MINISTRY OF HEALTH SERVISE OF UKRAINE

ZAPORIZHZHYA STATE MEDICAL UNIVERSITY

THE CHAIR OF MICROBIOLOGY, VIROLOGY, IMMUNOLOGY

INTERACTION OF MICROORGANISMS
WITH THE ENVIRONMENT

Learning guide on the subject "Microbiology" for the 2nd and

3rd year English media students of the International Faculty

ВЗАЄМОДІЯ МІКРООРГАНІЗМІВ З
НАВКОЛИШНІМ СЕРЕДОВИЩЕМ
Навчальний посібник з мікробіології для студентів
II-III курсу міжнародних факультетів

Zaporizhzhya

2015
UDC 579.26:616.31
BBC 52.64
I-69

Guidelines ratified on meeting of the Central methodical committee of

Zaporizhzhya state medical university (protocol numbers 5 (20.05.2015) and it is
recommended for the use in educational process for foreign students.

REVIEWERS:
Melnikova O.V., senior lecturer of the chair of pathophysiology department., Ph.D.

AUTHORS:
Voitovich A.V., assistant of the chair of microbiology, virology and immunology,
candidate of Biological Sciences.
Yeryomina A.K., senior lecturer of the chair of microbiology, virology and
immunology, candidate of Biological Sciences.
Kamyshny A.M., the heat of the chair of microbiology, virology, and immunology,
doctor of medicine.
Sokolovska I. A., senior lecturer of the chair of microbiology, virology and
immunology, candidate of Medicine.

Interaction of microorganisms with the environment : learning guide

on the subject "Microbiology" for the 2nd and 3rd year English media students of
the International Faculty / A. V. Voitovich [et al.]. – Zaporizhzhya: [ZSMU], 2015.
– 71 p.

Microecology a new scientific and practical fields of biology and medicine, located
at the intersection of microbiology, ecology, epidemiology and hygiene, which is important
for the further study of clinical microbiology.
The data symbiotic relationships between organisms themselves and with the human
body. Paid much attention to positive and negative effects on human health microbiota
different environmental habitats and habitats of various human body. The basic methods of
sanitary microbiology and virology.
 TABLE OF CONTENTS

The relationship of microorganisms among

themselves and with the environment 4

Sanitary microbiology 7

Methods microbial decontamination 49

Sanitary virology 56

Related protocols by topics microecology

Sanitary microbiology 71
THE RELATIONSHIP OF MICROORGANISMS AMONG THEMSELVES
AND WITH THE ENVIRONMENT
The relationship of organisms with each other and with the environment
ecology studies (greek. oikos - house, logos - concept, doctrine). This term
suggested in 1866 by E. Haeckel. Microecology examines the relationship between
man and the environment of microorganisms and symbiotic relationship with their
own microbiota. Range of interests is at the crossroads microecology
microbiology, ecology, epidemiology those hygiene.
Sizes microbial ecosystems are very diverse. This could be, for example,
pond, lake or root system of the tree. There are some small ecosystems as oral
cavity or portion of the intestine. Within each species for ecosystem can describe
his residence. This notion of "ecological niche" reflects not just a place of
microorganisms in space, and its function or population of microorganisms in the
community of organisms. Each species or population in this community performs a
specific function, which is caused by the need for food, mobility, modes of
reproduction, biochemical, structural features, beyond tolerance to environmental
conditions. It may or may not be any kind to perform a specific function in a
particular ecosystem depends on the totality of its properties. The degree of
adaptability of species to changes in environmental conditions called ecological
valence. Environmental valence type of microorganisms also called its ability to
colonize environment that is characterized by certain changes in environmental
factors. With a variety of mechanisms recycling power supplies and energy and
pronounced adaptation to external influences, micro-organisms can live where
other forms of life possible. The natural habitat of most of the microorganisms -
water, soil and air. The number of microorganisms that live on plants and in
humans and animals, much less. Widespread microorganisms associated with them
move easily through the air and water; including surface and bottom of freshwater
and salt water, and a few centimeters of topsoil are full of microorganisms that
destroy organic matter. Fewer microorganisms colonizing the surface of plants,
skin and hair, as well as some internal cavity of animals (gastrointestinal tract,
4
upper respiratory tract). In areas of residence microorganisms form microcenosis.
Each microbial community in a particular form cenosis autohtonnye specific
microorganisms (from the greek. autos - one, chthon - country), usually they occur.
In natural biocenoses (soil, water, air) and rozmnozhayutsya survive only those
microorganisms, which contributes to the environment; their growth stops as soon
as environmental conditions change. When the favorable conditions they give rise
to new clones of microorganisms.

Microorganisms as symbiotic partners

Since time immemorial, have developed complex relationships between
microorganisms with higher organisms flora and fauna, on the one hand, and the
environment - on the other. Some of coexistence of two different organisms,
including microorganism microorganism called symbiosis. Participants called
symbiosis symbyontamy. Symbiosis is characterized by different types of biotic
relationship with respect to the cells of their host and to each other.
Mutualism – is a form of coexistence when both symbionts - host
microorganism and get mutual benefit. In mutualism coexistence creates favorable
conditions for both partners, that is mutually beneficial symbiosis. Bacteria
produce many kinds of vitamins B6, B12, and vitamin C. An example is the
coexistence mutualizmu plants with rhizobia, which feed on plant juices with
substances (such as legumes - peas, vetch), and plants, in turn, use nitrogenous
compounds, synthesized by rhizobia that are retainers nitrogen.
Commensalism – is a form of cohabitation when one of symbionts (in this
case the microbe) lives at the expense of the owner, uses his protection, but the
owner does not cause any harm. In commensalism partnership can be beneficial to
one of the organisms without harmful effects on the other. Commensal microbes
(staphylococci, streptococci) inhabiting as normal microbiota skin and mucous
membranes of humans and animals. However, we must admit that commensalism
in this case is a relative concept, because among the pathogenic microbiota is such
that under certain conditions can cause severe disease.
5
 Parasitism – is a form of coexistence when mikroorhanizmy- parasites feed
on host tissue components, thus causing him harm, causing an infectious disease
and can not exist without it. Such organisms are called pathogenic. Thus, the
environment of the parasite is host organism to which the parasite adapts during
evolution. This environment directly affects the parasites as well as parasites affect
host. Environment in the usual sense influences such parasites are mediated
through the host. Many micro-organisms entering the human body, can not
influence each other, ie the interaction between them, such a situation is called
neutralism. When neytralizme partners (microorganism and microorganism) can
not give each other any effect.
Antagonism – is the opposite effect, mutual opposition. Antagonism of
microorganisms - is a complex relationship, when the joint development of
bacteria populations of one species or within the same species suppress the
development of other, sometimes completely destroying them. Antagonism of
microorganisms is widely used to prevent and treat various diseases, especially
gastro-intestinal diseases. For example, many strains of E. coli can inhibit growth
and kill streptococcus, staphylococcus, salmonella. Antagonistic relationships
between organisms are of great practical interest. Antagonism of microbes in the
soil have watched L. Pasteur (1870), I. I. Mechnikov (1905) observed antagonism
between lactic acid bacteria and putrid. Merit II Mechnikov is that he laid the
foundations of the doctrine of antagonism of microorganisms, which now
developed into a doctrine of antibiotics.
Synergy – is identical physiological processes of different microbial
associations, which resulted in an increase in end products.
Satellizm - a stimulation of the growth of a microorganism products other
life, which then becomes his companion.
Among the various representatives of the world of microorganisms have
evolved different forms of symbiotic relationships. Mutually beneficial
relationships developed between aerobic bacteria that live in the soil, in the
department of colon and other substrates. Aerobic bacteria use the oxygen present
6
in the soil, thereby creating favorable conditions for the development of anaerobes.
In turn, anaerobes decompose cellulose to form organic acids, which are the source
of energy for aerobic bacteria. With a huge number of microorganisms that live in
nature, only a small portion of disease. During the centuries of evolution some
species of microbes adapted to extract resources from food inanimate nature, this
time remain svobodnozhyvuschymy, other gradually adapted to coexistence with
animals or plants and through their get nutrients. In the evolution of parasite
adaptation to the host was through specialization, including through the acquisition
of the ability parasite in certain tissues, such as the causative agents of brucellosis
parasitized in the placenta, salmonella - in the lining of the small intestine. In this
case the tropyzme parasites, ie, the ability to selectively affect mainly certain
organs and tissues. The variable can be spatial relationship between symbyontamy.
If one symbiote is out of another cell, then talk about ektosimbioze, and if within
cells - the endosymbioz. Called symbionts of larger host. In infectious diseases, the
interaction of different types of microorganisms causes the development of so-
called associated infections. Associated infections caused by two or pain
pathogens. Association microorganisms - a community of different species existing
in natural or artificial conditions.

SANITARY MICROBIOLOGY
Sanitary microbiology - direction of medical microbiology, studying the
flora of the environment and its impact on human health. The main tasks of
sanitary microbiology:
study microbiocenosis, in which there are microorganisms pathogenic to
humans;
development of methods of microbiology external environment,
microbiological standards and measures to improve the health of the environment.
Practical sanitary microbiology uses two main methods of evaluation of
sanitary-epidemiological status of the environment: direct detection and
identification of pathogens indirect signs of their presence in the environment.
7
Microorganisms which can indirectly judge the possible presence of pathogens in
the environment called sanitary and demonstration (SDM). Main characteristics of
sanitary-indicative microorganisms:
SDM must constantly live in the human or animal and always stand out in
the environment;
SDM should not breed on environmental objects;
Duration of survival SDM in the environment must meet the length of
survival of pathogens;
Methods of identification and differentiation of SPM must be simple and
reliable.
The presence of MS is determined by two methods:
• direct count of the number of bacteria;
• sowing on nutrient media.
Number of SDM expressed in credits and indices.
Title SDM – the smallest amount of test material (in ml) or number of weight (in
grams), which is present at least one individual MS.
SDM index - the number of MS, found a certain extent or amount of the
object.
To identify the general microbial contamination determine the overall
microbial count (OMC) by counting all microorganisms (grown on nutrient media)
in 1 g or 1 ml substrate.
The estimation of population microbiota carried out by determining the level
of bacterial contamination in the unit studied habitats:
+ - very weak growth (growth of single colonies - 10 on the cup with the
environment) that is less than 10³ colonies / units;
++ - poor growth (10-25 colonies), which is 10³ - 5 • 10³ colonies / units;
+++ - moderate growth (from 50 to 100 colonies), which is 104 - 106
colonies / units;
++++ - a massive growth (continuous lawn colonies, beyond counting)
which is 109 colonies / units.
8
 Calculate the ratio of quantitative dominance microbiota (d) using the
formula:
d = Pi x 100%
where Pi - the fate of species (strain) bacteria, calculated as ni / N, ni - the
number of bacteria of this type, N - total number of bacteria in the sample is
washed away.
Expect index constancy of microorganisms in the composition of the
microbiota, as the ratio of the number of microorganisms of this strain to the total
number of samples tested. Depending on the value of the index constancy
microorganisms distributed constant (index of permanence within which 50% or
more), more (the value of the index constancy within 25 - 50%) and casual (index
constancy which are within 25% and below).

Soil microbiota.
The soil consists of inorganic chemicals and organic compounds that result
from the death and decomposition of living organisms. Soil living organisms in the
soil together constitute biocenose. Contained in the soil living organisms
(including bacteria) are living a phase of soil. It includes microorganism and
microorganisms, both animal and vegetable origin.
Microorganisms that live in the soil are divided into two types:
- autochthonous microorganisms (bacteria resident, resident microbiota), ie
germs that are unique to a particular type of soil;
- allochthonous microbes (transient microbiota), ie microorganisms which
under normal conditions do not occur in the soil.
Microorganisms in the soil and water in developing colloidal films covering
solid particles, especially in the capillary and gravitational water that fills the pores
between the mineral particles of the soil and contains dissolved organic and
inorganic materials.
In the living soil:

9
 1. Algae (green, blue-green and diatoms). They are ubiquitous, especially in
the surface layers of soil. The most important environmental factor governing the
spread of algae is moisture, although they are able to withstand long periods of
drought. Morphological diversity of algae is very large, but they are microscopic in
size, thread-like form and are composed of a single cell. The most numerous blue-
green and green algae. Their number in 1 g soil can reach 100 thousand.
2. Mushrooms. They can be divided into three groups: yeasts and yeast,
mold, including filamentous fungi, basidiomycetes. Yeast and yeast-like fungi was
common in ordinary soil, so their role and importance in the life of the soil are
small. Mold and basidiomycetes more numerous in the soil, especially in
basidiomycetes forest soils, where they cause the formation of mycorrhizae. Fungi
can live in conditions of partial anaerobiosis, but aerobioz stimulates their
development. The number of fungi in the soil surface layer of 8 thousand. 1
million. 1 g, and biomass - from 1000 to 1500 kg / ha. The most favorable reaction
medium for mushrooms - acidic (pH 4.0).
3. The bacteria (spore-forming bacteria, spirochetes, mycobacteria,
psevdomonady, azotfyksuyuchi and nitrifying bacteria, archaea). At the bacteria
cultivated soils surpass all other groups of microorganisms, both in number and in
their diversity. The number of bacteria in 1 g soil ranges from 300 thousand. To 95
mln. And even up to 4 billion. In the fertile soil total biomass of bacteria reaches
500 kg / ha and more. Bacteria are divided into heterotrophs and autotrophs.
Heterotrophs energy use and carbon enclosed in complex organic substances.
Autotrofy use the energy released by the oxidation of minerals, extracting carbon
from carbon dioxide and nitrogen - mineral compounds. Most of the soil bacteria
belonging to heterotrofsd, that demand for its existence hotovs orhanichns
rechovynb.
In relation to oxygen soil microorganisms are divided into aerobic (requiring
for their existence free oxygen) and anaerobic (not requiring for its existence
oxygen free). The greatest value in soil with nitrogen-fixing bacteria able to

