Onderstepoort Journal of Veterinary Research, 60:197-204 (1993)

Ultrastructure of the epithelium of the rumen,

reticulum and omasum of
grey, white and black Karakul lambs

H.B. GROENEWALD
Department of Anatomy, Faculty of Veterinary Science, University of Pretoria,
Private Bag X04, Onderstepoort, 0110 South Africa

ABSTRACT
GROENEWALD, H. B. 1993. Ultrastructure of the epithelium of the rumen, reticulum and omasum of grey,
white and black Karakul lambs. Onderstepoort Journal of Veterinary Research, 60:197- 204
Mortalities due to digestive disturbances occur in homozygous grey and white lambs after they have
reached weaning age. Milk-filled, distended rumens, due to malfunctioning of the oesophageal groove,
are found 24 h after birth. Scanning electron microscopical studies revealed that milk caused sloughing
of the luminal cells in the forestomachs of the affected lambs, while no sloughing of cells was apparent
in control black lambs. The purpose of this study was to compare the ultrastructure of the forestomach
mucosa of grey, white and black Karakul lambs; to determine whether the sloughing of luminal cells was
evident in sections; and, if possible, to find a reason for the desquamation of the cells. Samples of the
forestomach of grey, white and black Karakul lambs were prepared routinely for electron microscopy and
studied with a Phillips electron microscope. In all the lambs the mucosa of the forestomach was a
stratified squamous epithelium consisting of a stratum basale, stratum spinosum and stratum corneum.
In the grey and white lambs the luminal cells of the stratum corneum were electron dense, non-nucleated
and vacuolated. Sloughing of luminal cells was observed. In the black lambs no sloughing of cells was
evident and the luminal cells were moderately electron-dense, nucleated elements. Desquamation of the
luminal cells in the affected lambs revealed the underlying layer with its exposed desmosomal attachment
sites. This explained the differences in the appearance of the luminal cells in the three groups of lambs
as revealed by the scanning electron microscope.

INTRODUCTION (Contescu & Leagu 1941). Impaction of the aboma-

sum was found in grey Karakul lambs by Hartwigk
The Karakul industry has long been adversely af-
(1950). He suggested that the condition was due to
fected by mortalities of homozygous grey Karakul
a reduced ability of the abomasum to contract.
lambs. Mortalities in grey lambs of various breeds
have been described by a number of authors. Con- Homozygous grey Karakul lambs are born appar-
tescu & Epureanu (1939) reported a mortality rate ently normal, but at weaning age they develop pot-
of 30,3% in grey Zurkana lambs under the age of bellies, become weak and emaciated, and die (Nel
10 months. At 5- 6 months of age most of the grey & Louw 1953). On post mortem examination Nel &
Zurkana lambs died owing to digestive disturbances Louw (1953) found large, thin-walled rumens and
impacted abomasa in affected lambs. Nel (1965)
Received 14 June 1993- Editor described enlarged forestomachs in newly born grey

