Research

Our research activities are focused on four major areas namely the embryo biotechnologies, animal genomics, proteomics & system biology and stem cell biology. Faster multiplication of superior germplasms of dairy animals by the application of hand-made cloning technology remains one of the prime areas of our current research. Identification of superior animals utilizing the candidate genes/proteins related to economic and physiological traits of dairy animals ensuring faster genetic gains in animals and development of the selection criteria for dairy animal is the other major area.  Fundamental knowledge of one gene synthesizes many proteins is being investigated at the centre to assess the economic traits of dairy animals by utilizing in vitro embryo production system followed by the molecular investigations. Exploring the reproduction traits and disease resistance qualities of Indian dairy breeds are the current interests of the research groups. Proteomics is another branch of the functional genomics, responsible for improvement of dairy production, productivity and processing areas. With the introduction of high-throughput and cutting-edge technology of mass-spectrometry for proteomic studies, the researchers at center are generating the proteomic profile of sub-clinical mastitis and matritis diseases. Early detection of pregnancy utilizing PAGs and miRNA are also being investigated at the center.  Development of recombinant proteins as the intermediate technologies for fulfilling the needs of the animal and dairy industry forms another major area of research at ABTC. Many such useful recombinant proteins such as LIF, GDF-9, Lectoferrin, SSLP and amino-peptidases have been developed in the recent past. Researchers at the centre are also engaged in promoting development of designer animals and humanized dairy milk from animals by utilizing CRISPER and TALEN based genome editing tools. In the era of high usage of antibiotics in dairy industry, stem cell technology has emerged as an alternative to treatment with antibiotics for diseased udder, skin and hoof wounds. Mesenchymal stem cell technology has been developed at the centre and routinely utilized for treatment of animal diseases.

The state of art laboratories equipped with modern and high end equipments and tools are available at ABTC to carry out all research activities. The laboratories are equipped with equipments like LC-MS/MS, Flow cytometer with sorter, Bioreactors, DNA sequencer, Real-time PCR, Micromanipulator and range of incubators and freezers. A specialized bioinformatic facility is available to cater the need of researchers for studying the niche areas of bioinformatics related to animal biotechnology.

Current Research Project

List of externally funded projects (as on March 6, 2019)

1. 1. Deciphering the circulating miRNAs from terminal stage pre-implantation embryos and placentomes for early detection of pregnancy in buffalo ( DBT, Govt. of India: Rs. 52.2715 lakhs; Duration: 4.8.2016 to 3.7.2019)

(Project Leader: Dr. T.K. Datta, Co-Leaders:  Dr. Rakesh Kumar and Dr. A. Kumaresan)

1. 1. Synthesis, characterization and effect of graded levels of nano-selenium supplementation on the performance of broiler chicken (ICAR-NASF: Rs. 42.727 lakhs; Duration: 1.6.2017 to 31.5.2020)

(Project Leader: Dr. A.K. Mohanty, Co-Leader:  Dr. Sudarshan Kumar)

1. 1. Genetic variability of milk protein and its characterization by proteomic approach in Indian goats (ICAR-NASF: Rs. 34.45080 lakhs; Duration: 1.6.2017  to 31.5.2020)

(Project leader: Dr. S. De)

1. 1. Conservation of Indigenous Pig of Assam through Handmade Cloning Technique (DBT, Govt. of India: Rs. 40.59 lakhs; Duration: 2.1.2017 to 1.1.2020)

(Project Leader: Dr. M.K. Singh)

1. 1. Global transcriptome and miRNA analysis for deciphering reasons for low cloning efficiency in buffalo (SERB, DST, Govt. of India: Rs. 46.78 lakhs; Duration: 22.3.18 to 21.3.21)

(Project Leader: Dr. P. Palta, Co-Leader:  Dr. M.K. Singh)

1. 1. Production of multiple copies of elite buffalo bulls using animal cloning technology (ICAR-NASF: Rs. 264.2964 lakhs; Duration: 31.3.2018 to 30.3.2022)

(Project Leader: Dr. P. Palta; Co- Leaders: Dr. M.K. Singh, Dr. S.S. Lathwal, Dr. Subhash Chand)

1. 1. Genome editing of MFGE8 and S100 genes in bovine mammary epithelial cells to understand their role in milk production (SERB, DST, Govt. of India; Rs. 62.538 lakhs; Duration: 21.3..2018 to 20.3.2021)

(Leader:  A. K. Mohanty; Co- Leaders: Dr. Sudarshan Kumar, Dr. J.K. Kaushik, Dr. Malakar, Dr. Satish Kumar)

1. 1. Improving the usability of buffalo spermatozoa by sperm surface remodelling and immune acceptance in female reproductive tract (ICAR-NASF: Rs. 174.60 lakhs; Duration: 1.7.2018 to 30.7.2021)