10
assimilate molecular nitrogen (Azotobacter, Nitrobacter, Mycobacterium and
others), and spore-forming bacillus genera Bacillus and Clostridium.
Soil microorganisms involved in the process of soil formation, soil
purification, circulation in the nature of nitrogen, carbon and other elements. In the
soil there are all conditions for microbial growth, a sufficient number of organic
and mineral substances for their food, suitable humidity and reaction medium,
protection from direct sunlight, oxygen. Quantitative and species composition of
microorganisms in the soil due to it containing organic matter, moisture, pH,
temperature, weather conditions, method of treatment. As the number of organic
matter in the soil, usually increases and the number of microorganisms. Organic
matter is a breeding ground for most soil bacteria. The total stock of soil organic
matter reaches 1 400 tonnes per hectare, of which most of the surface layer is in
(30 cm) of soil. Main part of soil organic matter - the remains of animal and plant
tissues. The live weight of microorganisms in 1 hectare of soil (fertilized) exceeds
5.6 tons. The richest microorganisms black, chestnut soils sirozemy and specially
treated soils. The number of bacteria in 1 g of soil sometimes reaches several tens
of billions. Poor microbiota sand, rocks and soil devoid of vegetation. The most
numerous organisms in the upper 5-15- cm layer, at a depth of less than 20-30 cm
and the minimum number at a depth of 30-40 cm. However, the bacteria were
found in the soil even at a depth of 5 m. Soils rich in bacteria, biologically active
pain . Between the fertility of soil and content in it of microorganisms is a definite
relationship. Calculations showed that for every hectare of marginal soils account
for 2.5-3 tons of microbial mass vysokorodyuchyh - up to 16 tons. The number of
microorganisms in 1 g soil can range from 1-3 • 20-25 • 106 to 109.
The maximum number of microbes found in the soil at a depth of 10-20 cm.
Since the depth of 1-2 m, their number is declining dramatically. This is because
with the deepening of the soil organic matter content decreases and oxygen
necessary for life aerobic bacteria. The number of microorganisms in the soil
increases in the direction from north to south, and in the spring of their number
increases significantly, peaking before the summer, fall; winter - dramatically
11
reduced. Typical soil bacteria include Bacillus subtilis, Bacillus mycoides, Bacillus
mesentericus, Bacillus megatherium, and thermophilic bacteria and other
microorganisms that are sometimes 80-90% of the soil microbiota.
Pollution and soil purification.
Soil populated areas contaminated solid and liquid garbage, human and
animal secretions, their dead bodies, the remains of plants, domestic and industrial
wastewater. Together with organic contamination in the soil gets a lot of
microorganisms. Especially dangerous in epidemiological sewage slaughterhouses,
meat processing plants, companies in processing leather, wool, which may contain
pathogenic bacteria. In this connection, the soil may be a factor of transmission of
infectious agents. Because the soil contamination can occur saprophytic and
pathogenic microbes raw food, feed. Therefore scum entering the soil shall be
subjected to purification and disposal. The duration of survival of pathogenic
bacteria in the soil depends on biological characteristics and habitat conditions.
The longest living spore-forming bacteria - pathogens tetanus, botulism; bacilli of
anthrax spores may persist for decades. Under favorable conditions micro-
organisms in the soil not only survive, but for a long time (weeks, months or even
years) retain virulence properties.
Classification of soil pathogens:
- Pathogenic microorganisms live permanently in the soil (eg, the agent of
botulism). The bacteria into the soil with feces of humans and animals, their spores
stored in it indefinitely.
- Pathogenic spore-forming microorganisms for which the soil is secondary
reservoir (eg, anthrax). The bacteria into the soil with feces and other secretions of
sick animals, and with the corpses of dead animals.
- Pathogenic microorganisms into the soil with secretions of humans and
animals and are kept for weeks or months. In this group are not different
microorganisms form spores. The main factors that lead to rapid death of
microorganisms - inability to sporogonic and antagonistic properties of soil
microbiota (competition for energy and food).
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 The duration of survival of pathogens in soil depends on the biology of the
pathogen, moisture content and appropriate nutrients, pH, temperature, presence of
microbes-antagonists, bacteriophages. In wet soils survival 2-4 times longer than
dry. Asporogenous bacteria killed rather than sporoobrazuyuschye. Pathogenic
microbes asporogenous survive in the soil little time dysentery pathogens - from 10
days to 9 months; vibrio cholerae - from 10 days to 4 months; typhoid bacteria -
from 14 days to 10 months; tularemia bacteria - from 10 days to 2.5 months;
Mycobacterium tuberculosis - from 3 to 7 months or more; Brucella - 2 to 3
months. Nesporoobrazuyuschyh survival in the soil promotes microbial agent,
along with getting enough nutrients (faeces, sputum, pus and so on. D.) The
favorable physical and chemical environmental conditions, lack of microbes-
antagonists. The most dangerous is soil contaminated with feces of patients with
intestinal infections. Pathogens dysentery, cholera, typhoid fever, salmonellosis,
enterovirus diseases enter the body with contaminated ground vegetables, fruits
and other foodstuffs. A direct correlation between the level of morbidity with
intestinal infections and poor sanitary conditions of the soil caused by its poor
clearance. We describe a number of water outbreaks of intestinal infections caused
by contaminated soil had runoff and sewage. In the soil home to many fungi. Some
of them, such as fungi of the genus Fusarium, falling on crops and other plants in
its development process, produce toxic substances. When used bread baked from
grain threshing and later affected the fungus Fusarium sporotrichiella, there
toxicosis in humans, known poisoning "drunk bread". Mushrooms of the genus
Aspergillus (A. flavus, A. fumigatus), parasitic on ground nuts, cereals and fodder,
may also form toxic substances - aflatoxin. When eating foods contaminated
aflatoksynov there is severe poisoning, characterized by necrotic liver, kidney,
hemorrhagic inflammation of the digestive tract.
Soil health assessment for microbiological parameters.
In assessing the sanitary soil into account the results of chemical,
microbiological and helmyntolohycheskoho research. Microbiological studies
conducted for the health assessment of soil characteristics purification processes,
13
assessment methods biotermal soil and waste disposal, in determining the
suitability of land for construction, as well as epidemiological and epizootic
surveys to determine ways of soil contamination, duration of survival of
pathogenic microbes in it and t. e. Depending on the task using short or full
sanitary-bacteriological analysis of soil. Short sanitary and microbiological
analysis involves determining GMC, titles bacteria of Escherichia coli,
enterococci, Cl. perfringens, thermophilic bacteria, nitrifying bacteria. The results
indicate the presence and extent of fecal contamination. A brief analysis of the soil
carried out during the current sanitary surveillance of soil.
Full sanitary-microbiological analysis includes identification of all
parameters of a short analysis and total saprophytes, OMCH and percentage of
spore microorganisms, aerobic bacteria that destroy tissue, bacteria
ammonyfykatorov. Also, examine the toxicity to soil microorganisms. Full
analysis carried out in the implementation of preventive sanitary inspection, initial
inspection when choosing a site for the placement of individual objects.
Sanitary value of soil microbial number can not be considered without
taking into account the features of different types of soil. For example, black soil
microorganisms contain much more than podzol. Therefore, when determining the
total number of bacteria in the soil is necessary to compare the results of
unpolluted soil microbial count of the same type. Research on the direct detection
of pathogenic microbes in soil is carried out only in the investigation of outbreaks
of infectious diseases. As indirect indicators of possible soil contamination by
pathogenic bacteria use sanitary indicative microorganisms: bacteria of
Escherichia coli, CI. perfringens, bacteria of the genus Proteus, thermophilic
bacteria. The presence of soil bacteria of E. coli indicates faecal contamination it.
Detecting CI. perfringens in the soil as it indicates faecal contamination. The soil
layer once enriched bacteria of E. coli and CI. perfringens. After 4-5 months
marked dying E. coli, a CI. perfringens is found in titer of 0.01. Consequently, CI.
perfringens have sanitary indicative value only if the titer is determined in
conjunction with the circle captions and other indicators. Fresh or old faecal
14
contamination of the soil can be determined by the ratio of vegetative forms of CI.
perfringens and spore forms of the microbe. Detection of soil bacteria of the genus
Proteus indicates contamination of organic substances or animal feces of people.
Thermophilic bacteria are indicators of contamination of soil with manure,
compost. In pure soil thermophiles do not show.
Methods for determining the composition and activity of soil
microorganisms.
To evaluate the use of soil biota indicator of biological activity of the soil.
The biological activity of the soil is determined in the following ways:
- Estimates of the total number of soil microorganisms. Due to imperfect
methods, this method gives conditional determination, rough characterization of
the biological activity of the soil.
- Determination of the number of individual physiological groups of
microorganisms, such as nitrifying bacteria.
- Determination of soil carbon dioxide is released - the main biochemical
method for determining the biological activity of the soil. The more intense carbon
dioxide from the soil, the more place it biological processes, better conditions for
the cultivation of crops and their potential yields above.
Of carbon dioxide from the soil surface layer of the atmosphere called soil
respiration. The intensity of soil respiration depends on the properties of
hydrothermal conditions, nature of vegetation farming practices. Bold carbon
dioxide in the soil increases its cultivated due to the activation of biological
processes and improving the conditions of aeration. Reduction of carbon dioxide
ground (reduced biological activity) may impair the flow of oxygen into the soil,
which in turn promotes the formation of toxic substances. To examine soil health
assessment tests to identify pathogens. To adequately assess soil has special
significance selection indicator microorganisms.
Evaluation of fecal contamination of soil and its limitations is held on the
following parameters:
- For bacteria group E.coli index (number of bacteria of E. coli in 1 g soil);
15
 - For perfrinhens captions (least amount of soil, which turns CI.
perfringens);
- For a titer enterococci.
All Enterobacteriaceae most long stored in the soil E. coli, because its
content is judged by the presence in the soil of other Enterobacteriaceae.
Thermophilic bacteria into the soil with rotted manure or compost, so they should
exercise to ascertain the nature and limitations of organic soil contamination. Fresh
manure, sewage usually contain many bacteria group E.coli, but few thermophilic
bacteria. As the decomposition of organic matter increases the number of
thermophiles. The appearance of nitrifying bacteria indicates the development of
self-cleaning process, as they complete the cycle decomposition of nitrogen-
containing compounds, converting ammonia into nitrogen. In fresh fecal
contamination nitryfikatoriv will not, as a substrate for their development is
missing. During the microorganisms that decompose organic matter, ammonia is
formed, which stimulates the development nitryfikatoriv. In fresh fecal
contamination of soil indicating high titers bacteria group E.coli nitryfikatoriv
credits at low and relatively high content of vegetative forms of CI. perfringens.
Identification of enterococci always indicates fresh faecal contamination,
whatever the other indicators. The purpose of the sanitary-microbiological study of
soil:
sanitary evaluation of soil settlements and new areas for settlement and
placement of buildings;
issues of water supply and wastewater settlements;
sanitary evaluation of soil contaminated with chemicals;
control processes of self-purification of soil that has undergone biological
contamination;
Soil epidemiological survey to determine the ways of contamination.
Sampling was carried out with a square section (at least 5x5 m) with 5 points -
from every angle and center square ("envelope method"). Sample number 1 kg
taken in aseptic conditions with a depth of 20-30 cm. The frequency of monitoring
16
depends on controlled objects, but at least 1 time per year. In the study of the
dynamics of self-purification of soil in contaminated areas taking samples during
the first month after the contamination week, in the coming months - 1 once a
month during the growing season to the active phase of self-purification.

Microbiota water.
In the seas, rivers, lakes and other bodies of water as well as groundwater
contains a large number of species of microorganisms. The combination of all the
microorganisms that inhabit ponds, denoted by the term "mykrobyalnoy plankton."
Microbiota of natural waters largely depends on their origin. There are fresh and
sea water. Fresh waters are divided into superficial, including flow (rivers,
streams) and standing (lakes, ponds, reservoirs), groundwater (soil, groundwater,
artesian) and atmospheric (rain, snow). Study groups engaged in Water
Hydrobiology. The growing shortage of fresh water on Earth makes pay serious
attention on the development of the ecosystem in the pond and processing of water
pathogens entering the body of water contamination. Water - the natural habitat of
microbes, the bulk of which comes from soil, dust settling from the air, waste,
sewage and industrial livestock facilities and others. A lot of micro-organisms in
open ponds and rivers, they are often found in muddy sediments of oceans, seas,
swamps, mineral waters. Their invention as the surface layers, and to a depth of 10
thousand. Meters. Living organisms in hot springs. The process of photosynthesis
occurs to them at 75OC and alkaline waters bacteria survive at 100°C. Quality of
living microorganisms in water depends mainly on the properties of the water flow
to its wastewater and industrial waste. By constantly living microorganisms in
water include Azotobacter, Nitrobacter, Micrococcus, Pseudomonas, Proteus,
Spirillum and others. Deep groundwater, key, artesian water is almost free of
microorganisms. Slightly soiled microbiota precipitation, as snow and water
capture most germs the air with dust and after rainfall especially clean air.
Character microbiota water features determined by the particular aquatic
environment.
17
 Microbiota water form two groups: autohtonous (actually water) and
allochthonous (come outside at pollution) microorganisms. Autochthonous
microbiota - a set of microorganisms that permanently live and breed in water.
Microbial composition of water resembling soil microflora, which faces water
(bottom and coastal soil). Allochthonous microbiota - a set of microorganisms that
accidentally hit the water and kept it relatively short time.
The proportion of microorganisms in open water variyuyuye widely,
depending on the type of reservoir, the degree of pollution, changing weather
conditions, seasons. Water Microorganisms play an important role in the
circulation of substances splitting organic products of animal and plant origin and
providing nutrients other organisms living in the water. The source of water
pollution in rivers often serve domestic and industrial waste water. In open water
most of the bacteria comes from the soil. Therefore, in lakes, ponds, rivers highest
content microbiota noted in the coastal zone. In the water live all known groups of
microorganisms, but the most significant component of population water - bacteria.
As you know, tsytoplazmatycheskaya membrane of bacteria has the ability to
transfer active through the cell wall of nutrients. This bacteria can consume
nutrient substrate present in minute concentrations (1-5 mg / d). Microbes mineral
compounds to oxidize organic matter in large quantities fall into the water. The
degree of contamination, including pathogens, may be an obstacle to the use of
water. Therefore, any water source must expose sanitary microbiological
assessment.
Self-cleaning water is caused by several factors:
fast current of water, leading to a decrease in the concentration of organic
substances;
bactericidal action insolation;
mineralization of organic compounds by microbes;
existence food chain: bacteria - simple - insects - ryba- animals - man;
adsorption of solid particles of sludge;
adsorption of the plants;
18
 the influence of volatile plants.
Water is a factor in the transmission of many infectious disease pathogens.
In open water, especially in disadvantaged areas on infectious diseases, detect
intestinal pathogens and natural focal infections. In the sediments of ponds and
lakes often live pathogens botulism. Pathogenic microorganisms reservoirs may be
included in the food chain and transferred them to different groups of animals,
birds and fish. We know that waterway transmitted typhoid fever, bacterial and
amebic dysentery, cholera, leptospirosis, polio, hepatitis A and E and other
illnesses. When the sanitary-bacteriological analysis of water determined:
the total microbial count (total number of microorganisms in 1 ml);
pathogenic microorganisms;
number bacteria group E. coli as an indicator of the degree of fecal
contamination.
Advanced determine titer CI. perfringens, index bacteriophage and Giardia
cysts. The presence of pathogens determined by epidemiological indicators.
Among the facilities subject to microbiological controls, the most important place
is given to research water. In accordance with the existing regulations are subject
to control:
drinking water (central and local supply);
water swimming pools;
water open water;
wastewater;
purified water to prepare drugs;
water for preparing injection solutions and eye drops.
The warning surveillance exercise:
1. in matters of water and sewage populated areas;
2. In assessing the sanitary pools, beaches, places of collective recreation.
Current sanitary inspection shall:
1. in assessing the quality of drinking water supply of settlements;

