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ThesisItem Open Access DIAGNOSIS AND THERAPEUTIC MANAGEMENT OF CANINE PYOMETRA FOR RESTORING BREEDING EFFICIENCY(COLLEGE OF VETERINARY AND ANIMAL SCIENCES, MANNUTHY, THRISSUR, 2018-05-09) UNNIKRISHNAN M. P.; M.O.KurienA study was undertaken to evaluate the efficacy of different therapeutic protocols in the treatment of canine pyometra, based on clinical, haematobiochemical, B-mode ultrasonographic and Doppler ultrasonographic evaluation. Combination treatment using mifepristone @ 2.5mg/kg b. wt. bid orally for five days, along with incremental doses of dinoprost from 10 to 50 µg/kg b. wt. tid; 48 h after initiation of mifepristone, till complete uterine evacuation was superior for therapeutic management of canine pyometra in terms of clinical recovery and future fertility. Haematology recorded anaemia, leucocytosis with neutrophilia and lymphopenia in patients with pyometra, which resolved after treatment. Serum biochemistry revealed marginal rise in blood urea nitrogen, high C-reactive protein and rise in progesterone values, which reduced to normal by day 15 of treatment. Early reduction in progesterone was noticed in cabergoline-based group. B-mode ultrasonography revealed uterine horn distension with anechoic to hypoechoic contents and thickened wall, which reduced to normal by day 15 of treatment. Delay in uterine resolution was noticed in cabergoline-based group. Doppler ultrasonography revealed increased uterine blood flow velocity (peak systolic velocity: 0.69 ± 0.05 to 0.75 ± 0.07; end diastolic velocity: 0.27 ± 0.02 to 0.31 ± 0.01) and decreased resistive index (0.59 ± 0.04 to 0.62 ± 0.06), which returned to normal after treatment. Bacteriological isolates from anterior vagina revealed E. coli (52.63 %), followed by Staphylococcus spp. (37.74 %), Streptococcus spp. (5.66 %) and Pseudomonas (3.77 %). Escherichia coli and Pseudomonas spp. were sensitive to amikacin, whereas Staphylococcus spp was most sensitive to ceftriaxone-tazobactam. Streptococcus spp. was 100 per cent sensitive to cephalosporin. Hyper-salivation was the most common side effect of prostaglandin treatment, exhibited in 31.25 per cent of dogs. Time taken for complete uterine evacuation varied between 5.88 ± 0.40 and 8.25 ± 0.73 days, with more in cabergoline-based group. Fertility studies revealed better cyclicity and conception in 75 and 66.67 per cent, respectively of mifepristone-dinoprost combination treated dogs, whereas overall cyclicity and conception was 68.75 and 54.55 per cent, respectively. Overall recurrence of 28.13 per cent was recorded after medical treatment with minimum recurrence (12.5 %) in mifepristone-dinoprost treated dogs.ThesisItem Open Access EARLY THERAPEUTIC MANAGEMENT OF PSEUDOPREGNANCY IN GOATS WITH PROSTAGLANDIN F₂ ALPHA(COLLEGE OF VETERINARY AND ANIMAL SCIENCES-MANNUTHY,THRISSUR, 2018) DEVALAL K; METILDA JOSEPHThe study was undertaken from May 2017 to April 2018 at the clinical unit under Department of Animal Reproduction, gynaecology and Obstetrics of University Veterinary Hospitals, Mannuthy and Kokkalai and Goat and Sheep farm, Mannuthy by ultrasonographical screening of does with an objective of diagnosis and therapeutic management of pseudopregnancy in goats at an early stage (Group I) and to compare the conception rate with that of normally cycling does (Group II). Group I animals (with anaechoic uterine sacculations separated by tissue trabeculae) were treated with double regime of PGF₂α analogue on day of detection of pseudopregnancy and 12 days later and response to treatment and oestrus characters were studied and were subjected to natural service on induced oestrus on consecutive days, whereas group II animals were served on the first observed oestrus after selection. Of the 1162 animals screened, the occurence of pseudopregnancy was found to be 1.98 per cent only. Using ELISA technique, serum progesterone levels were estimated and the mean± SE progesterone levels on the day of diagnosis of pseudopregnancy in group I was 6.98±0.56 ng/mL and that of pregnant animals on the same stage postservice in group II was 8.04±0.56 ng/mL and they differed non-significantly. All the animals responded well to the double regime with PGF₂α. Confirmation of pregnancy was assessed by ultrasonography on day 45 postservice. With the first natural service, the conception rate in group I was 71.42 per cent whereas in group II was 90 per cent. The present study revealed that ultrasonography can be effectively used for early diagnosis of pseudopregnancy in goats and double regime treatment with PGF₂α analogue and breeding on consecutive days on induced estrus are effective in management of pseudopregnancy by minimising the chances of recurrence and improving the further