197
Ultrastructure of the epithelium of grey, white and black Karakul lambs

Karakul lambs and came to the conclusion that di- to compare the ultrastructure of the mucosa of the
gestive tract abnormalities are already present at rumen, reticulum and omasum in grey, white and
birth. Studies by Langlet (1949) and Nel (1965) black Karakul lambs. Therefore it was possible to de-
proved this lethal factor to be a genetic disorder. In termine whether there were any differences between
contrast, lvanenko (1949) postulated that mortalities the three groups with respect to the structure and
attributed to the lethal gene in grey Karakul lambs composition of the various cell layers; whether the
were, in fact, due to a delicate constitution and that sloughing of the luminal cells described by Groene-
affected lambs could be saved by careful rearing. wald & Booth (1992) was evident in sections; and
whether there was a reason for the desquamation
Only lambs homozygous for the grey colour are af-
of the cells.
fected. These animals can be identified at birth by
the lack of pigmentation of the tongue, palate and
ears (Nel & Louw 1953). Theoretically, should there
MATERIALS AND METHODS
be offspring from the homozygous lambs, all of them
would be grey. Heterozygous grey lambs are unaf- Five 24-h-old grey, white and black Karakul lambs
fected and are used for breeding. The ratio of the were slaughtered and samples were taken from cor-
offspring of the heterozygous lambs is one homozy- responding areas of the rumen, reticulum and oma-
gous grey, two heterozygous grey, and one black sum . Two newborn grey lambs were slaughtered be-
lamb. Twenty-five percent of the lambs are therefore fore they had suckled and matching samples of the
affected by the lethal factor and die before reaching forestomach were taken. Grey and white lambs with
breeding age, and 25% are black. Only 50% of the unpigmented tongues, palates and ears were specifi-
progeny can therefore be used for the production of cally selected and black lambs were randomly selec-
grey pelts, resulting in a relative scarcity of this com- ted.
modity.
The samples were rinsed in phosphate-buffered sa-
Homozygous white Karakul lambs are born with the line (PBS) with a pH of 7,4. Small blocks of tissue
same lethal factor. However, they do not become were immersion-fixed in 4% glutaraldehyde in Millo-
emaciated and develop pot-bellies so soon, and they nig's phosphate buffer for at least 24 h at 4 oc. The
survive for a longer period. They are typical "poor blocks were subsequently rinsed in Millonig's phos-
doers" and, although they usually reach sexual ma- phate buffer, post-fixed for 1 h at room temperature
turity, they eventually die. Black Karakul lambs are in similarly buffered 1 % osmium tetroxide and given
unaffected and were used as controls. two final buffer washes. The samples were dehydrat-
ed through a graded ethanol series (25 %, 50%,
The outermost component of the ruminant forestom- 75%, 96% and 100% x 2- 10 min per step), cleared
ach mucosa consists of a stratified squamous epithe-
in propylene oxide and embedded in Polarbed 812
lium (Hyden & Sperber 1965; Prins 1967; Lavker,
epoxy resin. Semi-thin sections (0,5 ~-tm) were cut
Chalupa & Dickey 1969; Henrikson 1970a; Henrikson from each sample to determine suitable areas for
1970b; Lyford 1988; Schnorr & Hild 1974; Tam ate
ultra-thin sectioning. Thin sections (0,1 ~-tm) were cut
& Kikuchi 1978; Ramkrishna & Tiwari 1979; Fath EI-
with a diamond knife on a Reichert OmU4 ultramicro-
Bab, Schwartz & Ali 1983; Liebich & Scharrer 1984; tome, stained with uranyl acetate (Watson 1958) (30
Hofman & Schnorr 1982; Amasaki & Daigo 1988).
min) and lead citrate (Reynolds 1963) (4 min) , and
There is general agreement that this epithelium con- examined with a Philips 301 or CM1 0 transmission
sists of a stratum basale resting on a basal lamina,
electron microscope operated at 80 kV.
a stratum spinosum, a stratum granulosum and a
stratum corneum (Hyden & Sperber 1965; Prins
1967; Lavker et at. 1969; Henrikson 1970a; Henrik-
RESULTS
son 1970b; Lyford 1988; Schnorr & Hild 1974; Ta-
mate & Kikuchi 1978; Ramkrishna & Tiwari 1979; The epithelial component of the mucosa of the ru-
Fath EI-Bab eta/. 1983; Liebich & Scharrer 1984; men, reticulum and omasum of the three groups of
Hofman & Schnorr 1982; Amasaki & Daigo 1988). lambs was a stratified squamous epithelium (Fig . 1).
The epithelium consisted of a stratum basale, stra-
Scanning electron microscopy revealed differences
tum spinosum and stratum corneum (Fig. 1). A dis-
in the appearance of the luminal surface of the ru-
continous stratum granulosum was observed.
men, reticulum and omasum in affected grey and
white lambs as compared to black lambs (Groene- The cuboidal basal cells displayed a large, round
wald & Booth 1992). The luminal surface of the grey nucleus and were characterized by the presence of
and white lambs had a weathered appearance due numerous mitochondria (Fig. 2). Polyribosomes, a
to sloughing of the surface cells. The appearance of rough endoplasmic reticulum, a Golgi apparatus and
the cytoplasmic processes on the cell surface also fine filaments were also present (Fig. 3). The api-
differed in the affected and control lambs (Groene- cal and lateral cell membranes were wavy with
wald & Booth 1992). The purpose of this study was cytoplasmic processes and intermittently arranged