19
 2. in assessing the health status of surface water to establish the impact of
biological contamination on the ability of water to cleanse itself;
3. When controlling the disinfection of wastewater;
4. epidemic indications to identify the possible transmission of infectious
diseases (sanitary spend detection demonstration and pathogens).
For water sampling is used as reusable and disposable sterile utensils.
Multiple made from materials that can withstand dry heat treatment and
autoclaving. Capacities for water intake closed tight plugs and protective cap foil
or thick paper. Before taking samples of tap water wipe swab moistened with
alcohol, and burn, and then 10-15 minutes drained stagnant water in the pipes and
then selected a sample for study.
Analysis carried out immediately after sampling. If you need to transport
water stored at 1-5º C and analyzed not later than 2-6 hours after its intake.
1. Determination of the total number of microorganisms in the water.
The total microbial count of water is determined by culturing bacteria contained in
the samples on solid media. Depending on the alleged contamination of the
reservoir before planting prepare Tenfold dilution of initial samples in sterile tap
water. Table 1 shows recommended for crop breeding water depending on the
degree of contamination (amount of each dilution for further planting in IPA is 1
mL).
Table 1.
Recommended for crop breeding water depending on the degree of contamination
(each volume for crop breeding is 1 mL)
Type studied water Recommended dilution water
Tap water and water from artesian wells 1 ml of water without dilution
Pure water (water wells, springs, etc., 1 и 1:10
the water of swimming pools)
Open water, not contaminated effluents 1; 1:10 и 1:100
Pure water bodies in places of mass 1:10 и 1:100

20
bathing
Open water contaminated with sewage 1:10; 1:100 и 1:1000
Heavily contaminated wastewater 1:10000; 1:10 0000 и 1:100 000

For dilutions are a number of tubes containing 9 ml of sterile water.

Investigated water in volume of 1 ml are making the first tube, getting dilution of
1:10, then this transfer tubes 1 ml to the next, etc. To prepare each dilution using a
new sterile pipette. From these dilutions made in 1 ml of water in 2 Petri dishes for
calculating the average. and pour 15-20 ml of melted and cooled to 45ºS IPA. The
content of the cups thoroughly mixed in a circular motion, moving them on the
table surface. After solidification of agar, a cup is placed in an incubator for 24
hours at 37ºC. Colonies of bacteria grow on the surface of the culture medium
(aerobes) and its depth (anaerobic). Calculates their total amount and calculate the
total microbial count. If pre-diluted water, the resulting amount is multiplied by the
degree of dilution and the result is the number of microorganisms in 1 ml of source
water. The total microbial count in 1 ml drinking water should not exceed 50.
2. Identification of bacteria of Escherichia coli group.
Sanitary representative microorganisms in water, as well as in the soil, is a
group of bacteria Escherichia coli. They also called koliformni. This group
comprises facultative anaerobic members of the family Enterobacteriaceae. All
have palochkovydnu form, do not form spores, Gram-negative,
oksidazoonehatyvni, decompose lactose to acid and gas. Note the temperature at
which the most actively manifested saccharolytic properties bacteria of Escherichia
coli group. Most are lactose positive 24-48 hours at 37ºC. These bacteria include
the general). A distinctive feature termotolerants bacteria is that they decompose
lactose to acid and gas at a higher temperature - 44ºC for a short time - 24 hours.
Detection indicates faecal contamination of fresh water. E. coli bacteria are a group
of different methods. The most common method of membrane filters.

21
 To determine bacteria of Escherichia coli group use this method Zeitz filter
device, which before the research wipe swab dipped in alcohol, sterilize calcination
and set on Bunsen flask.
Then the device is placed nitratsellyuloznyy or acetate cellulose membrane
filter of pore diameter less than 0.45 microns. Such filters are pre-sterilized by
boiling. Elected to study the volume of water passed through the filter attaching the
device to the vacuum pump. If several water samples analyzed, for each use
separate membrane filter. Before new tests filtration apparatus sterilized. After
passing through these water filters is placed on the surface of Endo medium in
petri dishes, placing them in a nutrient medium filter side up. Cups then incubated
in a thermostat at 24 hours 37ºC. The structure of the environment Endo are
lactose, indicator and therefore constitute bacteria of Escherichia coli group her red
colonies with a metallic sheen. Count the number of colonies, prepare them smears
and Gram stain and check oksydaznuyu activity. Oxidase-negative bacteria that
decompose lactose to acid and gas discovered in the filter can give a positive
response about the presence of water bacteria of Escherichia coli group. In the
analysis of drinking water bacteria of Escherichia coli group calculate the amount
contained in 100 ml.
For differentiation common bacteria of Escherichia coli group and
termotolerants bacteria of Escherichia coli group each colony grown on the filter
sown in two test tubes with lactose environment. One of the tubes is heated to 44ºC
pre-order in order to inactivate bacteria general. Then this tube incubated at the
same temperature for 24 hours. The second tube of sowing put in a thermostat at
37ºC 48 hours to verify the presence of common bacteria of Escherichia coli group.
The polluted water open water pre-diluted, as indicated in the table. 1 and is
used to filter the amount of not less than 10 ml. Further research is carried out as
described above.
To determine the range of water samples often use two-phase fermentation
method.

22
 The first phase (first day) - make crop on the environment Eykman (glucose-
peptonne environment) with floats for gas collection and crops pose a thermostat
(incubated) at 43OC 24 hours.
For sowing small volumes of water used Eykmana diluted medium (1%
peptone; 0,4% NaCl, 0,5% glucose). For sowing large volumes - Eikman
concentrated medium containing 10-fold concentration of the main components.
Concentrated environment Eikman used for analysis of water. Do took two water
samples 100 ml flasks of 10 ml of medium and ten samples of 10 ml of water in
tubes with 1 mL of medium. Therefore, the amount sown water - 300 ml: 2 flask of
100 ml and 10 tubes of 10 ml.
The second phase (2nd day) - make reseeding on Endo medium of the flasks
and tubes where there is growth of the colonies. Symptoms of E. coli growth in the
medium Eikman - diffuse turbidity and gas formation. Crops were incubated at
37OC for 24 hours.
The third stage (3rd day) - see crops on the environment Endo. Signs of
growth of E. coli on Endo medium - smooth formation of colonies of red color
with metallic luster. Conduct microscopic confirmation of E. coli: from suspicious
colonies make paint smears and Gram; gramme observed under the microscope "-"
small sticks.
Carry out biochemical confirmation E. coli - cytochrome oxidase test. If
there cytochrome oxidase - blue paper for 1 minute. E. coli - oksidazonehatyvna.
Oxidase test to distinguish E. coli from gram but oksidazopozytyvnyh bacteria
family Pseudomonadaceae. If detected in smears gram "-" small sticks that are
oksidazonehatyvnymy, the analysis is considered positive (conclusion: detected E.
coli).

Microbiota air
Air microbiota depends on the microbiota of soil and water, where bacteria
with the dust and droplets of moisture in the atmosphere are captured. Air -
unfavorable environment for the proliferation of microorganisms. Lack of
23
nutrients, sunlight and drying cause rapid death of microorganisms. Consequently,
in the air constantly there are processes of self-purification. The composition of the
microbiota air is very diverse - a pigment saprophytic bacteria (micrococci,
sartsiny), actinomycetes, molds, yeasts, etc. The greatest number of air containing
microorganisms major industrial cities. Air same fields, forests, meadows and
expanses of water, the removal of settlements is different comparative purity.
Undergoes significant changes microbiota air depending on the season. The
maximum number of microbes found in the summer, and the minimum - in winter.
Microbiota indoor air is more diverse and relatively stable. Among the
microorganisms dominate human inhabitants of the nasopharynx, including
pathogenic species that are released into the air by coughing, sneezing or talking.
The main source of air pollution pathogenic species - bacteria. The level of
microbial contamination depends on the population density, movement of people
activity, the sanitary condition of the premises, ventilation, ventilation frequency,
method of collection, and so the degree of illumination. D.
Microorganisms in the air in a state of aerosol. Aerosols - colloidal system
consisting of air, liquid droplets or solid particles, and includes a variety of
microorganisms. The size of aerosol particles ranging from 10 to 2000 nm. When
sneezing may form up to 40,000 droplets. There are three main phases of bacterial
spray:
drip phase consists of bacterial cells, surrounded by water and salt tablets.
Diameter of about 0.1 mm. Length of stay in the air is a few seconds;
shallow nuclear phase particles formed by the drying of the first phase. In
this phase particles have the smallest size, easily moving streams of air are
continued in limbo. That's how most pathogens spread airborne infections;
phase "bacterial dust" consists of large bistro deposited particles forming
dust that could rise into the air.
Pathogenic and opportunistic microorganisms in the air come from drops of
human saliva or animals when talking, coughing, desquamation of epithelial cells
in the skin. Transmitted through the air:
24
 bacteria - Mycobacterium tuberculosis, diphtheria, pertussis, spore forms of
bacteria etc.
viruses - pathogens of acute respiratory infections (varicella, influenza,
parainfluenza, and others.);
mushrooms of the genus Aspergillus, Mucor, Penicillium and others.
To assess the sanitary condition of indoor air determine the total microbial
count and the number of sanitary indicative microorganisms which are hemolytic
staphylococci, α- and β- hemolytic streptococci. If necessary, for example, surgical
hospitals, maternity hospitals additionally determine the presence and amount of
Pseudomonas aeruginosa and others. Opportunistic gram-negative bacteria -
pathogens of nosocomial infections.
The number of bacteria and the number of sanitary indicative
microorganisms determine their quantitative content in 1 m3 (1000 liters) of air.
Currently, there are many methods and devices for air sampling and their research.
The most simple and affordable for sanitary-bacteriological study is sedimentation
and air aspiration techniques.
Koch sedimentation method based on spontaneous microorganisms settling
under gravity on the surface of the culture medium open Petri dish.
To determine the total microbial number two petri dishes with sterile MPA
left open for 10-30 minutes. Then they close, nadpysuyut and incubated in a
thermostat at 37ºC within 24 hours. Then crops can withstand 24 hours at room
temperature to detect fungi. Thus, after 48 hours count the total number of colonies
that grew on the cups. Comes from the fact that for 5 min. the surface 100sm2
dense medium are deposited bacteria with 10 liters of air (Omelyanskiy V.L.), or
use the data in Table 2.
Table 2
Calculation of microorganisms in 1 m3 of air
The diameter of Area cups, sm2 The coefficients for calculating the
the cup, sm number of microorganisms in 1 m3 of air

25
8 50 100
9 63 80
10 78 60
11 95 50
12 113 45

Example: in the cup with a diameter of 10 cm 40 colonies grew, so, the

number of microorganisms in 1 m3 of air is 40 x 60 = 2400.
To identify sanitary-indicative microorganisms using special culture media:
for staphylococci - yolk-salt agar (exposure 15 min) to hemolytic streptococci and
staphylococci - blood agar (exposure 10-15 minutes) for mushrooms - medium
Saburo (crops stand 3- 5 days at 20-22 ° C).
Aspiration method is based on the air stream of impact on the surface of the
culture medium to which microorganisms settle. It is conducted using the
apparatus Krotov or its modern versions. They consist of a hub for air sampling,
micromanometer and electric motor. The phone Krotov node sampling is mounted
in a metal casing and a centrifugal fan, ground clamps for installation of the Petri
dish, cover with pleksyhlaza, which cut wedge gap for air intake. On the
playground set open Petri dish with nutrient medium, close lid and include motor
vehicle. Turning the centrifugal fan air is sucked through the gap wedge and force
hits the surface of the culture medium in which microorganisms are deposited
uniformly distributed on it. Rotation speed adjustable Petri dishes, allowing you to
pass different amounts of air per minute, which is fixed micromanometer. After a
specified exposure time turn off the engine, a petri dish with sowing air is
removed, close and put in an incubator.
To determine the total microbial numbers using MPA, the speed of air
passing through the apparatus 25 liters / min. 4 min of exposure, which ensures
sedimentation of microorganisms on the amount of at least 100 liters of air. To
identify Staphylococcus aureus using the yolk-salt agar, hemolytic streptococci and

26
staphylococci - 3-5% blood agar and exposure time increased to 10-15 minutes,
providing bacteria with sowing 250-300 liters of air.
Sowing air spend two petri dishes with agar IPA and grow 48 hours (24
hours in a thermostat at 37ºC, then stand 24 hours at room temperature). Petri
dishes with blood agar ynkubyruyut 37ºC in a thermostat at 24 o'clock. Count the
number of colonies that grew and the data transfer for 1 m3 studied air.
Example calculation: on one Petri dish with counting colonies detected 246,
the second - 254, an average of 246 + 254 = 250 colonies. The device rotating Petri
dish for 2 minutes at 25 l / min. There were missing 50 liters of air. Thus in 50
liters of air contains 250 microbes in the general microbial count in terms of 1 m3
of air (250 • 1000): 50 = 5000 bacteria.
The study of quality of the microbiota carried by conventional techniques:
from the colonies make smears, Gram stain, isolated pure culture, which identify.
In the study of air additionally determine spore-forming anaerobes. For this
purpose, make planting air volume of 200-300 liters per petri dish with iron-
sulphite medium is incubated in a thermostat at 37ºS 24 hours. To detect mold
sowing air Saburo have on the environment and cultured 3-5 days at 20-22 ° C.
Acceptable levels of bacterial contamination of air environment of different areas
of medical institutions and pharmacies are presented in Table 3.

