Assessment of genetic variability among Indian sheep breeds using mitochondrial DNA cytochrome-b region

Assessment of genetic variability among Indian sheep breeds using mitochondrial DNA cytochrome-b region - A. D. Sawaimul, M. G. Sahare, S. Z. Ali, A. R. Sirothia and Satish Kumar

Veterinary World, 7(10): 852-855

   doi: 10.14202/vetworld.2014.852-855

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Abstract

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Aim: The present study was conducted to estimate genetic distance, the phylogenetic relationship, and time of divergences using mitochondrial DNA (mtDNA).

Materials and Methods: The total 216 unrelated samples were collected from native breeding tract of six Indian sheep breeds. The genomic DNA was isolated and screened for restriction enzyme polymorphisms for cytochrome b (Cyt-b) region of mtDNA with seven restriction enzymes.

Results: The genetic distance among sheep breeds was ranging between 0.02833 and 0.0946. The phylogenetic analysis revealed that Malpura and Chokla were found closer relationship forming distinct cluster followed by Deccani individual were clustered with Nellore sheep, whereas Nali and Sonadi were distant to each other having separate cluster. Estimated divergence time among Indian sheep breeds were ranging about 1.41-4.73 million years ago (MYA) with an average of 3.063±0.27 MYA. It showed that Malpura and Sonadi sheep revealed highest divergence time as 4.73 MYA whereas Malpura and Chokla show the lowest as 1.41 MYA.

Conclusion: In conclusion, the restriction fragment length polymorphisms-polymerase chain reaction (RFLP-PCR) of the Cyt-b region of mtDNA is suitable and cost effective tool for estimating the genetic variability, phylogenetic relationship, and time of divergence among Indian sheep breeds. These findings will help to formulate proper breeding strategies for conservation and utilization of sheep breeds.

Keywords: genetics-diversity, mitochondrial DNA, restriction fragment length polymorphisms-polymerase, chain reaction, sheep breed.

Polymerase chain reaction amplification and cloning of immunogenic protein NAD-dependent beta hydroxybutyryl CoA dehydrogenase gene of Clostridium chauvoei

Polymerase chain reaction amplification and cloning of immunogenic protein NAD-dependent beta hydroxybutyryl CoA dehydrogenase gene of Clostridium chauvoei - Saroj K. Dangi, Ajay P. Singh, Satyaveer S. Dangi, Prasad Thomas, Santosh K. Gupta, Rajesh K. Agarwal and K. N. Viswas

Veterinary World, 7(10): 848-851

   doi: 10.14202/vetworld.2014.848-851

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Abstract

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Aim: The present study was aimed at polymerase chain reaction (PCR) amplification and cloning of NAD-dependent betahydroxybutyryl coenzyme A dehydrogenase (BHBD) gene of Clostridium chauvoei.

Materials and Methods: C. chauvoei was cultured and confirmed by 16-23S rDNA spacer region primers. The primers for nad-bhbd gene ofC. chauvoei were designed to aid in cloning into pRham-N-His SUMO-Kan vector, and nad-bhbd gene was amplified by PCR. The amplifiednad-bhbd gene was purified and cloned into pRham-N-His SUMO-Kan expression vector. The recombinant plasmid was transformed into E. cloni 10 G cells and the clone was confirmed by colony PCR using the pRham-SUMO-NAD-For and pRham-SUMO-NAD-Rev primers and also by sequencing.

Results: PCR amplification of nad-bhbd gene yielded a product length of 844 base pairs which was cloned into pRham-NHis SUMO-Kan vector followed by transformation into E. cloni 10G chemically competent cells. The recombinant clones were characterized by colony PCR, sequencing, followed by basic local alignment search tool (BLAST) analysis to confirm the insert.

Conclusions: Immunogenic protein NAD- dependent BHBD of C. chauvoei was cloned and the recombinant clones were confirmed by colony PCR and sequencing analysis.

Keywords: black quarter, Clostridium chauvoei, NAD-beta-hydroxybutyryl coenzyme A dehydrogenase.