198
 H.B. GROENEWALD

FIG. 1 The stratified squamous epithelium of the reticulum in a FIG. 3 A basal cell in the rumen of a black lamb. Polyribosomes
grey lamb. The stratum basale (B), stratum spinosum (S) (P) , a rough , endoplasmic reticulum (R) , a Golgi appara-
and stratum corneum (C) are visible. Note the reduced tus (G) and mitochondria (M) are evident (39000 x)
intercellular space (arrow) in the more distal level of the
stratum spinosum (1300 x)

FIG. 2 Basal cells in the omasum of a grey lamb. The nuclei are FIG. 4 The slightly folded basal surface (long arrow) of the cel ls
round (N) , and numerous mitochondria (M) and fine is visible. The basal lamina (short arrow) is situated be-
filaments (F) are evident. The intercellular space is narrow tween the basal cells (B) and submucosa (S) (2950 x)
(long arrow) and numerous desmosomes (short arrow)
are apparent (5200 x)

desmosomes linked neighbouring cells (Fig. 2). The were present between the keratinocytes and, whi le
intercellular space was narrow (125 nm) (Fig . 2). no Langerhans granules could be found, these cells
The basal surface of the cells was slightly folded were most probably Langerhans cells (Fig. 7) .
(Fig . 4) . The epithelium was separated from the
connective tissue by a typical basal lamina (Fig. 5) The cells of the stratum spinosum contained more
which appeared as an electron-dense structure filamentous material than those of the stratum ba-
separated from the basal cell membrane by a less sale (Fig. 6). The proximal cells bordering the stra-
electron-dense space (Fig . 5) . The basal lamina was tum basale were polygonal to spherical in shape (Fig.
slightly folded and hemi-desmosomes were observed 7) and many finger-like processes projected from
along the basal cell membrane facing the basal their surfaces (Fig. 6) . The processes seemed shor-
lamina (Fig. 5). Micropinocytotic vesicles were ob- ter than those of the basal cells and desmosomes
served along the basal cell membrane (Fig. 5). No were evident where the processes of adjacent cells
occluding junctions were observed. Dendritic cells made contact (Fig. 6). At this level the intercellular

199
Ultrastructure of the epithelium of grey, white and black Karakul lambs

FIG. 5 The basal lamina (short arrow) in the omasum of a black FIG. 7 The stratum spinosum in the reticulum of a white lamb.
lamb. Hemidesmosomes (D) and micropinocytotic vesicles Note the spherical cells (C) and prominent intercellular
(long arrow) are evident. Note the electron-dense ap- spaces (arrow). Dendritic Langerhans cells (L) are evident
pearance of the basal lamina and the space (B) between (1650 x)
the basal lamina and the cell (28500 x)

FIG. 6 Cells of the stratum spinosum in the omasum of a grey FIG. 8 The cytoplasm of a cell in the stratum spinosum in the
lamb. Note the abundant filamentous material (F) and reticulum of a white lamb. Mitochondria (M), polyribo-
mitochondria (M). Cytoplasmic processes (P) and des- somes (P) and a rough endoplasmic reticulum (R) are
mosomes (arrows) are conspicuous (11500 x) evident (28500 x)

spaces were prominent (1000 mm) (Fig. 7). Mito- Several layers of electron-dense non-keratinized
chondria were abundant (Fig. 8) and the cells also cells (Fig. 9) was characteristic of the stratum cor-
contained polyribosomes, a rough endoplasmic reti- neum in all the lambs. The cells contained abundant
culum and a Golgi apparatus (Fig. 8). At the more tonofibrils (Fig. 10), while no organelles could be
distal level of the stratum spinosum, the cells be- positively identified. A trilaminar cell membrane sur-
came flattened, they were more compactly arranged, rounded the cells (Fig. 11). The luminal surface of
and the intercellular space was reduced (400 nm) these cells showed finger-like cytoplasmic projections
(Fig. 1). There was an increase in the amount of fila- (Fig. 12) which were covered by an amorphous, fuzzy
mentous material in the cells. Fewer mitochondria coating (Fig. 11). The intercellular spaces were nar-
were observed, but free ribosomes were still present row (95 nm) (Fig. 13) and numerous desmosomes
in large numbers. The cells contained relatively little were evident (Fig. 13). The stratum corneum of the
rough endoplasmic reticulum. Keratohyalin granules omasum consisted of more cell layers than that of
were observed in the stratum granulosum (Fig. 12). the rumen and reticulum (Fig. 13).

