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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.

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20133
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Subject: Veterinary Parasitology × End date: 2013 × Start date: 2013 × Type: Thesis × Thesis Type: M.V.Sc. ×
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    ThesisItemOpen Access
    Transcriptome Analysis of Paramphistomum cervi of water buffalo (Bubalus bubalis) using next generation sequencing
    (AAU, Anand, 2013) CHOURASIA, REETIKA; PATEL, P. V.
    Rumen flukes are economically important parasites (Platyhelminthes: Trematoda: Digenea) that attack livestock adversely thereby affecting their productivity. In spite of its economic importance, molecular biology of the Paramphistomum cervi and its interaction with its hosts is still unknown. Advances in transcriptomic and bioinformatics provide biologically relevant insights into parasites, their developmental stages and their relationships with their hosts at the molecular level. The present study elucidates the first transcriptome and gene expression profiling of the adult stage of Paramphistomum cervi using next-generation (high throughput) sequencing and advanced in silico analyses. Expression level for predicted proteins of Paramphistomum cervi of buffalo were determined and classified based on homology, gene ontology and pathway mapping. These findings are expected to provide new insights into the genetic architecture and pathophysiology of Paramphistomum cervi and for the development of improved interventions for disease control. It will also facilitate a more fundamental understanding of Paramphistomes biology, evolution and the host-parasite interplay. Moiphological characteristics of adult fluke were identified as conical shape, elongate, curved ventrally, with evenly curved dorsal and ventral borders. Cuticle is provided with prominent tubercules/papillae on anterior l/3rd to half of the body. Tubercles are more extensive ventrally. Acetabulum is subtemiinal. hitestinal caeca have 7 nearly identical bends with ventrally directed temiinal part. Testes are tandem, oval or angularly oval or spherical and are deeply lobed. Gross examination of affected rumen showed, anaemic rumen with atrophied, degenerated and sloughing tips of villi. Removal of flukes revealed marked knobs at the attachment sites. Histopathology of rumen revealed proliferation of epithelium in the vicinity of flukes, along with villous atrophy and infiltration of macrophages and eosinophils. Transcriptome analysis of adult stage of Paramphistomum ceni was carried out at Department of Animal Biotechnology. Total RNA was extracted from parasites using TRIzol® (Invitrogen, UK)/ RNeasy® mini kit and mRNA isolation from the total RNA was carried out by using mRNA isolation kit. The quality and quantity of RNA and mRNA checked by running the sample on NanoDrop ND-1000 spectrophotometer. Concentration of RNA of adult fluke was 2,608 ng/µl and mRNA was 100 ng/µl. The cDNA library was constructed using the Ion Total RNA-Seq Kit v2. According to Qubit®Fluorometer, concentration of cDNA was 1.19 ng/µl and based on Aligent 2100 Bioanalyzer concentration of cDNA is 1.25 ng/µl.
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    ThesisItemOpen Access
    Diagnosis of Tropical Theileriosis in cattle and buffaloes using advanced molecular tools
    (AAU, Anand, 2013) KUNDAVE, V. R.; PATEL, P. V.
    The study on "Diagnosis of Tropical Theileriosis in cattle and buffaloes using advanced molecular tools" was carried out to effectively diagnose Theileria annulata by Poljmierase Chain Reaction and its quantification by real-time PCR assay in infected and carrier animals. Bovine tropical theileriosis caused by Theileria annulata is a tick-borne disease, associated with high morbidity and mortality rate in the livestock and pose a great threat to the farmers and dairy industry in India. The diagnosis by microscopic examination, has low sensitivity and it is difficult to detect the piroplasms in the carriers, while the PCR based assays are more sensitive. In this study a total of 116 samples, were collected from infected as well as apparently healthy cattle and buffaloes, 74 samples (63.79 per cent) were positive for Theileria annulata by PCR, which includes 15 samples that were positive by giemsa staining. The primers were designed to amplify the Tamsl gene encoding the 30- kDa major merozoite surface antigen of T. annulata. A product size of 430-bp afi:er amplification by Polymerase Chain Reaction (PCR). Highestprevalence was recorded in cattle above 5 years of age (82.35 percent) and the lowest prevalence was recorded in calves less than 1 year of age (14.28 per cent). A SYBR Green-based real-time PCR assay was carried out to quantify the load of parasites in positive samples. The parasitic load ranged from 1000 to 34,00,000 and 300 to 29,000 per microlitre of blood in cattle and buffalo samples, respectively, indicating the sensitivity of the diagnostic assay and also the degree of infection in the infected as well as carrier animals. The clinical signs suggestive of tropical theileriosis were more prominent in the acute phase of infection, which was also characterized by high levels of parasitaemia while their occurance in low level was found in the blood of carrier animals. The study undertaken suggests that the difficulties faced in detection and differentiation of Theileria piroplasms by conventional staining method could be overcome by the molecular methods like PCR. Real-time PCR assay could be used as a sensitive and accurate method to detect and quantify the parasitic load in the blood of cattle and buffaloes. PCR assays are advantageous since it has the ability to detect the disease in carrier animals and animals in chronic phase of theileriosis.
