Isolation and adaptation of bovine herpes virus Type 1 in embryonated chicken eggs and in Madin–Darby bovine kidney cell line

Research (Published online: 27-02-2016)

20. Isolation and adaptation of bovine herpes virus Type 1 in embryonated chicken eggs and in Madin–Darby bovine kidney cell line -Devprabha Samrath, Sanjay Shakya, Nidhi Rawat, Varsha Rani Gilhare and Fateh Singh

Veterinary World, 9(2): 222-225

   doi: 10.14202/vetworld.2016.222-225

Abstract

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Aim: Objective of the present study was to isolate bovine herpes virus Type 1 (BHV-1) from semen of infected bull and to adapt it onto embryonated eggs and Madin–Darby bovine kidney (MDBK) cell line. Further, the virus was identified by agar gel immunodiffusion (AGID) test.

Materials and Methods: Semen samples were collected from five BHV-1 positive bulls previously confirmed for the presence of antibodies against BHV-1 using avidin-biotin enzyme linked immunosorbent assay test. The virus from semen samples was adapted in chorioallantoic membrane (CAM) of 11-day-old embryonated chickens eggs and in MDBK cell line. The presence of BHV-1 in infected CAM and cell culture fluid was confirmed by AGID test.

Results: Virus infected CAM showed edema, congestion and thickening at first passage level. Small foci ranged from 1 to 2 mm in diameter, scattered all over the membrane were observed at first passage. More severe changes were observed in CAM after serial passaging. The large pock lesions, round in shape with opaque raised edge and depressed gray central area of necrosis ranged from 3 to 5 mm in diameter were developed at fourth passage. Blind passages in MDBK cell culture were made. The MDBK cell line at second passage level showed characteristic cytopathic effect viz. rounding of cells with shrinkage, followed by aggregation or clumping of cells which progressed rapidly and appeared as “bunch of grapes” at 72 h post inoculation. Few cells become elongated when compared with uninfected controls. A homogenate of CAM with distinct pock lesions and infected cell culture fluid developed precipitation line within 48 h against specific anti-BHV-1 immune serum by AGID test.

Conclusion: BHV-1 was easily adapted in CAM of chicken embryos and in MDBK cell line. Virus infected CAM and cell culture fluid showed precipitin band by AGID test.

Keywords: Bovine herpes virus Type 1, chorioallantoic membrane, cytopathic effect, Madin–Darby bovine kidney cell line, pock.

Effect of extended photoperiod during winter on growth and onset of puberty in Murrah buffalo heifers

Research (Published online: 27-02-2016)

19. Effect of extended photoperiod during winter on growth and onset of puberty in Murrah buffalo heifers - Ashwani Kumar Roy, Mahendra Singh, Parveen Kumar and B. S. Bharath Kumar

Veterinary World, 9(2): 216-221

   doi: 10.14202/vetworld.2016.216-221

Abstract

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Aim: To investigate the effect of extended photoperiod on growth rate, hormonal levels, and puberty in Murrah heifers.

Materials and Methods: About 14 Murrah buffalo heifers were divided into normal day photoperiod (NDP; n=7) and extended NDP (ENDP; n=7) groups. The ENDP group was exposed to 4 h of extended photoperiod with artificial light (160 lux) after sunset for 3 months during winter.

Results: Group, age and group-by-age interaction effects on plasma glucose concentrations were non-significant (p>0.05). A significant effect of age on non-esterified fatty acids (p<0.05), cholesterol (p<0.01), and triglycerides (p<0.05) concentrations was observed. Group and group-by-age interaction effects on plasma T3, T4, leptin, 17 β estradiol, prolactin and melatonin concentrations were non-significant (p>0.05) while significant (p<0.05) age effect on T4, leptin and melatonin concentrations was observed. With respect to the circadian pattern of melatonin and prolactin, the group, time and groupby-time interaction effects were non-significant (p>0.05). Average daily gain and dry matter intake of heifers were nonsignificant between the NDP and ENDP groups but were comparatively higher in ENDP group. By the end of the experiment, 6 out of 7 heifers attained puberty in ENDP group in comparison to 4 out of 7 in NDP group.

Conclusion: Extending the photoperiod by artificial light for 4 h during winter season resulted in better growth rate and early onset of puberty in Murrah buffalo heifers.

Keywords: buffalo, leptin, melatonin, metabolites, photoperiod, prolactin, puberty.

Comparison of different diagnostic techniques for the detection of cryptosporidiosis in bovines

Research (Published online: 27-02-2016)

18. Comparison of different diagnostic techniques for the detection of cryptosporidiosis in bovines - H. K. M. Rekha, G. C. Puttalakshmamma and Placid E. D’Souza

Veterinary World, 9(2): 211-215

   doi: 10.14202/vetworld.2016.211-215

Abstract

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Aim: Aim of the present study was to compare different methods, viz., Sheather’s sugar flotation (SSF), Ziehl-Neelsen (ZN), Kinyoun’s acid-fast method (KAF), safranin-methylene blue staining (SMB), and negative staining techniques such as nigrosin staining, light green staining, and malachite green staining for the detection of Cryptosporidium spp. oocysts in bovines.