(Project leader: Dr. T.K. Datta, Co-Leader: Dr. Rakesh Kumar, Dr. J.K. Kaushik)

1. 1. Molecular Markers for Improving Reproduction of Cattle and Buffaloes (Bill & Melinda Gates Foundation, USA: Rs. 648.38 lakhs; Duration: 31.07.2018 to 31.07.2023)

(Project Leader: Dr. T.K. Datta; Co-Leaders: Dr. Rakesh Kumar, Dr. Dheer Singh, Dr. Suneel Onteru, Dr. R.K. Baithalu, Dr. A.K. Mohanty, Dr. Sudarshan Kumar, Dr. T.K. Mohanty)

1. 1. Targeted immobilization of Y-bearing spermatozoa and modulation of oviduct milieu for skewing sex ratio towards female offspring in dairy cattle (ICAR-NASF: Rs. 11.34 lakhs; Duration: 01.08.2018 to 31.07.2021)

(Project leader: Dr. Rakesh Kumar; Co-leader: Dr. T.K. Datta)

1. 1. CRISPR/CAS9 guided functional analysis of genes regulating early embryonic survival in buffalo (ICAR-NASF: Rs. 62.4431 lakhs; Duration: 01.08.2018 to 31.07.2021)

(Project leader: Dr. Dhruba Malakar ; Co-leader: Dr. Satish Kumar)

1. 1. Development of early pregnancy diagnostic assay through discovery of biomarkers in cattle and buffalo (DBT, Govt. of India: Rs. 114.70725 lakhs; Duration: 10.5.2018 to 9.5.2021)

(Project leader: Dr. A.K. Mohanty ; Co-leader: Dr. R.K. Baithalu, Dr. T.K. Mahanty, Dr. Sudarshan Kumar)

1. 1. Semen sexing in cattle  (ICAR: Rs. 239.5 lakhs; Duration: 1.4.2017 to 31.3.2021)

(Project leader: Dr.  A.K. Mohanty; Co-Leader: Dr. Sudarshan Kumar)

PDF Projects:

• Title; Quantitative profiling of buffalo milk fat globule membrane, a promising nutraceutical for infant formula milk (Woman Scientist Scheme of DST). Project Workers; A. Turan, J.K. Kaushik, Duration; 2017-20.
• Title; Profiling of milk proteome in different species of farm animals  and comparative evaluation of host defense proteins (NPDF project). Project Workers; A. Chopra, A.K. Mohanty, Duration; 2016-18.
• Title; Protein profiling of semen of cross-bred cattle bulls for identification of biomarkers for high semen freezability. (SERB-NPDF). Project Workers; S. Kalra, A.K. Mohanty, Duration; 2017-2019.
• Title; Pesticide exposure and sperm fertilizing ability: A study on Bovine Model. (SERB-PDF). Project Workers; Shivani Chhillar and T.K. Datta. Duration; 2018-20.
• Title; Delineating genome wide methylation dynamics in cows of native and exotic origin: An approach to understand heat stress adaptation. (SERB-PDF). Project Workers; Preeti Verma and A.K. Mohanty. Duration; 2018-20.
• Title; Development of methods for detection of betacasomorphin-7 in biological samples, (urine or blood) by aptamers. (ICMR-RA). Project Workers; Abhishek and A.K. Mohanty. Duration; 2018-21.           