Table 3
Acceptable levels of bacterial contamination of indoor air treatment facilities
depending on their grade of cleanliness and functionality
Class of cleanliness The total number Number of Number of
of colonies of colonies of S. colonies of mold
microorganisms in aureus in 1 m3 of and yeasts 1 dm3
1 m3 of air air air
Class А (Super No more 200 There should be There should be
purity)
Class B (clear) No more 500 There should be There should be
Class C No more 750 There should be There should be

27
 (conditionally
clean)
Class G not standardized not standardized not standardized
(contaminated)

Notes:
Class A - operating, delivery rooms, aseptic boxes wards for premature
babies;
Class B - procedural, dressings, perioperative, intensive care wards and
rooms, children's Chamber;
Class B - Chamber of patients, review, ordynatorski, material, barns clean
linen;
Class D - hallways, stairways, sanitary rooms, toilet rooms, dirty laundry
and temporary storage of waste.

Microbiota of the human body

In humans live about 500 species of microorganisms that form its normal
microbiota. Microorganism and its microbiota in normal conditions in a state of
dynamic equilibrium (eubioza), which has developed in the course of evolution.
The open habitats, which communicate with the external environment is - skin
located to the glottis respiratory tract, mouth, gastrointestinal tract, mucous
membranes gas, nose, anterior urethra, vagina. They colonized by microorganisms,
mostly bacteria.
Protozoa and viruses are far fewer species. Normally free from
microorganisms - blood, cerebrospinal fluid, synovial fluid, bone marrow,
abdominal cavity, pleural cavity, the uterus.
As already mentioned above the natural microbiota any habitat depending on the
value of the index is divided into permanence resident (or constant), optional (or
more) and transient (or random). If the permanent members of the microbiota
contains specific to this habitat, it consists of random individuals listed outside.

28
Thus, in the gastrointestinal tract may be extraneous organisms caught from food
or drink. Skin most commonly kontaminiruyutsya random microbiota from the
environment. In the trachea, bronchi, lungs, esophagus can also occur transient
microbiota.
Permanent microbiota particular habitat is relatively stable in composition.
However, the structure and components of the physiological role of
microorganisms is not equivalent. Therefore, regular microbiota distinguish two
factions: oblyhatnuyu and optional. Obligate microbiota is the main component of
any mykrobotsenozov, it prevents colonization habitat random microorganisms
involved in the fermentation process, ymmunostymulyatsyy, that perform
protective functions or normofiziolohicheskie. Share obligate microbiota in healthy
biological community composition several times higher than the proportion
optional fraction. For example, the concentration of bifidobacteria in the colon
reaches 109-1012 colony forming units (CFU) - the number of colonies growing in a
nutrient medium at sowing 1 g or 1 ml of the material. In determining the
concentration of microorganisms take into account that each live bacterial colony
forming cell).
Optional microbiota is less than residents of habitat, the maximum
concentration of individual representatives not exceed 103-105 CFU / g. If
permanent microbiota manifests itself mainly brodylnoy activity (ie splitting of
carbohydrates with the formation of acid products) is optional fraction very active
in putrid processes.

Composition normal microbiota of the human body

Microbiota of skin.
The skin is the biggest area of the human body, available for regular contact
with environmental pathogens. The composition of the resident microbiota skin are
Gram-positive saprophytic bacteria - staphylococci, micrococci, nonpathogenic
corynebacteria. To include transient microbiota sartsiny gram, Staphylococcus
aureus, fungi of the genus Candida, molds. Thus, the composition of the skin
29
microbiota represented as aerobic and anaerobic species. The main areas of
colonization - surface dead cells of the epidermis, the mouth of the hair follicles,
sebaceous glands. At one cm2 of skin can be 10 thousand. 1 million. The bacterial
cells. Bacteria break down the secrets of the sebaceous glands to unsaturated fatty
acids, with a shift towards the acidic pH. Acidic reaction medium and metabolic
products of Representatives normal microbiota have unfavorable conditions for
pathogenic bacteria that are on the surface of healthy skin die quickly (within 5
minutes). With the weakening of defense reactions of microorganism on the skin
increases the number of Gram-negative bacteria, including Escherichia coli (E.
coli).
Microbiota of the upper respiratory tract
Most colonized the upper respiratory tract, are anatomically adapted for
deposition of bacteria from inhaled air. Rezydentnaya microbiota the nasal cavity
and nasopharynx represented by gram-positive streptococci viridans and
nehemolytycheskymy, peptostreptokokki, micrococci, staphylococci, lactobacilli.
From here hramnehatyvnyhmikroorhanizmiv live non-pathogenic and anaerobic
neyseriyi asporogenous sticks - bacteroides.
Microbiota of the gastrointestinal tract.
Mouth - one of the most populated areas of the human body, there appears
some 300 species of microorganisms (Table 4). In the mouth living representatives
of all morphological forms of bacteria: cocci, bacilli, spirochetes form and
protozoa, fungi, viruses. High oral contamination promote its anatomical features -
the presence of gingival pockets, folds mucosa, interdental spaces - a large number
of nutrients, alkaline environment, sufficient supply of oxygen.

Table 4
Microbs landscape oral cavity healthy
Мікроорганизми %
Streptococci 100

30
 Lactobacilli 90,3
Staphylococci 40,7
Fungi genus Candida 25,7
Bacteroides 21,0
Corynebacteria 13,1
Neisseria 6,9
Veilonella 5,3
Lepthotrichia 4,5
Fusobacteria 3,5
Actinomycetes 3,0
Spirochaetes 2,7
Micrococci 2,0
Simplest 0,9

To obligate microbiota relates primarily bulk Gram-positive cocci,

represented heterogeneous group streptococci. This group includes Str. salivarius,
Str. sanquis, Str. mutans. They differ in their ability to ferment carbohydrates and
form hydrogen peroxide. The shift towards the acidic pH leads to decalcification of
the enamel. Important as the ability to synthesize streptococci from sucrose
polysaccharides. This part of the molecule Glucose is converted to insoluble
dextran, which promotes the formation of dental plaque. Streptococci found in the
mouth in different proportions, depending on diet, oral hygiene, age and other
factors.
Gram-negative anaerobic cocci are born Veilonella. They are well laid
lactate, pyruvate, acetate and other carbohydrates to carbon dioxide and water. By
catabolism of lactic acid formed streptococcus, veylonelly can provide
protyvokaryoznoe action.
Gram-positive rods are in the mouth comes Lactobacillus. They decompose
carbohydrates to form lactic acid, while maintaining viability at low pH. The most

31
commonly in the mouth of healthy individuals are L. casei, L. acidophylius, L.
salivarius.
Gram-negative anaerobic and microaerophilic bacteria often belong to the
family Bacteroidaceae. They ferment sugar to gas and peptone - with the formation
amino acids. For this family includes three types: Bacteroides, Fusobacterium,
Leptotrichia. The most common B. melaninogenicus and B. gingivalis. They are
characterized by low saccharolytic activity, but glucose decompose to form a
mixture of acids and pH is quite high. These types are permanent residents gingival
pockets. The presence of proteolytic enzymes in bakteroydov is of great
importance in the pathogenesis of periodontal diseases.
Reed presented Fusobacterium fusiform sticks. They form a peptone glucose
or lactic acid. Fuzobakterii live in gingival pockets in association with the
spirochete.
With family Actinomycetaceae oral most common genera Actinomyces and
Bifidobacterium. The first fermented carbohydrates to form acidic products
without gassing. The final products of glucose cleavage are acetic, lactic, formic
and succinic acid. Have weak proteolytic activity. Actinomycetes are on the
mucosa of the mouth, form the stroma of tartar and part of the plaque. Along with
this, they are in the cavities of teeth in pathological gingival pockets in the ducts of
the salivary glands.
In the mouth there are bacteria genus Corynebacterium. A characteristic
feature is their ability to reduce the redox potential, thus creating conditions for the
growth of anaerobes. If periodontal diseases are found in association with
fuzobakteria and spirochetes.
Spirochetes that live in the mouth, belong to three families: treponemy
presented oral species T. macrodentium, T. denticola and T. orale. They differ
from each other in the formation of lactic, acetic and other acids and carbohydrate
utilization. Boreliyi presented oral B. buccalis, often found in association with
fuziformnymi bacteria.

32
 In the mouth there mycoplasma - rale M. and M. salivarius. They
hydrolyzuyut arginine, which does not ferment glucose and differ in some
biochemical characteristics.
Of all the factors that determine the nature and state of the microbiota of the
mouth, saliva is crucial. The most important factors in this respect saliva is the
intensity of its formation, viscosity, content of mineral components, ion potency,
buffer properties, pH, major metabolites, presence or absence of salivary gases,
organic composition (particularly amino acids, polysaccharides, vitamins, purine,
pyrimidine) , antibacterial properties (lysozyme, secretory antibodies, white blood
cells).
At 1 mg of plaque, according to different authors, contains from 5 to 800
million. Microorganisms. Microorganisms plaque divided into two groups: 1 -
acidophilus bacteria, which include species are able to grow in an acidic
environment; 2 -proteoliticheskie microorganisms producing proteases.
The first group includes milk - sour streptococcus, lactobacillus,
actinomycetes, leptotrihiy and Corynebacterium. Streptococci, Corynebacterium
and actinomycetes may develop in basic environments. In this case, because of its
ability to synthesize lactic acid they quickly neutralize the environment. Among
acidophilus bacteria is atsidohennym that are able to synthesize sucrose with a
large number of lactic acid (sometimes vinegar).
All of plaque streptococci are divided into groups: Str. salivarius, Str. mitis,
Str.mutans. Str. salivarius easily determined morphologically form colonies that
formed on the gelatin containing 5% sucrose: large slimy colonies that contain
large amounts of Levante. These streptococci in plaque found in small quantities,
but a lot of the mucous membranes and saliva.
Str. mitis make up the bulk of streptococci isolated from plaque. They are very
heterogeneous, are a group of viridans streptococci and have weak biochemical
activity. Only a few strains of Str. mitis able to synthesize extracellular
polysaccharides. Str. sanguis is the second quantitative content in plaque.

