Dr Anand Laxmi
Dr Anand Laxmi
Dairy Cattle Physiology National Dairy Research Institute Karnal-132001, HARYANA
Current Research Area
Reproductive physiology and cell culture
Major Research Accomplishments :
Externally funded project with Department of Biotechnology:
The cell cultures could be transfected with IFN-t-EGFP gene construct by nucleofection method. The efficiency of the transfected gene was monitored for green fluorescence which was observed to be 55-60% and confirmed by FACS to be 50-55%. The transfected gene was expressible from second to sixth passage as confirmed by RT-PCR. The protein fraction extracted from crude cell lysate and further obtained on immuno-precipitation with EGFP monoclonal antibody could be visualized on SDS-PAGE after silver staining .. The protein exhibited some characteristics which have been assigned for IFN-t such as antiproliferative effect on lymphocytes ,antibacterial activity. The cells transfected with IFN-t and subjected to SCNT to enucleated oocytes were competent enough to develop to 16-32 stage hybrid bovine embryos. Hence, it can be reported that the Sahiwal fibroblast cell lines were developed, which were competent for studies on gene expression/nuclear transfer.
In collaboration with Dairy Extension division:
As per the technical programme, seven villages were selected and conducted 46 infertility and veterinary aid campaigns. A total of 126 animals were identified with functional C.L and were treated under various protocols for synchronization & FTAI programme. Rest of the animals were also diagnosed for various causes and given appropriate treatments accordingly. Out of total 126 animals, 81 (64.28%) animals conceived. The overall pregnancy rate was 64.40 in cows and 64.17% in buffaloes. .
The project work involved regular organization of Kisan Sangoshthi and Infertility and Veterinary aid campaigns with active participation of local farmers. The involvement of farmers in the management and assessment of field trials helped to sensitize them with cases of reproductive disorders and timely precautionary steps to be taken by them to reduce these problems. Farmer Livestock Schools (FLS) were organized to sensitize the farmers about reproductive management. A Learning module on reproductive management was developed and used to educate farmers. Participants’ perception about FLS was measured in terms of best aspect, disappointment factors, suggestion for improvement, participants’ area of interest for future FLS and willingness of participants to facilitate school.
Guidance to Students :
- Dr. P.G. Nair Award in 2004.
- In Molecular Techniques in 2002
- In Cell culture technique in 2003
Patents, Technology, Methodology, Genetic Stock, Variety, etc.:
- Different ways of gene transfection namely lipofection,and electroporation have been optimised for transfection of EGFP gene construct in to eukaryotic Sahiwal fibroblast cells. These cells could be successfully propagated for studies on gene expression / nuclear transfer. Hence the technique of cryopreservation for cell culture in medium with 10% DMSO was effective for Sahiwal fibroblast cells also.
- The IFN-t gene construct with a promoter which was successfully expressed in eukaryotic cells was used for further, transfection of recombinant IFN-t-EGFP gene by superior method of nucleofection. The recombinant IFN-t –EGFP fusion protein could be successfully expressed in vitro, which is of importance in the field of reproduction.
- Development of EIA procedures for analysis of hormones in plasma definitely is cheap, cost effective and no costly instrument is required. It is less hazardous than radioimmunoassay .Assisted in development of EIA for Luteinizing hormone and Growth hormone.
- Estrus synchronization program was developed at the divisional level . In post partum animals upon detection of functional C L, PGF2a was administerd ,GnRH was administered 60h post PGF2a and A.I. at fixed time i.e. 72h post PGF2 a injection, and transferred to field in collaboration with Extension Division which resulted in conception rate ranging between 65%-67% in bovines. It was compared with the Cosynch and PGF2a synchronization programs.
- Anand Laxmi N., Gunjan G.,Neelam G. and Prakash B.S. (2008) Evaluation of long term fibroblast cell culture system for cytokines and other parameters in Sahiwal cattle. Int.J. Biochem.and Biotech.4:207-221.
- J Gupta, A Laxmi, Om Vir Singh and Ashutosh (2008) A comparative study on evaluation of three synchronization protocols at field level in both cattle and buffaloes.Livestock Res. Rural Devp. 20:175
- Laxmi N.A. and Prakash B.S. (2007) Comparative profiles of different lipoprotein cholesterol parameters and Growth hormone during hot humid and winter season. Livestock Prod. Sci. 115: 294-300.