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Kerala Veterinary and Animal Sciences University, Wayanad
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ThesisItem Open Access ASSESSMENT OF FERTILITY AND FECUNDITY IN FEMALE DOGS BY SONOGRAPHIC, HORMONAL AND GENOMIC STUDIES(COLLEGE OF VETERINARY AND ANIMAL SCIENCES MANNUTHY, THRISSUR, KERALA VETERINARY AND ANIMAL SCIENCES UNIVERSITY, 2022-12-09) NIKHILA E.S; Dr. Magnus Paul KPopularity of dog breeding is increasing worldwide. The commercial canine breeding mainly aims to attain maximum number of litters during the life time of a female dog. Low fecundity is a major issue faced by commercial breeders. The present study was undertaken for the assessment of fertility and fecundity in female dogs by sonographic, hormonal and genomic studies. A total of sixteen fertile female dogs aged two to six years presented to TVCC, Mannuthy during the period from February 2021 to November 2021 for breeding advice and subsequent pregnancy diagnosis forms the subject of the study. All the animals were subjected to exfoliative vaginal cytology (EVC) to assess the stage of oestrous cycle and subsequent Doppler assessment of ventral perineal artery haemodynamics. Mean cornification index (%) of the animals under study was 89.5 ± 1.81 and 88.2 ± 2.13 in high fecundity and low fecundity groups respectively. Early pregnancy diagnosis was carried out on day 21st of dioestrum. Progesterone estimation was also done on the same day in all the animals to relate with foetal number. Based on the foetal number on radiography animals were grouped in to two of eight bitches each. Group I with greater than three foetuses and group II with three or less than three foetuses. No significant difference was observed in the Doppler parameters of ventral perineal artery during different stages of oestrous cycle and between the groups. However, a gradual increase was observed in the ventral perineal artery resistivity (RI) and pulsatility index (PI) between the stages of oestrous cycle. This might be due to the vasodilatory effect of oestradiol. Serum progesterone profile in the high fecundity and low fecundity was not statistically significant. Mean serum progesterone concentration in high fecundity and low fecundity group was 44.60 ± 3.39 and 42.25 ± 5.13 respectively. Prediction of litter size by ultrasonography on day 21of dioestrum was successful in 75 per cent (n=6) animals in group II compared with 25 per cent (n=2) in group I. Partial characterization of LHCGRgene revealed a novel polymorphism in the exon-1 in high fecundity group. The sequence analysis of amplicon of exon-1 of LHCGR gene showed 99 per cent similarity with the reference sequence (GenBank Accession No:NC_0518141). Two SNPs at 815th (g.815 C>T) and 872th (g. 872 A>G) base pair of LHCGR was identified. The sequence analysis of amplicon of exon-2 of LHCGR gene showed 99 per cent similarity with the reference sequence (GenBank Accession No:NC_0518141). One SNP at 175th (g.175 C>T) base pair of LHCGR was identified. From the present study it could be concluded that blood flow in the ventral perineal artery do not having any correlation with the number of ovulations as well as the number of pups born. This was also evident in the progesterone levels as the level of progesterone was not influenced by the ovulations or the number of CLs. Further investigations are warranted to elucidate the effect of novel SNPs on fecundity in dogs after confirming the influence by extensive association studies.ThesisItem Open Access IMMUNOHISTOCHEMICAL LOCALISATION OF PROGESTERONE, OESTROGEN AND KI-67 RECEPTORS IN GENITAL TRACT TUMOURS OF FEMALE DOGS(COLLEGE OF VETERINARY AND ANIMAL SCIENCES MANNUTHY, THRISSUR, KERALA VETERINARY AND ANIMAL SCIENCES UNIVERSITY, 2022-11-24) KODANGE OMKAR TUKARAM; Dr. Shibu SimonThe objectives of the current study involved assessment of expression of progesterone and oestrogen receptors in the genital borne neoplasms by immunohistochemistry, correlation of steroid receptor status with proliferation marker (Ki-67) and their