ETIO-PATHOLOGICAL AND PROINFLAMMATORY CYTOKINE EXPRESSION STUDIES ON ENDOMETRITIS IN BUFFALOES 2512
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Date
2018-04
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JAU,JUNAGADH
Abstract
The present inestigation was carried out to study the prevalence of
endometritis, its bacterial etiology / their antibiotic sensitivity, pro-inflamatory
cytokine profiling /their role in understanding the pathogenesis of endometritis
leading to early diagnosis and pathomorphological changes on slaughter house
materials collected from slaughter house at Junagadh. The buffaloes were
comparatively aged ones and a total of 110 genitalia were collected and processed in
Pathology Department for fulfilling various objectives mentioned above.
The prevalence of endometritis was recorded in 81% (89/110) slaughtered
buffaloes and it was also categorized as acute (25.45%), subacute (20.90%) and
chronic (34.55%) based on pathomorphological feature.
Acute endometritis was characterized by severe congestion along with marked
stromal edema, degenerative changes and focal denudation of luminal epithelium,
focal hemorrhage in sub epithelial zone, infiltration of inflammatory cells
predominantly polymorphonuclear cells and mononuclear cells in lamina propria,
infiltration of mononuclear cells in glandular lumina and peri-endometrial glands.
Subacute endometritis consisted of denudation of luminal epithelium, congestion,
stromal edema and focal haemorrhagic spot, infiltration of mononuclear cells in
lamina propria, glandular lumina and around the atrophied endometrial gland,
glandular dilation, hyperplasia of mucosal epithelium, atrophy of endometrial glands
and thickening of blood vessels. The main features of chronic endometritis were
desquamation of mucosal epithelium, infiltration of mononuclear cells and plasma
cells in sub epithelial zone, dilatation of endometrial glands with degenerative
changes, infiltration of mononuclear cells in glandular lumina and periglandular
region with narrowing of glandular lumina, perivascular and periglandular fibrosis
leading to severe thickening of blood vessels resulting in narrowing of their lumina
and transformation of endometrial epithelial cells into low cuboidal against the
normal columnar epithelium. Beside the above histopathological lesions three uterine
samples revealed adenomyosis and twenty two genitalia showed metritis.
The bacterial cultural examination was carried out on all the110 uterine swabs
collected from slaughtered buffaloes. Fifty six (50.90%) uterine samples showed the
growth of various bacteria and remaining 54 swabs (49.10%) were found to be sterile.
Out of the 56 uterine samples 50 swabs (89.28%) showed single isolate and remaining
06 (10.72%) exhibited mixed infection. Bacterial isolates comprised of
Staphylococcus spp. (11.82%), Escherichia coli (8.18%), Micrococcus spp. (8.18%),
Corynebacterium spp. (7.27%), Fusobacterium spp. (4.55%), Pseudomonas spp.
Abstract…
(2.73%), Bacillus spp. (1.82%), Streptococcus spp. (0.91%) and mixed infections
consisted of two different types of isolates in different combinations (E. coli +
Corynebacterium spp. – 2.73%, E. coli + Staphylococcus spp. – 1.82% and E. coli +
Bacillus spp. – 0.91%).
The Escherichia coli and Staphylococcus spp. isolates (24.19%) topped the list
followed by Corynebacterium spp. (17.74%), Micrococcus spp. (14.52%),
Fusobacterium spp. (8.06%), Pseudomonas spp. (4.84%), Bacillus spp. (4.84%) and
Streptococcus spp. (1.61%).
Overall antimicrobial susceptibility of 62 bacterial isolates showed highest
sensitivity against Chloramphenicol (83.9%) followed by Gentamicin (80.6%),
Levofloxacin (77.4%), Oxytetracycline (77.4%), Ceftriaxone/Sulbactam (69.3%),
Cefoperazone/Sulbactam (61.2%) and Amoxicillin/Sulbactam (33.9%).
Subclinical and clinical endometritis were adjudged on the basis of
endometrial cytology and white side test for cytokine profiling. The endometrial
cytology showed more than 5% polymorphonuclear cells (PMNs) among 200 cell
counted in 45 (40.90%) cases out of 110 sample. Eighteen genitalia out of 110
genitalia having mucopurulent or purulent discharge and positive on white side test
were taken as clinical endometritis. A total of 36 categorized genitalia were divided
into three groups namely, Group-I (n=12; Normal), Group-II (n=12; Clinical
Endometritis / CE) and Group-III (n=12; Subclinical Endometritis / SCE). Further,
each group was subdivided as follicular (n=6) and luteal (n=6) stages of the ovarian
cycle based on gross morphology of ovaries.
The expression of CD14, TLR-4 and IL-1β encoding gene in endometrium of
buffaloes with subclinical endometritis were 1.87, 2.71 and 3.48 fold, while samples
with clinical endometritis showed only 2.90, 4.76 and 8.02 fold for CD14, TLR-4 and
IL-1β, respectively, though numerical value of expression was higher in subclinical
and clinical endometritis but it was at par with control. The expression profile for IL-
6, IL-8 and TNF-α mRNA for subclinical endometritis were 11.95, 14.87 and 12.95
fold while for clinical endometritis, values were 18.17, 37.97 and 28.83 fold for IL-6,
IL-8 and TNF-α mRNA, respectively, which were significantly higher in subclinical
and clinical endometritis as compared to apparently normal samples at follicular
phase. In the luteal group, the subclinical endometritis sample showed 2.13, 6.43,
14.52, 22.38, 18.64 and 2.82 fold respectively, for CD14, IL-1β IL-6, IL-8, TNF-α
and TLR-4 mRNA. Except TLR-4, the values of expression were significantly higher
when compared with apparently normal. Whereas, the expression values of clinical
endometritic uteri were 4.01, 7.15, 17.20, 45.55, 52.58 and 32.95 fold respectively,
for CD14, TLR-4, IL-1β, IL-6, IL-8 and TNF-α mRNA which were significantly
higher as compared to apparently normal uteri.
Based on present study it was concluded that: i) the prevalence of endometritis
in slaughtered buffalo was 81% and most common bacteria involved were E coli and
Staphylococcus which were highly sensitive to Chloramphenicol, ii) the mRNA
expression values of different cytokines (CD14, IL-1β, IL-6, IL-8 and TNF-α) might
represent a potential marker gene to detect endometritis in buffaloes, iii) chronic
endometritis was the major problems seen in slaughtered buffaloes and as these
animals cannot reproduce, caused economic loss to livestock owners / farmers and
this is why such animals are sold for slaughtering purposes.
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Keywords
VETERINARY PATHOLOGY