Age and lactation specific disposal pattern in Sahiwal cattle and influence of various genetic and non-genetic factors

Age and lactation specific disposal pattern in Sahiwal cattle and influence of various genetic and non-genetic factors - A. Upadhyay, D. K. Sadana, A. K. Gupta, A. K. Chakravarty, S. Dash, M. K. Das, Anushree M. and P. R. Shivahre

Veterinary World, 7(10): 842-847

   doi: 10.14202/vetworld.2014.842-847

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Abstract

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Premature disposal of female calves before reaching milch herd and undesirable disposal of lactating cows are the major constraints in achieving larger herd size. During the early lactations, younger cows are supposed to give higher milk yield and undesirable disposal of early calvers, thereby, greatly hampers profitability of a dairy farm. Knowledge of the incidence of disposal along with reasons in various age groups and at various parities is essential to identify which age group or parity is more vulnerable for disposal. Moreover, knowledge of various genetic and non-genetic factors associated with disposal of animals may also be helpful in developing breeding and management strategies to reduce the incidence of disposal. In most of the studies, it was found that major reasons of disposal of dairy cattle were mortality among female calves and involuntary culling among adult lactating cows. Maximum mortality in female calves was observed during earlier ages and pneumonia, gastro-enteritis and debility were major reasons of female calf mortality. Whereas, most of the adult cows left the herd, due to teat and udder and reproductive problems. Moreover, indigenous breeds were found to be more adapted to Indian tropical climatic conditions in comparison to crossbred and exotic cattle breeds.

Keywords: culling, disposal pattern, heritability estimates, mortality, Sahiwal cattle.

Electrocardiographic and hemato-biochemical effects of two balanced anesthetic protocols in dogs

Electrocardiographic and hemato-biochemical effects of two balanced anesthetic protocols in dogs - Anubhav Khurana, Adarsh Kumar, Sandeep Kumar Sharma and Amit Kumar

Veterinary World, 7(10): 835-841

   doi: 10.14202/vetworld.2014.835-841

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Abstract

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Aim: The purpose of this study was to compare the electrocardiographic (ECG), hematological and clinico-biochemical effects of two balanced anesthetic protocols in dogs.

Materials and Methods: A total of 20 clinical cases of dogs, randomly divided into two groups of 10 animals each were made part of study. All dogs were premedicated with injection atropine sulfate @ 0.04 mg/kg body weight (b. wt.) subcutaneously followed 15 min later with injection butorphanol tartarate @ 0.2 mg/kg b. wt. intravenous (IV). Subsequently after 10 min premedicated with injection diazepam @ 0.5 mg/kg b. wt. IV (Group DP) or injection acepromazine maleate @ 0.015 mg/kg b. wt. IV (Group AP) followed by injection propofol “till effect” IV for induction of surgical anesthesia. The animals were immediately transferred to halothane in oxygen. Observations recorded in dogs included ECG recordings, hematological and clinico-biochemical observations at various time intervals.

Results: No arrhythmia was observed in any animal pre-operatively and intra-operatively in any of the groups. Significant fall in packed cell volume (PCV) and total erythrocyte count occurred in DP group in early phase, whereas only PCV decreased significantly in AP group. Biochemical parameters were non-significant in both the groups.

Conclusion: Both diazepam-butorphanol-propofol-halothane and acepromazine-butorphanol-propofol-halothane are safe with respect to their ECG, hematological and biochemical effects in clinical cases.

Keywords: acepromazine, butorphanol, diazepam, dog, propofol.

Comparison of standard lactation curve models using fortnightly milk records in Frieswal cattle

Comparison of standard lactation curve models using fortnightly milk records in Frieswal cattle - Amit Kumar Dohare, B. Singh, Med Ram Verma, Bangkeng Perme, Vijay Bahadur Sharma, Neha Gupta and Shashank Kshandakar

Veterinary World, 7(10): 831-834

   doi: 10.14202/vetworld.2014.831-834

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Abstract

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Aim: The aim was to compare standard lactation curve models using fortnightly milk records in Frieswal cattle.

Materials and Methods: A total of 2904 fortnightly milk yield (FMY) records from 132 Frieswal cattle maintained at Military Farm, Bareilly, Uttar Pradesh were taken for study. The Wood (WD), Morant and Gnanasakthy (MG), Mitscherlich x Exponential (ME), and Wilmink (WK) models were fitted on average FMY (AFMY) by nonlinear regression using statistical package SAS 9.3 version. The goodness of fit of models was judged by the adjusted coefficient of determination (Adj. R2) and root mean square error.

Results: The AFMY ranges from 127.09 kg (first fortnight) to 110.04 kg (last fortnight) with peak fortnight yield of 189.51 kg and peak period at fourth fortnight. Predicted peak yield by different models ranges from 182.7 to 190.2 kg. The herd average milk yield was predicted with a high degree of accuracy (Adj. R2>92%) by all models with the maximum accuracy (Adj. R2=99.20%) obtained by ME model followed by MG (Adj. R2=98.8%) and WK model (Adj. R2=96.0%).