200
 H.B. GROENEWALD

FIG. 9 The stratum corneum in the reticulum of a white lamb. FIG. 11 The luminal surface with cytoplasmic projections (C) of
Note the non-nucleated, electron-dense luminal cells (C) a cell in the stratum corneum in the rumen of a grey
with vacuoles (V) in the cytoplasm. The cytoplasmic pro- lamb. Note the trilaminar cell membrane and the amor-
cesses on the cell surface (arrows) are stubby (661 0 x) phous, fuzzy coating (arrow) (39000 x)

12

FIG. 10 The cytoplasm of a cell in the stratum corneum in the FIG. 12 The luminal surface of a cell in the stratum corneum in
rumen of a black lamb. The absence of organelles and the reticulum of a black lamb, with numerous long cyto-
abundant tonofibrils (arrows) are apparent (39000 x) plasmic projections (arrows). Keratohyaline granules are
evident (8900 x)

The appearance of the luminal cells of the stratum The structure of the luminal cells of the stratum cor-
corneum in the grey and white lambs differed from neum in the grey lambs that did not suckle, cor-
that in the black control lambs. In the black lambs responded to that of the cells in the black lambs.
these cells were moderately electron-dense, nucle-
ated elements (Fig. 14), and no sloughing of the cells
was evident. In the grey and white lambs the luminal DISCUSSION
cells were electron dense, non-nucleated and vacuo-
lated (Fig. 9). Sloughing of the surface cells was The basic structure of the stratified squamous epi-
observed (Fig. 15). thelium of the rumen, reticulum and omasum in the
grey, white and black Karakul lambs differed very
The cytoplasmic processes on the surface of the lu- little from that described by various authors in rumi-
minal cells were fewer in number and stubbier in the nants (Hyden & Sperber 1965; Prins 1967; Lavker
grey and white lambs (Fig. 9) than the same pro- et a/. 1969; Henrikson 1970a; Henrikson 197Gb;
cesses in the black lambs (Fig. 12). Lyford 1988; Schnorr & Hild 1974; Tamate & Kikuchi

201
Ultrastructure of the epithelium of grey, white and black Karakul lambs

13 15
FIG. 13 The stratum corneum in the omasum of a grey lamb.
Narrow intercellular spaces (long arrows) and numerous FIG. 15 The stratum corneum in the omasum of a white lamb.
desmosomes (short arrows) are evident (5200 x) Sloughing of the luminal cells (arrows) is evident (5200 x)