clinical management. The study was conducted in 12 postpubertal bitches between two to twelve years of age, presented at University Veterinary Hospitals, Kokkalai and Mannuthy. The bitches presented to either of the University Veterinary Hospitals, Kokkalai and Mannuthy with suspected genital neoplasia confirmed based on symptoms, vaginal cytology, ultrasonography and radiological evaluation were included in the research. The incidence of neoplasia was the highest (50 %) among the age group of 5-8 years in comparison with other groups and the nulliparous bitches were twice in number when compared to multiparous bitches. The breeds of afflicted canines consisted of Pomeranian (n=3), Labrador retriever (n=3), Rottweiler (n=2), English cocker spaniel (n=1), Dachshund (n=1), Nondescript (n=1) and Dobermann (n=1). No breed predisposition was recorded in the study. The symptoms recorded were stranguria (58.3 %), aberrations of oestrous cycle (16.6 %), haemorrhagic vaginal discharge (91.6 %), faecal tenesmus (50 %), perineal swelling (33.3 %), visible protruding mass (66.6 %) and inappetence (58.6 %). Clinicogynaecological examination revealed that bitches afflicted with pedunculated and sessile tumours were equal in numbers. The intra-luminal and extra-luminal masses accounted for 25 per cent and 75 per cent, respectively. The cytological evaluation of exfoliated cells in the samples confirmed dioestrus in three and anoestrus in nine animals. Exfoliated neoplastic cells were only observed in canine transmissible venereal tumour and vaginal cytology was unremarkable for other tumours. All haematological values were within the normal reference range. Ultrasonographic evaluation of all the neoplasms indicated a heterogenous echotexture. Doppler evaluation of the tumours revealed unidirectional perfusion in all except two types of tumours; the canine transmissible venereal tumour (CTVT) and squamous cell carcinoma (SCC) which had an arbitrary perfusion. Ultrasonographic screening of the vital abdominal organs indicated absence of metastasis to other organs. Similar observations were made with radiography of abdomen and thorax. Episiotomy was performed for extraluminal tumours while local excision was performed in all intraluminal tumours except CTVT. A chemotherapeutic regimen involving intravenous infusion of vincristine sulphate at 0.025 mg/kg weekly was given in CTVT and SCC until complete remission of the neoplasm. The tumours werehistopathologically confirmed as myofibroma (n = 6), leiomyoma (n = 1), CTVT (n = 3), SCC (n = 1) and fibroma (n = 1). Masson’s trichrome staining confirmed connective tissue involvement in myofibroma, leiomyoma and fibroma. The combined scores for ERα expressions in myofibroma, leiomyoma and fibroma were 5, 5 and 7, respectively. The combined scores for PR expression in myofibroma, leiomyoma and fibroma were 5, 5 and 2, respectively. The expression for ERα and PR was absent in CTVT and SCC. The combined score for Ki-67 expression myofibroma, leiomyoma, fibroma, CTVT and SCC were 3, 4, 2, 7 and 8 respectively. Spearman’s rank correlation between the scores of immunohistochemistry staining for the expression of ERα, PR and Ki-67 in these tumours was highly significant (p < 0.01). The correlation between ERα and Ki-67 had a significant negative correlation (p < 0.05). The correlation between PR and Ki-67 was not significant (p > 0.05). A target selective therapy in a histologically confirmed leiomyoma with oral mifepristone therapy (25 mg/day for 12 weeks) revealed 70 per cent reduction in tumour volume.This result suggests probable use of hormone modulation therapy in PR expressing genital tumours. The study also suggests that a decline in ERα expression may concurrently upregulate Ki-67 receptor expression in genital tumours indicating a shifttowards malignancy by the tumour. The differential expression of these proteins should be considered for the stratification of patients while designing therapeuticThesisItem Open Access INTRAUTERINE OZONE TREATMENT FOR SUBCLINICAL ENDOMETRITIS IN CROSSBRED DAIRY COWS(COLLEGE