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CLINICAL, HISTOPATHOLOGICAL AND IMMUNOHISTOCHEMICAL CHANGES IN VAGINAL HYPERPLASIA IN DOGS
(COLLEGE OF VETERINARY AND ANIMAL SCIENCES MANNUTHY, THRISSUR, 2019-08-16) ANILA BABU; Bibin Becha
The study was undertaken with the objectives of the objectives of clinico gynaecological examination, haematological studies, exfoliative vaginal cytology, ultrasonographic assessment of ovarian status, histopathological studies and immunohistochemical localisation of oestrogen receptor-α in normal oestrus dogs (Group I) and in dogs affected with vaginal hyperplasia (Group II). The overall occurrence of vaginal hyperplasia was found to be 1.02 per cent. Occurrence was more in Labrador retriever breed (48.94%), in dogs of one to two years age group (43.62%), nulliparous dogs (61.70%) and was observed during proestrus (35.11%) and oestrus (60.64%) stages of ovarian cycle in intact dogs. Presence of varying degrees of vaginal mass (86.17%), followed by vaginal discharge (25.53%) were major symptoms reported. Physiological parameters like rectal temperature, heart rate, respiratory rate and colour of mucous membranes of these dogs were within reference ranges. Clinical examination of prolapsed vaginal tissue of hyperplastic dogs revealed 83.33 per cent as Type II and 16.67 per cent as Type III vaginal hyperplasia with 50 per cent dry and pale prolapsed vaginal tissue. The mean anuclear cell index in group I and II dogs were 63.17±6.11 and 63.33±6.24, respectively. Mean recorded follicular diameter (mm) of left and right ovaries were 3.90±0.40 and 4.60±0.50 among Group I dogs, whereas in Group II it was found to be 4.70±0.50 and 4.20±0.20, respectively. Mean recorded follicular number of left and right ovaries are 3.67±0.49 and 3.50±0.42 among Group I dogs, whereas in Group II it was found to be 2.68±0.55 and 3.66±0.55 and they differed non-significantly between the groups. There was no ovarian abnormalities could be observed in Group I and Group II dogs during presented day of examination. The oestradiol levels in Group I and II were 74.11±10.52 and 82.95±11.56 pg/mL, respectively and they differed non-significantly. On histopathological analysis, oestrogenic response of the vaginal epithelium was found to be more evident in Group II than Group I dogs. The mean thickness of stratum germinativum, stratum granulosum and stratum corneum of Group I dogs were measured as 31.42±2.73, 13.93±0.73 and 32.86±6.32µm, respectively. The respective values for Group II dogs were obtained as 27.11±1.45, 13.69±1.03 and 29.77±4.10µm and no significant difference was observed between groups. The proportional score of ER-α in the vaginal epithelium was found to be significantly high (p<0.05) in Group II (3.00±0.25) than Group I dogs (1.66±0.21) and that in the stroma was found to be same (2.50 ± 0.22) in both groups. It was concluded that ER-α expression was more in vaginal epithelium of vaginal hyperplasia affected dogs.
Open Access
PREDICTION OF WHELPING DATE IN DOGS BASED ON VAGINAL EXFOLIATIVE CYTOLOGY, VAGINOSCOPY AND PERI-OVULATORY SERUM PROGESTERONE ASSAY
(COLLEGE OF VETERINARY AND ANIMAL SCIENCES, POOKODE WAYANAD, 2019-08-20) NIKITA NITIN, PATIL; Chacko, Leeba
The study was designed to assess the efficacy of vaginal exfoliative cytology (VEC), vaginoscopy and peri-ovulatory serum progesterone assay in predicting the whelping date in dogs. VEC and vaginoscopy were performed in 43 animals with the history of proestrual bleeding on all days till the onset cytological dioestrus (D1). Serum progesterone (P4) level was estimated on alternate days till the value reached four ng/mL or higher and two breeding were recommended. Eighteen whelped dogs were selected randomly for the current study. On VEC, mean ± SE cornification index peak was 71.63 ± 2.55. On vaginoscopy, mean ± SE period of shrinkage with angulation and period of declining shrinkage with rounding out of mucosa was 5.22 ± 0.31 and 2.89 ± 0.26 days, respectively. Mean serum P4 values in ng/mL on the day of LH surge and ovulation were 2.53 ± 0.10 and 4.99 ± 0.14, respectively. Whelping was predicted on 57th, 61st, 65th and 63rd day from D1, day of maximum angulation (S5), day of LH surge and day of ovulation, respectively. When calculated from D1, 22.22 per cent animals delivered on 57th day. The accuracy of prediction of whelping date within ±1 and ±2 days was 83.33 and 100 per cent, respectively. When calculated from S5, 22.22 per cent animals delivered on 61st day. The accuracy of prediction of whelping date within ±1, ±2, ±3 and ±4 days was 44.44, 77.78, 94.44 and 100 per cent, respectively. When calculated from the day of LH surge, 33.34 animals delivered on 65th day. The accuracy of prediction of whelping date within ±1, ±2 and ±3 days was 66.67, 83.34 and 100 per cent, respectively. When calculated from the day of ovulation, 27.78 per cent dogs delivered on 63rd day. The accuracy of prediction of whelping date within ±1, ±2 and ±3 days was 66.67, 83.34 and 100 per cent, respectively. Significant correlation was found between serum P4 value on the day of LH surge (R2 = 0.867) and gestation length. However, no significant correlation was found between serum P4 value on the day of ovulation (R2 = 0.461) and gestation length. Two gestational equations were also derived from the observations. In conclusion, VEC was the most accurate method for prediction of whelping date. Whereas serum P4 values were found to be more accurate than vaginoscopy.