Materials and Methods: A total of 455 fecal samples from bovines were collected from private, government farms and from the clinical cases presented to Department of Medicine, Veterinary College, Bengaluru. They were subjected for SSF, ZN, KAF, SMB and negative staining methods.

Results: Out of 455 animal fecal samples screened 5.71% were found positive for Cryptosporidium spp. oocysts. The species were identified as Cryptosporidium parvum in calves and Cryptosporidium andersoni in adults based on the morphological characterization and micrometry of the oocysts.

Conclusions: Of all the techniques, fecal flotation with sheather’s was found to be more specific and sensitive method for the detection of Cryptosporidium spp. oocysts. Among the conventional staining methods, the SMB gives better differentiation between oocysts and yeast. Among the three negative staining methods, malachite green was found sensitive over the other methods.

Keywords: bovines, Cryptosporidium, Kinyoun’s acid-fast method, modified Ziehl–Neelsen, oocysts, safranin-methylene blue staining, SSF, diagnostic methods.

Evaluation of Emblica officinalis fruit powder as a growth promoter in commercial broiler chickens

Research (Published online: 27-02-2016)

17. Evaluation of Emblica officinalis fruit powder as a growth promoter in commercial broiler chickens - A. P. Patel, S. R. Bhagwat, M. M. Pawar, K. B. Prajapati, H. D. Chauhan and R. B. Makwana

Veterinary World, 9(2): 207-210

   doi: 10.14202/vetworld.2016.207-210

Abstract

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Aim: The present study was conducted to evaluate the dietary addition of Emblica officinalis (Amla) fruit powder as a growth promoter in commercial broiler chickens.

Materials and Methods: An experiment was conducted on 135 commercial broiler chicks (Ven-Cobb 400 strain) divided into three groups with three replicates of 15 chicks each. Three treatment groups were as follows – T1: Basal diet as per BIS standards; T2: Basal diet supplemented with 0.4% of E. officinalis fruit powder; and T3: Basal diet supplemented with 0.8% of E. officinalis fruit powder.

Results: The average body weights at the end of the 6th week were significantly higher (p<0.05) in groups T2 and T3 compared to group T1. Feed intake, feed conversion ratio and feed cost per kg live weight production were similar among the treatment groups. The net profit per bird was the highest in group T2 (Rs. 19.22/bird) followed by group T3 (Rs. 17.86/bird) and the lowest in group T1 (Rs. 14.61/bird).

Conclusion: Based on the results of the present study, it was concluded that dietary addition of E. officinalis (Amla) fruit powder had a positive effect on growth performance and net profit per bird in commercial broiler chickens.

Keywords: broiler chickens, Emblica officinalis, feed conversion ratio, growth performance.

Assessment of motion and kinematic characteristics of frozen-thawed Sirohi goat semen using computer-assisted semen analysis

Research (Published online: 20-02-2016)

16. Assessment of motion and kinematic characteristics of frozen-thawed Sirohi goat semen using computer-assisted semen analysis -Mukul Anand and Sarvajeet Yadav

Veterinary World, 9(2): 203-206

   doi: 10.14202/vetworld.2016.203-206

Abstract

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Aim: The aim was to determine the motion and kinematics characteristic of frozen-thawed spermatozoa in Sirohi goat using computer-assisted semen analysis.

Materials and Methods: A study was carried out in Sirohi buck. Semen collection was made biweekly from each buck with the help of artificial vagina. A total of 12 ejaculates were collected from two bucks (six ejaculates from each buck). Freshly collected semen was pooled and later evaluated. The pooled semen sample was extended with standard glycerolated egg yolk tris extender and later subjected to a process of cryopreservation. The motion and kinematic characteristics of spermatozoa were studied during freez-thawing process.

Results: Significantly (p<0.01) higher value of live percent, hypo-osmotic swelling test, and acrosomal integrity were recorded in neat semen followed by diluted and frozen thaw semen. The proportion of spermatozoa showing slow progression were the highest in the neat and diluted semen followed by rapid and non-progressively motile, while a reverse pattern was observed in the frozen thaw semen where the proportion of non-progressively motile spermatozoa were significantly (p<0.01) higher followed by slow and rapid progression.

Conclusion: This study showed that the best results for motion, vitality, plasma membrane integrity, and acrosome status were obtained in the neat semen followed by diluted and frozen thaw semen. Further, the process of cryopreservation results in a shift of motility from slow to non-progressive in the post-thaw semen with a significant decrease in the path velocities when compared to neat and diluted semen. Hence, it can be concluded that freezing-thawing process reduces the motility and kinematic characters spermatozoa and may be an important factor affecting the fertilizing ability of spermatozoa resulting in poor conception rate after insemination in goats.