OF VETERINARY AND ANIMAL SCIENCES MANNUTHY, THRISSUR, KERALA VETERINARY AND ANIMAL SCIENCES UNIVERSITY, 2022-10-13) H NEELAKANTA NIKSHEPA NITHISH; Dr. C. JayakumarThe study was undertaken at University Livestock Farm and Fodder Research and Development Scheme, Mannuthy with the objective of assessing the therapeutic efficiency of intra uterine ozone therapy in management of sub clinical endometritis (SCE) and its influence on the reproductive performance in postpartum (PP) cross bred dairy cows. The study assessed the efficacy of intra-uterine ozone therapy with and without PGF2α analogue in the treatment of SCE in crossbred dairy cows. Efficacy of treatment was evaluated through endometrial cytology, microbiological evaluation (CFU), endometrial histopathology and uterine sonography findings before and after treatment in SCE positive animals. Also, the diagnostic competence of these techniques in SCE was also undertaken. Twenty four cows with normal calving and confirmed with SCE on 30th Day Postpartum (DPP) was randomly divided in to three experimental groups after their first exhibited oestrus on 45th DPP viz,Group I - Intrauterine ozone therapy with 3300 mcg for two days from the onset of oestrus, Group II - Intramuscular administration of 500 mcg cloprostenol on 10th day of oestrus and Group III- Intrauterine ozone therapy with 3300 mcg for two days from the onset of oestrus followed by intramuscular administration of 500 mcg cloprostenol on 10th day of oestrus. The (mean±SE) per cent of PMN cells before and after treatment were 5.12± 0.56, 5.25 ± 0.56, 4.81 ± 0.56 and 1.06 ± 0.21, 1.68 ± 0.21, 1 ± 0.21, respectively for three treatment groups. The (mean±SE) total score of histopathology samples before and after treatment were 6.25 ± 1.57, 5.88 ± 0.89, 6.75 ± 0.79 and 2.75 ± 0.52, 4.75 ± 0.92, 3.13 ± 0.58 in Group I, II and III, respectively. Both endometrial cytology and histopathology yielded significant difference before and after treatment, except in Group II with regard to histopathology. In CFU and transrectal ultrasonography (TRUS) findings, though the values were non-significant before and after treatment, a positive variation in both parameters could be noticed envisaging a usefulness of treatment in all the groups. The overall conception rate to Artificial Insemination in four consecutive oestrous periods was 62.5 per cent. The average of number of inseminations (2.38 ± 0.37, 2.88 ± 0.35 and 2 ± 0.42) and days to conception (89.66 ± 5.59, 100 ± 10.49 and 85.83 ± 8.78 d) varied in treatment groups, but without significant difference. The study determined that intra uterine O3 therapy alone or in combination was effective in management of SCE in PP cows signified by a decrease in PMN leucocytes, histopathology scores and satisfactory PP reproductive performance indices. The study concluded the usefulness of intra uterine ozone therapy as a novel, alternate and safe PP reproductive management strategy to improve reproductive health and efficiency in cattle herds with increased incidence of SCE.ThesisItem Open Access FOLLICULAR DYNAMICS AND EXPRESSION PROFILE OF B-CELL LYMPHOMA-2 GENE IN MALABARI DOELINGS SUPPLEMENTED WITH MORINGA OLEIFERA LEAVES(COLLEGE OF VETERINARY AND ANIMAL SCIENCES MANNUTHY, THRISSUR, KERALA VETERINRY AND ANIMAL SCIENCES UNIVERSITY, 2022-06-22) AMRITHA ARAVIND; Dr. Shibu SimonThe study was conducted to assess the effect of of M. oleifera leaves on reproductive performance in Malabari doelings. Apparently healthy Malabari kids (n= 32) of age three months were selected and randomly allocated to two different treatment groups. Control group (T1, n= 16) was fed with complete feed as pellets and experimental group (T2, n=16) was given complete feed in which 50 per cent of concentrate was replaced with M. oleifera leaves on dry matter (DM) basis. The feeding trials were continued till two oestrous cycles were completed. The mean initial (T1-8.00 ± 0.50 kg; T2- 7.89 ± 0.55 kg) and final body weights (T1-18.73 ± 0.89 kg; T2- 19.95 ± 0.89 kg) did not differ significantly whilst showed a linear