Open Access
EVALUATION OF SEMEN QUALITY ON LIQUID STORAGE OF SPECIFIC FRACTIONS OF LARGE WHITE YORKSHIRE BOAR SEMEN
(COLLEGE OF VETERINARY AND ANIMAL SCIENCES, POOKODE WAYANAD, 2019-08-09) MALATI NAIK; Harshan, Hiron M.
The present research was carried out to assess the quality of Large White Yorkshire (LWY) boar semen in specific fractions within an ejaculate and to assess the keeping quality of fractions of LWY boar semen stored in liquid state at 15C. Two fractions of the semen were taken for the study, initial 10 mL of sperm rich fraction (SRF) formed F1/Group I, rest of the sperm rich fraction formed F2/group II and mixture of half of F1 and F2 each formed group III. A total of 30 ejaculates were used for the study as fractions from four adult healthy LWY boars (16 in Phase I and 14 in Phase II). In phase I, differences in fresh semen characteristics of F1 and F2 were evaluated with respect to pH, concentration, seminal plasma protein content and SDS PAGE profile were analysed. Significant differences were observed between fractions with respect to pH and sperm concentration. The sperm progressive motility and total protein content of semen did not differ significantly between F1 and F2 fraction. An average number of 13.5 ± 0.51 bands were observed in F1, 12.92 ± 0.44 bands in F2 and 12.78 ± 0.23 bands in the sperm free fraction. The proteins observed varied in the molecular weight from 18 - 244 kDa, 8 – 242 kDa and 9 – 248 kDa in F1, F2 and sperm free fraction, respectively. In Phase II of study, semen fractions were analyzed for their keeping quality at 15C after being extended in BTS as group I, group II and group III. Semen was extended with BTS to a final concentration of 50 × 106 sperms/mL and gradually cooled to 22°C, held at same temperature for three hours. The semen was subsequently cooled to 15°C and stored for 72h and evaluated at 4, 24, 48 and 72 h of storage. The pH of semen was significantly (p < 0.01) higher in group II at all time point intervals. The progressive motility of semen was significantly (p < 0.05) higher in group I at 24h, 48 h and 72 h of storage. The sperm viability was significantly higher (p < 0.05) in group I at 72 h of storage alone. The sperm abnormality, acrosome integrity and rapid HOS response was significantly higher in group I at 24 h, 48 h and 72 h of storage alone. Sperm plasma membrane integrity showed a trend of being higher in group I at 72 h of liquid storage (p < 0.1). The cholesterol values at 0h and 72h was significantly higher (p < 0.01) in group I than other groups. The study revealed that initial 10 mL of SRF had showed better preservability at 15C than rest of SRF.
Open Access
STORAGE QUALITY OF LIQUID SEMEN OF LARGE WHITE YORKSHIRE BOAR SUPPLEMENTED WITH CHOLESTEROL LOADED CYCLODEXTRIN
(COLLEGE OF VETERINARY AND ANIMAL SCIENCES, POOKODE WAYANAD, 2019-09-04) T., SHYLESH; Harshan, Hiron M.