33
The most interesting type of milk - sour streptococci are Str. mutans due to its
pronounced kariyesohennymi properties.
Among acidophilus bacteria of dental plaque, 15% are filamentous form
(actinomycetes, lactobacilli and leptotrihiy). Actinomycetes form levaniny;
lactobacilli do not form extracellular polysaccharides, except Lactobacillus casei,
which can form some capsular polysaccharides; leptotrihiy do not produce
polysaccharides; the second group of bacterial plaque up anaerobes that use food
proteins and amino acids. Number of anaerobic bacteria in plaque decreases in the
use of sucrose and increases in the use of maltose.
In non-carious dental plaque make the most of veyllonelly, neyseriyi and
lowest -spirohety. In carious dental plaque bacteria are the main proteolytic
ristelly. In the above microbiota in dental plaque found and other microorganisms,
particularly yeast-like fungi, difteroyidy, staphylococci. At all stages of the dental
plaque it is dominated by streptococci.
The bacteria are found mainly in three areas:
1) in dental plaque on teeth crowns, and in case of caries - a cavity;
2) hinhivalnyh furrows;
3) on the back of the tongue, especially the back of its departments.
According to different authors, the number of bacteria in saliva range from
43 million to 5.5 billion in 1 ml, an average of 750 million to 1 ml. Microbial same
concentration in the plaques and hinhivalniy furrow almost 100 times higher.
About half of the residents are optional and obligate anaerobic streptococci, which
includes in its membership Str. mutans, Str. mitis, Str. sanguis peptostreptokokky.
R-hemolytic streptococci is not part of the resident flora. Different types of
streptococci occupy a niche, such as the largest number of enterococci were found
on the back of the tongue and hinhivalniy furrow, Str. mutans in plaque usually
located on crown.
The other half of the resident flora consists of veyllonel (about 25%) and
dyfteroyidiv (about 25%). Staphylococci, lactobacilli, flagellated bacteria,
spirochetes, leptospira, fuzobakterii, bacteroides, neyseriyi, spiral shape, yeast,
34
other fungi, protozoa found in the mouth in much smaller quantities. Although
these organisms are always present in the mouth, they are never so well
represented as streptococci, veyllonelly, diphtheroids. These data indicate that it is
necessary to distinguish between major and minor members of the resident
microbiota.
Uneven microbial density different biotypes mouth, indicates the presence of
space-reproductive groups of microorganisms. The greatest microbial density, high
ecological significance pathogenic flora and smallest species diversity index
suggests plaque most important in the epidemiological sense.
Among the resident microbiota composition of the oral microbiota dominant form
Str.salivarius, Str.sanguis and lactobacilli. Their combination define individual
tsenotip, and with it the distinctive features of a particular ecosystem. By number
of dominant microbiota in tsenotypi divided into polidominantni, monodominant
and adominantni. Individual tsenotyp microbiota form plaque streptococci (Str.
Salivarius, Str. Sanguis) and lactobacilli, so their presence in tsenotypi is
paramount.
Tsenotyp plaque healthy individuals, which is dominated Str. salivarius, Str.
sanguis and lactobacilli normotsenozu refers to the first order, which is the most
physiological.
The appearance in microbiocenosis Str. mitis and changing the proportions
between the main representatives tsenotipa characterizes normotsenoz second
order.
Tsenotypi presence in healthy individuals and opportunistic pathogenic
species (Str. Mutans) staphylococci, fungi of the genus Candida) refers to
normotsenozu third order, is regarded as dysbiotic reaction.
The characteristic allows oral microbiota present its structure in general and
grasp the main trends in the considered states. In fact, this collective image of the
oral microbiota, which can vary from individual to individual cases. Meanwhile, in
practice there is a need of clinicians in the assessment of such individual
ecosystems.
35
 Since Str.salivarius prevails as the index of constancy, and on population
levels of biotope and has a high antagonistic activity to most other bacteria of the
oral cavity, its presence in tsenotypi biocenotic determines the nature of the
relationship. Therefore tsenotipy, which include the salivary streptococcus, were
attributed to tsenotipiv first order. The balance of the ecosystem observed at oral
tsenotypi consisting of Str.salivarius, Str.sanguis, lactobacilli and often achieved at
the expense of dominant. However microbiota met where found along with other
dominant species within the taxonomic community structure. Their expression
probably reflects the change in proportions between tsenotypa happens within
normotsenoza because not accompanied domination unusual types and causes
exceeding the threshold density of bacterial populations.
Options, which took place Str.salivarius, Str.sanguis, made up the second
group - tsenotipom microbiota of the second order.
The indicators of decompensation microbiota or dysbiosis is or reduce
species diversity index lower than 1,71 lg CFU / h, or the appearance of unusual
microorganisms for plaque microbiota of healthy people, such as Str. pyogenus,
enterobacteria, peptostreptokokki and others.
Study tsenotipa individual options and their ecological characteristics can
create estimation algorithm oral microbiota. It can help you determine the five
states of ecosystems eubioza first order designating quantitatively and functionally
balanced normotsenoz to dysbiotic reaction - community compensated quantitative
or qualitative imbalance. Two state normotsenoza first and second order reflects
the effectiveness of substitution missing components tsenotipa. Existing options
normotsenoza are gradations of violations biocenotical relations of harmony
(normotsenoz first order) by discomfort (normotsenoz second order) to disharmony
(normotsenoz third order or dysbiotic reaction). The criteria for the latter are not
randomly selected species and the presence of unusual species diversity index
below 1,71 lg CFU / h. These figures indicate a breach spatial and functional
structure of ecosystems when homeostatic mechanisms lose their ability to return
to its original level and it goes on controlled state.
36
 The study of oral microbiota.
To study the qualitative and quantitative composition of the oral microbiota
studied plaque, mucous cheeks, gums, palate, the surface of the tongue and oral
liquid.
Plaque is going to study a sterile excavator. The resulting material is
weighed on an analytical balance with subsequent dilution of 1: 100 to 1: 1000 and
crop nutrient media.
Collecting the material from the mucous membranes of the tongue and held
a sterile cotton swab from an area 1sm2 and subsequent hanging on nutrient media.
Oral fluid collected in a sterile tube, 0.1 ml investigated.
To study the oral microbiota, the following nutrient medium: 5% blood agar
for counting the total microbial contamination, the yolk-salt agar - for
staphylococci, broth and sugar "Mitis Salivarius Agar" - for streptococci, vegetable
-molochni environment for lactobacilli, Saburo with polimeksyn - for fungi of the
genus Candida, Wilson - Blair for anaerobic, environment Endo - for
Enterobacteriaceae. Crops stai 24 hours in an incubator, the medium Saburo about
5 days. Identification of selected strains carried on the basis of morphological,
cultural and biochemical characteristics of bacteria according to the determinant D.
Berg.
Quantifying the density of populations of various environmental groups
carried out by counting CFU in one gram of plaque, 1 ml. oral liquid per 1 cm2
surface of the tongue and mucous membranes of the cheeks, gums and palate.
General characteristics of the population of different habitats microbiota of the oral
cavity is presented in Table 5.
Esophagus and stomach in healthy people has no permanent microbiota. The
esophagus of healthy people not contain microorganisms or very little (Candida
albicans, Actinomyces israelii). In the stomach, got accustomed yeast (C. albicans,
C. tropicalis); sarcine (S. ventriculi); campylobakteria (Campylobacter fetus, H.
pylori); rarely find lactobacilli, staphilo- and streptococci.

37
 Table 5
Population levels of oral microorganisms, CFU / units.
Biotope oral Microorganisms Fungi genus CFU
cavity Candida
Gr+ Gr-

dental plaque 8,50х104 4,69х102 4,09х102 8,59х104

Oral liquid 7,52х104 3,12х102 3,17х102 7,53х104

The surface of the 7,58х104 1,68- 102 4,80х102 7,64х104

tongue
The mucous 1,14х104 1,38х102 3,17х102 1,15х104
membrane of the
cheek
Gums 2,70х103 0,73х102 2,24х102 2,72х103
Palate 1,12х103 0,60х102 1,86х102 1,14х103

In the small intestine is 105-108 organisms per 1 ml content. Here are

bifidobacteria, lactobacilli, clostridia, enterococci. In the colon there is the largest
number of microorganisms. 1 g of feces to 1012 contains microbial cells. About
95% of all microorganisms constitute asporogenous anaerobic bacteria. The main
representatives microbiota of the colon are:
Gram-positive anaerobic bacillus (bifidobacteria, lactobacilli, eubacteria);
Gram-positive anaerobic spore-forming bacillus (Clostridium perfrinhens);
Gram-negative anaerobic bacillus (bacteroides); Gram-negative facultative
anaerobic bacillus (Escherichia coli and similar bacteria with them - Klebsiella,
Proteus);
anaerobic gram-positive cocci.
In smaller quantities are fusobakteria, Proteus, veylonelly, staphylococci,
Pseudomonas aeruginosa and yeast-like fungi.

38
 The most numerous and diverse microbiota of the colon. The bulk of its
weight up anaerobic microorganisms: Bifidobacterium spp., Bacteroides spp. The
share of these two families account for 96-99% of all bacteria that inhabit the
colon. Here vegetate significant number of Escherichia spp. , Enterococcus spp. ,
Lactobacillus spp. The residual flora of the colon up numerous species of
Clostridium, Staphylococcus, Proteus, Candida, Enterobacter, Pseudomonas,
Veillonella, Peptococcus, Peptostreptococcus, Actinomyces and others. Total more
than 260 described species of bacteria. In some people in the gut are enteroviruses,
which are in violation of the resistance of the organism can cause various diseases.
In some cases, the stool can detect various types of protozoa.
Persistent disruption of normal microbiocenosis called dysbiosis. It is
changing the very structure avtoflory and proportion of its individual
representatives: a significant reduction of the normal microbiota species until their
complete disappearance or appearance of a large number of those who normally
rare. This is mainly staphylococci, gram-negative bacilli, fungi Candida and
Clostridium.
The need to investigate intestinal dysbiosis occurs when long-term diarrhea,
which do not emit pathogenic enterobacteria, after the transfer of intestinal
infections with a long period of convalescence, prolonged antibiotic therapy,
malignant tumors before surgery on abdominal cavity in premature newborns and
in diseases difficult to treat ( enterocolitis, ulcerative colitis, cholecystitis, etc.).
Material from the stomach and small intestine are using special probes or capsules
that open in a certain department tract and closed after sampling. Recently for this
purpose and widely used Fibergastroscopes hastroduodenoskopy that allow to the
analysis not only of the stomach or bowel contents, but they mucosa biopsies.
Material from the sigmoid and rectum are tampon tube Tsimana,
kolonofibroskopom or rektoromanoskopom.
In the diagnosis of intestinal dysbiosis exploring cal. Its made of pre-
weighed bottles in an amount of 0.5-1.0 g without preservative and transported to

39
the laboratory within 2 hours of collection. An appointed stool sample is diluted
with a special buffer from 10-1 to 10-12.
From 10-3-10-6 dilutions on 0,1sm3 sown on medium VSA (to identify
staphylococci), blood agar (for enterococci and detection of hemolytic forms),
Saburo (for mushrooms), Wilson-Blair (for clostridia).
From 10-5-10-8 dilutions on 0,1sm3 sown on Endo medium (for
Enterobacteriaceae), MPC-2 and MRS-4 (for lactobacilli) and dilutions of 10-7-
10-10 of 1.0 cm3 Blauroka sown on the environment, which is poured high column
(for bifidobacteria), and special protection for Bacteroides.
For detection of pathogenic enterobacteria native liquid feces, or from
dilutions 10-1, bacteriological loop wreaked on the environment Endo,
Ploskyryeva and bismuth-sulfite agar. 1-2 10-1 sm3 of breeding sow to enrich the
environment (Muller, selenitovyy magnesium or broth). The composition and
method of production of culture media is given in the instructions for diagnostics
dysbacteriosis.
Crops for growing facultative anaerobic bacteria were incubated at 37 ° C
for 24-48 hours, bifidobacteria - 48 h, anaerobic - 4-5 days in anaerostatah
mushrooms - 96 hr at 28-30 ° C. After identification of isolated cultures carried out
calculations of microorganisms of a group of 1 g of faeces.
Urinary tract microbiota.
Parenchyma of the kidneys, ureters, bladder, urine, uterine cavity and
fallopian tubes in healthy people free of germs. In the outer part of the urethra and
genitals of men and women are Mycobacterium smegmatis, a small amount of
staphylococcus, streptococcus, peptokokki, peptostreptokokki, Corynebacterium,
bacteroides, fuzobakterii, fungi genera Candida, Torulopsis, Geotrichum.
In the vagina of healthy women predominate lactic acid bacteria (bacillus
Doderlayna) dyfteroyidy and gram Comma variabile. Much less show strepto-,
stafilo-, peptokokki and Clostridium. In 15-20% of pregnant women found
Streptococcus agalactiae, very dangerous for babies. The presence of Gram-
negative bacteria are a consequence of fecal contamination.
40
 Portia first morning urine (3-5 cm3) are in sterile dishes from the mid
urination, and no later than 1 hour crop carried out to quantify the microbiota or
calibrated platinum loop (diameter 2 mm) on agar in petri dish, or 30 cup capacity
cm3, the walls of which are breeding ground area of 12.5 cm2. Environment pour a
measured amount of urine, then it is poured (method Neycheva). After incubation
count the number of colonies crops and determine the number of bacteria in 1 cm3
of urine. The critical (dangerous) level of bacteriuria - 105 or more.
The material for the study of vaginal microbiota and determine the degree of
purity of vaginal contents are Volkmann spoon, spatula or zholobopodibnym probe
from the rear vaginal vault and put a thin layer on a glass slide. Smear record 10
minutes in a mixture of Nikiforov, stained by the Gram method and examined
under immersion system.
Pregnant women define four degrees of vaginal secretion purity:
first - single desquamated epithelial cells, many sticks Doderlayna no white
blood cells;
second - epithelial cells, sticks and Doderlayna Comma variabile, isolated
white blood cells;
third - rarely sticks or Doderlayna Somma variabile, many leukocytes
available coccoid microbiota;
fourth - none Doderlayna sticks and Comma variabile, many leukocytes
available hnoyeridna microbiota.
The first and second degree of purity found in healthy women considered the
norm; third and fourth - a pathological condition characterized genitals and
sanitation needs before delivery.
Age-related changes in the microbiota.
A child is born sterile, but passing through the birth canal, captures
accompanying microflora. forming microbiota carried out through contact with
microorganisms newborn and the mother of the environment and health as defined
environment in which the child was born, the type of feeding. Up to 3 months of
life the child is normal microbiota similar to the microbiota adult. First, after the
41
birth of the child mouth inhabit aerobes, which after teething replaced anaerobes.
When breastfeeding gut microbiota basis of the child are bifidobacteria and
lactobacilli. When artificial feeding in premature and weak children disturbed
reproduction of bifidobacteria, and increasing the number of gram-negative
bacteria (E. coli) and cocci. These children often get sick.
The value of the normal microbiota of the human body.
Normal microbiota performs important physiological functions and is
involved:
in metabolism - regulation of intestinal gas in the cleavage of proteins,
lipids, nucleic, fat and bile acids;
regulation of motor functions in the intestine;
the synthesis of vitamins B, C, nicotinic, folic acid;
in the detoxification of endogenous and exogenous toxic products;
in the process of formation stimulation of the immune system in infants and
maintaining immune status in adults.
But the most important feature is its normal microbiota involved in
colonization resistance. Colonization resistance - a combination of antagonistic
properties of normal microbiota to prevent colonization of mucosal third party,
including pathogenic or opportunistic microorganisms. Antagonistic activity of
normal microbiota implemented through the following mechanisms:
1. The formation of acid products that inhibit the growth of competing
microorganisms (lactic acid, acetic acid). Wednesday Acid prevents reproduction
of pathogenic and putrefactive microbiota stimulates peristalsis;
2. Biosynthesis of substances with antibiotic activity (bactericins);
3. Competition bacteria in food substrates;
4. The competition area for adhesion to epithelial cells.
Violation of normal microbiota.
In various diseases disrupted quantitative and qualitative value of
Representatives normal microbiota that promotes growth of pathogenic and