increase in both the groups. Daily DMI and haematological parameters (Hb, Haematocrit, TEC, TLC) did not vary significantly between groups. In T2, the mean total protein was significantly higher (p < 0.05) while mean ALT remained significantly lower (p < 0.05). Post weaning mean progesterone profile remained less than 1ng/mL until fifth month of age but showed significant increase during sixth month (T1- 0.24 ng/mL, T2- 0.44 ng/mL; p < 0.05) and elevated more than 1ng/mL during seventh month in T2 (T1- 0.72 ± 0.22 ng/mL, T2- 1.14 ± 0.29 ng/mL). First behavioural oestrus was observed during sixth month age (T1-12.50 %; T2- 31.25 %) without any significant variation between groups. On ultrasound examination of follicular dynamics in two consecutive oestrous cycles, significantly higher number of small, medium and large follicles could be observed in T2 (p < 0.01). Typical wave patterns of one to five wave cycles could be noticed in Malabari doelings in peripubertal period regardless the experimental groups and short cycles were characteristic of first oestrous cycle in both the groups. The first interwave interval of five wave cycle and ovulatory wave of four wave cycle was longer in T2. The two groups did not differ significantly with regard to diameter of largest follicles in each wave, day of appearance of largest follicle, day of ovulation, number and size of ovulatory follicles. The size of corpus luteum, blood flow assessed by pixel count and mean progesterone values were higher during luteogenesis of first oestrous cycle in T2 (p < 0.05). Significantly higher behavioural and physiological score and oestrus intensity could be observed in T2 (p < 0.01). Length of oestrous cycle and duration of oestrus did not vary significantly between the groups. Following follicular aspiration (n=6), significantly higher (p < 0.05) culture grade (A and B) oocytes could be observed in T2 (85.71%) than T1 (43.75%). BCB staining revealed, significantly higher (p < 0.05) number of oocytes with developmental competence in T2 (3.83 ± 0.48) than T1 (2.17 ± 0.48). Follicular health assessed by relative expression of antiapoptotic gene BCL-2 showed no significant variation between the groups. Thus, the present study authorizes the effective and safe means of replacement of 50 per cent concentrate ration of goats with M. oleifera leaves without affecting the growth, dry matter intake and haematobiochemical profiles in Malabari doelings and it improved the reproductive performance by advancing the puberty, folliculogenesis, luteogenesis and enhanced developmental competence and quality of oocytes during peripubertal period.ThesisItem Open Access ASSESSMENT OF MYOMETRIAL CONTRACTILITY, GENE EXPRESSION OF HORMONE RECEPTORS IN UTERUS AND FOETO-MATERNAL VELOCITY INDICES IN CANINE UTERINE INERTIA(COLLEGE OF VETERINARY AND ANIMAL SCIENCES MANNUTHY, THRISSUR, KERALA VETERINARY AND ANIMAL SCIENCES UNIVERSITY, 2022-05-19) Magnus Paul K; Dr. C. JayakumarThe present study was designed to assess the uterine contractility characteristics, gene expression of hormone receptors and the change in velocity indices of maternal and foetal arteries in canine uterine inertia. Dogs presented (n=22) with dystocia were classified as complete primary uterine inertia CPUI (n=9), partial primary uterine inertia PPUI (n=6) and foetal cause of dystocia FCD (n=7) based on the cause of dystocia, contractile state of the uterus as well as the stage of progress of the foetus. All the selected dogs were subjected to ultrasonography with Pulse wave spectral Doppler analysis, radiography, sigmoidoscopy, non-invasive tocodynamometry, haematology, blood gas, electrolyte and metabolite analysis and serum hormonal profile analysis. Caesarean section was performed in all the dogs, and after the removal of the foetus, fullthickness uterine biopsy samples were taken and frozen in RNA-later. Apgar scores of all the foetuses were recorded at five minutes, two hours and 24 h. Relative expression of genes regulating myometrial contraction as well as hormonal receptor genes were performed using