk, he Is usually not suspioious of

off-flaTors being presosit* fhia psyohologioal faatxxra is perhaps more
favorable toirard milk oonsne^tlon than for the consoaptlon of zasay of tlM
other so-oalled essential foods*

iSarket milk has the endorsement of noat of the leading physiolaxxs of

the nation* It has consistently gained faror with nutrition specialists

and Tarious other health authorities* It is generally assmed that all

noimal milk is palatable to the consumer* As suggested above, fortunately,

milk is usually palatable; however, there are many faotors eonnooted wi^

the production and eare of milk which have a tendenoy to eanse abnorml

undesirable flavors* These off-flavors have been attributed to abnormal

oonditiona of the cow, iatproper feeding methods, and earaless hundHwg of the
milk*

Bie problem of preventing and controlling off-flavors vdiioh are same-

times tx^arted to milk is sonsidist oomplex, and yet undesirable flavors are
eeldma reported by the oonstimer* fhia is probably due to the seientifls

knowledge and effort en^loyed by the produoer and distributor to reduce the

intmasity of the flavor to eueh a point that it is not notioeable* Sens-

times the oonsuoer fails to notice the undesirable flavoars which are

actually present in the milk* Nevertheless, an investigation to include a

study of the factors influsnoing the flavors of milk should prove very
beneficial for both the milk industry and the milk consuming public*
Mathods of Seorlng.

The swthoda anployed for seorlng the «aiQ>las of milk for flavor in

this Investlgatioa were similar to those reoojoooadod hy the Rules Coaraittoo

for the "Students' National Dairy Produots Judging Contest Asx. illustra

tion of the student score eard used in 19Sd is giTsn in Figure 1« The

ailk in all of the trials on flavor iras scored on a eoaparative basis using

the student score card for a guide* The method of sampling will be

indicated later in this section* In order to make this a ocng>erative study

of s«BS of the different factors which affect the flavor of milk, a number

of preliminary trials were oondueted so that good, medium, and poor

flavored samples could be selected for the ezperimental work* These

preliminary trials with sai^les from the different milk patrons shipping

to the nnlversi'ty Creamery ware helpful in selecting: (1) the best flavored

milk being received at the time of the experiment, (2) a typical silage

flavor, and (3) en unclean flavored milk* These series of saiiQ>les were

selected for the purpose of studying the effect of aeration on the flavor

of milk*

Influence of Aeration*

ttmy investigators have found that aeration during the cooling process

helped to reduce the off-flavors inparted to milk la^ oertain feeds*

Aeration may consist of allowing warm milk to flow over a surface

tubular cooler containing circulating water, or water and brine as the

cooling agents* This practice rests on the theory that whean warm lallk la

exposed to the air in thin fllme, the air has a tendency to rssiove some of

the feed flavors which ere oftma present*

 MUX SCORS CARD

For Students* National Contest in Judging Dairy FrodiBOts

Contestant No. Sanple No*

Grades
Criticisms
Studmt Omeial Score Criticism

yiecror
FlaTor (25) « Bitter Flat
Cardboard Garlic or onion Rancid
Cooked High acid Sallqr
Sediiomt (ID) Cosy mty Uhclean
Disinfectant Metallic Weedy
Feed Musty
Bottle and cap (5)

Sediment
Placing
Score cottons as photograph in U* S*
Depairteant of Agriculture Ciroiilar S84 *
Grade on oritioism
Bottle aaad Gap

Absorbent cap paroteotor Lip not protected

Total Grade Chipped mouth Lip partially protected
Dirty bottle Loose cap cover
Leaky cap Not full

8~701S U* S. Goverxment Printing Office

Figure 1* Student score Card.

 68

Zn thXa investIgatIon» seniplea of milk wore obtained from tso different

patron*B ebo were shipping to the IMiversity Creamery* Speoial effort was

made to seleot samples from the best flavored milk being received at the

time the ezperiment was started and from a relatively poor flavored milk*

These trials were oonduoted during July 1936, at whioh time most of the

milk being received at the creamery had a slight feed flavor* The same

patron*8 milk was used for san^les througd^out each series* The data shown

in Tables 71 and 711 were obtained from samples and duplicates which had

been taken from the warm cans of morning's milk, divided, and cooled Iqr

two different methods* The temperature of the milk when received was

between 75^ and 60^ F. Part of the milk was put in regular quart milk
bottles whioh were izmiediately subaerged in ice water end cooled to 40° F*
Mother portion of milk fzxsa the same can was poured over the tubular cooler

(Figure Z) until Ihe tao^erature at the bottom of the cooler had reached

40° F*, at whioh time samples were taken* All of the saoples were placed in
the milk storage room and held at 40° F* until they were ready to be scored*
The tima required for cooling the samples in glass bottles surrounded

by cold water from the origixul taBU>erature to 40° f* with the tsECQ)erature
of Me water from 34° to 36° F* ranged from 1 to 1^ hours* This is
representative of the time required to cool milk in cans in electrically

operated units where the water is agitated around the oan* The tiros

required to cool the milk over the tubular cooler with a circulation of

larine at 80° 7* through Me coils was lees than one minute*

The ea&^les were scored for flavor during the training period for Me

dairy products judging team* The students wers not aware of the treatment
 69

Cooling Iqr Aoratlon

figure S* Surfaee Tubular Cooler

 70

wlxioh had bean giToa the eaioplee* They were« however, requested to plok out

duplioatee, or determine diffeareuoea where poesible* When disagreement as

to Booree to be given to a ssBQtXe existed, averages were reeorded provided

the differenoes did not snount to more than one point* When the differenoee

aeoTuted to more than one point, a seeond seore was seoured by passing the

sample arotind after aeveral others had been eoored*

TABUS VI

sooras for good flavor® milf coolrd by diffyrot methods

Coolixig Method

In bottle Ae3?ated

Trial So ored SCOred

6 hr. 34 hr. 6 hr
. at hr.

1 Seore 22 21 22 22

2 Score 22 21.5 22 18

3 Soore 22.5 22 22 22
''

Average £2.16 21,16 22 20,66

The average soore at the six-hour period was &2«16 for the samples

oooled in the bottle as oooq^ared to S8 for the samples cooled over ^

aerator. At the S4-hour period the average score for the samples oooled in

the bottle was 21«lft as ocsitpared to 20«d6 for the samples cooled over the

aerator.

The resolts as given in Table VI indicate that there was very little

differanoe between the flavor soores of milk oooled in bottles without

agitation and that oooled over an aerator. The original milk was considered

good in flavor with only a slL#t feed flavor present* These results
 n

IMioata farther that eooling lollk of good flavor vithla one to t«o hoars

mus juet as satisfactory as to flavor aoore as cooling aore quioldy*

Preliiainary trials shoired that 8aiiq>le8 ooolod soon after dram soored

hii^iuKr in flavor at the 0 and 34~ho\ir periods than at the 48»>hour period*

The data In table VII show the effeots of aeratim of milk on a strong

feed flavor and slii^tly tmolmn in flavor when handled in a similar

manner to ths sables used in the above experiments*

TABLS VII

SOORIS FOB POOH FLAVOBSD MILK OOOLtD BY DIFFSH®1T MITHCfDS

Cooling Method
In bottle Aerated
Trial Soored Soored
0 hr* B4 hr* 0 hr* 84 hr.