The present study was carried out to assess the effect of supplementation of cholesterol loaded cyclodextrin on quality of liquid semen of Large White Yorkshire (LWY) boar stored at 15C. A total of 12 ejaculates were used for the study from four adult healthy LWY boars maintained at Centre for pig production and research, Mannuthy. The fresh semen characteristics were evaluated immediately after collection and semen was extended with Beltsville thaw solution to a final concentration of 120 × 106 sperms/mL. The extended semen was split into five groups and processed as Group I (without any supplementation); Group II (supplemented with cholesterol loaded cyclodextrin (CLC) at the rate of 0.75 mg/120 million sperms); Group III (supplemented with CLC at the rate of 1.5 mg/120 million sperms); Group IV (supplemented with CLC at the rate of 3 mg/120 million sperms) and Group V (supplemented with methyl- cyclodextrin at the rate of 1.5 mg/120 million sperms). The processed semen was gradually cooled to 22°C and held at same temperature for three hours. The semen was subsequently cooled to 15°C and stored for 72 h and evaluated at 4, 24, 48 and 72 h of storage for semen attributes. The progressive motility of semen was significantly (p < 0.01) higher in group III and IV at 4h of storage and in group III at 24h, 48 h and 72 h of storage. The sperm viability was significantly higher (p < 0.01) in group III at 4, 24, 48 and 72 h of storage. The sperm abnormality was significantly (p < 0.01) higher in MBCD treated group at 48 and 72 h of storage. The HOS response was significantly (p < 0.01) higher in group II and III at all stages of preservation. The acrosome integrity was significantly (p < 0.01) higher in group III at 24 and 48 h, whereas both group II and III had significantly (p < 0.01) higher values at 72 h of preservation. The cholesterol content was significantly (p < 0.01) higher in group IV at 72h. The study revealed that cholesterol treatment of the spermatozoa with means of CLC at a rate of 1.5 mg/120 million sperms improved the quality of stored spermatozoa at various stages of storage at 15C till 72 h. It was also found that the supplementation of cholesterol as CLC could increase the amount of sperm cholesterol. Thus the positive effects of CLC on sperm characteristics during storage at 15C could be due to the supplementation of cholesterol to sperm membrane.
Open Access
POSTPARTUM RESUMPTION OF OVARIAN ACTIVITY AND ITS ASSOCIATION WITH SUBCLINICAL KETOSIS IN CROSSBRED COWS
(COLLEGE OF VETERINARY AND ANIMAL SCIENCES, POOKODE WAYANAD, 2019-08-20) MEGHASHAM, KATHANE SONAM; K., Promod
The aim of the study was Postpartum resumption of ovarian activity and its association with subclinical ketosis in crossbred cows. The work was conducted at Instructional Livestock Farm Complex (ILFC), Pookode and Livestock Research Station (LRS), Thiruvazhamkunnu during the period of September 2018 to May 2019. The age, parity, milk yield, peripartum and postpartum complications of the selected animals was recorded. Crossbred cows on day 30 postpartum, with the history of peripartum and postpartum complications were screened by rectal examination at 12 days interval (30, 42 54 66, 78 and 90) for the assessment of uterine involution and ovarian functional status of the animals. Crossbred dairy cows at 30 days postpartum were screened for clinical ketosis by Rothera’s test of urine samples. On day 45 postpartum, the animals without any clinical signs of ketosis was screened again for subclinical ketosis (SCK) using cow side BHB testing reagent strips. Blood samples were collected from animals of both the groups at 12 days interval (30-90 days postpartum). The following biochemical constituents in serum were analysed in both the groups at 12 days interval as per standard procedures using diagnostic kits. Total serum cholesterol, Total lipids, Blood urea nitrogen (BUN), Aspartate aminotransferase (AST), Serum glucose, BHB. Serum progesterone concentrations analysis was done in both the groups to assess the resumption of ovarian cyclicity. Out of 30 animals 40 per cent of animals showed slightly positive, 33.33 per cent of animals showed moderately positive and 8per cent of animals showed highly positive for Rothera’s test. Control animals observed no colour change due to absence of ketone bodies. Out of 30 animals 105 200µmol/L, 300µmol/L, 350µmol/L, 400µmol/L and 500µmol/L of BHB level were present in 13.33, 26.66, 3.33, 53.33 and 6.66 per cent of animals, respectively. In SCK and non SCK cows mean ± SE of glucose, total cholesterol, total lipid, AST, BUN and BHB were significantly (P < 0.01) varied at 12 days interval (30-90 days postpartum). Mean ± SE of glucose, total cholesterol, total lipid, AST, BUN and BHB in early, late and not resumed animals were non-significance at 12 days interval (30-90 days postpartum). Result indicated that ROA, measured by the appearance of the first significant progesterone rise ( > 1 ng/ ml in blood ) in early postpartum period was not associated with SCK .More than 1.2 to 1.4 mmol/L level of the BHB which indicates presence of SCK. BCS was significantly associated with SCK and ROA as odds ratios were higher. Biochemical parameters were associated with the development of SCK, but no association was observed with postpartum ROA.