face was slightly folded and free of interdigitations-

as described by Henrikson (1970b)-in lambs a few
days old. In adult sheep the width of the space be-
tween the proximal surface of the basal cells and the
basal lamina varies considerably (Henrikson 1970b) .
The narrow space between these structures, demon-
strated by Henrikson (1970b) in lambs a few days
old, was also corroborated in this study. The organ-
elles of the basal cells and the structure of the cell
membrane with its associated desmosomes did not
differ from that described by other authors (Hyden
& Sperber 1965; Lavker et a!. 1969; Lyford 1988;
Henrikson 1970a; Henrikson 1970b; Scott, Gardner,
Fulton & Mcinroy 1972; Tamate & Kikuchi 1978) .
Langerhans cells as described by Marshall & Steven
(1969), Gemmell (1973) and Gerneke (1980), were
observed in ruminants , but no Langerhans granules
could be demonstrated. No large, membrane-bound
FIG . 14 A nucleated (N) luminal cell with a less electron-dense vesicles (Lavker eta/. 1969; Lyford 1988) could be
cytoplasm (C) in the stratum corneum in the rumen of a
black lamb (661 0 x) demonstrated in the cells of the stratum basale in any
of the lambs studied. The presence of occluding
junctions in the stratum basale, noted by Scott eta!.
1978; Ram krishna & Tiwari 1979; Fath EI-Bab eta!. (1972), could not be confirmed in this study .
1983; Liebich & Scharrer 1984; Hofman & Schnorr The cells of the stratum spinosum with their associ-
1982; Amasaki & Daigo 1988). In the developing ru- ated organelles, filamentous material, large intercel-
men the basal lamina originates as a smooth struc- lular spaces and desmosomes, and the decrease in
ture, becomes undulated after 5 months of foetal age the number of organelles and increase in the amount
and is folded in adult sheep (Arias, Fernandes, of filamentous material within the more distal levels
Oreste, Baeza & Pavez 1991). The basal lamina of of this layer, confirmed the findings of Hyden & Sper-
the rumen in the 24-h-old lambs used in this study ber (1965), Lavkereta/. (1969), Lyford (1988), Hen-
was slightly folded and conformed to the contours rikson (1970a), Henrikson (1970b) and Tamate & Ki-
of the base of the overlying cells. kuchi (1978) . The mucous granules reported in
In adult sheep the proximal surface of the basal cells sheep by Lavker (1969) were not observed in the
stratum spinosum in any of the lambs.
resting on the basal lamina, displays a complex array
of cellular processes (Hyden & Sperber 1965; Lavker Keratohyaline granules in the stratum granulosum
eta/. 1969; Henrikson 1970a; Henrikson 1970b; Ta- and stratum corneum of the rumen have been re-
mate & Kikuchi 1978). In this study the proximal sur- ported by several authors (Prins 1967; Lavker eta/.

