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ThesisItem Open Access REPRODUCTIVE PERFORMANCE OF EARLY POSTPARTUM CROSSBRED COWS SUPLLEMENTED WITH BYPASS FAT AND SUBJECTED TO HORMONAL INDUCTION OF OESTRUS(COLLEGE OF VETERINARY AND ANIMAL SCIENCES MANNUTHY, THRISSUR, 2018-11-30) ANI S.DAS; Metilda JosephPostpartum infertility due to negative energy balance is a major problem among the high producing cows of Kerala, incurring huge economic loss. The present study was aimed to evaluate the effect of supplementation of bypass fat on the reproduction and production parameters of high yielding crossbred cows with or without postpartum hormonal induction of oestrus and its techno-economics. The trial was conducted with four experimental groups of six cows each (GI – control, GII- 200 g bypass fat 90 days, GIII – 200g bypass fat daily + Ovsynch protocol on day 45postpartum, GIV – Ovsynch protocol alone). The animals in GII and GIII were observed to be superior in milk production parameters viz. peak yield (P<0.05), and days taken for attaining peak yield. Cessation of post-patum lochial discharge and its characteristics were normal in all groups. The animals in GII and GIII showed earlier post-partum oestrus compared to others (P<0.05). As per rectal examination and ultrasonographic findings, more number of animals in GII and GIII showed early resumption of ovarian cyclicity. The number of days taken for complete involution of uterus and reduction of uterine wall thickness as per ultrasonographic examination were significantly lower in those animals fed with bypass fat. With respect to various serum biochemical parameters, blood glucose and total cholesterol were significantly higher (P<0.01) in those animals fed with bypass fat. Blood urea nitrogen, non-esterified fatty acids and triglycerides were significantly lower (P<0.01) in bypass fat fed groups. (GII and GIII). The mean circulating blood progesterone level and serum insulin concentration were also significantly higher (P<0.05) in the animals fed with bypass fat. The conception rate was highest in GIII followed by GII. The study indicated that it is ideal to supplement bypass fat before attempting hormonal induction of oestrus in early postpartum cows for enhanced reproductive efficiency. The techno-economic analysis of study showed that bypass fat supplementation is a financially viable, eco-friendly and socially acceptable method for improving the production and reproduction parameters of the cows, thereby augmenting the farm income.ThesisItem Open Access DETECTION OF BIOFILM FORMING BACTERIA AND THERAPEUTIC EFFICACY OF BIOFILM DISRUPTING AGENTS WITH ANTIBIOTICS IN ENDOMETRITIS OF CROSSBRED COWS(COLLEGE OF VETERINARY AND ANIMAL SCIENCES, POOKODE WAYANAD, 2020-07-24) LEEBA CHACKO; K. PromodThe present study was carried out to investigate the presence of biofilm forming bacteria in uterine samples of postpartum cows with endometritis and evaluate the efficacy of different therapeutic protocols incorporating biofilm disrupting agents along with the investigation for the presence of VF genes, Agn43aCFT073 and Agn43bCFT073, in E. coli isolates. Crossbred dairy cows (n=137) at 28-35 days postpartum, aged between three to eight years, parity between two to six without any postpartum reproductive complications were selected for the study. Screening for endometritis was carried out by the evaluation of vaginal mucus discharge using Metricheck® device and endometrial cytology by cytobrush technique. Postpartum cows with VMS score ≥ 1 or more than five per cent PMN cells in the EC smears by cytobrush technique were considered as positive for endometritis. Isolation and identification of bacteria and in vitro antibiotic susceptibility tests were performed by disc diffusion method. The antibiotic to which most of the bacteria were highly sensitive was used for the in vivo treatment for endometritis. Biofilm disrupting agents (EDTA-Tris and NAC) were also used for treatment incorporated with antibiotic or alone. Biofilm forming ability of isolates were assessed using microtitre plate assay. The E. coli isolates were screened for biofilm VF genes viz., Agn43aCFT073 and Agn43bCFT073 by PCR employing suitable primers. Sixty postpartum cows with endometritis were randomly allotted into five groups (n=12). Group I were treated with intrauterine infusion of ciprofloxacin to which most bacteria were sensitive, Group II were treated with 30 mL of 3.5 mM EDTA, 50 mM Tris, (pH 8.0) and ciprofloxacin, Group III were treated with 30 mL of 3.5 mM EDTA, 50 mM Tris (pH 8.0), Group IV were treated with 30 mL of 3.3 per cent NAC and ciprofloxacin and Group V were treated with 30 mL