1 Soors 17 18 17.9 18

8 Boors 18 17.9 18 18

3 Boors 17 18* 17 10.5*

Average 17.33 15.83' 17.5 17.5

^Criticized hi^ acid*

The average score for the &»hour period for the sasples that

cooled in the bottles vas 17*39 as ooopared to 17*9 for thorn ooolsd over

ths aerator* The samples cooled in the bottles had aa, average score of

19*^ as oonparsd to 17*5 for those eooled over the aerator at the 24-hoar

examination*

The results as given in Table VII Indieats that aeration la these

trials failed to remove muoh of the strong feed and unoloan flavof from
 72

lolllc irhloh ««» d«oi4adly poor la flavor. Thigr «how further that verjr

little change in flavor is brought about betvam the 6th and 24th-hour

eacsmination, ezeept in the third trial vhere an off«flavor waa present*

fhis was possibly due to aoid development whieh appc^^tly had taken plaes

at the 24th-hour examination*

A series of trials was conducted to datejsmlne the effect of aeration

on milk with a noticeable silage flavor. This milk was takan from the

tJniversity herd during October 1935. At that time eaoh of the cows was

being fed from 30 to 20 pounds of com silage per day at milking time*
They were also getting a regular grain mixture oonsiatlng of com and cob
meal, wheat bran, oottonseed meal, and all of the good hay they would oat.
This milk was taken from the morning milking which consisted of a mixture
from the Hoisteins and Jerseys Just as they were normalLy being milked*
The milk was diwia by maohine, after which it was poured into
40-quart cane* When the oans were about three-fourths full, the milk was
well mixed and the ten^erature taken and found to range frcaa 82® to 88^ F*
Half of this milk was set la the eleetrioall;sr operated Daniel's milk cooler
in the milk room at the bam and cooled to 40® F* Bie time required for
cooling was from 1 to hours. The other half of the milk, at the tem
peratures of 82® to 88® F., was carried to the creamery and cooled over the
tubular cooler to 40® F. This method required less than one minute from the
time cooling began* Hot more than twenty minutes elapsed from the time the
milk was first mixed until it was cooled over the aerator. Sauries were
taken in dt^lioate and scored as eaq)lained in the preeeding trials*
 73

TABLB Vin

SCOBSS TOR HSDIQU TLilYDRSD MHJC OOOLHD B7 DIFFSRBST

BIHTHODS

Cooling ;,fathod
In can 1 Aerated
Trial Scored Scored
3 hr. 24 hr. [ 6 hr. 24 hr.

1 Score 21.5 21.5 21.5 21.5

2 Score 21.5 21.5 1 22 21.6

3 Score 21.5 21 1 21.5 21

Average 21.5 21.33 21.36 21.33

The aTorage soora for the aansplea that vere cooled In the ean at the

elz-honr period was 21«5 ae oonpared to 21»d6 for the epistles that were

aerated and aeored at the siz-honr parlod. The aaaplea that were eeoled

in the ean and those cooled ower the aerator had the aaxne anrexage aeore of

21*33 when aeored at the 24-hour period*

The results aa giwen in Table Till indloate that aeration had wery

little effeet on the flavor of milk which originally had a decided silage

flavor. The scores and oriticiama were practically the same in the milk

just after it wae drawn, after it waa cooled in the can, and after it was

cooled over the tubular cooler.

 74

Paetaurlzation Temperatares.

Pastaurlwtion la unlTarssLLly rMOgoisad aa ona of tlio leading safe*

guards against iaillEi->boma epidemie diaaaasa* Apparently, the general publlo

la'in faror of pasteurized milk aa eridenoed by the rapid trend toward its

use* The milk distributor is interested in the adTanoement of pasteoriza-

tion beeause it isQ}roTes the keying quality of his milk and inereaaes milk

sales* One of the claims which has led to objeetion to pasteurized milk on

the part of some eonsumsrs is that it does not possess a natural milk flavor*

This may be attributed to the remoTal of scrbs of the flavors which are

actually present in the raw milk* Tot example, the heating may possibly

reduce the intensity of certain feed and weed flavors present in the raw

milk, thus leaving a flat taste, or the heating may possibly destroy staae

of the characteristic flavor properties of normal milk* Pasteurization,

evwi though it may in some eases inprove the flavor of milk, should be

aeocoDplished at relatively low temperatures if the pleasing adlk flavor is

to bs retained*

A series of experiments, consisting of six trials with each of the

three connon pasteurization teiq;>eratures, (148^ - 143^® > 145^), were used
in this investigation to determine the effect of pasteurization tacq;)eraturea

on the flavor of market milk* Theee experimenta were conducted in the

IlaiTsrsity Creamery, using the Cherry Burrell epray^type pasteurizing vat*

The milk sanplee used for these uperimsnts were secured from the regular

runs of the pasteurized milk as carrisd through the daily routine at the

oreamary* The only variations from the regular practios were the pasteuri

zation teiaperatureB* A "drip** sample was eaught at the preheatsr (Pig* 3}

 75

Figure 3* Milk Prsheater

whleh serred ae a Mntrol for ooDparing the flavor of the raw and pasteur*
Ized niilk* This aaEsple represented a ooiaposlte of the milk flowing into
the vat after it had been heated to a teoperature of 95® F. and waa
eeoured hy allowing the milk to drip through a olean rubber tubing whioh
extended from the end of the top eoll of the prdheater to a saiall*top
glass bottle whioh was kept la ioe water so that the milk was cooled as it

dropped into the bottle*

 76

of the paatourized milk iruro taknn from tka bottla*filling

attoklae after tkey had be«a held in the Tat for thirty mlautea at their

respeotlTe pasteurization t^eraturee* then partially oooled in the Tat to

to IS^ F*, and the oooling finished to 33^ to 40^ F« OTer a surface
tuhular cooler* These ses^lea with duplicates were held in quart nsllk

bottles in the milk storage room with the control samples which had be«a

transferred to quart bottles until they were ssored procedures elndlar

to those outlined under the heading ■*> Methods of Scoring*

TABLE IX

FLAVOR SCORES FOR MIUC SAMPLES PASTMIRIZED AT DIFFSREHT TEMPBIATUHES

Pasteurizing Raw Pasteurized

T«aperature
6 hr. 6 hr* 24 hr.

142° Score 21.9 22*23 22*1 —•

143^° Score 21.5 22 21.75

143® Score 21*5 22 22»

*ATerags of 3 trials.