Open Access
EVALUATION OF FERTILITY IN POSTPARTUM DAIRY COWS BY INDUCING OVULATION USING HCG IN PROGESTERONE-BASED OESTRUS SYNCHRONISATION
(COLLEGE OF VETERINARY AND ANIMAL SCIENCES, POOKODE WAYANAD, 2019-08-20) M.P., NAYANA; C.P., Abdul Azeez
The study was conducted to evaluate and compare the efficacy of 3000 IU of hCG for induction of ovulation in five day and seven day CIDR-based oestrus synchronisation protocols in postpartum crossbred dairy cows. Twenty-four apparently healthy, cyclic cows, which had completely involuted uterus were selected and randomly allotted into two treatment and one control groups (n=8 each). On day 40 postpartum, animals of Group I (5 day CIDR group) were inserted with CIDR followed by administration of GnRH on the same day. The CIDR insert was removed on day 5 and PGF2α was administered. After 36 h of CIDR removal, 3000 IU of hCG was given followed by TAI at 50 - 56 h after CIDR removal. In Group II (7 day CIDR group), similar protocol was followed as in group I, with the difference that CIDR was kept for 7 days. Animals in Group III (Control) were observed for occurrence of natural postpartum oestrus from day 40 to 90 postpartum and inseminated at detected oestrus. CIDR based oestrus synchronisation resulted in 100 per cent oestrus response in postpartum animals whereas only 37.5 per cent oestrus response was observed in control animals. Higher per cent of animals exhibited high intensity oestrus in both 5 day (75 per cent) and 7 day (62.5 per cent) CIDR protocols with 3000 IU hCG whereas oestrus signs of high intensity could not be observed in the natural postpartum oestrus. There was no significant difference in the mean time taken (h) for the onset of oestrus after removal of CIDR between two synchronisation protocols but the mean duration of oestrus (h) observed in treatment animals were significantly (p<0.01) higher than that observed in control animals. Pregnancy diagnosis was carried out by transrectal ultrasonography on day 25 post AI and later confirmed by rectal examination on day 60. The first service and overall conception rate after three consecutive AI were higher in 5 day CIDR group than in 7 day CIDR and control group. Days open was also higher in animals treated with 5 day CIDR treatment. There was no significant difference in serum progesterone concentration between the groups on the day of CIDR insertion or on day 40 postpartum, at the time of hCG injection, at the time of AI and at day 10 post AI. However, the serum progesterone concentration on the day of CIDR removal was significantly higher (p ≤ 0.05) in animals of 5 day than that of 7 day CIDR groups. Higher conception rate was found in the animals having serum progesterone concentration above 1 ng/mL at the time of CIDR insertion or on day 40 postpartum, below 0.5 ng/mL at the time of AI and above 4 ng/mL on day 10 post AI in all the groups. The present study can be concluded that interms of first service and highest overall conception rate, the five day CIDR-based protocol was superior to seven day CIDR-based protocol when 3000 IU of hCG was used to induce ovulation in oestrus synchronisation protocols. But both protocols gave better oestrus response with more high intensity of oestrus resulting in improved conception rates compared to control group
Open Access
PROGNOSTIC EVALUATION OF CANINE PYOMETRA BASED ON SERUM PROSTAGLANDIN METABOLITE CONCENTRATION AND HAEMATOLOGICAL CHANGES
(College of Veterinary and animal Science,Mannuthy, 2019) VIDYA V.K.; M.P. Unnikrishnan