fertility rate.ThesisItem Open Access SHORTENING OF INTEROESTROUS INTERVAL IN DOGS USING CABERGOLINE AT DIFFERENT STAGES OF ANOESTRUS(COLLEGE OF VETERINARY AND ANIMAL SCIENCES-MANNUTHY,THRISSUR, 2018) SMITTY JOHNSON; C. JayakumarThe exceptionality of canine oestrous cycle is that an obligatory anoestrus follows dioestrus in dogs making the interoestrus interval (IEI) unpredictable. The efficacy and safety of prolactin antagonist, cabergoline at two dose rates and at two different stages of anoestrus in reducing the interoestrous interval (IEI) was studied. Group I (n = 14) and group II (n =6) dogs were treated with cabergoline @ 5 and 0.6 µg /kg.b.wt/orally/ day starting from 90 ± 3 days of preceding proestrous, while group III (n =11) and group IV (n = 8) dogs were treated with high and low doses of cabergoline starting from 120 ± 3 days of preceding proestrus until induced proestrus or for 40 days. Group V (n = 11) consisted of anoestrous dogs that completed 90 ± 3 days of preceding proestrus and was persued through their natural oestrus during the study period. The response to treatment was complete, except for two dogs in group I ( 85.71% ) and one dog in group IV (87.55%) .The duration of treatment was significantly (p<0.05) shorter in dogs treated with 5 µg /kg.b.wt/ day (high dose cabergoline) when treatment was initiated on day 120 ± 3 days of preceding proestrus, in comparison to other treatment groups. In response to treatment, the interoestrous interval (IEI) also reduced in all the treatment groups. On the contrary, an increasing trend in IEI (228.90 ±15.47 days) was noticed in control group. A satisfactory conception rate was noticed in all treated groups in comparison to the control group. The results signify the merits of cabergoline in terminating the obligatory anoestrus and thereby reducing the IEI, irrespective of the dose and the outcome happens moderately quicker with treatment initiated at 120 days than 90 days. Induced oestrous cycles were normal with proestrus and oestrus parameters and in which the ovulation and hormone profiles identical to physiological cycles. Fewer side effects were noticed with cabergoline decreased with adaptation. Duration of treatment was inversely proportional to depth of anoestrus. No hyperprolactinaemia was noticed in dogs during anoestrus as the serum prolactin level was less than 0.5 ng/ml and thus concluded that the action of cabergoline was not associated with the reduction of prolactin level, but a direct stimulatory effect on gonadal axis and other dopaminergic effects. Though all treatment protocols were successful in inducing a fertile oestrus and decrease the IEI, the most recommendable treatment from this study is use of low dose cabergoline at 90 ± 3 days, as the duration of treatment was shorter with a maximum shortening of IEI and a good conception rate.ThesisItem Open Access INDUCTION OF WHELPING IN CANINE HIGH RISK PREGNANCIES WITH SMALL LITTER SIZE USING PROGESTERONE RECEPTOR ANTAGONIST AND PROSTAGLANDINS(COLLEGE OF VETERINARY AND ANIMAL SCIENCES-MANNUTHY,THRISSUR, 2018) RAHEEMA S.; C. JayakumarSmall litter size is considered high risk pregnancy in dogs from a probable dystocia due to prolonged gestation, uterine inertia and foetal over size. Consequently, induction of whelping becomes inevitable for a successful outcome. The aim of the study was to determine the efficacy and safety of progesterone antagonist, mifepristone; prostaglandin analogue, cloprostenol and natural prostaglandin, dinoprost tromethamine in combination with anterior vaginal administration of PGE1, misoprostol to induce whelping in canine high risk pregnancy with three or fewer foetuses. The induction response and course of whelping was investigated in 18 dogs radiographically confirmed with three or fewer foetuses under three treatment groups and also in six spontaneous whelping dogs which formed the control group. Treatment was initiated when no impending signs of whelping was evident, but attained a gestational age of 61±l days on sonographic measurement of foetal head diameter. Dogs in Group I was treated with mifepristone @ 5mg/kg body weight twice daily, orally for two consecutive days; Group II dogs were treated with a single subcutaneous administration of cloprostenol @ 2.5 μg/kg body weight and Group III dogs were treated with subcutaneous administration of dinoprost tromethamine @ 0.1mg/kg body weight. Twenty four hours following the institution of treatment, misoprostol @ 400μg was administrated in anterior vagina of all the dogs in the three treatment groups. Induction response was 100 per cent as whelping was successfully induced in all the 18 dogs belonging to three treatment groups. Consequently, the gestation length of the bitches in the induced group (61.67±0.42 