History

• History

Achievements

• A male cloned calf named ‘Rajat’, was born on July 23, 2014 by normal parturition. Rajat was produced through Hand-guided cloning using somatic cells isolated from the frozen-thawed semen of a highly ranked progeny-tested Murrah buffalo bull (MU-4393) which had died many years back.
• A female calf named ‘Deepasha’ was born on December 12, 2014 by normal parturition. Deepasha is a clone of a wild buffalo named ‘Asha’, that has been kept in semi-captivity at Chhattisgarh. The wild buffalo is an endangered species.
• A female cloned calf named ‘Apurva’ was born on February 5, 2015 through normal parturition. Apurva was produced using somatic cells isolated from urine.
• A female cloned calf named ‘Lalima’ was born on May 2, 2014 by normal parturition. Lalima is a clone of an elite Murrah buffalo (MU-5345) of NDRI Livestock Farm.
• Garima II, the cloned buffalo, delivered second female calf named ‘Karishma’ on December 27, 2014 through normal parturition.
• A female cloned calf named ‘Swarupa’ produced through ‘Hand-guided Cloning’ was born on August 1, 2015. Swarupa is a clone of ‘Karan-Kirti’, the highest milk producing Murrah buffalo at NDRI Livestock Research Centre.
• Transgenic buffalo and goat embryos containing human insulin gene were produced from transfected buffalo fetal fibroblasts.
• Germ line cell-specific genes were identified in buffalo and buffalo embryonic stem cell lines were differentiated towards germ lineage.
• VASA-transgenic buffalo embryonic stem cell lines were established and differentiated to germ lineage.
• Global transcriptome profiling was carried out in goat embryos at different stages of development, produced by in vitro fertilization, somatic cell nuclear transfer and parthenogenesis.
• Buffalo recombinant interferon-tau was produced.
• Cloning work at NDRI was highlighted as 9^th among top 20 science stories of 2014 by ‘Scientific India’, a science magazine.
• Cloned buffalo embryos were successfully produced using trophoblast cells, urine- and milk-derived somatic cells and lymphocytes as donor cells.
• Cloned blastocysts derived from fibroblasts, milk-derived cells, lymphocytes, trophoblast cells and those produced by in vitro fertilization were found to differ in their developmental competence, level of apoptosis, epigenetic status and expression level of many important genes.
• Buffalo trophoblast cell lines were developed from blastocysts produced by in vitro fertilization, Hand-guided cloning and parthenogenesis. A feeder-free in vitro culture system was developed which enabled their long-term culture.
• Treatment of buffalo donor cells with epigenetic modifiers was shown to improve the blastocyst rate and quality of cloned embryos
• Cloned female and male buffalo embryos were found to differ in their developmental competence, epigenetic status, sex-biased transcription patterns in X-linked genes and response to epigenetic modifiers. Aberrant X-linked gene expression occurred frequently in female embryos.
• Transgenic cloned buffalo embryos were produced containing pAcGFP-buSCD, a mammary gland specific expression vector, which is able to convert high level of conjugated linoleic acid in mammary gland epithelial cells.
• Global transcriptome and miRNA profiling was carried out in buffalo blastocyst-stage embryos produced by cloning and in vitro fertilization techniques to identify the shortcomings with cloned embryos. This will help in making strategies for improving the success rate of producing cloned calves in cattle and buffaloes.
• Transgenic buffalo spermatogonial stem cells were produced and used for homologous transplantation.
• Mammary lineage cells were produced by directed differentiation of buffalo embryonic stem cells.
• Novel non-coding RNAs were discovered in buffalo oocytes which may influence their development competence.
• Recombinant bovine lukemia inhibitory factor was produced successfully. Its use as a supplement for culture of bovine stem cells may help improve efficiency and reduce the cost.
• Direct application of mesenchymal stem cells isolated from adipocyte tissue was demonstrated to be effective in management of chronic hoof wounds in cattle and buffaloes.
• Treatment of human intestinal cells with recombinantly produced lactobacilli surface layer proteins could prevent pathogen binding to the extent of as high as 76%.
• Application of pulsed electromagnetic field to reconstructed embryos during Hand-guided cloning was shown to increase the blastocyst production rate and improve the quality of cloned embryos produced.
• Transgenic goat embryos, having GFP integration at Rosa 26 locus, were produced using one of the latest genome editing tools Transcription Activator-like Effector Nucleases (TALEN).
• Tetraploid complementation with putative parthenogenetic embryonic stem cells was successfully used for production of chimeric blastocyst-stage goat embryos.
• Treatment of cloned embryos with miR-145 inhibitor was shown to improve their developmental competence and quality, and increase histone acetylation and expression level of several pluripotency-related genes.
• A novel non-coding RNA (BOA 290U) was identified in buffalo oocyte and its association was established with expressed proteins. It is potentially important for imparting developmental competence to oocytes.
• The chronology of GVBD event was found to be associated with competence of buffalo oocytes. Holding oocytes temporarily at GV stage was found to be useful in enhancing their developmental ability.
• High throughput RNA sequencing of buffalo oocytes revealed genes which are significantly affected in ‘Good’ vs. ‘Bad’ oocytes. Affected pathways in inferior quality oocytes were identified.
• A battery of unique non-coding mi & pi RNAs was discovered in buffalo oocytes, which could be linked to their competence. Major genes under CLR and NLR category were amplified and sequenced completely.