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    ThesisItemOpen Access
    Transcriptome Analysis of Paramphistomum cervi in Sheep (Ovis aries) Using Next Generation Sequencing
    (AAU, Anand, 2013) VIJAYATA; Hasnani, J. J.
    Rumen fluke, Paramphistomum cervi, is responsible for production losses to the livestock (mainly sheep, goat and buffalo) and meat industries due to clinical disease, reduced weight gain and milk production, and deaths. Surprisingly, despite the massive economic loss, knowledge of the fiindamental molecular biology of Paramphistomum cervi is scanty. Currently, there is a need to focus on the development of new approaches for the prevention and control of Paramphistomosis in livestock. Paramphistomum cervi were found mainly in the rumen and were light pink in colour with a sucker at the tip of the cone and another sucker ventrally at the posterior end. The body of P. cervi was pear-shaped, slightly concave ventrally (conical) and convex dorsally. Caeca were wide, pursued a serpentine course and reached anterior margin of posterior sucker (acetabulum) with blind ends more dorsal than lateral. Genital pore was situated behind intestinal bifiircation. Histological peculiarities of the muscular organs such as a liorchis type pharynx, gracile type genital opening, Paramphistomum type acetabulum in median sagittal section and distinctly lobed testes, situated a little obliquely tandem were also supported the histological diagnosis. The present study explores, for the first time, the transcriptome and proteome of the adult stage of the rumen fluke, Paramphistomum cenn of sheep using Ion Torrent PGM sequencing and in silico analysis, and also provides a basis for applied outcomes such as the development of drug and vaccines against this neglected parasite of veterinary importance. Transcriptome analysis oi Paramphistomum cenn of sheep was carried out to study identification of differentially expressed genes. Relative levels of transcription determined for individual molecules, were also characterized on basis of homology, gene ontology and pathway mapping. Total of 2,944,061 reads were generated and after removing low quality reads 2,178,136 (73.98%) sequences remained. These 2,178,136 reads fi-om the cDNA library of sheep Paramphistomes were then mapped with reference file data. Out of 11,593 sequences, 11,588 sequences were mapped and classified based on homology searches, protein motifs, gene ontology and biological pathway mapping. BLASTx manual annotation revealed 11,582 sequences displaying significant similarity to known genes in NCBI (99.94% of total P. cervi sequences). Species distribution of BLAST hits based on the BLASTx analysis showed that most of sequences matched to species of Trematode parasites, mainly to Clonorchis sinensis. Schistosoma mansoni, Schistosoma japonicum, Schistosoma haematobium, Fasciola hepatica. A significant proportion (56.6%) of the transcriptome of Paramphistomum cervi was inferred to encode 6,562 conserved protein domains or family signatures. Amongst the InterPro domains identified 'protein kinase-like domain' (130, 2.9%) 'Protein kinase, catalytic domain' (109, 2.4%), 'WD40/YVTN repeat-like-containing domain' (91. 2.0%), 'armadillo-like helical' (74. 1.6%), 'armadillo-type fold' (72.1.6%)) and WD40-repeat-containing domain' (69, 1.5%)) were most abundant. KEGG annotated a total of 1,926 (16.6%) sequences were fiirther classified into 108 pathways by using the pathway identification tool in Blast2G0. Total 14 parent pathways were identified and categorized into four types, with metabolic pathways being the most abundant, followed by genetic information processing, environmental processing and organismal systems. Based on the annotation, based on conserved domains and protein families, total 6,527 sequences (56.32%)) were assigned to GO term. GO annotation of the Paramphistomum cervi predicted peptides revealed 4,473 different GO teniis, which were assigned to three main GO categories of biological process (2,645 GO terms; 59.1%), cellular component (644 GO terms; 14.4%) and molecular function (1,184 GO terms; 26.5%). The integration of transcriptomic and proteomic datasets generated herein sets the scene for fiiture studies, aimed at exploring the potential roles of molecules which may play at the host-parasite interface and for estabhshing novel strategies for the treatment or control of parasitic fluke infections. Recent transcriptome analysis of Paramphistomum cervi has identified novel genes and genes not previously reported for Paramphistomes, as well as the identification of the molecular mechanisms for host-dependent maturation, immune evasion, development and signaling.