42
opportunistic microorganisms. In this case, developing pathological process called
dysbiosis (dysbiosis).
Dysbiosis - a quantitative and qualitative change of the normal microbiota,
leading to the development or worsening of the pathological process. Causes of
dysbiosis:
gastro-intestinal tract infectious or non-infectious nature;
irrational use of antibiotics and chemotherapy;
inadequate (unbalanced) diet (especially in children 1 year of life)
malignant neoplasms;
surgery;
hormonal disorders;
immunodeficiency states.
Thus, dysbiosis - is not an independent disease, but a state body, which can
occur in patients with different diagnoses.
Examples dysbacteriosis:
1. Candidiasis lesions of the mucous membrane of the mouth - often in
infants.
2. Dysbiosis in infectious diseases - a condition in which sharply reduced the
number of obligate anaerobes and facultative anaerobes population increases,
resulting in the colon begin to dominate septic processes, increased flatulence,
increased bowel motility. There bloating, tenderness, diarrhea.
3. Dysbiosis vagina (vaginosis) - developing bacterial vaginosis in women at
lower production of estrogen. There lactic acid substitution resident flora
Gardnerella, staphylococci.
For the treatment of intestinal dysbiosis using drugs normal microbiota
(eubiotics) containing live bacteria - residents: bifidobacteria, lactobacilli, E. coli.
For example: bifidumbacterin, lactobacterin, colibacterin, bificol, Bifilakt. Applied
orally. Thus, the normal microbiota plays an important role in protecting the body
against pathogens. At the same time, representatives of the normal microbiota
under certain conditions also can cause inflammation of the microbial etiology. For
43
example, after suffering flu bacteria that live in the nasopharynx cock cause
pneumonia.
The emergence of diseases caused by the normal microbiota may be due to
the following reasons:
1. Penetration of microorganisms in unusual habitats for them - normally
sterile (blood, abdominal cavity, lungs, urinary tract);
2. Reducing reactivity. In immunosuppressed individuals with normal
microbiota representatives can cause severe disease with fatal consequences. For
example: generalized candidiasis - in patients with end-stage AIDS.
3. Against the background of some systemic diseases can dramatically
increase revenues into the systemic circulation endotoxin gram-negative intestinal
microbiota. Developed endotoksinovyy shock, multiple organ failure. However, no
microorganisms does not come from outside. Endotoksinemiya own microbiota
associated with microorganism. Such a process is possible in the pathology of
pregnancy, congestive heart failure, liver pathology. For example: hepatitis,
cirrhosis of the liver.
4. As a result of lypopolysaharyda hramnehatyvnyhmikroorhanizmiv
released additional amount of histamine, which can cause allergic conditions.
It should be noted also that some members of the normal microbiota used as
sanitary indicative microorganisms that indicate environmental pollution (water,
soil, air and food) discharge the person, which enables us to judge their
epidemiological danger. These microorganisms are, for example reside in the colon
Clostridium perfringens and Enterococcus faecalis.
Data on the pathogenicity of the bacteria to humans are presented in Table 6.

Table 6
Classes (Group) pathogenicity of bacteria
№ Species bacteria Disease

Бактерії

44
I group

1. Yersinia pestis Plaque

II group

1. Bacillus anthracis Anthrax

2. Brucella abortus Brucellosis

Brucella melitensis
Brucella suis

3. Francisella tularensis Tularemia

4. Legionella pneumophila Legionellosis

5. Vibrio cholerae 01 Cholera

Vibrio cholerae non 01

III group

1. Bordetella pertussis Pertussis

2. Borrelia recurrentia Relapsing fever

3. Campylobacter fetus Abscesses, sepsis

4. Campylobacter jejuni Enteritis, cholecystitis,

septicemia

5. Clostridium botulinum Botulism

6. Clostridium tetani Tetanus

7. Corynebacterium Diphtheria
diphtheriae

8. Helicobacter pylori Gastritis, peptic ulcer disease

9. Leptospira interrogans Leptospirosis

10. Mycobacterium leprae Leprosy

11. Mycobacterium tuberculosis Tuberculosis

45
 Mycobacterium bovis
Mycobacterium avium

12. Neisseria gonorrhoeae Gonorrhea

13. Neisseria meningitidis Meningitis

14. Salmonella paratyphi A Paratyphoid А

15. Salmonella paratyphi B Paratyphoid В

16. Salmonella typhi Брюшной тиф

17. Shigella spp. Dysentery

18. Treponema pallidum Syphilis

19. Yersinia pseudotuberculosis Pseudotuberculosis

20. Vibrio cholerae Diarrhea

01

21. Vibrio cholerae non 01 Diarrhea, wound infections,

septicemia

IV group

1. Aerobacter aerogenes Enteritis

2. Bacillus cereus Food poisoning

3. Bacteroides spp Lung abscess, bacteriemiya

4. Borrelia spp. Tick-borne spirochetosis

5. Bordetella bronchiseptica Para-whooping cough

Bordetella parapertussis

6. Campylobacter spp Gastroenteritis, periodontitis

7. Citrobacter spp Local inflammation

8. CIostridium perfringens, Gas gangrene

CIostridium novyi,

46
 CIostridium septicum,
CIostridium hiatolyticum,
CIostridium bifermentans.

9. Escherichia coli Enteritis

10. Haemophilus influenza Meningitis, pneumonia, laryngitis

11. Klebsiella pneumoniae Pneumonia

12. Mycobacterium spp. Mycobacteriosis

Mycobacterium
рhotochromogens
Mycobacterium
scotochromogens
Mycobacterium
nonphotochromogens
Mycobacterium rapid
growers

13. Micoplasma hominis 1 Local inflammation

Micoplasma hominis 2
Micoplasma pneumoniae

14. Proteus spp. Local inflammation, food

poisoning

15. Pseudomonas aeruginosa Sepsis, local inflammation

16. Salmonella spp. Salmonellosis

17. Staphylococcus spp. Food poisoning, sepsis,

pneumonia

18. Streptococcus spp Pneumonia, tonsillitis, arthritis,

septicemia

47
19. Vibrio sрр., Diarrhea, food poisoning, ranova
Vibrio parahaemolyticus, infection, septicemia
Vibrio mimicus,
Vibrio fluviales,
Vibrio vulnificus,
Vibrio alginolyticus

20. Yersinia enterocolitica Enteritis, colitis

If the deviation of quantitative and qualitative characteristics of

microorganisms (microbiota) in our body beyond where they can be offset by their
own physiological mechanisms of the body, developing a pathological condition
called dysbiosis. The factors that give rise to dysbiosis, often act the tendency of
stress, antibiotic and hormone therapy, allergy of the body, radioactive radiation
and frequent changes in climatic conditions. Bacteriological analysis of feces
shows the balance between the groups simbiontnyh (bifidobacteria, lactobacilli,
etc..) And pathogenic (pathogenic species of Enterobacteriaceae and Escherichia
coli, psevdomonad, microscopic fungi, etc.). Microorganisms. In some cases, if
disrupted the ecosystem of the small intestine, it is "pollution" small bowel
intestinal microorganisms have pathogenic properties. In severe cases, these
pathogens can spread beyond the intestine and settle in internal organs. The next
stage is accompanied by deep digestive disorders (splitting polysaccharides) and
biosynthetic (production of vitamins, essential amino acids, etc.). Functions.
Excluding these conditions under control and normalize the situation, will
experience increased reproduction in high mortality and some other species of
microorganisms. This could cause a negative impact on human activity by
enhancing the formation of toxic products.
To avoid the situation described above, you should: Avoid unnecessary use
of antibacterials; consume prebiotics that are substrates of their own food and
stimulate the intestinal microbiota of reproduction (yogurt, kefir, etc.); encourage

48
local and systemic immunity; used functional and even food. This includes eating a
large amount of dietary fiber (bran, vegetables, fruits) and foods fortified with
living cultures of microorganisms (milk mixture), enabling reproduction of their
own bifidobacteria in the gut (potato, rice broth, carrots, pumpkins, soybeans).

METHODS MICROBIAL DECONTAMINATION

Sterilization (from the Latin. Sterilis ~ sterile, free from bacteria) - the
complete destruction of vegetative and spore forms of microorganisms on all
subjects, materials in nutrient media.
Sterilize tools, dressing and stitching ma¬terial, operating clothes,
medicines. In microbiological laboratori¬yah - culture media, test tubes, pipettes,
flasks, Petri dishes and more.
Processing tools:
1. rinsed in running water;
2. soaked in detergent solution for 15 minutes;
3. washed in the same roz¬chyni 0.5-1 min;
4. rinsed with distilled water and running;
5. suhozharoviy dried in oven at 80-85 ° C to extinction moisture.
Test tubes, bottles, flasks close cotton plugs. Zahorta¬yut paper tubes in 25-
30 pieces, and petri dish - 4-5 pieces or placed in steryli¬zatsiyni box (biksy).
Pasteur pipettes and graduated from the wide end za¬tykayut cotton wool wrapped
in paper or placed in cardboard or metal canisters 10-15 pieces. Nutrient medium
in flasks, bottles, tubes plugs.
Types of sterilization:
a) physical (high temperature, radiation);
b) mechanical (cold);
c) chemical (dis. solutions and gases).
Physical methods

49
 High temperature sterilization. Efectivity characterized by (D) - time is
required at a given temperature to get a tenfold decrease in the population of
bacteria (90%). The value is measured, usually in minutes.
Flambie in the flame - Bacterial loops, tweezers, subject and cover lenses.
Boiling - 40 minutes in special sterilizers - surgical instruments, syringes, needles,
rubber tubes. To increase the boiling point and removing water hardness add 1%
sodium bicarbonate. The method does not provide complete sterilization, some
survive (clostridia).
Dry heat (suhozharova wardrobe) 160 ° C, 120-150 minutes / 180 ° C, 45-60
minutes (after reaching the set temperature). Sterilize glassware. Advantage - not
damaged glass, not metal corrosion vidbu¬vayetsya tools. Used for sterilization of
heat-resistant po¬roshkiv other substances. Disadvantages - quite a long period of
sterilization is charring and burning cotton jams and paper, which wrapped dishes.
Couple under pressure - the complete destruction of bacteria and spores. Achieved
action pairs, to which the pressure is higher than the boiling.
Flowing steam (100°C) is carried out in an autoclave with nezahvynchenoyu
cover. When heated steam penetrates between embedded objects. In this way the
medium is treated with carbohydrates. A single pair of action does not kill spores
used fractional sterilization 3 days in a row for 30 minutes. Disputes that are not
killed, to germinate next day and vegetative cells are killed by second and third
processing.
For substances not withstand 100 ° C (liquid protein, vitamins, some
medicines) - tindalisation - water bath 58-60 ° C for 5-6 hours consecutive days.
Pasteurization - heating the material to a single T below 100 ° C, destroying
vegetative forms microbs. The debate alive. Microorganisms remaining
significantly weakened. Widely used in the food industry. The heat treatment of
milk, beer, wine, various juices at 70 ° C for 30 min or at 80 ° C - 5-10 min.
Pasteurized products stored in the cold.
Autoclaving. A more effective method of sterilization effect than dry heat.
Structurally autoclave - Double-walled solid metal cylindrical pot with lid
50
(hermetically closed). The inner part - camera (for material). The air outlet valve
and pressure gauge (determines the working steam pressure in the chamber) with a
safety valve.
The irradiation. Apply different types of radiation. In the practice used for
this, electrons, gamma rays, ultrafiole¬tovi rays, radio-frequency radiation.
Electronic accelerator allows focusing the electrons into a narrow beam of high
power. Use for sterilization in industrial scale bandaging and surgical suture
material at the production. Disadvantage - the low permeability rays.
By gamma radiation sensitive vegetative and spore forms of various
bacteria, fungi, yeasts, viruses. Irradiation capacity of 2.5 Mrad - disinfection of
antibiotics, vitamins, hormones, plastic disposable equipment (Petri dishes,
syringes), bandaging and surgical suture materials, etc.
Ultraviolet radiation - neutralization of bacteria in the air of operating rooms,
wards, boxes, microbiological laboratories, etc. - bactericidal lamps of different
power - was 15, was 30 and others. Germs can be protected by numerous organic
substances, dust and other factors. Vegetative forms of bacteria in 3-10 times more
sensitive to UV radiation than disputes.
Methods intense radio frequency exposure. Difficulty - danger for staff
(interference communication systems, different frequent exposure).
Mechanical methods
Do not destroy substrates. Liquid and liquid medium (containing protein,
vitamins, antibiotics, carbohydrates, volatile substances etc). Used for treating
bacterial toxins, bacteriophages from microorganisms.
Mechanical (cold) sterilization is performed by filtration through finely
antibacterial or anti-virus filters. They provide with special materials permeated the
pores which have a different shape and are throuw filter tortuous. Filters can be
made with positively charged material, while bacteria have negative charge on the
surface also interact electrostatically with him, not just mechanically different
diameters due to bacteria and pores. To check the quality of filters using small test
bacteria (Serratia marcescens, or Pseudomonas aeruginosa). The filtrate vysi¬vayut
51
in growth medium and maintained at optimum temperature pro¬tyahom 5 days. In
the absence of growth of test bacteria can be used to filter sterilization. Membrane
or colloidal filters which are made of nitrocellulose, representing up to 3 discs with
a diameter of 5 mm. Filters Seitz - plates (discs or squares) thickness of 4-6 mm (a
mixture of asbestos and cellulose).
Disadvantages:
1. filtrates possible contamination by foreign substances popada¬yut it from
the filter (alkali, alkali metals, asbestos fibers);
2. asbestos because of its negative charge binds certain substances from the
fluid that is filtered. In addition, you should carefully check the filters before work
to not use those with mechanical deformation (cracks, nadlomy etc.).
Before the work affirms a special filter holder. In particular, azbes¬tovi plate
is placed between the bearings and cylindrical metal cabinet Seitz. Both of the
connecting screws. The collected filter is inserted into the rubber plugs Bunsen
flask with a side shoot. Fully vmon¬tovanyy wrapped in filter paper and sterilized
in an autoclave. The liquid for filtru¬vannya poured into metal cylinder side
connecting process of vaccum bulb pump to create a vacuum in the flask and
accelerate filtration. The filtrate in the flask is sterile.
Chemical methods
Sterilized products made of rubber and plastics - 6% hydrogen peroxide
solution, which dipped products for 6 hours at 18 ° C and 3 h at 50 ° C. You can
apply deksona solution with an exposure of 45 min at 18 ° C. After finishing
products wash twice in sterile distilled water, each time changing it, and
transferred to sterile forceps biks. Tools for endoscopy and automatic pipettes can
also alcohol.
Sterilization vapors of formaldehyde, chloroform, ethylene oxide, propylene
oxide, methyl bromide, ozone - disinfection of endoscopic instruments, electronic
equipment, plastic products, catgut etc. Effectivity proven mixture of ethylene
oxide and methyl bromide ratio of 1: 1.44. For sterilization use spetsi¬alni dense
vapor chamber, which is hermetically closed. For each factor operating modes
52
designed their sterilization. Once the gas mixture is pumped from the chamber and
replaced with sterile air. Subjects who were sterilized specified method is
recommended to use no earlier than 24 hours - to vydalyvsya all gas.
To test the effectiveness of sterilization autoclaves zas¬tosovuyut reliability
of chemical and biological control. There are chemicals with a specific melting
point: benzonaftol - 110 ° C, -115 ° C antipyrin, sulfur -119 ° C, benzoic acid -
120-122 ° C, mannose and urea - 132-133 ° C. It is at such temperatures often
perform sterilization. Chemicals placed in a glass tube, add a small amount of
aniline dyes (safranine, fuk¬syn or methylene blue), sealed and placed between
objects that sterylizu¬yutsya. Uniform color drug in color dye in the tube indicates
the proper temperature in an autoclave and therefore reliability sterilization. For
biolohich¬noho control sterilization in an autoclave containing special Biotest -
strips of filter paper, gauze, etc., which are bacterial spores from vido¬moyu heat
resistance, disputes the number of known and others. Decomposed in biksah to be
sterilized. After the end of the tubes with strips pour culture medium and incubated
at the optimum temperature. The lack of germination of spores of bacteria indicates
effective sterilization.