Table IZ represents an arerags of six trials using different pasteuri

zing temperatures as indicated* The pasteurized milk from eaeh of the

three standard teiag>eratureB scored a little hi^er in flavor than the

corresponding raw milk. The judges for these ssoaqrles were accustomed to

drinking pasteurized milk; howsTer, they were also aceustomsd to scoring

raw milk sai^les* With the difference between the raw and pasteurizsd

milk being so anall, the consumer would not likely notice any change* Scant

eoasuaere may notice a slight cooked flavor present in the pasteurized

 77

Muagplesj it Imd rai>lae6d a sli^t feed flavor in the rear ean^lM*

Preliminary ea^srinents in NoTemher 1935, oondaeted at Southern Dairies

and Arondale Fame Creamery, ocancieroial plants of l^xville, showed different

reetats from the aboye* Using water at 180*' F* for the heating meditim in
the 00il-type Cherry Burrell -rats, the results se^oed to indioate: (1) no

apparent ohange in flavor of milk pasteurized at IdS** F« for thiarty minutes}

(2) a cooked flavor whcui pasteurized at IdS^*' F* for thirty minutes, possibly
lowering the soore an average of *5 points; and (3) a decided cooked flavor

at 145*^ F* with at least a one-point cut in soore for cooked flavor*

These results, although the number of trials were not sufficient to

draw speoifio oonolusion, indicate that milk may be pasteurized at any of

-the above teaperatures in the spray-type vat without materially affeetlog

its fine fla-ror. They further indicate that relatively high pasteurization

teaq;>eratures have a tmdeooy to produee an uadaelrable eooked fla-ror tHam,

the coil-type vat is used with water at 190*' F* for the heating mediimu
 78

Creaming of M^laat Milk*

The length ot the layer of oream appearing on a bottle of milk when

it is dellwered to the oonstsaer may be Influenoed by eeweral different

faotore* Many of these factors can be o<mtrolled tlw milk distributor

In prooesaing plants, but aom of then are beyond his control. lor

axan^lot he cannot oontxol the breed of animals on the producing farm, yet

the breed sometimes Influences the creaming of milk. This cream layer is

of considerable ocmerolal importance. It is oommonly accepted by the

oonsucaer as an index to the riohnsse of the milk.

The faetors influeneing the eream layer of xoilk, which were studied

in this investigation, have to do with eome of the major problema

encountered in oosneroiel paeteurizing plente. The temperatures to whloh

milk ie heated during pasteurization and the time bottled milk is held in

storage are of vital importance fromi the standpoint of ^a resialtlng oreaa

layer on milk at the time of delivery.

Methods of Ifeasurlng.

The mstiiods oomBonly used for measuring the oreemlng of market milk

are: (1) those whloh reveal the length of the layer as It appsars on the
bottle, and (8) those which make use of graduate cylinders. Both of the

above methods were used In this Investigation. The esoplee for the

oreamlng toete were taken from the milk reoelved at the Ohlverslty Creamery

which constituted the regular pasteurized supply. This milk, as stated in

the foregoing section, consisted of a mlxt\ire from Eolstelns, Jerseys, and

grades. All of the milk was atasdsrdised to 0 butterfat as ihoim by tho

Babooek test*
 n

The »m eenplee iMsre seeured Itjr the "drip" nethod ae described in the

eeotion devoted to flavors. iStaiidardization vae aoooi^liidted by eeparatiag

the required amount of sldmllk and pouring it baok into the reoeiving

tank so that it eould be puo^d baok through the lines, thus giving the

"drip** test a butteirfat test of 4^ The pasteurized saDoyples vere seeured

from the isilk after it had been cooled to 130'^ > 135^ F. in the vat, then
potqped over a surfaoe tubular cooler and cooled to 33° • 40° F*
Meaearing In Bottle. ?lhan the creaming tests vere first begun, the

oreen layer vas measured as it appeared in the bottle* The saB^les vere

secured as described under methods of measuring, ^en the Bangles vere

ready to be issasured for cream voloms, the bottles vere carried frcaa the

storage room to the laboratory and placed on a level table, eaire being

given to avoid destroying the line of demarcation. By extending a pair of

calipers over the entire length of the layer and transferring this

measurement to a rule graduate In sirte^ths of an inch, the length of the

layer vas determined.

One feature vhich makes this method attractive is the volume of oream

indioated that msy be 8e«a by tlie consumer*

Measuring In (Graduated Cylinders* This method of measnriag the

erseoing of milk required the use of 100 co, olear glass eylindere

graduated to 1 eo* divisions* The cylinders used in this inveetigation vere

approximately seven inches in hei^t and of an. even diameter of about one

irrnh. Bjr using eylinders of this heii^t, the fat clusters rose about the

s«M distanee as they normally vould in a regular quart milk bottle. Other

features vhlch oaks this method attractive are that the reaolte are easily

interpreted, and they may be skpz^ssed in percentages*

 ao

Sflnq;>l0S and duy^lloatos for iiia croamirtg teets in the cylinders were

seewred as deeoribed aadar the mthods of measoring* The raw aanpXes vhloh

WBV9 used for controls were talosn fxom the bottle whioh represented the

eoBQ>o3its "drip" test* The pasteurised samples were taken from the bottles

as they cams from the fiUizig nashine* The milk was poured te<m the bottle

to the 100 ee« mark on the cylinders. All of the cylinders were placed on

a IsTsl shelf in the milk storage room end held at 40^ 7* until the
creaming tests were auide*

Pasteuriaation Tmgperatures.

Many inrestigators haws shown that pasteurisation at the tea^eratures

eommonly used in eoBBaereial creameries oausss a dsoreaae in cream woluae

when ooB^ared to the cream Tolume of the original raw milk* lijcst

authorltiea agraa that this dscrease may bs only a slight one wh«i propsiT

control methods srs praotiosd. This investigation Inoludsd a study of ths

three most ommon tan^eratures of pasteurization and their effects on the

creaming ability of milk. The trials on pasteurization were conducted in

the SOOogaUon Cherry Buirell qpray-type pasteurizer (Figure 4)* The basle

for OQBQ>ari8on was the ereem volume of ths raw milk.

ATerages were aseured from three trials using each of the following

processing series: (1) raw milk, (2) pasteurized at 143^ F., (3) pas
teurized at 143|^ F., and (4) pasteurized at 145*^ F. All of the pasteurized
sang>leB were held for thirty minutes at their respeotlve tenQteratures*

Of the three trials* not more than one-alxteenth of an inch variation was

found between the original and the duplicate sauries* Not more than two«

sixteenths of an inch variation was fCund between the individual trials in

any one of the above series*

 m.

»
m

>

■V Asw r Mm

Flgtir* 4« The Cherry Burrell Sprey-type Pasteiirizer*

 88

TABLE X

CPEMDJG OF MILK PASTTOJRIZED AT DIFFEREOT TKMPSRATORSS

AVERAGE OF 3 TRIALS

Creaming Time

6 hours 24 hours

cream ▼olume cream Tolums

Haw (Controls) 3 3/16 inches 3 9/16 inches

Pasteurized at 142® F* 3 8/16 inches 3 12/16 inches
Pasteurized at 143^® F* 3 6/16 inches 3 10/16 inches
Pasteurized at 145® F* 3 SAA inches 3 7/16 inches