of 3.3 per cent NAC as intra uterine infusions. In the present study, vaginal mucus discharge was collected from 137 dairy cows and among that 20.44, 45.26, 15.33, 10.95 and 8.03 per cent cows exhibited VMS no discharge, zero, one, two and three, respectively. Cows with purulent vaginal discharge (PVD) had lowest reproductive performance at 35 days postpartum was a better criterion for the diagnosis of endometritis. Out of 137 animals, 47 cows exhibited VMS ranging from one to three at 31.58 ± 0.13 dpp were diagnosed positive for CE and the prevalence recorded was 34.31 per cent. The prevalence of subclinical endometritis detected by cytobrush technique in the present study was 10.95 per cent. The overall prevalence of endometritis by VMS and EC at 31.58 ± 0.13 dpp was 34.31 and 43.79 per cent, respectively EC detected more number of cows with endometritis than VMS. The results of the present study indicate that by measuring the VMS and EC (five per cent cut off) at 31.58 ± 0.13 dpp predicted animals with endometritis. Hence, it could be concluded that the cytobrush samples were superior and this method of sample collection is more consistent and reliable in postpartum dairy cows. The most frequently isolated bacteria in the present study were E. coli, Staphylococcus spp., Bacillus spp., Streptococcus spp., Enterobacter spp., Enterococcus spp. and Pseudomonas spp. Heirarchichal cluster anlaysis of antibiotics under present study revealed highest sensitivity for ciprofloxacin followed by enrofloxacin. In the present study, biofilm forming ability was detected in 80.52 per cent of bacterial isolates on microtitre plate assay. Maximum number of isolates (77.42%) exhibited moderate biofilm forming ability while, 8.06 and 14.52 per cent exhibited strong and weak biofilm forming ability, respectively. No significant (p>0.05) was noted in the biofilm forming ability of different bacterial isolates. None of the E. coli isolates recorded the VF genes, Agn43aCFT073 and Agn43bCFT073. Highest recovery rate (100%) and conception rate (83.33%) was noticed after treatment with NAC and ciprofloxacin intrauterine. There was no significant difference (p>0.05) was noticed between the treatment groups in the conception rate. Recovery from endometritis was more with combination biofilm disrupting agents and antibiotics than using antibiotics or biofilm breaking agents alone. Disruption of biofilm resulted in better penetration of antibiotics which aid in recovery. Collection of vaginal mucus discharge using Metricheck® device and endometrial cytology samples by cytobrush technique help detection of endometritis during early postpartum period under field conditions. It was concluded that resistance to antibiotics could be attributed to the presence of biofilm forming bacteria which form a layer and resist the action of antibiotics. Biofilm disruption aid in better penetration of antibiotics and hence, recovery. Further studies in a wider population in different dosage regimens and combinations needed to be undertaken to elucidate the efficacy of NAC for uterine infections.ThesisItem Open Access COMPARATIVE EFFICACY OF BROMOCRIPTINE, CABERGOLINE AND THYROXINE IN INDUCING OESTRUS IN BITCHES(College of Veterinary and Animal Sciences, Mannuthy., 2008-05-30) G. AJITKUMAR; Dr. T. SreekumaranDetailed survey on breeding, feeding and management of 1721 dogs belonging to 817 owners was conducted in different parts of Kerala State. Among the dogs surveyed, 518 (30.10%) were male and 1203 (69.90%) were female. Breed-wise details of dogs maintained by individual owners revealed German shepherd dog (20.74%) as the most popular breed of dog followed by Labrador retriever (18.13%), Dachshund (11.39%), Rottweiler (9.12%) and Spitz (5.52%). The percentage of local non-descript dogs was found to he 14.64 only. The prevalence, nature and magnitude of anoestrus among the female dogs were evaluated based on breeding details. Haematological, hormonal, mineral and cholesterol profile of anoestrous hitches were estimated. Data on breeding revealed that 92.93 per cent of hitches were allowed to breed and the remaining 7.07 per cent were kept as virgin throughout their lifetime. The age at first breeding in majority of the hitehes (57.85%) was between 1 to 2 years with an average of 16.30 months. The average number of oestrus periods and the number of breeding per year were 2.14 ± 0.24 and 1.47 ± 0.32 respectively. The average duration of proestrual bleeding was found to he 10.87 ± 1.24 days. Most of the hitches (51.97 %) were bred twiee in one oestrus and the period of acceptance varied from 4 to 22 days with an average of 8.94 ± 2.18 days. The incidence