202
 H.B. GROENEWALD

1969; Henrikson 1970b; Lyford 1988; Tamate & Ki- the form of nipple-like cytoplasmic processes-on
kuchi 1978; Gerneke 1981; Hofman & Schnorr the surface of the underlying cells. This observation
1982). A discontinous stratum granulosum and ker- was confirmed in the present study and may explain
atohyaline granules were observed in the lambs the differences in the cytoplasmic processes observed
studied. Amasaki & Daigo (1988) declare that in the between the grey, white and black lambs described
bovine, ruminal surface cells develop keratohyaline by SEM (Groenewald & Booth 1992). The sloughing
granules from the fifth month of gestation onwards. of the cells in the grey and white lambs is probably
Henrikson (1970b) describes a non-keratinized epi- due to the presence of milk in the rumens of these
thelium in foetal, 12-h-old and 3-d-old lambs, and lambs (Groenewald & Booth 1992). The underlying
states that by the seventh day the epithelium resem- cells with the former desmosomal attachment sites
bles the keratinizing epithelium of adult sheep. Ram- on their surface are then revealed.
krishna & Tiwari (1979) also describe a non-keratin- It is concluded that the different cell type seen on the
ized epithelium in the foetus of the goat. luminal surface in black lambs (swollen, clear cells) ,
The cells of the stratum corneum in the lambs stud- as compared to that on the luminal surface in grey
ied contained abundant tonofilament bundles and and white lambs (flattened, electron-dense cells),
remnants of nuclear material similar to that described simply reflects the loss of surface cells by the latter
in ruminants (Hyden & Sperber 1965; Lavker et at. due to the presence of milk in the forestomach. The
1969; Lyford 1988; Henrikson 1970a; Henrikson desquamation of the surface cells revealed the un-
1970b; Tamate & Kikuchi 1978). derlying cell layer with its exposed desmosomal at-
tachment sites. This also explains the differences in
The superficial layer of the stratum corneum in adult the appearance of the luminal cells in the three
sheep had flattened, non-nucleated, electron-dense, groups when studied with the scanning electron mi-
vacuolated surface cells (Hyden & Sperber 1965; croscope.
Prins 1967; Lavker et at. 1969; Henrikson 1970a;
Lyford 1988; Tamate & Kikuchi 1978; Hofman &
Schnorr 1982). In contrast, the luminal layer of the REFERENCES
stratum corneum of the developing rumen displayed AMASAKI , H. & DAIGO, M. 1988. Morphogenesis of the epithe-
larger, less electron-dense, nucleated cells (Arias, lium and the lamina propria of the ru men in bovine fetuses and
Soledad Fernandes & Cabrera 1979; Liebich & neonates. Anatomia Histologia Embryologia , 17:1 -6.
Scharrer 1984). The luminal cells of the stratum cor- ARIAS, J.L., SOLEDAD FERNANDES, M. &CABRERA, M. 1979.
neum of the grey and white lambs did not differ in Distribution and nature of desmosomes in the bovine devel-
appearance from those of older animals (Hyden & oping ruminal epithelium . Acta Anatomica, 105:309- 313.
Sperber 1965; Lavker et at. 1969; Lyford 1988; Hen- ARIAS, J.L., FERNANDES, M.S., ORESTE, P. , BAEZA, S. &
rikson 1970a; Tamate & Kikuchi 1978), and showed PAVEZ, V. 1991 . An ultrastructu ral biochemical analysis of the
epithelial-mesenchymal interface of the ruminal mucosa during
evidence of sloughing. However, in the grey lambs development. Anatomischer Anzeiger, 172:165- 176.
that were not allowed to suckle, and in the black
CONTESCU , D. & EPUREANU, S. 1939 . Beitrag zur Vererbung
lambs, the luminal cells were larger, less electron der grauen Farbe bei den Zurkanaschafen. Zeitschrift fur Tier-
dense and nucleated. They contained abundant tone- zuchtung und Zuchtungsbiologie, 44:211 - 218.
filament bundles although no other organelles were CONTESCU, D. & LEAGU , A. 1941. Neue Beitrage zur Vererbung
observed. No sloughing of the cells was evident. der grauen Farbe bei Zurkanaschafen. Zeitschrift fur Tier-
Arias et at. (1979) described these cells in bovine zuchtung und Zuchtungsbiologie, 50:172- 176.
foetuses as "clear" cells. These clear, luminal cells FATH EL-BAB, M.A. , SCHWARTZ, R. & All , A.M.A. 1983.
were also described by Liebich & Scharrer (1984) Micromorphological studies on the stomach of sheep during
in 1-week-old lambs. prenatal life. Anatomia Histologia Embryologia, 12:1 3 9- 153.
GEM MEL, R.T. 1973. Langerhans cells in the ruminal epithelium
The surface of the luminal cells in the stratum cor- of sheep. Journal of Ultrastructural Research , 43:256- 259.
neum possesses cytoplasmic projections which give GERNEKE, W.H. 1980 . The origin and significance of the Lang-
the cells a granular appearance when seen by scan- erhans cell granules. Journal of the South African Veterinary
ning electron microscopy (Scott & Gardner 1973; Association, 51:137- 142.
Groenewald & Booth 1992). The cells are connected GERNEKE, W.H. 1981 . The nature of membrane-coating and
to adjacent cells by means of desmosomes. Scott & keratohyali n granules in the rumen epithelium. Proceedings
Gardner (1973) described the desmosomes in the of the Electron Microscopy Society of Southern Africa . Durban:
11:103.
surface epithelial cells of the stratum corneum of
adult sheep while Arias et at. (1979) reported on the GROENEWALD, H.B. & BOOT H, K.K. 1992. Scanning electron
microscopy of the mucosal surface of the forestomachs and
distribution and nature of desmosomes in the devel- abomasa of grey, white and black Karakul lambs. Onder-
oping ruminal epithelium in the bovine foetus . Ac- stepoort Journal of Veterinary Research , 59: 167- 174.
cording to Scott & Gardner (1973) the desmosomes HARTWIGK, H. 1950. Futteranschoppung im Labmagen als Letal-
eventually break down to allow the surface layer to faktor bei grauen Karakulschafen. Berliner und Munchener
slough off, leaving the former attachment sites- in Tierartzliche Wochenschrift, 8:161 - 164.