Ivtoasuremsnt in t}i« l3ottl«

The results as shcnm ia Table Z indicate that milk pasteurissA in lAis

Cherry Burrell spray-type vat has a smaller cream -VDlone than raw milk at

the eix-hour examination* In general* the hi^er the pasteurization tern*

perature* the SEoaller the cream Toluas* They fturther indicate that the

saaq>lss pasteurized at 142^ and 143^^ F* hare a larger cream Tolaas than
the eorreeponding race samples at the 24-hour examination. Ebeever* samples

pasteurized at 145** F* shoved a smaller eream voluias thsea Ibe corresponding

raw samples at the 24-hour examination*

Influence of Storage*

The time elapsing between the bottling and delivarlng of markat milk

is of considerable importance* The time relationship for raw milk is

different frcna that of pasteurized milk* Raw milk shows its maxiaattaa cream

layer more quickly after bottling than pasteurized milk. The cream layer

on raw milk after reaching a naxliTBum has a tendency to shrink with age*
 83

This shrinkags has bssa attriteitad to a higher oonoentration of the fat in

the oream* The cream layer on pasteurized milh, even though it is usually

a little shorter than that of raw milk aooording to Tables X and XI, did

not shrink as rapidly as that of raw milk*

The data presented in Tables X and XI show the relationship of the

different taEQ>erature8 of pasteurization to the remilting eream layer, and

the relationship between storage and the change in the wolume of cream*

TABLE XI

ojmmm of milk pastsorizsd at DiFFSRsrn? TaaPSRAmRss

A7SRA0B OF 6 TRIALS

Creaming Time

6 houra 24 hours

cream volume cream volume

Raw (controls) Id oc* 1 14 CO*

Pasteurized at 142® F, 14 oe* 1 16 00*
Pasteurized at 143^® F, 14 CO* 1 15 CO*
Pasteurized at 145® F, 13 cc* 1 14 CO*
Msasurament in 100 oo* graduated eylinders

The data in Table U indicate that raw milk has a larger eream toliate

at 6 hours than at 24 hours. Milk pasteurized at 142**, and 145** F*

bad a larger eream wolums at 24 hours than at 6 hours* Milk pasteurized at

142** - 143^^ F« had a larger eream woltias than raw aiilk at 24 hours. The
raw milk at 6 hours had a greater cream wolxnae then any of the pasteurized

milk. The milk pasteurized at 145** F* had the smallest eream volume of the
group; it was the same as raw milk at 24 houra*
 64

Bacteriology of ?terkBt ?311k«

The bacteria la loarkst milk largely detezmlae Its kseplag qualities*

Tbe original bacteria Ibund at the time the milk is drawn are not consid

ered as Bignifleant as those resulting tram multiplication* This may be

attributed to the relatiyely low counts and to the types of organisnai

present in normal freeth milk* Roweyer, milk known to contain only a few

tubercular or typhoid organisms would be considered very dangerous for use

as a human food* There are a number of sources from which bacteria may

enter milk and each of them should be safeguarded against disease-producing

types of organisms* This investigation covers a study of some of the

fundamental requirements which are necessary to produce, process, and

distribute milk with a relatively low bacterial count> and to destroy

disease-prodxioing types of bacteria*

Methods of Analysts*

There are four popular mathods used for estimating the numbMr of

bacteria in milk* Baeh smtiiod has been very valuable to the market milk

indiistry* The measurement of their practical value is influenced largely^

hy the quality of the milk being handled and the desired information*

Some milk contains several different types and species of baoteria*

Raeh of ^e different types In milk has peculiarities which require various

prsperation and treat»aent for detenoixiing their presence. Thus, negative

results for a given type are not at all unoonsoon when employing ecuoe

mathods; whsreas, a change in methods or a oombinatlon of methods may give

entirely different results.

 Both tlMi agaar plate eoimtisg aethod aad tbe aathylene bloe reduotaae

test have been used very extensively for ailk analyses* These teo aathods

were eonipared In this Investigation* The data were seoured from a eeries

of tsrials with the use of eaeh xaathod on milk from the same souroe in order

to get the oorrelation between the two methods* The proeedmres are given

later in this seotlon*

The agar plate oounting method was used for determining the influenee

of eooling on the baeterial eontent of raw milk in this investigation*

This lasthod was also used for studying ths sffioieney of pastsurization*

Preliminary rsduotase tests on pasteurized milk gave inconsistent results*

Some investigators believe that the organism responsible for the produotion

of the enzyme, ald<diyde reduotaee, is killed during pasteurteatlon.

Aooordingly, the color change upon which the reduotaae method is based would

not be brought about, evtai thou^ there weere baeteria present in largs

numbws# The inoonsist results found by using the reduotase method for

tests on pasteurized milk may have been partly due to the relatively Imog

time required for deoolorization* Only a few of the ssagtles deoolorized

in eight hours* Some of the sauries in the preliminary triale were held

until they had eoagulated end the blue eolor still had not

disappeared*

The baoterial analyses for the series of ezperiments oondueted on

sterilization of cans were limited to the Agar plate smthod* The ehief

objeot of these e:i$«Brii!^t8 was to detemdne the effloiemy of seats of tiie

different methods of sterilization* The plate method was used for

analyzing the baoterial content of the rinse water which was used in -^e eans*
 ae

TheM studiaa wero intended for oatlffiatixig tha total ntnabar of baotoria

presant and not a partioular group. HovaTnr, It is genorally agraad tliat

BK>8t of tha bacteria presant in milk are mambera of tba S. laotis group.

Agar Plate Counting. This is probably the moat eonoaoa method uaed for

aatimating tha number of baeteria la milk. It has proved to be» for the

moat part, a very eatisfaotory index to the quality of milk. principle

of -^s method ia based on the growth of indiTidual oella or organiama into

comparatively lar^ colonies. The tern "counting" is perhaps a little

misleading because the individual baeteria are not actually counted. The

oount or estimation la made on the eoloniee whieh are presutoably prodiwed

from a single cell. Some of the unfavorable oritlolsa for the plate method

has resulted from the theory that naay types of baeteria appear in pairs,
and lAen coloniss are produced frcaa their growth, each colony may easily

represent the growth of two bacteria. This is not at all is^ssible.

A colo^ may result from several individual cells and be eounted as thou^

it wen a single baoterium. However, if consistent results are obtained

from the plate method, It stiU has some very distinct advanteges over

other methods. It ia tha ind^, or approximation of numbers, whiOh helps

to satablish atendarda for grading milk according to bacterial content.

The material and equipnmnt used in the egar plate method includei

1. Dilution bottles in which 99 eo« water oan be steriliaed.

8« A supply of 1 00. pipettea for mldjog the dilutions; with one

.1 oc. graduation.

S. A liquifiable solid media for growing the eoloniee. (Beef extraot

nutrient agar)
 87

A steam pressure autoolare for sterillaljag 99 ce« water blanlos,

aad melting egar*

5« A dlry heat oven for sterillziag the plates a:i^ plpettee«

Aa automatie teiaperatare eontxolled ineuhator in whieh to grow

the colonies*

7« A solution of lysol for disinfeoting the worUng table*

In addition to the above equiiment a wax peooil for marking the plates and

a eounting glass are desiTable for part of the work*

The procedures followed in this investigation were similar to Ihose

reeomnendod by the Uhited States ?ublla Isalth Ssrvioe in the Standard

Methods of Milk Analysis* Briefly, they were as follows:

General: Tha agar wma made cMoording to standard methods* Shen the

plates were ready to be potjcrod it was asltsd and cooled back to about 100®
7* The dilution blanks were sterilized at l&-pounds pressure for one hour*