A study was undertaken for prognostic evaluation of canine pyometra, based on serum prostaglandin metabolite (PGFM) concentration and haemato-biochemical changes. Female dogs presented to University Veterinary Hospitals, Kokkalai and Mannuthy, with history and clinical signs suggestive of pyometra were utilised for the study. Sixteen dogs were selected after detailed clinico-gynaecological and sonographical examination for confirmation of pyometra and treated with mifepristone @ 3.5 mg/kg body weight bid for five days and cabergoline @ 5g/kg body weight once daily orally for five days, along with supportive fluid and antibiotic therapy. Detailed clinical, haemato-biochemical, hormonal and sonographic studies were conducted on day 0, 3, 7 and 14 of treatment. Six apparently healthy dogs in diestrus were selected as control group and same combination of mifepristone and cabergoline was administered to all dogs. Clinical, haemato-biochemical and hormonal studies were conducted on day 0 and 14. Among the dogs selected for study, proportion of nulliparous dogs and those belonging to the age group of six years and above were 56 per cent each; all dogs were in the luteal phase of oestrous cycle. The symptoms and clinical signs exhibited by the pyometra affected dogs were suggestive of the systemic involvement of the disease, more prominent being anorexia, dullness, vaginal discharge, vomiting, polyuria and dehydration. Of the total pyometra cases, 25 per cent were closed-cervix type. Physiological parameters were not altered due to pyometra. B-mode ultrasonography revealed enlarged uterus, filled with fluids of varying consistency, ranging from anechoic to hyper-echoic; significant reduction in uterine horn diameter (36.60 ± 3.20 to 12.20 ± 1.40 mm) and uterine wall thickness (6.00 ± 0.44 to 3.98 ± 0.23 mm) during the course of treatment was recorded among pyometra affected dogs. Uterine contents were present in all the cases on the day of admission; complete uterine content evacuation was achieved by day 14 in 87.50 per cent of treated dogs. Majority of bacteria (76.00 per cent) isolated from the anterior vaginal swab of pyometra affected dogs was Gram negative bacteria, sensitive to gentamicin and resistant to cefotaxime. Haematology recorded anaemia (TEC, Hb, VPRC values were 4.09 ± 0.28×106 /mm3 , 9.89 ± 0.66 g/dL and 25.43 ± 1.66 %), leucocytosis (36.46 ± 4.51×103 /mm3 ) with neutrophilia (76.64 ± 1.95 %) and increased band cell per cent (5.38 ± 0.75 %) in dogs with pyometra. The BUN and creatinine values as well as platelet counts were found to be within normal range. Serum PGFM concentration in pyometra affected dogs was not significantly different from control animals, either before (35.52 ± 6.79 vs 19.41 ± 3.62 pg/mL) or after treatment (22.62 ± 3.52 vs 21.89 ± 3.83 pg/mL). Serum progesterone concentration was indicative of dioestrus in all dogs, irrespective of groups and didn’t vary significantly between pyometric and normal, dioestrous dogs (25.13 ± 3.47 vs 22.27 ± 3.59 ng/mL). After treatment, the values significantly reduced in both the groups (7.95±1.04 vs 8.21±1.20 ng/mL). Mean duration required for initiation and completion of uterine evacuation, clinical recovery and duration of treatment were 2.60 ± 0.22, 12.30 ± 0.95, 12.60 ± 0.96 and 19.60 ± 1.25 days, respectively. Cystic structures in the ovary as well as permanent damage in the endometrium were noticed after OHE, among the dogs which were refractory to medical treatment (12.5%). Oophoritis, caused by leakage of uterine contents into ovarian bursa was also recorded in the study. Recurrence was recorded among 25 per cent of medically treated cases, during the period of surveillance of six months. Changes in TLC, band cell per cent, lymphocyte per cent, thrombocyte count, TEC, Hb and VPRC could be useful diagnostic or prognostic markers in canine pyometra. Feasibility of serum PGFM measurement as a tool for diagnostic and prognostic evaluation of canine pyometra needs further studies involving more number of dogs.
