Animal Biotechnology Center
Animal Biotechnology Center
Jain T.,Animal Biotechnology Center |
Sachdeva G.K.,National Dairy Research Institute |
De S.,National Dairy Research Institute |
Datta T.K.,National Dairy Research Institute
Current Trends in Biotechnology and Pharmacy | Year: 2016
Terminal deoxynucleotidyl transferasemediated dUTP nick end labeling (TUNEL) is used widely for detecting the apoptotic status in biological cells of diverse origin. Reproducing the TUNEL procedure with specialized cells like cumulus oocyte complexes (COCs) poses technical challenge in view of mammalian oocytes which undergo very dynamic transitions in the course of oocyte maturation within a short time. The current work describes a standardized TUNEL protocol where the process of fixation and permeabilization were optimized using different concentrations of paraformaldehyde and Triton X-100 with sodium citrate, respectively. Effectiveness of the procedure was validated at different time intervals of oocyte culture using known oocyte stimulators. The procedure was further validated in the in vitro produced early embryos. Consistent and specific signal could be obtained using the procedure depicting the apoptotic status of cumulus cells and early embryos at different cell stages in buffalo. © 2016, Association of Biotechnology and Pharmacy. All rights reserved.
Manjari P.,ICAR NDRI |
Kapoor S.,ICAR NDRI |
Hyder I.,NTR University of Health Sciences |
De S.,Animal Biotechnology Center |
And 2 more authors.
Biological Rhythm Research | Year: 2017
Implantation is a critical process wherein the dam accepts the semi-allogenic embryo, characterized by certain vital immunological changes involving certain factors and cytokines. The current study was undertaken in Sahiwal cows to analyze the changes in certain selective immunological components of the body during peri-implantation period. Post-insemination the study was performed for a period till day 40 and the plasma was used to estimate the levels of progesterone, interleukin 1β (IL-1β), IL-2, IL-4, IL-6, and leukemia inhibiting factor (LIF) using Bovine specific ELISA Test Kits. The phagocytic activity and mRNA expression of progesterone-induced blocking factor (PIBF) were also analyzed. It was observed that in pregnant cows, the expression of PIBF increased from day 16th onward and remained high throughout the study period whereas the phagocytic activity was higher in non-pregnant cows all through the study period. The IL-1β, IL-2, and IL-6 were significantly higher in non-pregnant cows from 16th to 18th day post-AI as compared to P cows. On the other hand, IL-4 and LIF showed a significantly higher concentration in pregnant cows during peri-implantation period. Hence, it can be concluded that early embryonic survival is dependent on interplay of certain critical immunological factors in the body. © 2017 Informa UK Limited, trading as Taylor & Francis Group
Prashant Chaudhary P.,Animal Biotechnology Center |
Sirohi Kumar S.,Animal Biotechnology Center |
Kumar S.,Animal Biotechnology Center
Asian Journal of Animal Sciences | Year: 2011
The present study has been planned to standardize a simple and effective method for the isolation of good quality as well as quantity of methanogenic DNA in total genomic DNA from rumen liquor of Bubalus bubalis. Methanogens are a diverse group of organisms found in anaerobic environments such as anaerobic sludge digester, wet wood of trees, sewage, rumen, black mud, black sea sediments, etc which utilize carbon dioxide and hydrogen and produce methane. Methanogens exhibit great diversity in cell envelopes, ranging from simple, non rigid surface layers consisting of protein or glycoprotein subunits to a rigid "pseudomurein" sacculus, analogous to eubacterial murein. Methanogens having different chemical composition of cell wall and known for tough cell wall which is difficult break to isolate good quality and quantity of genomic DNA. Various DNA extraction methodologies have been used but problems are most often encountered in terms of low DNA yields and quality of DNA for further application. Method of DNA isolation based on guanidine thiocynate lysis buffer was compared with commonly used phenol chloroform method and commercial kit based methods, results showed that modified protocol generated high molecular weight genomic DNA while the other two methods resulted in considerable DNA degradation. Further, the isolated genomic DNA was tested for downstream applications such as PCR and Real-Time PCR using methanogens specific primers. In both the cases, the genomic DNA isolated by our protocol was comparatively better than rest of two protocols tested. © 2011 Knowledgia Review, Malaysia.