All of the pipettes and plates need were sterilized la the dry heat oven at

SCO® y* for two houre*

I« Dilutione of 1 to 100 were made by pipetting X ee« of the sample of

milk into 09 oe* of sterile water*

2* "Stom thie dilution, *1 ec* and X co* reepeotively were transferred

to plates, giving duplicate plates for eaoh sample in different dilutions*

3* The warm agar sas poured Into the plate and the two Edxsd gently
and placed on a level table until the agar had aoUdlfied, after which they
were inverted and plaeed in the electrically heated oven and Ineubated at

37® C* (98*6® T*) for d8 hours*

4* The eoimta were made and averages taken from the count recorded

for the 1 - 100 and X • 1000 dilution*

 88

Th* above prooedure givee a genaral summary of the methods used for

all of the plate oouxitlzkg in this investigation ezoept In the trials

oondueted on sterilization of eens* There were no dilutiozis made for

plating the rinse solutions talEsn from the cans. The procedures were the

same as those outliiuid for milk senplSNe except 1 co. of the rinse solution

was plated in duplicate and the average tai»n from the tso*

Soioe of the advantages olaimsd for the plate xastbod aret (1) it

indicates the growth of the living organisms} dead or inactive organisms

ere not considered Important, (2) dilutions may be made to seoiQ!<e estimatea

of different grades of milk, and (3) the method has been more xmifoznally

standardized for milk analysis.

imong the disadvantagee for the plate method may be: (1) the ezpanse

of the material and equipment, (S) lesa generally useful for hi£^ eount milk,

(3) the incubation tesperatures are differmit from those at whioh milk ie

generally held, (4) resulte are not obtained very quickly, (5) not all of

the bacteria grow under the conditions of the test, and (6) not much

knowledge of the type of organiame present is given.

Methylene Blue Reduetase. The nethylene blue reduetase test was used

in this investigation to dstsrmine its relation to the plate laethod ae to

the bacterial action In milk. The principle of the test is bassd on ths

notion of an enzyme formed from baeteria cox methylene bine dye solutions.

The action of this enzyme aldehyde reduetase produces a color change, or

coB^lete deoolorization. This change in color and the time required ie

said to be directly related to the aotivitiee of the bacteria#

 89

*!!&« redootastt lasthod r«qttires tli« tisft of a eoa^aratlvaly largi

of mLllc, whioh apparantly la favorable toward Its aecuraoy* Haay

Investigators have found a very olose agreement between the SGa{q;>Iea and

duplioatea* Some arguments for this method are that it is siiqple and

inexpensive, the results are usually obtained in a e(»Q>aratively short ttast

and the nmthod pexmits the use of a follow test* (l^e fermentation test).

The xoaterial and equlpaant used for mahing the reduotase tests aare shown

in Figure 5*

r.

jkn.

-V .

M' i|
1^
\

3
Figure 5* Material and Squipment TTmid For ISfiaklng The Reduotase Test.
 90

A list Of th« aatttrial and oqalpaeiii aaad for m)slAg th* roduBtaaa

tost aa shoim in Hgara 5 aro:

1* A supply of sterile test tubes

SlU A 10 eo» pipette for lasasuring tbe aiillc

8« AX 00* pipette for aeasuring the laathylene blue solution

4« An inoubator equipped with a water bath

5* A standard Mthylene blue solution made by diesolwlnig one tablet

of dye in 50 eo» of sterile water and diluting this to SCO eo«

Note) The tablets may be obtained from the National Aniline and COittBloal

CompanyI Ine>» Hew York*

16
Dhe procedures followed were similar to those used by EUenberger

and others. Methods for securing milk samples used for the rsduetass tMt

for making the o<»^arison were similar to those practiced by the dairy

inspector* The samples were secured directly from the can* The trials

were planned so that the sasoples for the reductase test could be taksn at

the same time the milk inspector from the IQ30X7ille Bureau of Health took

samples. The milk inspeotor was taking sanyjles at least once each month

during the entire year of 1936 in which the emperSment was eonduet^*

SsBplss were taken from the milk of each of the patrons shipping to

the Dhiversity Creamery* A 10 eo* pipette was used for all of the

sampling just after the city ixiapaetor had taksn his sample. In ease a

patron had more than one can* the eaogile was secured from the differmt cans

in proportion to the milk in each. The pipette was first rinsed with a

sanple of the milk which was to be seeured, then a pipette full was drosn

and transferred to a sterile test tube. ifh»n all of the esnplee war* talnm
 91

th»7 W9r« earried to tho lalioratoxy and 1 oe« of tba standard lasthjrlexui

blue solution vas added* After thorotmb mixing they were ineuhated at

F« until the blue oolor had disai^earod.

fable XII shows the relation of the naaber of baoteria per ee« in

thottscEQids to the reduotion time in hours* The tubes for the reduetaee teat

were ineubated for a aaxingaa tta» of 8 hours and examined and reoorded at

15»minute intervals* Sinae the test has been ireeommended by authorities

primarily for milh with hi^ oounts or short reduotion time* the tubes

retaining the blue color for 8 hours and over were recorded as deeolorieing
at 8 hours* The results are recorded in thousands of bacteria in round

numbers and hours, half ood quarter hours for the reduotion time* These

trials were eonduoted for the aatire year of 11KS6 using a sasq^le from oaeh

patron once each month* It was found that sens of the patrons did not

ship rsgolsrly, so only the sasoples from ten of the regular shippers were

recorded in the results*

The avaregee as idioea in Table XII indioate that there ie a fairly

close correlation between the reduetase method and the agar plate counting

method* In general it may be said that the shorter the reduotion time the

higher the baoteria oount*

The averages taken frcm each patron*s eoliam, as given in Table ZCI,

and ehown on a graph (Figure 6), indioate that the coeffielent of verlatl(m

is hi^iar then the noimal aXlowanoe for experimental error* For exsixple,

a diagonal line from S50 in the baoteria column to 7*8 in the reduotion

column should fall very close to the mean* The variation from this mean

may be due, in part, to experimental error and, in part, to unknown faotoxe

which distinguish oolor reduotion from the number of baoteria present*
 O
O
• • • t a
1 1
# H
1 1 1 « 1 r• s
1 S
"at""
-»a CD Bacteria o
ss a
CD sa to
o
i § 8 1 8 IS 8 s 8 per eca
nortaP
»
• Reduction
at 09 1 a 03 SI a> 0> ® ffi Tim hrsa
SI SI
0> H 01
Jaoteria
8 itk 8 to ta O to
8 8 8 per 00•
to
g
o>
• ^eduotion
00 (D 09 SI XlH CD 03 03 CB ® rime hrsa
ImI ^oteria
8 1^ tD SI 8 8 *■ 8 b 9er oca
CA
§ 1
>3
• Eduction
1
OI a> 09 09 e» a CD ® 1 03 ® ® ® rime hrsa
H •<a CD
8 o> bacteria
% to 8 SI o O H to o
i 8 s H
)er 00a iD Q
at
»
at 09 09 09
leduotioa
OI ► -o m 03 s ® g- W
fltae hrsa
H CD
<1 0*
ti M )aoterla
I y ^
ift \ § B <o 1 at S CA
ler eoa CD
o
•
at
Haduetion
09 CD CD
I ® 03 ® ®
!i ^
3
li H M
-0
Bacteria
i 1 H to g 8 8 8 CD CD to iD>
per 00a at
'S
• 1 Reduction Q
at • 09 ® 01 ® at ® ® ® ® O
® a rime hrsa
a SI S3 SI
Bacteria
8 to H 8 i 8 8 s § to H
per 00a s
51
at
•
SI 09 at
deduction
a ® 0> 09 CD ® ®
rime hrsa
N S9 SI SI M
ht H 1 H Baeteria
SI 1 09 OI to <D 1 s
8 8 8 8 H
per 00a
a
at
m
i ^eduetion
(a 1 09 (D ® A ffi 1 s (J>
{8* 03
rime hre*
S 0>
s CD Baeteria
8 1 H to 8 O 8 to § to H per 00a to
o>
a =leduotion ca
1 09 ® 1^ CD #> m 03 01 CD ® rime hrsa
CD CD Saoteria
H i» «R 1 to 8 8 8 to 8 01 ® H
;ier oc*
O
SI
a leduotioa
cn 09 09 1 o> ® 01 o o s a m ® !?ime hrsa
26
 98