Methods for sterilizing medical objects

Items sterilization method of sterilization
Dry heat glassware, oil, tools, needles, powders
Wet steam Solutions for parenteral administration tools, environment, rubber
stoppers
Filtering medium, which can not withstand heat, with protein, some vitamins,
amino acids, gases, liquids, ointments, oils with low viscosity
Ethylene oxide anesthesia equipment, catheters, diagnostic equipment, prosthetics,
laboratory equipment, auxiliary surgical supplies, optical instruments, plastic
products, packaging materials
Ionizing radiation powders, dressing material, blood collection tubes, brushes,
ointments for burns, centrifuge cups, suture, surgical clothing
53
 Disinfection - a set of measures for full, partial or selective destruction of
potentially pathogenic to human pathogens in various environmental objects to
prevent transmission of infection from a source to a susceptible organism.
There are four stages of disinfection (the difference in sensitivity to
disinfectants): A, B, C, D.
Level A - destruction asporohennyh forms of microbes, rickettsia,
mycoplasma.
Level B - the liquidation of fungi, some viruses, bacteria, with improved
stability (staphylococci, mycobacteria).
Level C - causative agents of especially dangerous infections (plague,
cholera, typhus, melioidosis, glanders).
Level D - spores of microorganisms and protozoa.
Events disinfection in clinics, microbiological, and other laboratory
virusolgy include the effect of physical and chemical factors. It is activities such as
the burning of used bandaging material, waste, garbage burning in the flame of the
burner, the effect of dry heat, autoclaving at different modes using ultrasound
boiling objects with surface active substances, desinfect air through UV exposure.
Takes elementary events - wet cleaning, washing, cleaning, vytripuvannya
blankets, sheets, etc. Moreover disinfection measures include the use of chemicals
- disinfectants, which impose certain requirements:
1) antimicrobs broad-spectrum effect;
2) high water solubility, the ability produce with water or air active and
stable suspensions, emulsions, aerosols;
3) the ability not to lose antimicrobial properties in the presence of
contaminants in the environment organic;
4) low toxicity;
5) allergy lack of action;
6) absence of harmful effects on subjects which they are processed;
7) the availability of raw materials, which are produced disinfectants, its low
cost and so on.
54
 These disinfectants include alcohols, aldehydes, quaternary ammonium
compounds. The most commonly found hallogen use drugs. These include 0,2-
1,0% bleach, which produce ex tempore 10% illuminated solutions of the
substance; 0,2-1,0% solutions hlo Ramin-V or T; 5% aqueous solution of calcium
hypochlorite; 0.05-0.1% solution tryhloizotsyanurovoyi acid (dykonitu); 0.1-0.2%
solution sulfohlorantynu.
Oxidizing represented 1-10% hydrogen peroxide solution, phenols and their
po¬hidnymy - 3-5% solution of Lysol, carbolic acid, phenol.
The group prepa¬rativ of heavy metal salts include sodium mertyolyat, sublimate.
2-3% solution of formaldehyde, cresol 3-10% solution and others.
Usage gaseous disinfectants - 40% aqueous solution of formaldehyde,
mixtures of ethylene oxide with carbon dioxide (1:10) and ethylene oxide with
bromyidmetylom (1: 1).
Allocate current and final disinfection. Conduct current disinfection to
reduce microbial contamination in the foci of infection (bed linen, underwear,
towels, mattresses, pillows, furniture, rugs, dishes, tools, appliances, air separation,
wastewater, etc.).
This surface tables, windows, ceilings, walls, furniture and wash wipe
disinfectant solutions, bed linen and other washed in these solutions. Beds,
mattresses, pillows treated in special kame¬rah thermochemical methods,
upholstered furniture - by special aerosols dishes - disinfectant solutions.
Treatment and discharge of wastewater pro¬vodytsya thermal and chemical
methods. Indoor air can be disinfected by passing through special antibacterial
filters, like they do in isolation wards hnotobiolohichnoyi or tanning ultraviolet
pro¬menyamy it. Medical Instruments, appliances first cleaned, disinfected, and
then, if necessary, sterilized by known methods.
The final disinfection is to destroy pathogens infektsiy¬nyh diseases in the
room where the patient was infectious, and objects - after discharge from hospital
infection, conversion of physical separation to infectious and so on. To ensure care
holding dezinfikuyuchyhzahodiv should:
55
 a) zabespechyty external and internal departments kon¬trol disinfection of
sanitary and epidemiological stations and laboratories of medical institutions,
which carried out a visual, biological, chemical and other methods;
b) to control bacteriological identifying the source of infection in indicator
bacteria. When intestinal diseases - E. coli drop in infections, tuberculosis -
staphilococi in health care settings - opportunistic mikroorhan¬izmy.
Bacteriological control is 1 time in a month - once a quarter, depending on the rank
of the laboratory;
c) conduct sampling control samples (10-30 pieces) not earlier than 30-45
minutes after disinfection; washings area shall not be less than 200 cm2;
d) take sterile wipes and cotton swabs zasiva¬ty on nutrient media with all
the rules in order to aseptic prevention contamination outsider microbiota.

SANITARY VIROLOGY
The subject of health Virology is the study of various pathogenic viruses to
humans in the environment (water, soil, air, food and so on.), Development of
methods for identification and effective measures to sanitation facilities
environment.
In nature, viruses occupied all ecological niches and are part of all
ecosystems, ranging from a drop of water (microekosystem) to the biosphere
(global ecosystem). Obviously, there is no cell in the genome which would no
prophage provirus and mammals. In this regard, the isolation and identification of
viruses difficult to find cell cultures free from viral genetic structures. In fact, the
genetic material of viruses has become part of the genetic stock of all organisms.
However, to identify it in eukaryotic cells is much harder than in prokaryotic,
including the induction of lysogenic bacteria with ultraviolet rays or DNA-tropic
substances.
By raising the issue of integration viruses should be noted that it has long
gone beyond the scientific interests of individual researchers and received more
practical. Viral conversion, such as bacteria zatrudnyaye identification and
56
diagnosis of infectious diseases associated with it malignant degeneration of cells,
the occurrence of slow viral infections, autoimmune and some other pathological
processes.
Temperate phages and viruses integration animals and humans, transferring
alien genes play an important role in the evolution of their owners. They can
inactivate genes of cells or promote their expression enter, delete or move
vstavochni sequence (IS-elements), to facilitate recombination processes and
recombine with masked defective viruses that are in the cellular genome.
Integration viruses are transmitted to descendants of animals and humans
transovarian way through the egg, and the subsidiaries of individuals bacteria - in
amitotychnomu division. Turning to autonomous status, integration viruses get
capacity for replication and the effect on the cells indistinguishable from
infectious, ie have a cytopathic effect. Since then, however, we can not conclude
that ecological systems infectious viruses violate the natural relationships of
organisms. Instead, they stabilize the ecosystem. Situated in symbiosis with
organisms, infectious viruses regulate populations at a level that emerged in the
course of evolution. Virulent phages, such as purified water from excess bacteria
and algae, and animal viruses align recurring bursts of excessive increase in the
number of rodents and insects (voles, rats, locusts). In antagonistic symbiosis with
the host viruses as pathogens purified populations of genetically defective
individuals that are a powerful factor of natural selection, as well as antigens -
provoking organisms to continuous improvement of mechanisms of immunity. The
body is resistant individuals antigenic alteration viruses, parasites and honed their
mechanisms of adaptation and survival.
Violation of established relationships in symbiosis virus-host based on the
principle of unequal stability, can lead to the collapse of the system. It released the
ecological niche which, as the comparative analysis of infectious diseases last 30 -
40 years, immediately filled by another, equally dangerous parasite.
Thus, the task of sanitary virological service is systematic virological
examination sanitary wastewater in urban wastewater treatment plants, water,
57
water bodies used for drinking and household water supply, drinking water sources
centralized water supply, soil zemlerobnyh fields of irrigation, air hospital, food
and t. e. These studies to monitor the circulation of viruses pathogenic to humans
in the environment.
Indication of viruses in the environment consists of several stages:
1) the concentration of viral agents of the environment;
2) transportation of samples to the laboratory;
3) identification of viruses using cell cultures and laboratory animal or
chicken embryos;
4) identification of selected agents.
Transportation, isolation and identification of viruses carried by
conventional virological techniques specific to health and virology problem is only
the concentration of viruses from the environment, which requires the development
of special instructional techniques.

Sanitary water Virology

The main reason for the availability of water for human pathogenic viruses is
the contamination of human feces. In human faeces found more than 100 different
viruses, some of them belonging to the family pikornavirusiv, Reovirus and
adenovirus, have high thermal stability and a long time can remain viable. A
number of viruses resistant to conventional disinfectants, including chlorination,
and can be detected in wastewater at a great distance from the source of
contamination. In water contamination at its human faeces found the same viruses
in feces.
The largest allocation of intestinal viruses occurs in summer and autumn due
to the increase in the number of intestinal diseases. However, outbreaks of
gastroenteritis caused by rotavirus, usually occur in winter and early spring.
Massive selection of enteroviruses intestines of healthy people and patients
infected with HIV virus causes significant pollution of wastewater, and their their