Conclusion: The ME model provided best fit for FMY data in Frieswal cattle followed by WK and MG model, whereas WD model fitted least.

Keywords: Frieswal cattle, fortnightly milk yield, lactation curve model, peak yield.

Thermoregulatory and adaptive responses of adult buffaloes (Bubalus bubalis) during hyperthermia: Physiological, behavioral, and metabolic approach

Thermoregulatory and adaptive responses of adult buffaloes (Bubalus bubalis) during hyperthermia: Physiological, behavioral, and metabolic approach - Alok K. Wankar, Gyanendra Singh and Brijesh Yadav

Veterinary World, 7(10): 825-830

   doi: 10.14202/vetworld.2014.825-830

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Abstract

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Aim: The study was planned to evaluate the indigenous animal adaptive capabilities during optimum temperature versus heat stress (HS).

Materials and Methods: Four adult buffaloes were exposed at 25°C, 30°C, 35°C, and 40°C for 21 days at every treatment in environmentally controlled chamber and physio-biochemical variation and animal behavior was observed.

Results: The study revealed significantly increased rectal temperature, respiration rate, water intake, sodium, reactive oxygen metabolites, cortisol, aspartate aminotransferase, and alanine aminotransferase while, pulse rate and thyroid hormones decreased during thermal stress. Panting, restlessness, salivation, and sweating were higher during HS while, rumination and urination contrastingly lowered.

Conclusion: The results reflect the impact of hyperthermia both acute and chronic, on the animals forcing various physiobiochemical, endocrinal, and behavioral changes for acclimatization during a stressful period aimed at maintaining homeothermy.

Keywords: acclimatization, behavior, endocrinal, heat stress, panting, physio-biochemical.

Prevalence of Rotavirus in shellfish from Southern Kerala

Prevalence of Rotavirus in shellfish from Southern Kerala - Vysakh Mohan, Shriya Rawat, K. M. Lokesh, H. V. Mohan, D. Avinash Reddy, Ashok Kumar and K. N. Bhilegaonkar

Veterinary World, 7(10): 821-824

   doi: 10.14202/vetworld.2014.821-824

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Abstract

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Aim: To study the prevalence of Rotavirus in shellfish from Southern Kerala.

Materials and Methods: The shellfish samples after processing was concentrated using proteinase K. RNA was isolated from the concentrated samples using phenol chloroform method. Rota viral RNA was detected using reverse transcriptionpolymerase chain reaction.

Results: A low prevalence of 2.5% (5/200) was obtained during the study. Rotavirus was detected in 2 out of 60 brown mussels (3.33%), 2 out of 70 yellow clams (2.86%) and 1 out of 70 black clams (1.43%).

Conclusion: Low prevalence of Rotavirus was obtained in our study. A more extensive study need to be conducted to estimate the prevalence of enteric virus in shellfish.

Keywords: concentration, proteinase K, reverse transcription-polymerase chain reaction, Rotavirus, shellfish.

Effect of uterine immunomodulation on hematobiochemical parameters in cyclic non-breeding cows

Effect of uterine immunomodulation on hematobiochemical parameters in cyclic non-breeding cows - Saraswat Sahoo, Debendra Narayan Mohanty, Srinibas Das and Arpita Padhy

Veterinary World, 7(10): 816-820

   doi: 10.14202/vetworld.2014.816-820

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Abstract

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Aim: To study the effect of uterine immunomodulation on hematobiochemical parameters and total immunoglobulin concentration in cyclic non-breeding cows.

Materials and Methods: Twenty-one repeat breeding cows around Bhubaneswar area were screened by white side test to detect and treat the endometritis and were assigned to three treatment protocols with an equal number of seven animals in each group. Cows in control group were administered with 50 ml of normal saline while treatment Group I animals were given single intrauterine infusion of 20 ml of fresh colostrum and treatment Group II animals received nonpathogenic Escherichia coli in 10 ml sterile saline. The blood samples were collected from all the experimental animals, and hematobiochemical parameters and total immunoglobulin concentration were estimated.

