Thumbnail Image

Anand Agricultural University, Anand

Anand Agricultural University (AAU) was established in 2004 at Anand with the support of the Government of Gujarat, Act No.(Guj 5 of 2004) dated April 29, 2004. Caved out of the erstwhile Gujarat Agricultural University (GAU), the dream institution of Sardar Vallabhbhai Patel and Dr. K. M. Munshi, the AAU was set up to provide support to the farming community in three facets namely education, research and extension activities in Agriculture, Horticulture Engineering, product Processing and Home Science. At present there seven Colleges, seventeen Research Centers and six Extension Education Institute working in nine districts of Gujarat namely Ahmedabad, Anand, Dahod, Kheda, Panchmahal, Vadodara, Mahisagar, Botad and Chhotaudepur AAU's activities have expanded to span newer commodity sectors such as soil health card, bio-diesel, medicinal plants apart from the mandatory ones like rice, maize, tobacco, vegetable crops, fruit crops, forage crops, animal breeding, nutrition and dairy products etc. the core of AAU's operating philosophy however, continues to create the partnership between the rural people and committed academic as the basic for sustainable rural development. In pursuing its various programmes AAU's overall mission is to promote sustainable growth and economic independence in rural society. AAU aims to do this through education, research and extension education. Thus, AAU works towards the empowerment of the farmers.