Wu Y.F.,Animal Biotechnology Center
Bing du xue bao = Chinese journal of virology / [bian ji, Bing du xue bao bian ji wei yuan hui] | Year: 2013
To study the proliferation characteristics of PPV in differently infected way and the variance of concentrations in different cells. A strain of porcine parvovirus(PPV) was adapted to PK-15 cells, and a Real-time fluorescent quantitative PCR (FQ-PCR) assay was developed based on the specific region of the NS1 gene of PPV to quantify the PPV. The FQ-PCR was used to measure the viral concentration of virus-infected cells by simultaneous or step by step inoculation and plot one-step growth curves. The proliferation characteristics of PPV strain in different cells lines (HeLa, MDBK, PK-15 ,ST, F81, BHK-21 and Marc-145) was also compared. The results showed the PK-15 cell -adapted strain of PPV produced CPE after 12 passages, and maintained stable CPE at the following 10 messages. The one-step growth curve showed that the virus concentration of simultaneous inoculation was higher than that of the step-by-step inoculation, and the proliferation cycle of step-by-step inoculation was shorter. The proliferation ability of PPV strain in different cells showed that CPE appeared first inPK-15, followed by ST, HeLa and MDBK, and the virus concentration was highest in ST, followed byPK-15, MDBK and HeLa. NO proliferation was observed in F81, BHK-21 and Marc-145 cells. These findings lay a material foundation for the basic researches on PPV and the development of vaccine.
Ranjan R.,Animal Biotechnology Center |
Bhong C.D.,Animal Biotechnology Center |
Parmar S.,Animal Biotechnology Center |
Joshi C.G.,Animal Biotechnology Center
Indian Journal of Animal Research | Year: 2015
The Nramp1 (Natural resistances macrophage protein1) gene or recently renamed Slc11a1 (solute carrier family 11 member1) is a member of a large family of genes coding for metal ion-transporting proteins. Nramp1 gene plays a critical role in innate immunity favoring bacterial killing by macrophages in addition to its influence on adaptative immunity. The aim of the present investigation was to identify the genetic variations in the 3’UTR (Untranslated region) of Nramp1 gene in the Gaolao breed (Bos indicus) cattle, using the technique PCR-SSCP and by sequencing. PCR-SSCP (Single Strand conformational polymorphism) of 440 bp amplicon of Nramp1 gene revealed four common SSCP patterns in Gaolao breed. The chi-square test revealed that difference between observed frequencies of different patterns in Gaolao was non-significant at 5% level of significance. The sequence analysis of Nramp1 region did not revealed sequence variation. The sequence analysis of Nramp1 region bare only two sequence variation in four SSCP patterns. This indicates that this region is either heterologous or SSCP could be independent of sequence variation. © 2014, Indian Journal of Animal Research. All Rights Reserved.
Gupta R.,Lovely Professional University |
Pramanik T.,Lovely Professional University |
Gupta R.,Animal Biotechnology Center
Der Pharma Chemica | Year: 2016
Biologically active La (III), Sm (III), Gd (III) and Dy (III) complexes were synthesized by using Furan-2-carboxylic acid (FCA) as ligand and were characterized by elemental analysis and spectral measurements. Coordination of the ligand atoms to the metal ion was deducted by IR spectral data. The in vitro antibacterial screening of the free acid and its metal complexes has been carried out against Pseudomonas and bacillus. Antifungal activities of all the synthesized compounds were screened for in vitro growth inhibitory activity against Aspergillus flavus and Aspergillus niger, Aspergillus fumigatus by using the cup plate method. Antimicrobial activities of the ligand increase on coordination with the metal ion and show more promising activity than the corresponding free acid, and the standard control antibiotics. Such increased activity of the metal chelates can be explained on the basis of Overtone's concept and chelation theory.
De S.,Animal Biotechnology Center |
Mir N.A.,Dairy Cattle Physiology
International Journal of Dairy Technology | Year: 2014
Nowadays, studies about the anti-obesity potential of probiotics are of growing interest. Lactobacilli are one of the well-studied probiotics owing to their preventing effect on metabolic disorders. This study was undertaken to access the anti-obesity effect of probiotic dahi containing Lactobacillus casei NCDC 19 on C57BL/6 mice. Feeding of probiotic dahi showed reduce body weight gain and epididymal fat weights. Moreover, blood glucose, plasma lipids and expression level of leptin were reduced and caecal bifidobacteria counts and adiponectin expression levels were significantly increased. It can be concluded that feeding of probiotic dahi containing L. casei NCDC 19 showed potential anti-obesity effects. © 2014 Society of Dairy Technology.