B«aAlae fnm left to ri^t on the graph (figore 6), the indeagee for

patrons 6 and 8 ere extrecaely aontrery to the theoretical oorrelation for

the teo methods. The other patrons* Indexes are probably within the nsonual

range Of experimental error*

The eorrelation for the indlrldual months (Hgore f) Is naioh closer

than the aTorages taken from the year as shown in (Flgnre 6). The range

of baeteria counts as taken fztna aweragea in Table XII is from 6.5

thoosand to 530 thousand. The average range for ^ reduction time ie

from 4*9 hours to 8*0 hours. According to ths data secured from the

areragee, (Tigure 7), the relation between the methylans blue reduetioa

method and the baet«nrial plate oount is very olose in oases where the

plate eount is over 9 thousand*

Tor exanple, the months of lune, Ihly* August, and teptember.

(Figure 7), have a very high average baoterial indooc* The proportional

reduetion time for the months listed above is relatively eonsistent with

the baoteria eounts. For the months not mentioned above, the baoterial

IndeaMs are oonparatively low. The average reduetion time was not as

oonslstwat nor w«re the otxrrelative indexes as close ae they were for the

four months with high beetnrial indases.

The ten patrons* milk, as shown in the Table, represents the different

aamples used in this investigation. Saoh patron*s milk was kept separate

and recorded by the KUoxville Milk Produoers* oontraot ntmber xmtil all of

the data were Kittaarl2sed* The results in tie Table indicate that for some

months during ths year, or some seasons of the year, the variations between

the methylene blue reduetase test and the egsr plate oount were greati^ than

the others*
 94

Bactfilia.
per c;0«:

(Th|35asam
425

375

350

325

300

275

250 v3

225

(I-

200

175

150

125 (t)

100

25

tl06 ifa; JiRiiir)3j

M T,.
Publiiih« c.4s,t^.%li nAC '•<> '•» '•* '
Figure 6* Correlation Between Bacteria Ootmts in thousands per cc«,
and reduction time in hours.
 95

Averages for Each Month

Bactei^aj!o^n-tf^ Reduction Hme
in t.i fdlh;Hoursi-:

850
#15 ;E (lOt

000 Z7777 at on llQU ^.25

750 .50

700 .75

: 7
650 bIIi.OO

600 7- ;ii».25
75 /
/
/
550 75 u50
/5
>5
75 i.75
500
/5^
75
450 3..00

400 i t; fu25
55

55
?■ Z !(i.50
350 /
z z
/ z
z
300 (i.75

250 Z ".00
Z
Z
Z
200 ".25

Z Z
cz Z
150 : z z z 7.50
Z z z
; Z Z z
z z
z z
z z z .75
100 z z z
z z
z
z
z z
50 Z 8.00
Z
z z z
z z
z ; Z

^,_7uSe ep .
: ■Jz lipz i pz
Oct. JSiov. i-iec.

Figure 7. Correlation between bacteria counts in thousands per cc.

and reduction time in hours.
 n

Iirfliifflioe of Cooling.
Baotaria that are usually responaihle* for tha souring of toiUc

apparently hara an optlom growth taoparature of 08P to 100** 9oam of

tha other factors which influsnca bacterial growth are moisture and oxjgim*

Tharaforai tha conditions in milh at tha time it la drawn from tha eow are

said to be almoat ideal for bacterial growth because the tenparatu3Hi»

oxygen, and moisture are near the optinum requirements. Wum the millc is

held at tenperaturas around tha optiaum range for two to four hours, the

bacteria counts indleate that a wary rapid growth has taJaen place} whereas,

whan milk is cooled to ralatiwaly low tesparaturas ianediataly after it ie

drawn, tha bacterial growth is retarded*

This inwastigation ineludad a eariee of six trials made by using milk

soon after it was drawn at tha tmiwersity dairy bam. Tha milk was carried

from the bam in 10-gallon cans and cooled to different temperatures owar tha

surface tubular cooler shown in Figure 2.

Control samples for makiag the bacteria plate counts were taban fvom

the oans Just before cooling the milk. The other saoplas in each aeries

ware cooled as indicated in Table ZIII. The maxiiiswi elapse between the time

the milk was drawn end when it was cooled was not more than thirty minutes.

This part of the inTeetlgatlon was planned eo as to repraaant cosnon

practices on warious dairy farms* For axsepls, the batter milk producers

usually cool their milk to 40^ F* inmadiataly after it is dmwn* This

praotics usually rsquirss a meehanical refrigeration unit or tha purohaaing

of loa* It is quite obwious that cooling the abowa methods should help

to retard bacterial growth* There are also Boma producers who use spring
 99

init«r or voll water to eool their milk. The afirage tengpevature range to

which milk can normally he cooled throu^ut the year in most sections of

the Uhited States^ using water as the cooling agent, would probably bo

fnna 60^ F. to 70^ F« The aeries moled to 70^ F* was selected to irepresent
coolizig practices by the producer who is trying to produce good milk with a

inlnina Investaent for cooling equipment* In addition to tiie above

mentioned classes of producers, there are some who deliver milk to processing

plants without cooling it at all* They try to make deliveries soon after

milking to avoid cooling* Seme producers deliver twice eaeh day* These

conditions may be represented by the trials in which no cooling was done*

The same supply of milk was usod for the three different temperature

series* A sample was taken for baeteria plating, them the can of milk was
o
divided into three separate portloim, one of which was cooled to 40 F*,

anothar to 70*' F*, end the othar was not cooled* All smplts were txtuuH
fensd to quart milk bottles idiich had been washed and steamed for handling

eommeroial market milk* Duplicate eaHaplee were held fma eaeh trial for

eaeh teiQ>erature, one for the sixth and one for tho S4-hour plating* The

caps were earefully placed on the bottles by hand in sueh a way as to avoid

eontaainatlon*

The saddles whieh were cooled to 40® F* were held in the milk storage
room at that temperature; those cooled to 70® F* were held in a water bath
at that tsmperatiire; and those i^ieh were not eooled were held in a room

with a tos^erature of 68® to 78^ F., representing the tampearature of tke

average milk house* These samples were removed and plated at the periods

indicated in Table HII using the regular agar plate method with two

dilutions.
 99

TABUS ZIII

IHFLII3N0S OF COOLIRa OH BACT^IAL GHOWTH « A7ERAGE OF SIX TRIAIB

Platlag Time After Cooling

Original Count 6 hours 1 84 hours
Average number of bacteria p«r co«

Cooled iBBaediatelF
to 40° F, 9,500 81,000 30,000
Cooled isnedlately
to 70® F. 9,500 09,000 418,000
Rot cooled 9,900 458,000 over 1 millioa