Browse

20082
Reset filters
Author: NISHA × Thesis Type: M.V.Sc. ×
Reset filters

Search Results

Now showing 1 - 2 of 2
  • Thumbnail Image
    ThesisItemOpen Access
    MOLECULAR CHARACTERIZATION AND DIVERSITY OF RUMEN PROTOZOA IN BUFFALO
    (AAU, Anand, 2008) NISHA; Pandya, P. R.
    Rumen harbors diverse types of microbes mainly bacteria followed by protozoa, fungi and yeast. Bacterial population is as high as 10 to power 10 and protozoa population is 10 to power 5 to 10 to power 6 per ml of fluid, found in the rumen and has a profound effect on nutritional and physiological processes in the host. The ability and characteristic of rumen microbes differs. Some are good at digesting concentrate while others at roughage. The study of rumen ecology involves their relationship with each other and the host. Much of the pioneering studies on the rumen microbiota are based on microscopic examination and anaerobic culture techniques. But the polymorphic nature and the difficulty of cultivating the microbes have hampered effective assessment of ruminal ecology by these methods. Newer molecular approaches are available to identify and characterize the eukaryotes and prokaryotes that are based on detecting highly conserved gene regions-16S rRNA and 18S rRNA, respectively for bacteria and protozoa. India possesses more than 50 per cent of world buffalo population. The present study was aimed to determine genetic characterization of rumen protozoa in buffalo and their phylogenetic classification, based on sequence comparison of 18S rRNA gene of rumen protozoa. Three adult buffaloes were maintained on standard dietary regimen as per ICAR (1998) feeding standards, composed of green and dry roughage and compound concentrate mixture, continuously for three weeks. Samples of Rumen liquor (about 500 ml) were collected from three buffaloes at 2, 4 and 6 hrs. after feeding by a suction pump using a flexible stomach tube. Samples were filtered through four layers of cheesecloth. The strained rumen liquor was used for the study. The protozoal DNA was isolated following enzyme-chemical lysis method. The DNA stock samples were quantified using Nano-drop spectrophotometer at 260 and 280 nm using the convention that one absorbance unit at 260 nm wavelength equals 50 µg DNA per ml. The Ultra violet absorbance was checked at 260 and 280 nm for determination of DNA concentration and purity. Quality and purity of DNA were also checked by agarose gel electrophoresis. Protozoa specific primers viz. Euk-82F 5' AAA CTG CGA ATG OCT C -3' and Medlin Be 5' TGA TCC TTC TGC AGG TTC ACC TAG -3' targeting 18S rRNA gene were used for amplification of DNA. The amplified product was visualized as a single compact band of expected size under UV light and revealed amplicons of 1346 bp size when documented by gel documentation system. The PCR products after purification were ligated in pDrive vector followed by transformation into competent cells (DH5-a strain) of E.coli. One hundred fifty white recombinant colonies were obtained out of which 124 were randomly selected, revived on another plates and screened for expected insert by colony PCR. Recombinant colonies were inoculated in Luria Broth for 16-18 hrs. Plasmid extraction from overnight culture was carried out by alkaline lysis method. The concentration of the plasmid DNA was determined and was subjected to automated DNA sequencing on ABI PRISM® 310 Genetic Analyzer (Applied Biosystems, USA). Sequencing was carried out using BigDye® Terminator v3.1 Cycle sequencing kit. Sequences obtained were in the range of 250 to 850 base pairs. Sequences after cleaning (removal of primer and vector sequence) were searched against BLASTn database to find similarity matches. Analysis of the 18S rRNA sequences of protozoal species from ruminal fluid revealed predominance of family Isotrichidae {Dasytricha and Isotricha) and Ophryoscolecidae. Phylogenetic analysis was performed using DNADIST and DNAPARS (Parsimony method) program of PHYLIP package (Felsenstein, 1989). Phylogenetic tree was constructed by neighbor-joining method (Saito and Nei, 1987). Out of 124 clones, 21.7% showed similarity with Ophryoscolex spp., 7.2% showed similarity with Cycloposthium spp., 18.5% with Trichostomatidae, 1.6% with Haptorians and about 51% clones remained as unclassified protozoa which are supposed to be new. In DNADIST analysis, two clones (1.6%) from 124 sequenced clones fall in Haptorian group. The AVCRPN62 clone showed 99% similarity with the Enchelyodon spp. while AVCRPN86 showed 99 % similarity with Arcuospathidium spp. In DNAPARS analysis, AVCRPN62 clone showed 95% similarity with Arcuospathidium spp. and was clustered separately in subcluster II of Cluster III. Twenty seven clones (21.7%) clustered separately with Ophryoscolex spp. (21 clones) and Spathidium spp. (6 clones). Members of this group showed 82% similarity in DNADIST analysis. No clones showed similarity with this group in DNAPARS analysis. Nine clones (7.2%) were showing 94% similarity with Cycloposthium spp. which belonged to Cycloposthiidae family and were grouped in Cluster IV in DNADIST analysis, while 24 clones were showing similarity with Cycloposthium spp. and were grouped in subcluster 1 in Cluster III in DNAPARS analysis. Twenty three clones (18.5%) out of 124 clones were showing similarity with Trichostomatidae and were grouped separately in Cluster V in DNADIST analysis, while 36 clones were showing similarity with Trichostomatidae and were grouped in subcluster IV of Cluster III in DNAPARS analysis. Members of Cluster V showed 77% similarity in DNADIST analysis. This cluster was again divided in four sub clusters:Australian clade, Entodiniomorphs, Dasytricha spp., Isotricha spp. Sixty three clones (51%) clustered separately which were showing no significant similarity with cultured as well as uncultured representatives. Clones in this group can be considered as novel clones. Taxonomic classification of these clones showed that the maximum no of clones show similarity to Isotricha spp., Dasytricha spp. and uncuhured rumen protozoa. All the sequences were submitted to Genbank and are available with the accession numbers EU796091- EU796211 in EMBL, GenBank and DDBJ Nucleotide Sequence Databases.
  • Thumbnail Image
    ThesisItemOpen Access
    “Molecular characterization and Diversity of Rumen Protozoa in Buffalo
    (Anand Agricultural University, Anand, 2008) NISHA; Dr. P.R. Pandya
    Rumen harbors diverse types of microbes mainly bacteria followed by protozoa, fungi and yeast. Bacterial population is as high as 1010 and protozoa population is 105 to 106 per ml of fluid, found in the rumen and has a profound effect on nutritional and physiological processes in the host. The ability and characteristic of rumen microbes differs. Some are good at digesting concentrate while others at roughage. The study of rumen ecology involves their relationship with each other and the host.