Yadav A.,Animal Biotechnology Center |
Singh K.,Animal Biotechnology Center |
Singh M.,Animal Biotechnology Center |
Saini N.,Animal Biotechnology Center |
And 5 more authors.
Reproduction in Domestic Animals | Year: 2013
Contents: For investigating the effects of physiologically relevant heat shock, buffalo oocytes/embryos were cultured at 38.5°C (control) or were exposed to 39.5°C (Group II) or 40.5°C (Group III) for 2 h once every day throughout in vitro maturation (IVM), fertilization (IVF) and culture (IVC). Percentage of oocytes that developed to 8-cell, 16-cell or blastocyst stage was lower (p < 0.05) and the number of apoptotic nuclei was higher (p < 0.05) for Group III > Group II > controls. At both 8-16-cell and blastocyst stages, relative mRNA abundance of stress-related genes HSP 70.1 and HSP 70.2 and pro-apoptotic genes CASPASE-3, BID and BAX was higher (p < 0.05) in Groups III and II than that in controls with the exception of stress-related gene HSF1. Expression level of anti-apoptotic genes BCL-XL and MCL-1 was also higher (p < 0.05) in Groups III and II than that in controls at both 8-16-cell and blastocyst stages. Among the genes related to embryonic development, at 8-16-cell stage, the expression level of GDF9 was higher (p < 0.05) in Group III than that in controls, whereas that of GLUT1, ZAR1 and BMP15 was not significantly different among the three groups. At the blastocyst stage, relative mRNA abundance of GLUT1 and GDF9 was higher (p < 0.05) in Group II than that in controls, whereas that of ZAR-1 and BMP15 was not affected. The results of this study demonstrate that exposure of buffalo oocytes and embryos to elevated temperatures for duration of time that is physiologically relevant severely compromises their developmental competence, increases apoptosis and affects stress-, apoptosis- and development-related genes. © 2013 Blackwell Verlag GmbH.
PubMed | Animal Biotechnology Center
Type: Journal Article | Journal: Acta veterinaria Hungarica | Year: 2012
In order to detect infectious bursal disease virus (IBDV), bursal tissue was collected from 10 IBD-suspected birds from a 30-day-old, IBDV-vaccinated commercial broiler chicken flock of 2000 birds exhibiting clinical signs suggestive of infectious bursal disease (IBD). The presence of IBDV was confirmed by partial amplification of the VP2 gene by reverse transcription and polymerase chain reaction. Isolates were identified as very virulent strains of IBDV (vvIBDV) by nucleotide sequence analysis. The comparison of the VP2 nucleotide sequences among the isolates revealed the presence of single-nucleotide polymorphisms in the VP2 gene of IBDV in the same flock. The comparative analysis indicated that these viruses were genetically close to the vvIBDVs previously detected in India. Our analysis provided information about the existence of vvIBDV in Central India.
PubMed | National Dairy Research Institute and Animal Biotechnology Center
Type: Journal Article | Journal: Cytokine | Year: 2015
The aim of our study was to optimize growth and induction parameters, for expression and large scale purification of functionally active buffalo interferon tau, and to study its possible impact on in vitro blastocyst development. The buffalo interferon-tau gene (BuIFN-T1) bearing gene bank accession No. JX481984, with signal sequence, was obtained through polymerase chain reaction (PCR) from bovine early embryos and was cloned into pJET vector. After being verified, the fragments without signal sequence, were inserted into the expression vector pET-22b and the recombinant plasmid was induced to express the recombinant protein in a prokaryotic expression system. The recombinant BuIFN-T was confirmed by SDS-PAGE and Western blot and subjected to three steps of large scale purification using His Affinity chromatography, Anion Exchange chromatography and Gel Filtration chromatography. The purified recombinant BuIFN-T protein was validated by mass spectroscopy analysis. To examine the effect of recombinant BuIFN-T protein on developmental competency of buffalo embryos, purified recombinant BuIFN-T protein was added to in vitro embryo culture medium (at concentration of 0, 1g/ml, 2g/ml, 4g/ml) for 9days. Addition of recombinant BuIFN-T (2g/ml) significantly improved the rate of blastocyst production, 45.55% against 31.1% control (p<0.01). Here we conclude that the recombinant BuIFN-T was successfully purified to homogeneity from a prokaryotic expression system and it significantly increased the blastocyst production rate in buffalo. These findings suggest a potential impact of IFN-T in promoting embryonic growth and development.