• Basal expression pattern of important RLR, NLR and CLR genes was compared in immune and non-immune buffalo tissues. A large number of splice and transcript variants were observed to be present under RLR, NLR and CLR category in different types of buffalo tissues.
• Potential protein biomarkers have been identified in urine and serum of cows for early detection of pregnancy 16 days post artificial insemination.
• A proteomics-based method was developed for detection of A1/A2 milk of cattle and buffaloes
• Candidate microRNAs, which were uniquely expressed during early pregnancy (30 days) in buffalo, were identified.
• Specific miRNAs associated with maternal and fetal placentomes were identified in early pregnant buffaloes. A step towards detection of pregnancy by miRNAs.
• Comparative sequence analysis of β-defensins from different species by bioinformatic analysis revealed that buffalo β-defensins have unique conserved functional motifs although their sequences are quite different from those of other closely related mammals.
• Increased expression of sperm coat protein, beta-defensins 126 and 129 was detected in epididymis of buffalo testes.
• Eight novel variants of goat kappa casein variants were revealed from seven different Indian goat breeds.
• Functional characterization of Milk fat globule membrane epidermal growth factor 8 was performed by gene silencing using shRNA. It was observed that MFGE8 is essential for maintenance of cell shape and morphology during lactation in farm animals.
• A total of 1600 proteins were identified in bovine urine by using LC-MS/MS for the first time. Various methods were optimized for extraction of proteins from bovine urine.
• Three potential urine protein biomarkers (bovine allergen, Mannan-binding lectin serine peptidase 2, Alpha-1-microglobulin, Glutaminyl-peptide cyclo-transferase) were identified for detection of early pregnancy (~ 20 days) in cows. Three proteins namely vitamin-binding protein, apolipoprptein A and complement 3 were up-regulated in bovine serum during pregnancy (16^th to 45^th day).
• Differential transcript analysis in fetal cotyledon of early pregnant buffalo (~2 months) revealed that prostate-specific antigen is over-expressed during early pregnancy.
• Recombinant leucyl aminopeptidase of Lactobacillus species was expressed in biologically active form.
• Buffalo hepatocytes were successfully cultured for over a week on several extracellular matrices and feeder fibroblast cells.
• Methods were standardized for in vitro differentiation of buffalo mammary epithelial cells for expression of milk proteins (casein).
• The function of MGP-40, a mammary-specific protein which protects the loss of mammary epithelial cells during mammary gland involution via activation of STAT-3 phosphorylation, was deciphered.
• Buffalo oviductin was observed to have a positive effect on several sperm quality-related parameters such as motility, viability, acrosomal integrity and capacitation. Oviductin was found to increase cleavage rate and blastocyst formation during IVF.
• Salivary proteome of Sahiwal cows revealed 3000 proteins, which is the highest number reported hitherto.
• A stably transfected COS-1 cell line was established which expressed recombinant buffalo leukemia inhibitory factor.
• Recombinant buffalo GDF9 (57 kDa) was shown to enhance cleavage and blastocyst rate during production of IVF buffalo embryos.
• Recombinant mucus binding protein (rMub), S-layer protein (rSlp) of lactic acid bacteria were shown to preclude the binding of enterotoxigenic coli (ETEC) with human enteric cell lines Caco-2 and HT-29.
• Recombinant leucyl aminopeptidase and prolyl aminopeptidase of lactobacilli origin were expressed, their physicochemical properties studied and their specific activity was shown to be better or comparable to that of commercially available Corolase enzyme.
• Proteins differentially expressed in milk whey of A1 and A2 milk were identified for analyzing their health implications
• New definition of precious pashmina fibers was established with its unique proteomic compositions. Key proteins involved in Pashmina fibre growth and development were identified by analysis of Pashmina fibre and skin proteome.Around 7000 proteins were identified in the skin proteome of Pashmina goats
• A proof-of-concept was developed for the detection of pregnancy in cattle, as early as 35 days, using an antibody against Pregnancy Associated Glycoprotein-7 (PAG7).
• A LC-MS method was developed for identification of A1 and A2 beta caseins in milk of Karan Fries and Sahiwal cattle. The method has been validated in more than 200 cows.
• More than 6800 proteins were identified for the first time in milk of Sahiwal cattle.
• 1371 differentially expressed genes and 757 differentially expressed proteins were identifies during lactogenic differentiation in buffalo mammary epithelial cells and their possible role in lactation was studied.
• A stably transfected COS-1 cell line was developed which expresses recombinant bovine Leukemia Inhibitory Factor .
• Mesenchymal stem cells used successfully to permanently cure hoof wounds in cattle and buffaloes.
• Mouse allogenic and cattle xenogeneic mesenchymal stem cells were used to successfully heal fractured tibial bones of mice within 5 weeks.
• Recombinant buffalo sperm lysozyme like protein 5 (SPACA5) was produced using Coli expression system.
• More than 12000 proteins have been profiled in Buffalo mammary epithelial cells, and 6 novel proteins were found to be involved in lactogenesis.
• Three different isoforms of buffalo pregnancy associated glycoproteins (PAG-1, PAG-2 & PAG-18) were produced in coli expression system.
• An innovative microfluidic device was developed for partial enrichment of live and motile sperm cells.
• Mesenchymal stem cells were successfully used to cure mastitis and metritis in cattle.

Courses Offered

• Master's courses
• Ph.D. Courses

Technologies

• PATENTS
• TECHNOLOGIES

Alumni

Contact us