The resolts Meur«d fxm the series of six trials* using thive different

eooling tss^retures* indicate that prtngit oooXing to relatireiy lov tsoH

peraturss is esswxtiaX for pxodusing miXk with a zumber of baoterla*

TabXe ZIII shows that froa an arerage of six triaXs* miXk with an

origlnaX count of 9,500 wh^ cooXsd izDn^diately to 40^ F« and held 6 hours
contained an average of 21,000 haeteria per oc« This aeries had a variation

of frcm 10,000 to 60,000 basteria per oc-

The average count froa the aaisples cooled to 70^ F« at the 6-hotir
plating was 89,000 bacteria per oc., and at tiM S4»hour plating the average
count was 418,000 bacteria per oe*

The aveimge count fvem the sen^iles which were not cooled but were held

in a room at 68^ to 78® T, at the 6-hour plating was 458,000 bacteria per oc,
and at the £4-hour plating, over one million baoterla per oc* The estimates

from each of the six trials on the 84—hour plating of the samples not cooled

were over one million bacteria per eo«

 99

Efflelenoy of Fasteurlzatloa.

The peroentage of baetorial deorMUM duo to pastourizatioa, or

offlolonoy of paoteurlzation, is of vaat iB^rtanoo to the pasteurizod milk

industTy* Health authorities require a relatiToly lov haeterial oount for

pasteurized milk. In order for the milk plant operator to market pasteur*

ized milk with low eouuts* hie eqalpmmit and methods must be effioient.

This inTOBtigation oonsisted of a series of six trials for eaeh of the

o iO o
following pasteurization tempMratures, 142 , 149^ , and 148 F*, as

indieated in Table XTf* IRie experiments were eondueted, using the regular

pasteurized milk at the Ihiiwersity Creaioery, with the raw drip samples for

controls* Baeterial analyses were made by the agar plate method at the

6 and 24-hour periods. All of the assqplee were plated in duplicate using

1 - 100 and 1 - 1000 dilutions. The eounts were reeorded in ronad thousands.

TABIS XI7

HFnCIHRCY OF PASTBDRIZATION

Awerage of 6 Trials

Plating Tims After Pasteurization

Haw 6 hours 24 hours Average %

(Controls) affloienoy
Bacteria per oc.

Pasteurized at 142^ F. IBS S 7 97

Pasteurized at 143i^ F. 126 2 8 98

Pasteurized at 148^ F. 200 4 4 98

Of the series of six sasqples pasteurized at 142 F., the original oount

when raw varied from 60 to 500 thousand, and after pasteurised, the renge
 200

mw trcm 1 to 6 tSiousamd ot tho 6«hour period, aad 1 to 25 tbottaoad at

the 84->hottr period. ^Rioae pastetirized at X43i-^ P. varied from 86 to

228 thotiaend alum raw; from 1 to 5 tliouaand plated at the 6-hoiar

period; and I to 5 thousand at the 24>bour period after pasteurization.

Those pasteurized at 145^ F. varied when raw from 60 to 600 -tiioussod;

1 to 10 thousand at the O^hour period; and 1 to 10 thousand at the

24-hour period after pasteurization.

The results in Table XX7 indioate that from an average of six trials,

eaeh of the different pasteurization t<Rq>eratures had a very high

peroentage of effioienoy. These figures were obtained hy getting an

average from the six trials of the raw saiiq>le8 and getting the per oent

of deorease using an average of the six trials as taken from the 6 and

&4-hour plating periods. For illustration, the average for the six

trials in the raw milk, in the 143|-'* F. series, was 126 thousand. The
eounts on the pastetirized seogtles at the 6 and 24-hour plating periods

were 2 and 3 respeetively, or an average of 2.5. The per east of

effioieney may be ealculated as follows:

126 - 2.5 or 123.S X 100• 98-. effioieney.

126
The other seriee were ealoulatad In a similar manner.
 101

Sterlllaatlon of Gang*

Procedure t The cans ue^ in this investi^tioB ware regular lO-'gallon

milk oana in use at the UhlT^rsity Oreamncy* Son of the ewos had not beau

washed slnoe their usei some of them had been imished and steamed and

allowed to set in the wash rooa for 8eT<«KLl hours

| end some of them had

been waedied the day before thay were used for the es^^eriments* the lids

were rmaowed from the cans because the steaming apparatus did not prorlde

for stemalng the lids*

Baeteria oounts wers enured by plating 1 eo« portions of 600 eo* of

sterile water whicdi had been used to rinse eaoh oma* the water used for

riuse solutions was sterilised in qusoct milk bottles for a period of one

hour at 15->pounds steam pressure in an soitoolawe* The differeaees in the

beetsrial oountsi aa ehowa in Tables 27 and XV1« were detezmined by ftret

rinsing the eans whleh were to be etariliaed with 500 eo* of sterile water

and 1 ec« of this solution plated* *nie differenees between the baeteria

eounts for the separate platings oonstitute the effloleney of sterilization*

Steam steriliaatloa^* The cane were first rinsed with 000 ee* sterile

water which was poured back into the bottle to be plated later* "nis cans

were next steamed orer a jet with approzimtely 60->pouads bollar eteem

pj^BSsure for one minutet accurately timed by a stop wateh* After steaming,

they were dried onwr a hot air (135® ?«) jet for X|- minute* After the oans
had cooled, another 500 ee* of sterile water was need to rinse the sens in

the same manner as hefox« sterilizing* The standard plate method was

for determining the number of bacteria per eo* In the rinse solutions*

The results were glren in baeteria per oc* from th# 500 oc* of rinse
 102

solutloa* vbA Bot tlie total mimbor of baoterla ia oaoh can.

Data fr^BB the six trials with steam sterilization are giwan in Table

XV* All of the plates mere Incubated for 4B hours before the eotants ware

made* This was also the incubation period for all of the azperiments on

sterilization that follow.

TABLE X7

NDlffiaSR 01 BACTERIA PE? CO. OF RDISB WATER BEFORE AND AFTER STERXLIZATION

OF CANS BY STEAM.

Colony Plate Count Before and After Steaming

Before After
Trial

I 6,200 Sft
2 3,300 74
3 9,000 23
4 1,000 70
5 23,000 11
ft 2,000 41

Average 7,417 42

The data in Table X7 show that steam killed the majority of the

bacteria present in the oaos. HoweTer, quite a number of bacteria

BurriTed after treatoient* The efficiency of sterilization by steea

according to these trials was over 9^. Swoe advantages for steam are

that it ie generally acceesible and is therefore inazp<nasiwe» and it la

aleo a very quiek method of eterilization.

Dry Heat Sterilization. The electrically heated Bseo sterilizer at

the dairy bam was used for these experiments. Each of a series of

10«gallon cans was rinsed with 500 oe. of sterile water before and after
 108

Btariliaatiozi th« atma an vlth stB«oi* Tha oaas were iuverted and plaeed In

the eleetrio cabinet» and the switch turned on* were left in this

eabinat tvtm one millcing until another which was a period of about 12 hours.