Keywords: computer-assisted semen analysis, cryopreservation, motility, semen, Sirohi goat.

Effect of management system and season on semen freezability in Jakhrana bucks

Research (Published online: 20-02-2016)

15. Effect of management system and season on semen freezability in Jakhrana bucks - Narendra Kumar, B. Rai, Showkat A. Bhat, S. D. Kharche, Chetna Gangwar, S. K. Jindal and Subhash Chandra

Veterinary World, 9(2): 199-202

   doi: 10.14202/vetworld.2016.199-202

Abstract

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Aim: The objective of the study was to determine the effect of the management system (intensive and semi-intensive) and season (autumn and winter) on semen freezability in Jakhrana bucks.

Materials and Methods: A total of 24 Jakhrana bucks of same body weight and age (BW=30 kg, age=1 year) were randomly allotted into two groups, viz., Group I (intensive system, 12 bucks) and Group II (semi-intensive system, 12 bucks). These two groups were statistically tested for their homogeneity with respect to age and BW. Semen was collected twice weekly using an artificial vagina during two seasons: autumn (September-November) and winter (December-February). A total of 240 semen samples (120 from each group and season) were evaluated for post-thaw motility (PTM), viability, abnormality, functional membrane integrity (hypo-osmotic swelling [HOS]) response and acrosomal integrity.

Results: The mean values of PTM and acrosomal integrity of spermatozoa were significantly (p<0.01) higher in Group II as compared to Group I. The mean values of viability and abnormality were also differed significant (p<0.05) between groups. However, the mean values of HOS response were found non-significant (p>0.05) between groups. The season showed a significant effect on all parameters except viability and HOS response. The PTM and acrosomal integrity of spermatozoa were significantly (p<0.01) higher in winter as compared to autumn season. Abnormality of spermatozoa was significantly (p<0.05) lower in winter season.

Conclusions: This study indicates that both management system and season influence semen freezability. The semen collected from bucks reared under the semi-intensive system and winter season showed better semen freezability characteristics.

Keywords: freezability, Jakhrana, management system, season.

Effect of feeding different levels of Azolla pinnata on blood biochemicals, hematology and immunocompetence traits of Chabro chicken

Research (Published online: 20-02-2016)

14. Effect of feeding different levels of Azolla pinnata on blood biochemicals, hematology and immunocompetence traits of Chabro chicken - Deepesh Bharat Mishra, Debashis Roy, Vinod Kumar, Amitav Bhattacharyya, Muneendra Kumar, Raju Kushwaha and Shalini Vaswani

Veterinary World, 9(2): 192-198

   doi: 10.14202/vetworld.2016.192-198

Abstract

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Aim: The present study was conducted to see the effect of feeding different levels of Azolla meal on blood biochemicals, hematology and immunocompetence traits of Chabro chicken.

Materials and Methods: The study was conducted on 160 Chabro chicks, which were randomly divided into four treatment groups each with four replicates of 10 birds. The first treatment (T1) served as a control in which basal diets was offered without Azollasupplementation while in T2, T3, and T4 groups, basal diet was replaced with Azolla meal at 5%, 7.5%, and 10% levels, respectively. A feeding trial was conducted upto 8 weeks. At the last week of trial, blood samples were collected randomly from one bird of each replicate and plasma was separated to estimate certain biochemical parameters, some blood metabolites, minerals and enzymes like alanine aminotransferase and aspartate aminotransferase (AST). Hematological parameters such as hemoglobin, packed cell volume, total leukocytes count and differential leukocytes count were estimated in fresh blood just after collection. The humoral immune response was measured against sheep red blood cells,and cell-mediated immune response was measured against phyto hemagglutinin lectin from Phaseolus vulgaris (PHA-P).

Results: The study showed that hematological profile of the Chabro bird was not affected by any treatment except heterophil and lymphocyte which was found higher in T2 and T3 groups and eosinophil was found higher in a T3 group than control. Blood glucose, creatinine, cholesterol, total protein, albumin, uric acid, and triglycerides were found similar in all the groups and within the normal values for broiler chicken. Liver enzymes and macro mineral content in blood were found similar in all the treatment groups and within normal physiological range. Although AST was found higher in 10% replacement group than control, the value was within normal range for broiler chicken. Although antibody titer was found similar in all the experimental groups in the present study, cell-mediate immune response (response to PHA-P) was found higher in 5%, 7.5%, and 10% replacement groups than control(p<0.05).

Conclusion: Similar blood biochemical parameters and higher cell-mediated immune response in Azolla replacement group indicated immune-modulatory effect of Azolla meal without any toxicity.

Keywords: Azolla pinnata, blood biochemicals, hematology, immunocompetence traits.