203
Ultrastructure of the epithelium of grey, white and black Karakul lambs

HENRIKSON, R.C. 1970a. Ultrastructure of ovine ruminal epithe- NEL, J.A. 1965. The effects of the sub-lethal factor on the internal
lium and localization of sodium in the tissue. Journal of Ultra- organs of the grey Karakul. Proceedings of the South African
structure Research, 30:385-401. Society of Animal Production, 1965: 282-285.
HENRIKSON, R.C. 1970b. Developmental changes in the struc- NEL, J.A. & LOUW, D.J. 1953. The lethal factor in grey Karakul
ture of perinatal ruminal epithelium: Basal infoldings, glycogen lambs. Farming in South Africa, 5:169-172.
and glycocalyx. Zeitschrift fur Zellforschung, 109:15- 19.
PRINS, K.R. 1967. Eienskappe van karkasse en spysverterings-
HOFMANN , R.R. & SCHNORR, B. 1982. Die functionelle Mor- organe van suiplammers onder intensiewe voedingskondisies.
phologie des Wiederkauer-magens. Stuttgart: Ferdinand Enke M.Sc.-verhandeling, Universiteit van Pretoria.
Verlag.
RAMKRISHNA, V. & TIWARI , G.P. 1979. Histological and histo-
HYDEN , S. & SPERBER, I. 1965. Electron microscopy of the
chemical observations on the forestomach of goat during pre-
ruminant forestomach, in Physiology and digestion in the rumi-
natal life. Acta Anatomica , 103:292- 300.
nant, edited by R.W . Dougherty. Washington: Butterworth.
IVANENKO, V.I. 1949. The determination of viability of grey fur REYNOLDS, E.S. 1963. The use of lead citrate at high pH as an
lambs at birth. Animal Breeding Abstracts, 17:367. electron-opaque stain in electron microscopy. Journal of Cell
Biolology, 17:208- 212.
LANG LET, J. 1949. Genetische Beobachtungen an grauen Kara-
kuls. Zeitschrift fur Tierzuchtung und Zuchtungsbiologie, 58: SCOTT, A. , GARDNER, I. C., FULTON, D.R. & MciNROY, G.B.
248- 257. 1972. Tight junctions in the stratum basale of the ruminal epi-
thelium. Zeitschrift fur Zellforschung, 131:199-203.
LAVKER, A.M. 1969. Fine structure of mucous granules in the
rumen epithelium. Journal of Cell Biology, 41:657- 660. SCOTT, A. & GARDNER, I.C. 1973. Desmosomal attachment
sites and the surface of ruminal epithelial cells. Zeitschrift fur
LAVKER, A.M. , CHALUPA, W. & DICKEY, J.F. 1969. An electron
mikroskopische Anatomie, 87:573- 577.
microscopic investigation of rumen mucosa. Journal of Ultra-
structure Research, 28:1- 15. SCHNORR, B. & HILD, A. 1974. Electronmikroskopisch-zyto-
LIEBICH , VON H.G. & SCHARRER, E. 1984. Entwicklungs- chemische Untersuchungen an Zellmembranen der Hornzellen
bedingte Verahderungen von Struktur und Funktion des Pan- vom Vormagenepithel der Ziege. Anatomie, Histologie, Embry-
senepithels. Anatomia Histologia Embryologia , 13:25-41. ologie, 3:324-336.
LYFORD, S.J. 1988. Growth and development of the ruminant di- TAMATE, H. & KIKUCHI, T. 1978. Electron microscopic study on
gestive system, in The ruminant animal: Digestive physiology parakeratotic ruminal epithelium in beef cattle. Japanese Jour-
and nutrition, edited by D.C. Church. New Jersey: Reston nal of Veterinary Science , 40:21 - 30.
Books. WATSON, M. L. 1958. Staining of tissue sections for electron
MARSHALL, A.B. & STEVEN, D.H. 1969. Branching cells in the microscopy with heavy metals. Journal of Biophysical and Bio-
rumen epithelium. Journal of Physiology, 204:13- 14. chemical Cytology, 4:475-478.

204