Open Access
ENDOMETRIAL ECHOTEXTURE, CYTOLOGY AND BIOPSY IN POSTPARTUM SUBCLINICAL ENDOMETRITIS OF CROSSBRED DAIRY COWS
(College of Veterinary and animal Science,Mannuthy, 2019) GAYATHRI PRATHAP; Shibu Simon
The study was undertaken at University Livestock Farm and Fodder Research and Development Scheme, Mannuthy with the objective of comparing the efficacy of endometrial cytology, biopsy and ultrasonography in crossbred postpartum dairy cows for the diagnosis of subclinical endometritis (SCE). Twenty-four apparently normal cows without any periparturient complications were randomly selected so that at least 12 animals with SCE could be obtained. Transrectal ultrasonography (TRUS) was performed on days 20, 30 and 40 to record the ovarian status and echobiometry of uterus and cervix. Endometrial cytology (EC) and uterine biopsy (UB) samples were collected on days 30 and 40. The mean (±SE) percentage of polymorphonuclear (PMN) cells by EC were 1.81 ± 0.88 and 1.00 ± 0.64 in normal group and 18.00 ± 0.81 and 9.92 ± 0.59 in SCE group, respectively, on 30 and 40 days postpartum (DPP). The mean (±SE) percentage of total histopathological score were 11.09 ± 0.59 and 9.64 ± 0.51 in normal group and 17.69 ± 0.55 and 14 ± 0.47 in SCE group, respectively, on 30 and 40 DPP. Significant difference was noticed between EC and UB in normal and SCE groups both on 30 and 40 DPP. The SCE positive animals also had a fast clearance rate which was evidenced by the reduction in the inflammatory status between the days of 30 and 40 DPP in both EC and UB. Among the TRUS parameters cervical diameter was the least valuable one, though this could be due to smaller girth for cervix in crossbred cows used for study. It could be inferred that ultrasound findings alone has poor efficacy in diagnosing SCE but it could be used as a supporting tool along with cytological or histopathological studies. Both cytology and biopsy could be used as valuable diagnostic techniques in detecting SCE. Biopsy allowed more detailed information about the deeper layers of uterus and if samples are collected with care the future fertility is not affected. All the animals were inseminated during next observed heat. Animals found to be positive for SCE was treated with intrauterine infusion of 500 mg Cephapirin Benzathine 12 hours after insemination was done. There was no significant difference in the conception rates in SCE and normal groups in the present study.
Open Access
INFLUENCE OF DEXAMETHASONE AND EPIDERMAL GROWTH FACTOR SUPPLEMENTATION IN SYNTHETIC OVIDUCTAL FLUID MEDIUM ON IN VITRO BOVINE EMBRYO DEVELOPMENT
(COLLEGE OF VETERINARY AND ANIMAL SCIENCES-MANNUTHY,THRISSUR, 2019) RAKSHITHA P; Abhilash R.S.
The study was designed to assess the individual and combined effect of dexamethasone and epidermal growth factor (EGF) on in vitro bovine embryo development. Ovaries of abattoir origin from crossbred cattle of Kerala were subjected to study. All visible follicles of size 2-8 mm were aspirated and culture grade oocytes were subjected to IVM in TCM-199 medium and fertilized in fertTALP medium and embryos were cultured in mSOF. Presumptive zygotes in group I (n= 162) were treated with 0.1µg/ml of dexamethasone; Group II (n= 198) with 5ng/ml of EGF; Group III (n=199) with 0.1µg/ml of dexamethasone and 5 ng/ml of EGF and Group IV (n= 154) kept as control. Embryo developmental competence was assessed every 48 h and morphological evaluation and embryo viability (Hoechst 33342 staining) was performed on day 8. On day 2 significantly higher (p ≤ 0.05) fertilization and cleavage rate was observed in group II (53.04 ± 2.87 and 35.69 ± 1.02 %) and group III (50.34 ± 2.18 and 34.47 ± 1.98 %) compared to group I (36.33 ± 4.15 and 20.05 ± 3.46%) and group IV (33.00 ± 4.38 and 16.29 ± 4.64 %). On day 4 significantly higher 4 and 16 cell stage embryo was obtained in group II and group III than group I and IV. A significantly higher (p≤0.05) 8 cell stage embryo was observed in group III when compared to group I and IV. However there was no significant difference in 8 cell stage embryo was observed between group II and III. On day 6 and 8 morula/ blastocyst yield was significantly higher (p ≤ 0.05) in group II and III (9.06 ± 0.89 and 8.21 ± 2.21) than in Group I and IV (3.63 ± 1.29 and 2.66 ± 1.54). The proportion of good and fair quality embryos were recorded on day 6, group I (75 and 12.5%), II (52.6 and 31.6%), III (77.8 and 5.6%) and IV (28.6 and 57.1%). Higher per cent of morphologically good and fair quality embryos on day 8 were observed in group I (50 and 25%) and III (44.4 and 27.8%) than group II (10.5 and 15.7%) and IV (14.3 and 14.3%). Embryo viability assessed by Hoechst 33342 on day 8 revealed a higher per cent of viable embryos in group I and III (83.33 and 83.33%) when compared to group II and IV (50 and 16.67%). The present study revealed that supplementation of dexamethasone in mSOF improved the quality of embryos. The supplementation of EGF in mSOF improved the yield of embryos and combining dexamethasone and EGF in mSOF resulted in increased number of embryos with better quality.

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