the qPCR in the study groups. Tocodynamometric studies confirmed that dogs exhibiting less than 15 mm of Hg pressure and feeble or infrequent contractions without delivery for more than 30 min, could be considered as uterine inertia. Apgar scores in pups at five minutes after the surgery were poor in all groups, but the rapid improvement was noticed in two hours. Analysis revealed normal pH, High pO2, low pCO2, low HCO3⁻ and low base excess. Significantly lower Na+and Cl⁻ concentration in the FCD group, and lower iCa and anion gap in the FCD group were noted. Respiratory acidosis and metabolic alkalosis were evident in all animals, with the maximum level in FCD. Quantitative analysis of RI, PI and SVDV of the foetal abdominal aorta and the uterine artery did not differ significantly between the groups. Haematology revealed no significant difference between the groups, but lymphocytosis and anaemia were evident in all groups. The serum hormone profile revealed no significant difference between oestradiol and progesterone concentrations across the groups. However, oestradiol level was higher in all groups than previously reported levels in normal pregnancy. The present study concluded that ionised calcium deficiency, higher oestradiol concentration, upregulation of Erβ receptor gene and downregulation of MLCK gene contributed an additive effect to cause CPUI, along with the possible absence of oxytocin, which was indirectly revealed through the elevated Na+and Cl⁻concentration. Whereas the PPUI was found to be a myometrial defect due to extreme down-regulation of MLCK, MYHII and PKC genes, higher oestradiol levels also might have contributed to the development of the pathology.ThesisItem Open Access EVALUATION OF THE UTERINE FLUID AND ENDOCRINE CHANGES ASSOCIATED WITH METOESTRUAL BLEEDING IN DAIRY CATTLE(COLLEGE OF VETERINARY AND ANIMAL SCIENCES, POOKODE, WAYANAD, KERALA VETERINARY AND ANIMAL SCIENCES UNIVERSITY, 2022-09-05) NISARGA M; K. PromodThe present study was conducted with the objective to evaluate the changes in uterine fluid, serum oestrogen and progesterone profiles associated with metoestrual bleeding in dairy cattle. The study was conducted in Group I animals (n=12) which included cows with the historyof profuse metoestrual bleeding and Group II cows (n=12) without metoestrual bleeding. A total 24 cows were studied for the rheological properties of cervical mucus discharge (CMD), endometrial cytology and biochemical changes of uterine fluid during oestrus (day 0) and metoestrus (day 2). The serum progesterone and oestrogen concentrations on days 0, 2 and 10 of oestrous cycle were estimated by RIA. Per rectal examination revealed persistence of dominant follicle (58.33 per cent) on ovaries and high uterine tonicity during metoestrus in group I cows. Majority of group I cows exhibited high oestrus intensity and prolonged duration of oestrus. The Group I cows voided CMD with bloody opaque, thin/watery consistency and ‘nil’ fern pattern during metoestrus. While the Group II cows exhibited CMD with turbid, thick/viscous and atypical fern pattern during metoestrus. The mean ± SE pH of CMD of Group I cows (7.96±0.10) had significantly (p<0.01) higher values than that of cows group II cows (7.64±0.07) during metoestrus. The mean spinnbarkeit values of Group I cows (9.35±0.50 cm) was significantly (p<0.01) higher than that of Group II cows (7.75±0.30) during metoestrus. The mean serum progesterone concentration of Group I cows (1.61±0.55 ng/mL) was significantly (p<0.05) higher than that of Group II cows (0.36±0.07 ng/mL) on day of oestrus. The mean serum oestradiol concentration of Group I cows (6.85±0.53 pg/mL) was significantly (p<0.05) lower when compared to Group II cows (8.48±0.49 pg/mL). Cytology revealed the PMN cell per cent during metoestrus was significantly (p<0.01) higher in Group I cows (4.04±0.25 per cent) than in Group II cows (3.02±0.11 per cent). A significantly (p<0.01) higher LE activity of uterine fluid was observed in Group I cows (142.5±33.26 cells/μL) than that of Group II cows (7.5±2.26 cells/μL) during