58
resistance to adverse environmental factors causes prolonged survival in water.
Thus, waste water is the main reservoir of enteroviruses in the environment.
The presence of enteroviruses in the water district water supply represent a
danger polio epidemic and other enteroviral infections, gastroenteritis, hepatitis A
and may lead to sporadic cases and outbreaks of these infections.
The duration of storage of viruses in water increases significantly with
decreasing temperature. Thus, the polio virus remains viable in river and tap water
4OS at 90 days and at 37 and 20oC respectively 10 and about 49 days. The higher
the initial concentration of the virus, the longer he is in the water. Terms viability
of different viruses vary in wide limits. Most durable to external factors is
Coxsackie virus group A, less hardy - the polio virus, the shortest term
preservation of the viability of Coxsackie virus group B (30 - 50 days). Viruses
ECHO 7 kept much longer in water than the polio virus. Adenoviruses are more
resistant to external factors than the polio virus and ECHO. Adenoviruses
serotypes some retain their viability in water at 4OS for two years or more. To
adverse environmental factors most resistant group of enteroviruses is hepatitis A.
This virus for a long time can survive in water - from a few weeks to months.
There are outbreaks of hepatitis A in connection with the spread of infection
through raw kolodyaznu water. In polluted waters viruses remain infectious ability
much longer than in the net. Yes, seawater term viability viruses much shorter due
to the high content of various salts and the presence of iodine, which has a
virucidal effect. Certain virucidal effect produce different substances produced by
microorganisms, and dissolved chemical substances found in seawater. With the
number of microorganisms in experimental conditions longest term sustainability
in water saving different degree of contamination detected in intestinal coli phage
(over 10 months). It is a possible candidate for sanitary-indicative microorganisms.
The main objects of sanitary virological research is sewage, waste water treatment
and the stages of disinfection, water water bodies used as sources of water, tap
water, underground water sources, drinking water tap wrenches network
doochischennaya drinking water, drinking water Bottled water marine and fresh
59
water used for recreational purposes and water of swimming pools and water
parks.
Hygiene and virological studies of water bodies is carried out during
preventive and current state sanitary and epidemiological surveillance and
epidemic indications. Hygiene and virological studies of water consists of the
following stages:
• sampling for the study;
• preparation of samples for research (primary processing material);
• concentration of viruses in water samples;
• decontamination viral concentrate;
• virological study (virus isolation or detection of it antigens and fragments
of the genome).
As for methods of concentration of intestinal viruses in the water, the water
research be central water supply wells, open water and swimming pools. Research
wastewater study carried out for the purpose of circulating virus in the population
of the area, the degree of contamination of water with viruses, the efficiency of
sewage treatment plants and so on. Study of water surface and groundwater
conduct when any source of water for the central water supply, to assess the
sanitary condition of places of recreation and epidemiological indicators. Research
carried drinking water only epidemiological indicators.
Methods for concentration of viruses from water can be divided into 4
groups:
I. Physical (ultracentrifugation, filtration, ultrafiltration, flotation,
electrophoresis and electroosmosis).
II. Physico-chemical methods (Precipitation ethanol, ammonium sulfate,
aluminum sulfate, dvohvalentnymy cations Precipitation in isoelectric point viral
protein concentration poliyetylenhlikolem).
III. Adsorption methods (adsorption on a gauze pad, charcoal, natural
mineral sorbents - bentonite, askaninti and other ion exchange resins and
adsorption on aminoetoksyaerosyli, polimetylksyloksani, macro-porous glass etc.).
60
 IV. Biological methods (adsorption on yeast cells and other
microorganisms).
The method the concentration of viruses in water samples depends on many
factors: the degree of water pollution sensitivity methods, availability of
standardized adsorbent, appropriate equipment (eg cooling Ultracentrifuges or
special equipment for ultrafiltration) and the load on the lab. All work associated
with concentration and release viruses are carried out subject to the rules in full
compliance with the regulatory documents.
Sanitary-virologic monitoring of water pollution (sewage, surface and
underground water, drinking), usually in practical labs conducted in several
indicators (enteroviruses, rotavirus, adenoviruses, viruses hepatitis A, coliphage),
so it is advisable to use this method of concentration and use only for reagent
concentrations, which would allow the virus to determine several indicators
simultaneously. The most reliable method is a method of virus concentration
ultracentrifugation. Other methods are also used, including methods of
ultrafiltration, concentration methods using viruses and polyethylene glycol
adsorbtsionni methods - adsorption on a gauze pad and ion exchange resins. These
methods are simple, fast and efficient enough.
To isolate the virus infecting a cell culture or laboratory animals. Drinking
water is considered safe for viral infections if it contains at least one virus particle
to 1 liter.
Sanitary Virology of soil
Intestinal viruses can adsorb podzolic soils, but as a result of a number of
factors may desorbuvatysya and come back into the environment. In this way
intestinal infection can be transmitted through the soil and vegetables using virus-
infected sewage irrigation on the fields, orchards and gardens, sewage soil → →
→ vegetables man.
Intestinal viruses long kept on vegetables. Keeping their infectivity depends
on the type of plants, vegetation conditions, the type of virus and its initial
concentration. So at 6 - 10 ° C and polio virus type retained its infectivity for
61
radishes for 2 months. With vegetables fastest virus inactivation occurs on a
cabbage in phytoncide result of its activity. Infection can occur vegetables not only
by getting viruses on their surface, as well as by the virus of land in the land of
vegetables through the root system. It is therefore necessary to systematically carry
out sanitary-virologic research of water for irrigation, soil and field irrigation
products for the presence of intestinal viruses. Due to the rapid absorption of
intestinal virus particles of soil most likely localization of viruses in the upper layer
(0 - 25cm), but sometimes is important to identify viruses in deeper soil layers (75
- 100cm).
Research carried out on soil epidemiological indicators. Samples of soil (10 -
20g) are taken from a depth of 0 - 20 cm in several points target areas. The samples
are mixed and transported to the laboratory in a sterile plastic bag. Laboratory
tillage involves desorption of viruses from the soil surface uridynnu phase
(phosphate buffer, pH 8.2) and the concentration of liquid phase through filters or
using ammonium sulfate precipitation. The same method and treated sewage
sludge.
Sanitary Virology of air
Airborne transmission infentsiyi characteristic of respiratory diseases - the
most widespread infectious diseases. Viruses get into the air in aerosol droplets
phase as a result of sneezing, coughing, talking, and is composed of droplets of
different sizes, which consist of saliva, mucus and salt. At the highest
concentrations of the virus is in large drops that are less stable in aerosols and
quickly settle. A longer time in the air are small drops of viral aerosol. Drying
aerosol droplets accompanied by inactivation of viruses. Infection with respiratory
viruses almost always at the expense of phase aerosol droplet indoors. First of all
infected people with weakened immune systems that are too close to the sick
person. Less dangerous to inhale dried drops of which have inactivated viral
particles. The concentration of viral particles in the aerosol cloud is reduced by
dilution large volume of air and settling large aerosol droplets. However, the
presence of small aerosol droplets enables the virus to penetrate into the lower
62
respiratory tract. Infectious agent can be transported air flows over long distances -
dysyatky kilometers from the center of infection. Spread of the virus depends on
the wind speed, on the other hand rainy weather reduces the spread of viruses.
Resistant viruses in the environment, especially adenoviruses can pylevoyu phase
of an aerosol into the air flow repeatedly and for a long time to circulate in this
room. According resistant viruses in aerosols and the surface, they can be divided
into three groups: low stability viruses, such as paramyxovirus (parainfluenza
virus, especially respiratory syncytial virus), more resistant viruses hryppu
transmitted from sick people healthy only in the form of aerosol droplet phase and
resistant viruses such as adenoviruses and ECHO viruses that enter the body not
only in the form of droplet phase, but pylevoyi aerosol phase.
To study the microbiota air, such as bacteria and molds are various methods
and established a number of instruments for the concentration of microorganisms
from the air. Some devices used with appropriate modifications and concentration
of viruses from the air. Since viruses are usually found in indoor upovitri low
concentration which does not allow them to allocate necessary preliminary
concentration of viruses from the air. The most favorable conditions for catching
viral aerosol preserving infectious virus activity is created by concentrating them in
a liquid medium. Trapping liquid used as material for virus isolation or conduct
further concentration of viruses, adding to tubes with liquid catching 30% solution
of polyethylene glycol with a molecular weight of 4000 or 6000.
After centrifugation of the suspension removed the top layer of matter and
analyze virusvmisnyy bottom layer. In addition to capture fluids for concentration
of viruses from the air can be used membrane filters. With surface filters viruses
wash liquid medium with antibiotics. Determining the length of conservation
activity in infectious air of different respiratory viruses is a key issue in connection
with the airborne spread of respiratory infections. In experimental studies
determined that the duration of the infectious activity of the virus in aerosol state
depends on factors such as temperature and humidity, sunlight, humidifying liquid
composition, etc. Indoors major factor that affects the rate of inactivation of
63
viruses in aerosols, is humidity. The viruses are resistant to varying degrees to that
of relative humidity. Thus, influenza viruses, parainfluenza and respiratory
syncytial virus inactivated faster at high relative humidity and to a lesser extent - at
a low. Polio virus, viruses and adenoviruses ECHO more resistant in the air at high
relative humidity, but rather are inactivated at low relative humidity. Vesicular
stomatitis viruses and bark stable both at low and at high relative humidity, but
inactivated at average relative humidity.

Sanitary Virology household items

Houseware be intermediaries in carrying viral agents from an infected to a
healthy person. Especially great role of consumer goods in children's institutions
due to close contacts of children. Viruses can be isolated from swabs of various
household items in children's institutions and hospitals, toys, utensils, kitchen
tools, textiles, glass, wood, hands of staff.
Adenoviruses type 5 and 7 ECHO viruses remain infectious activity on some
household items more than 7 days, and parainfluenza viruses and respiratory
syncytial virus - for several minutes or hours. Adenoviruses and enteroviruses can
with everyday objects come second in the air and spread in aerosol form pylevoyi
phase. Research washings of consumer goods for viruses carried by
epidemiological indicators in kindergartens, nurseries, hospitals and other
institutions. Are washed away from the surface of objects with a sterile swab is
placed in a test tube with liquid (solution Hanks, lactalbumin hydrolyzate).
Tampon squeeze out the liquid and the virus was concentrated by filtration or
sedimentation using aluminum sulphate.
Hygiene and food virology
On the spread of viruses in food is much less known than the bacteria and
fungi for several reasons.
First, being obligate parasites, viruses do not grow on nutrient media.
Usually their growing use tissue cultures and chicken embryos.

64
 Secondly, viruses do not multiply in food, their concentration is lower
compared with bacteria and concentration techniques required for their discharge.
Despite a number of studies that focus on this issue, it is extremely difficult to
identify more than 50% of virus particles from products such as minced meat.
Third, work with viruses is not practiced in many microbiological
laboratories food industry. However, the use of polymerase chain reaction (PCR)
using reverse transcriptase (RT-PCR) to detect possible number of viruses in food,
especially in the tissues of oysters and clams. PCR laboratory diagnostics raised
food to a new level - the level of direct detection of extremely small
concentrations.
A number of viruses that are found in food, are able over time to preserve
their infectious activity. In particular, enteroviruses retain their infectivity in
mincemeat for 8 days at 23 - 24 ° C, which is independent of the deterioration of
the product. The use of foods containing neinaktyvovani viruses are the cause of
certain diseases. In particular outbreak of hepatitis have a close connection with the
consumption of certain food products. Because hepatitis A virus is transmitted by
fecal-oral route, the shellfish use of infected reservoirs leads to disease. There is
evidence on the relationship of hepatitis from eating salads and meat sandwiches in
the hotel and restaurant establishments. Approximately 8% of patients with
hepatitis B infection is the reason people water and food that contained a virus.
Noroviruses (genus Norovirus, family Caliciviridae) cause intestinal infections in
mammals, particularly gastroenteritis. Research carried out by food
epidemiological indicators. Viruses that are in liquid dairy products, concentrated
using polyethylene glycol with a molecular weight of 4000 or 6000. Processing
semisolid foods (cheese, cheese products, meat and fish prepared food, bread) and
solid (cereals, cheeses, meats and al.) is the extraction of viral particles in the
liquid phase, and from the virus extract precipitated using polyethylene glycol.
Data on pathogenicity viruses to humans are presented in Table 7.

65
 Table 7
Classes (Group) pathogenicity viruses

№ Species of microorganism Disease

I група

1. Filoviridae: Hemorrhagic fever

viruses Marburg and Ebola

2. Arenaviridae: Hemorrhagic fever

viruses Lassa

3. Poxviridae: Smallpox human

virus smallpox

II група

1. Togaviridae Encephalitis,
horse encephalomyelitis viruses encephalomyelitis,
fever

66
2. Flaviviridae: Encephalitis, entsefalomiyelyty.
Tick-borne encephalitis virus
complex: Hemorrhagic fever
Omsk hemorrhagic
fever, Japanese encephalitis, West Parenteral hepatitis,
Nile. Yellow fever virus. hepatocellular carcinoma liver
HCV .

3. Bunyaviridae, Encephalitis, encephalomyelitis,

Gen. Bunyavirus : meningoencephalitis, fever.
Complex California encephalitis
Gen. Nairovirus:
Crimean hemorrhagic fever virus. Fever, myositis, arthritis

Fever with meningeal syndrome,

encephalitis

4. Rhabdoviridae, Rabies
gen. Lyssavirus :
rabies virus.

5. Hepadnaviridae: Parenteral hepatitis

hepatitis B and D virus

6. Retroviridae: AIDS,
Human immunodeficiency virus T - cell leukemia human
(HIV-1 HIV 2)
virus T cell leukemia human
(НТLV)

7. Unconventional agents: Kreytsfeld-Jakob disease, Kuru,

Pathogens slow neuroinfections scrapie.
(prions) 67
III група

1. Orthomyxoviridae : Influenza
influenza viruses
А, В and С

2. Picornaviridae: Poliomyelitis,
gen. Enterovirus : enteral hepatitis
wild polio virus strains
hepatitis A and E

3. Herpesviridae: Herpes simple

herpes simplex virus types I and Chickenpox, shingles.
II,
zoster-herpesvirus cytomegalovirus,
herpes virus type 6 (HBLV- infectious mononucleosis,
HHV6) Berkita lymphoma,
virus cytomegalovirus nasopharyngeal carcinoma.
Epstein-Barr virus

IV-група

1. Adenoviridae: Acute respiratory viral

all types of adenoviruses infections, conjunctivitis.

2. Reoviridae, Rhinitis, gastroenteritis.

gen. Reovirus : human
retroviruses Gastroenteritis, enteritis.
Gen. Rotavirus : human rotavirus.

3. Picomaviridae, Acute respiratory viral

gen. Enterovirus : infections, polyneuritis.
Coxsackie virus group A and B
ECHO viruses Serous meningitis, diarrhea,
enteroviruses types 68-71 polyneuritis, uveitis

68
 gen. Rinovirus : Serous meningitis,
120 types of human rinoviruses conjunctivitis.

4. Coronaviridae Acute respiratory viral

Coronaviruses infections, enteritis
human

5. Paramyxoviridae: Pneumonia
Human parainfluenza virus type 1- Pneumonia, bronchitis,
4 bronchiolitis,
respiratory syncytial virus (PC- parotitis
virus) measles
paroatyta virus epidemic, conjunctivitis
measles virus

6. Togaviridae Rubella
gen. Rubivirua:
rubella virus

7. Rabdoviridae, gen.: Vesicular stomatitis

vesicular stomatitis virus

Contamination viruses vegetables possible using infected sewage irrigation

on the fields, orchards and gardens. For desorption of viral particles from the
surface of vegetables pour phosphate buffer (pH 8.2), vzbovtuyut, illuminate the
liquid by centrifugation and the virus was concentrated from the supernatant as
described above.
Study of the nature of viruses to date suggest that viruses are not organisms,
even small, as any, even the smallest organisms such mycoplasma, rickettsia and
chlamydia have their own protein synthesizing system. Viruses - independent
genetic structures that can function only in cells that have varying degrees

69
depending on the cellular synthesis of nucleic acids and are totally dependent on
protein synthesis and cell energy systems and are able to evolve.
Thus, viruses are a diverse and large group of many non-cellular forms of
life that are not microorganisms and merged into the realm of Vira. Viruses studied
within virology, which is a separate discipline, which has its object and methods.
Considering the relationship between viruses organisms must be borne in mind that
they are special symbionts that parasitize the genetic and biochemical levels, but
despite this, the viruses have their own history, independent of the evolution of
organisms in which they reprodukuyutsya. As pro- and eukaryotes, viruses
inherent genetic continuity, the ability to play and variability caused by mutations
and recombination.

70
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71