days in Group I, 61.17±0.54 days in Group II and 61.83± 0.60 days in Group III) was significantly shorter than that of the spontaneously whelping bitches (64.33±1.50 days). The mean interval from first treatment to whelping was slightly lesser in dogs administered with mifepristone (35.58±3.54 h) when compared to cloprostenol and dinoprost tromethamine treated groups (36.00±2.79 h and 36.58±1.22 h respectively). The mean duration of whelping was shorter in dogs of Group II (78.00±12.90 min) followed by Group III (102.00±21.06 min) and Group I (152.50±49.66 min). However the expulsion phase length in the induced group did not differ significantly from that of control group. A decreases in serum progesterone concentration was noticed after treatment in all the dogs of treatment groups indicating the efficiency of the drugs in initiating the parturition. However, in mifepristone treated dogs the circulating progesterone at whelping was still higher (2.71±0.41 ng/ml) when compared to group II and group III (0.60±0.14 ng/ml and 0.90±0.18 ng/ml respectively). An increasing trend of serum cortisol levels were noticed in all the dogs after the treatment. Majority of the spontaneously whelping dogs required medical and surgical assistance to complete the whelping as against unassisted vaginal delivery in majority of the dogs in which whelping was induced. Moreover, no side effects were noticed for any of the treatment provided. The number of puppies born alive in the three induced groups were 12 out of 13 (92.3%), 7 out of 10 (70%) and 9 out of 11 (81.8%) respectively, while in the spontaneously whelping dogs, only 7 out of 13 (53.8%) were born alive. However the neonatal survival rate upto two weeks of birth did not differ significantly between the four groups. The study revealed the merits of medical induction of whelping in canine high risk pregnancy with small litter size as evident from a cent per cent induction response within a relatively short and predictable period and also from the high puppy survival at birth. A high number of puppies born alive at birth and a decreased medical and surgical assistance for completing the whelping process makes induction of parturition using mifepristone with anterior vaginal administration of misoprostol a better choice than cloprostenol and dinoprost tromethamine.ThesisItem Open Access FREEZABILITY OF BOAR SEMEN SUPPLEMENTED WITH SODIUM DODECYL SULPHATE IN LOW OR HIGH SPERM CONCENTRATION DOSES(COLLEGE OF VETERINARY AND ANIMAL SCIENCES, POOKODE WAYANAD, 2018) MEGHA WILSON; Hiron M. HarshanBoar semen has conventionally been frozen with egg yolk based extenders containing Orves Es Paste (OEP). Sodium dodecyl sulphate (SDS) is the major component of OEP, which interacts with egg yolk lipids for improving their availability for spermatozoa. In case of boar semen, high concentration packing has been preferred as post-thaw damage are low with it. The objective of this study was to compare the post-thaw attributes of boar semen, when extended with extender supplemented with and without SDS and packed at high and low sperm concentration. Four ejaculates each were collected and cryopreserved from four adult healthy Large White Yorkshire boars (total 16 ejaculates). The fresh semen was evaluated for its volume, colour, pH, sperm concentration, sperm progressive motility, sperm viability, sperm abnormality, sperm acrosome integrity and hypo osmotic sperm swelling (HOS) response. The collected ejaculate was extended 1:1 extended which was then extended with lactose egg yolk extender (LEY) to either contain 1500 over 90 min and further extended with LEY with glycerol in such a way as to form four groups, namely group I (1000 × 106 sperm/mL), group II (1000 × 106 sperm/mL with 0.2% SDS), group III (200 × 106 sperm/mL) and group IV (200 × 106 sperm/mL with 0.2% SDS). The pre-freeze sperm motility values did not show any significant difference between boars or treatment. Pre-freeze sperm viability, abnormality, acrosome integrity, HOS response differed significantly between the boars (p<0.01), but did not differ between the groups. Post-thaw sperm attributes, except for sperm abnormalities, showed better values in groups supplemented with SDS. Better values were also observed for these post-thaw traits when semen was packaged at higher concentration. Incorporation of SDS or varying sperm concentration in packing did not influence post-thaw abnormal sperm counts. The results revealed that, there is an improvement of post-thaw boar sperm characteristics with supplementation of SDS into egg yolk based extender and with high sperm concentration packing.ThesisItem Open Access FERTILITY MANAGEMENT IN CROSSBRED CATTLE EXHIBITING PROLONGED OESTRUS USING PGF2α ANALOGUE AND OVSYNCH PROTOCOL(COLLEGE OF