metoestrus. In conclusion, the oestrus characteristics, the pH and spinnbarkeit value of CMD, serum progesterone and oestrogen concentration, PMN cell per cent and LE activity of uterine fluid during metoestrual bleeding was significantly varied. The overall conception rate in these cows (33.33 per cent) was found lower than that of the cows without metoestrual bleeding (83.33 per cent)ThesisItem Open Access CRYOPROTECTIVE EFFECT OF BUCK SEMEN SUPPLEMENTED WITH DIMETHYLACETAMIDE AND L-GLUTAMINE IN A TRIS BASED EXTENDER(COLLEGE OF VETERINARY AND ANIMAL SCIENCS, POOKODE, WAYANAD, KERALA VETERINARY AND ANIMAL SCIENCES UNIVERSITY, 2022-12-23) KANIMOZHI T.M; Dr. Leeba ChackoA study was designed with the objective to assess the cryoprotective effect of dimethylacetamide (DMA) addition to Tris based buck semen extender and to evaluate the synergistic effect of DMA and L-glutamine on post-thaw semen quality. Ten ejaculates were taken from four Malabari bucks. After preliminary evaluation, pooled samples was extended in four different extender including control, 3 per cent DMA, control with L-glutamine and DMA with L-glutamine. Pre-freeze and post-thaw evaluation including sperm kinectics was studied. Post thaw motility, viability, HOST and acrosome integrity (56.26 ± 3.54; 48.25 ± 0.75; 59.5±1.13; 59.88 ± 2.51 per cent), respectively was significantly higher (p<0.05) and low MDA values (2.22 ± 0.15 μmol/mL) in control with L-glutamine group was noticed whereas DMA group had lower post-thaw motility, viability, HOST and acrosome integrity (32.19 ± 3.28; 35.13 ± 1.17; 41.25 ± 1.52; 43.00 ± 1.17), respectively and high MDA value (4.11 ± 0.25 μmol/mL). From the present study it was concluded Lglutamine had synergistic effect with glycerol and manual freezing of Malabari buck sperm with DMA was not feasible comparing to glycerol.ThesisItem Open Access PREDICTING PROGNOSIS OF CANINE PYOMETRA BY EVALUATING RENAL DYSFUNCTIONS USING SPECIFIC BIOMARKERS AND ITS THERAPEUTIC MANAGEMENT(COLLEGE OF VETERINARY AND ANIMAL SCIENCES, POOKODE, WAYANAD, 2022-08-31) AISHWARYA RRetrospective study conducted on canine reproductive disorders for past three years and occurrence of pyometra was analysed. Out of the 704 reproductive cases, incidence of pyometra was 12.21 per cent. Among pyometra cases, open and closed cervix pyometra were 72.90 and 27.90 per cent, respectively. The prospective study was conducted in 24 open cervix pyometra cases and were subjected to three treatment protocols with eight animals in each group. Group I: treated with inj. Dinoprost tromethamine @ 0.2 mg/kg S/C, BID for 5 days, Group II: treated with tab. Mifepristone @ 2.5 mg/kg,PO, BID 5 days and Group III: combination of inj. Dinoprost tromethamine @ 0.2 mg/kg S/C, BID for alternate days and Tab. Mifepristone @ 2.5 mg/kg, PO, BID for 5 days. Ultrasonographic examination conducted on day 0, 5, 10 and 20 revealed statistically significant decrease (P<0.001) in the uterine horn diameter from 36.75 ± 1.10 mm (day 0) to 12.51 ± 0.91 mm (day 20). Haematobiochemicalparameters on day ‘0’ revealed regenerative anemia indicated by reduced haemoglobin and total erythrocyte values and DLC values indicated neutrophilia and lymphocytopenia and on day 20 revealed a significant increase with respect to Hb, PCV and TEC values. DLC indicated significant decrease in neutrophil count and a significant increase in lymphocyte count. Serum estimation on different days of observation revealed a significant decrease (P<0.01) in BUN values and a marginal decrease in the creatinine values. Urinalysis revealed no significant changes with respect to colour, pH, specific gravity, crystals and casts in urine on different days of observation. Biochemical tests of urine with respect to UPCR and UGGT conducted on different days of observation revealed no significant changes in UPCR values, however, a highly significant (P<0.01) decrease in the mean UGGT values ranging from 31.00 ± 2.63 (day 0) to 12.75 ± 0.86 (IU/L) (day 20) observed in all the three groups.Present study concluded that, UGGT could be