During sterilization, the tenp«rature want to aj^roxtmately 212® F. at

tirtiieh point the switch was cut off by a thenaostat* The tanQ>eratu3re

remained high tor at least one hour and thmi went down gradually, riowewer,

the cans were still warm at the end of 12 hours.

The data for these trials are given in Table X7I* The hlg^ counts
obtained before sterilinlng were only approximate.

TABLS m

NOMBER OF BAOTFRIA PBR OC. OF RINSE WATER BEFORE AND AFTER STERILIZATiaif
OF CANS BY mr HEAT

Oolony Plate Oount Before and After Sterilization

Trial Before After

1 320,000 169
s 21,000 2
s 236,000 20
4 76,800 5
0 134,400 280
6 102,000 45

Average 151,500 87

The data in Table xVi indicate that the effioienoy of sterilisation

in these trials was over 99.9^. The above results show that dry heat is

also a very effective method of sterilization. Tha initial counts in

these cans were relatively hi^. That, however, should not have affected

the results enough to cause any marked inaccuracy.

 XM

Chemloal StertXization* Itoea of the most oonnm& eheoleaX stttrllizers

for dairies were used for steriXizixtg the oaus In this aeries* fhea

referring to ehemieaX sterilization or sterilizers) they are generally

called ohXorine sterilizers because ehXorine prodooes the germioidaX aotion.

fhe trade naiaes for the sterilizers used were as follows: Diversol,

H-T-H, and B-KV

the oans were treated as outlined in the above prooedure* Several

trials were run using solutions of eash produet eontaining 80 parts per

nlllion, 100 parts par laillioa, and aoo parts per million of available

ehlorine* the oans w<ne rinsed with sterile water and plated for controls*

they were next rinsed with one gallon of the sterilizing solution made fron

Idle different chlorine eompounds of known strength* After thoroui^

exposure to the chlorine solutions, the cans were well drained before they

were rinsed with sterile irater* they were then rinsed and 1 oo* of the

rinse water was plated* the data lediown in table X7II were secured by using

solutions containing 50 P*P*M* of available chlorine*

TAHU X7II

HDMBiatl Oy BAOtSRlA PSR 00. OF 500 CO. STBRILB RINS3 WJUmt HSPOSS AM) AJT®
SfSRILIZINO WITH OHIORIRS SOLDTIOUS CONtAITOIG 50 PARtS PER MILLION
OF AVAILABLE GKLORBJE

Average Colony Count Before and After treatment

Sterilizers trials Before After

B-K 8 1,612 1
H-t-H 8 4,400 6
Dlversol 4 4,750 4

Average of the three 3,578 4

1 ..-t: -.i - .'.' •

 lOd

The aTerage effioienoy of sterilization, aooording to the trials shown

in Table X7II, was a little less than 99«9^« S&oh of the ehssBioaX
sterilizers used in these trials was apparently very satisfactory. The

dlffsrenoe in results for the three sterilizers used was not great enough to

be oonsidered ta^rtant.

One trial for each sterilizer containing 100 P.P.M., and one trial for

each sterilizer containing 200 P.P.M. of aTailable chlorine was made. The

data indicate that the efficiency for the higher oonoentrations was not

much greater than the lower ones where a thorough exposure was gLwen.

The above results indicate that chemical sterilization is a very

effective method of sterilizing milk cans. In all the trials with ohesnieal

sterilization the percentages of efficiency were greater than those obtained

from steam and equal to those obtained by dry heat methods. These results

further indioate that in sterilizing milk oane, using 50 parts per million

of available chlorine was just as afficient as 100 or 200 parts per million.

The differences in results probably were not great enough to warrant the use

of more than SO parts per million in sterilizing cans. Btowever, there are

some disadvantages connected with chlorine sterilization whioh steam and

dry heat do not havO. Chemical products are usually more expensive. They

do not retain their strength unless properly stored, and they sometimes

produce a chlorine flavor in the milk. This flavor is considered very

objectionable. However, this flavor can be preiranted if the ohemieals eat

the utensils are handled properly.

 m

GoncluBloae*

resiats obtained from the reeearoh in this inveatlgation indicate

that the following oonolueiona may be justifiable}

!• Aeration, to the extent used in these trials, did not recooTe the
feed and weed flavors from milk. There was no evidence that the flavor

was it^roved by aeiwition over the flavor of that milk cooled by other

methods*

2* Pasteurization at t«e^eratures of 142^ to 145^ F. inqproved the

flavor of milk slightly* feed flavor was not as noticeable in the

milk after it had beea pasteurized* However, there was usually a sli^t
cooked flavor prs8«at in the pasteurized milk*

3. Pasteurisation at temperatures of 142^ to 145^ F* for 30 minutes

caused a sli^t decrease in cream volxitae o<mipared to raw milk at the
O O
6-hour creamizig period* Saaqples pasteurised at 142 to 143i^ F* had a

largwr cream volume than raw control saoqples at the 24-hour creaming
period. Saoplas pasteurized at 14^ F* had a smaller cream volume than .
raw samples at the 24-hour creaming period*

4* Pasteurissation apparmitly retards the rising of cream* In general,

the higher the tmaperatuxee of paatetxrlzation the slower the rising of the

cream*

5* Raw milk had a larger cream volume at the 6-hour eraaming period

than at the 24-hour creaming period* Pasteurlssad milk had a latger voltnte
of cream at the a4-hoar period than at the 6-hour period*
 107

6» A very elosa oorrelatlon iias ftnmd between the methylene blue

reOuetaee teat and the ager plate iMthod for estizoating the baeterlal

eontent in raw milk. The plate method wae apparently more aatiafaotory for

determining the ttuiat>er of baeterla in pasteurised milk*

7. Cooling milk to 40^ F* iiaaaediately after it wae drawn trtm the oow
iretarded the baeterlal growth throughout the following 24 hours. Cooling

to 70® F* helped to retard the baeterlal growth for the 0 hours after the
milk was drawn then rapid multiplioation took place between the 6 and

Sd-hour periods*

S* Thilure to cool milk resulted In a wexy rapid baeterlal growth

before the end of the d-hour period* ISillions of baeterla were present at

the end of the S4-hour period*

o o
9* Pasteurisation at tenpexatures of 142 to 145 ?• for 30 minutes

killed a Wary high peroentage of the baeterla in milk* There apparently

was wary little differeme between the effloiency of pasteurisation at

148®, 143|5?, and 145® F.

10* Steam, dry heat, and ohemieal sterilization were all very effective

methode for killing the bacteria in milk oene* Steam wae apparently the

most eeonomloal and ohaimioals were probably the quiekast of the methode

used in this inwestlgation. Dry heat was very effeotiwe and may be

reoQiaaended where oonditions will warrant its use*

 108

^Iblloffpayhy

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6. Brew, Jaaes D., Dairy Short Course Lecture, Uiolverslty of Tennessee,

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 1D9

14a* Doreraau, S* MiohlgsA Ag* Ixp* sta., J'oumal Dairy Sei«ioa»

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 no

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%
 Ill

44. Parbitt, S. H., flilk Plant Monthly, p. 66; January 1936.

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 112

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