of failure of conception, abortion, stillbirth and pseudopregnancy were 18.87, 5.72, 15.56 and 13.69 per cent respectively. The neonatal problems encountered in 13.88 per cent of hitches included agalactia / hypogalactia / mammitis (5.29%), cannibalism (2.65%), puppy fading syndrome (2.09%), puerperal tetany (1.98%) and rejection of young one by dam / poor mothering ability (1.87%). Among the hitches surveyed, 134 (11.14%) were identified as in the stage of anoestrus, 65.67 per cent in primary and 34.33 per cent in secondary anoestrus and the average age was 22.68 and 34.34 months respectively. The average II duration of secondary anoestrus was 11.71 months. Analysis of breed-wise prevalence revealed that it was high in breeds such as Dachshund (18.18%), Labrador retriever (16.74%), Great Dane (16.67%), Dalmatian (15.38%) and Doberman pinscher (14.00%) and was lowest (2.44%) among local non-descript bitches. Data on body condition revealed that 42 per cent of the anoestrous bitches were with the ideal body score of five, 46 per cent with body score more than five (obese) and 12 per cent with body score less than five (lean). Analysis of haematological profile of anoestrous bitches revealed normochromic normocytic anaemia. The mean serum progesterone, prolactin and thyroxine levels in anoestrous bitches were of 0.57 ± 0.03 ng/ml, 0.75 ± 0.05 ng/ml and 1.80 ± 0.06 pg/dl respectively. The mean serum calcium and phosphorus levels in anoestrous bitches were 8.94 ± 0.24 and 3.48 ± 0.12 mg per cent respectively. The mean serum iron, copper, cobalt, zinc, manganese and cholesterol levels were found to be within the normal range. Oestrus induction trials were carried out in forty anoestrous bitches and the data obtained were compared with that of control bitches. Out of 10 animals treated in each group, five (50%) in Group 1 (bromocriptine @ 50 pg/kg. body weight), nine (90%) in Group 11 (cabergoline @ 5 pg/kg. body weight), eight (80%) in Group III (thyroxine @10 pg/kg. body weight) and seven (70%) in Group IV (thyroxine @ 5 pg/kg. body weight) evinced proestrual bleeding. The mean treatment onset to proestrus in Groups 1, II, III and IV was 28 ± 3.39, 13.44 ± 3.12, 24.50 ± 3.18 and 33 ± 2.21 days respectively. The average duration of proestrus in the treatment groups was 9.80 ± 0.86, 10.11 ± 0.68, 11.25 ± 0.88 and 10.71 ± 0.68 days respectively as against 9.70 ± 0.42 days in the control group. The average duration of oestrus in the four treatment groups and the control group was 7.60 ± 0.24, 8 ± 0.29, 8.5 ± 0.63, 7.85 ± 0.46 and 8 ± 0.39 days respectively. Fifty per cent of the animals treated with bromocriptine and 10 per cent of the animals treated with cabergoline exhibited nausea and vomiting as Ill side effects, whereas none of the animals treated with thyroxine exhibited side effects. The mean serum progesterone and prolactin levels during anoestrus, on the first day of proestrus and on the day of first breeding did not differ significantly within and between the treatment groups and the eontrol group. The mean serum thyroxine level during anoestrus and on the first day of proestrus did not differ significantly within and between the treatment and the eontrol groups, whereas on the day of first breeding the mean thyroxine level reeorded statistieally significant difference between the treatment and eontrol groups, the lowest being 1.56 ± 0.18 pg/dl in Group I and the highest being 2.10 ± 0.13 pg/dl in Group IV. Animals treated with levothyroxine (Group 111 & IV) exhibited a slight increase in the mean level of serum thyroxine on the first day of bleeding as well as on the day of first breeding, but the values were found to be within the normal limits. The haemoglobin level and total erythrocyte eount were found to be increased on the first day of proestrus and on the day of first breeding in animals of Group 111 and IV. The mean serum phosphorus level on the first day of proestrus and on the day of first breeding in the treatment groups were found to eontinue at low levels as in the anoestrous stage. The mean serum copper level on the first day of proestrus exhibited statistieally significant difference between the treatment and eontrol groups. The mean serum level of ealeium, iron, cobalt, zinc and manganese during anoestrus, on the first day of proestrus and on the day of first breeding did not reveal statistieally significant difference between groups. The overall eoneeption rate in the treatment groups were 40, 70, 50 and 40 per eent respectively as against 70 per cent in the control group. The average gestation length and litter size did not differ significantly between the treatment and eontrol groups.