considered as a urinary biomarker as it is an enzyme located on proximal tubule membrane and the levels will be increased in acute renal injury and can be considered early prognostic biomarker for canine pyometra cases affected with kidney damage. Further, All the animals in group III recovered completely and no recurrence of pyometra was noticed during one month period ofobservation. PGF2α promote ripening of cervix and contraction of myometrium whereas, mifepristone blocks the progesterone receptors and causes opening of cervix resulting in complete evacuation of pus contents. The recovery rate for group I andgroup II were 75 and 87.50 per cent, respectively.ThesisItem Open Access ISOLATION AND IDENTIFICATION OF BIOFILM FORMING BACTERIA IN CLINICAL CASES OF CANINE PYOMETRA(COLLEGE OF VETERINARY AND ANIMAL SCIENCES, POOKODE, WAYANAD, 2022-09-15) SURAJ S. GOURAV; Leeba ChackoThe present study was conducted for the assessment of biofilm forming bacteria and presence of fimH gene from canine pyometra cases presented during the period of March 2020- July 2021 at Teaching Veterinary Clinical Complex, Pookode, Wayanad. Female dogs irrespective of age, breed, type (closed/open pyometra) and parity were screened for pyometra based on detailed history, physiological parameters, clinical signs and confirmed by trans-abdominal ultrasonography. Sterile swabs were adopted to collect pus samples from the anterior vagina of the affected dogs, which were later subjected to bacterial isolation and antibiogram studies.The present study included eighteen animals that revealed positive culture of E. coli bacteria screened by employing laboratory procedures. These eighteen dogs revealed haemoglobin level (mean ± SE) of 12.62 ± 0.86 g/dL, total leucocyte count (x103/µL; mean ± SE) of 41.9 ± 4.61 x103/µL, lymphocyte count (in per cent) of 13.38 ±1.74, monocyte count (in per cent) of 2.97 ± 0.15, neutrophil count (in per cent) of 84.84 ±2.38 and an eosinophil count (in per cent) of 2.81±0.28. The serum biochemical parameters such as BUN and creatinine (mean ± SE) recorded on the day of admission was 34.49 ±7.11 mg/dL and 1.97 ±0.43 mg/dL, respectively. On antibiotic sensitivity test (ABST), all the recovered isolates were found susceptible to cefotaxime-clavulanic acid and ceftazidime-clavulanic acid.The antimicrobial susceptibility (in per cent) of the isolates was found in the decreasing order as chloramphenicol (94.44) followed by gentamicin and ceftriaxone (83.33) each, cotrimoxazole and cefotaxime (77.77) each, ciprofloxacin (66.66), colistin (61.11), doxycycline (55.55) and ampicillin (50). The least sensitivity was observed against amoxicillin. Laboratory tests revealed 61.11 percent (11/18) of E. coli isolates as strong biofilm producers and 38.88 per cent (7/18) as weak biofilm producers when subjected to congo red agar binding assay. In tissue culture plate 89method, 88.88 per cent (16/18) isolates showed moderate biofilm forming potential, while 11.11 per cent (2/18) of the isolates revealed weak biofilm forming potential, none of the isolates showed strong biofilm forming potential. In 55.55 per cent (10/18) isolates, the presence of fimH gene was confirmed by PCR analysis. Amongst the eighteen dogs under study, seven dogs (38.88%) showed closed cervix pyometra and six dogs (33.33%) were unfit for medical management. Five (27.77%) clinically stable dogs treated with natural prostaglandin(Dinoprosttromethamine) and Inj. Amoxirum Forte® 300mg) at the dose rate of 20mg/kg intravenously for seven days showed complete evacuation of pus which was confirmed by ultrasonography.The uncontrolled and rampant use of antibiotics in combination with faulty diagnosis has resulted in increased number of canine pyometra cases with biofilm affection. Tissue culture plate method stands superior to congo red binding agar assay in detection of biofilms because, isolates can be efficiently differentiated as strong or moderate or weak biofilm producers. The present study formed the first and foremost study of biofilms in canines affected with pyometra in India.