VETERINARY AND ANIMAL SCIENCES, POOKODE WAYANAD, 2018) SHAKIR ARAFATH K.; Abdul Azeez C.P.The present research work was undertaken to evaluate the occurrence of prolonged oestrus as a cause for infertility among crossbred cattle and to evaluate the efficacy of double prostaglandin injection protocol and Ovsynch protocol for the treatment of prolonged oestrus so as to formulate an effective therapeutic regimen to improve the conception rate of animals exhibiting prolonged oestrus. Animals exhibiting oestrus for more than 36 h were grouped into three with eight animals each. The first group was treated with the hormone PGF2α analogue as per double prostaglandin injection protocol. The cows in second group were treated with the Gonadotropin releasing hormone (GnRH) and PGF2α analogue as per Ovsynch protocol. The cows in third group were not subjected to any hormonal treatment and AI done with cryopreserved semen 12 h after the onset of oestrus at 24 h intervals till the end of the oestrus. Blood samples were collected on the days of prolonged oestrus, at the time of different treatment protocol and at the time of AI for serum progesterone estimation. Administration of PGF2α during luteal phase increased the per cent of animals exhibiting high intensity of oestrus among animals having prolonged oestrus and better oestrual characteristics of oestrual mucus were noticed in induced oestrus compared to spontaneous oestrus. Suprabasal progesterone levels during oestrus and suboptimal levels during luteal phase were noticed among prolonged oestrus cattle and the levels were significantly different from that of animals having normal duration of oestrus. Double prostaglandin injection and Ovsynch protocol were effective to correct the endocrine imbalance and conception rate was also better among the treatment groups compared to control group. It can be concluded that suprabasal progesterone levels during oestrus and luteal insufficiency in the luteal phase existed among animals exhibiting prolonged oestrus can be truncated by induction of oestrus by using PGF2 administration in the luteal phase followed by timely insemination improved conception rate.ThesisItem Open Access EFFECT OF GELATIN BINDING PROTEINS (GBP) AND NON GELATIN BINDING PROTEINS (NGBP) OF SEMINAL PLASMA ON FREEZABILITY OF VECHUR BULL SEMEN.(COLLEGE OF VETERINARY AND ANIMAL SCIENCES, POOKODE WAYANAD, 2018) MAHALINGESHWARA H. S.; Abdul Azeez C. PBinder of sperm protein (BSP), represents the major class of proteins in seminal plasma of ungulates. The proteins have been reported to play a major role in multiple aspects of fertilization including capacitation and sperm reservoir formation. In addition, they have been reviewed to influence the cryopreservability of bovine semen. The objective of this study is to characterise Fn2 proteins in Vechur bull semen, and to evaluate the effect of gelatin binding proteins (GBP) and non gelatin binding proteins (NGBP) of seminal plasma on freezability of Vechur bull semen. Semen collected from three Vechur bulls was centrifuged to separate seminal plasma. Gelatinagarose affinity chromatography was carried to isolate gelatin-binding proteins (GBP) and non gelatin-binding proteins (NGBP) from crude proteins. Isolated proteins were pooled separately dialysed against ammonium bicarbonate and lyophilized. SDS-PAGE of crude seminal plasma revealed presence of 15 bands. Out of which three were gelatin-binding proteins, in the molecular weight range of 14 to 16 kDa and one at 26 kDa. And the rest 12 bands were non gelatin binding proteins. For second phase of the study 18 ejaculates were collected from same three Vechur bulls, evaluated for fresh semen characters, pooled and washed with Tris buffer to remove seminal plasma and divided into four groups. Group I contain washed spermatozoa supplemented with NGBP (30 mg/ml) and GBP (30 mg/ml). Group II, Group III contain washed spermatozoa supplemented with NGBP (30 mg/ml), GBP (30 mg/ml) respectively, and Group IV contain washed spermatozoa without any seminal proteins. The semen in each aliquot incubated for 10 min. extended with TECY extender, equilibrated and frozen manually over liquid nitrogen vapour and stored in a cryocan. No significance difference was observed in sperm progressive motility and viability between the groups at pre-freeze and post thaw stage. Sperm abnormality per cent of was significantly (p<0.01) higher in group IV during pre-freeze and post thaw. Acrosome integrity, plasma membrane integrity and sperm membrane cholesterol were significantly (p<0.01) lower in group III at pre-freeze and post thaw stage. From the results it can be concluded that sperm supplimented with GBP at 30µg/ml reduced semen freezability, which can be ascribed to the deliterious effect of GBP.