Results: A high significant difference (p<0.01) was accounted in lymphocyte count of E. coli treated group within different days of sampling. Analysis of variance recorded a highly significant difference with neutrophil percent in E. coli lavaged cows. In colostrum treated group monocyte count showed a significant difference (p<0.01) between 0 and 14th day of sampling. The analysis of hematocrit values did not show any significant difference apart from the erythrocyte sedimentation rate parameter in the colostrum infused group with the highest significant (p<0.01) variation being observed between 7th and 14th day of sampling. The analysis of aspartate amino transferase values in the colostrum lavaged cows revealed a significant difference, but that of alanine amino transferase values did not show any significant difference. Comparison of immunoglobulin values for different days in all the treatment protocol revealed a highly significant (p<0.05) difference within various days of sampling.

Conclusion: In the present study, the local immunomodulation by different agents have been highlighted and which indicated potentiation of uterine immunity by different drug that might serve as a new direction of treatment to uterine diseases. The scope of research in the future should be widened by considering a larger population for validation.

Keywords: colostrum, cyclic non-breeders, hematocrit values, non-pathogenic Escherichia coli.

Detection of African swine fever virus from formalin fixed and non-fixed tissues by polymerase chain reaction

11. Detection of African swine fever virus from formalin fixed and non-fixed tissues by polymerase chain reaction - P. D. Luka, A. R. Jambol and B. Yakubu

Veterinary World, 7(10): 811-815

   doi: 10.14202/vetworld.2014.811-815

Abstract

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Aim: Formalin fixing and paraffin embedding of tissue samples is one of the techniques for preserving the structural integrity of cells for a very long time. However, extraction and analysis of genomic material from formalin fixed tissue (FFT) remains a challenge despite numerous attempts to develop a more effective method. The success of polymerase chain reaction (PCR) depends on the quality of DNA extract.

Materials and Methods: Here we assessed the conventional method of DNA extraction from FFT for African swine fever virus (ASFV) detection. The modified conventional method gave a higher quality DNA when compared with commercially available DNA extraction kits (QIAamp® DNA Mini Kit, DNeasy® Blood and Tissue Kit, and ZR Genomic DNA™ Tissue MiniPrep).

Results: An average A260/A280 DNA purity of 0.86-1.68 and 3.22-5.32 μg DNA/mg for formalin fixed and non-fixed tissues, respectively using a conventional method. In a reproducible and three times repeat PCR, the ASFV DNA expected product size of 278 bp was obtained from the DNA extract of the conventional method but not from the DNA extract of the commercial kits.

Conclusion: The present study has demonstrated that the conventional method extracts ASFV genome better than commercial kit. In summary, the commercial kit extraction appeared not suitable to purify ASFV DNA from FFT. We, therefore, recommend that the use of the conventional method be considered for African swine fever DNA extraction from FFT.

Keyword: African swine fever, detection, formalin fixed, non-formalin fixed, polymerase chain reaction.

Research (Published online: 12-10-2014)

10. Association study of genetic variants at single nucleotide polymorphism rs109231409 of mannose-binding lectins 1 gene with mastitis susceptibility in Vrindavani crossbred cattle - V. N. Muhasin Asaf, Bharat Bhushan, Manjit Panigrahi, Prashant Dewangan, Amod Kumar, Pushpendra Kumar and G. K. Gaur

Veterinary World, 7(10): 807-810

   doi: 10.14202/vetworld.2014.807-810

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Abstract

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Aim: The present study was undertaken to identify whether single nucleotide polymorphism (SNP) rs109231409 located on mannose-binding lectins 1 (MBL1) gene was associated with mastitis tolerance/susceptibility.

Materials and Methods: After grouping 100 Vrindavani crossbred cattle as mastitis positive and negative animals, they were genotyped using polymerase chain reaction (PCR)-restriction fragment length polymorphisms method. Gene and genotype frequencies of different patterns were estimated by standard procedure (POPGENE version 1.32, (University of Alberta, Canada) and statistical analysis was carried out by logistic regression methods using STATA 12 software (StataCorp LP, USA).

Results: The 588 bp fragment of MBL1 gene was amplified using PCR. PCR product was digested with ApaI restriction enzyme showed two distinct genotypes viz., GG (311 bp and 272 bp fragments) and GA (588 bp, 311 bp and 277 bp fragments). The gene, genotype frequencies, average heterozygosity, polymorphic information content and χ2 values for the locus rs109231409 was ascertained.

Conclusions: No significant association between SNP “rs109231409” with mastitis tolerance was found. Although there is a lack of association, further studies have to be undertaken in a large population in order to validate the impact of rs109231409 (g.855G >A) on mastitis tolerance.

Keywords: mannose-binding lectins 1, mastitis, polymerase chain reaction-restriction fragment length polymorphisms, single nucleotide polymorphism.