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ArticleItem Open Access A Serological Study on Porcine Parvovirus among Pigs in Chennai, Tamil Nadu(Haryana Vet., 2022-06) Lalrinkima; Hemalatha, S.; Nagarajan, K.; Raj, G. Dhinakar; Thangavelu, A.; Rao, G.V. Sudhakar; Balasubramanyam, D.; TANUVASArticleItem Open Access VIABILITY PCR TO DETECT THE MOST-PROBABLE-NUMBER OF VIABLE PROBIOTIC BACTERIA IN COMMERCIAL PREPARATIONS(TANUVAS, Chennai, 2022-01) Vasanthi, B.; Nirmala, K.; Tirumurugaan, K.G.; Rounak, Kumar; Alagumaruthanayagam, A.; Raj, G. Dhinakar; Raman, M.; TANUVASThe polymerase chain reaction (PCR) modification that enables molecular diagnosis and detection viable cells from diverse samples is based on the Viability PCR (V-PCR). In this study, we optimized V-PCR using a candidate L. plantarum probiotic strain and the technique performed efficiently in detecting live cells from an admixed suspension of live and dead cells. Application of the V-PCR on different probiotics strains (B. coagulans, L. plantarum and L. fermentum) also revealed a strong positive correlation in its performance across the strains tested even with an admixture of varied concentration of live and dead cells and different dilutions respectively (r=0.93 to 0.98). We obtained five and four commercial probiotics available for humans and animal use respectively from the market and tested for the recovery of total viable bacteria by agar pour plate method and also the count of viable bacteria by V-PCR. All commercial preparations when tested by the pour plate method recovered their listed viable counts except for the probiotic B and F, and C & G revealed lower counts (<1 to 2 log, and <3 to 4 log respectively) than the the manufacturerās claims.ArticleItem Open Access Antibiotic residue screening of milk samples using microbial inhibition assay: A preliminary study(2018) Sophila, J Ruth; Raj, G. Dhinakar; Kumanan, K.; Sarathchandra, G.; Vairamuthu, S.; TANUVASA preliminary study was conducted to detect the presence of antibiotic residue in milk samples collected in and around Chennai using the microbial inhibition assay with freeze dried vegetative form of Geobacillus stearothermophilus. All the samples were found to be negative for antibiotic residues. Before field testing, the LOD of the commonly used antibiotics namely Enrofloxacin, Ciprofloxacin, Neomycin and Oxytetracycline was determined to be 50, 25,100 and 25 ppb respectively This was much lower than the MRLs for each antibiotic indicating that it is a sensitive assay. The assay would be useful for screening bulk samples as a qualitative test before the confirmatory test using HPLC.ArticleItem Open Access A field study on the relationship between tick burden and deltamethrin resistance in Rhipicephalus microplus isolates of Indian household cattle(2021) Azhahianambi, P.; Harikrishnan, T.J.; Sagar, Sharath V; Rajeswaran, Abiraami; Raj, G. Dhinakar; Latha, Bhaskaran Ravi; TANUVASA total of 2809 ixodid ticks were collected by screening 4560 household cattle from six different agro-climatic zones of Tamil Nadu, a southern state of India. Among the ixodid ticks, Rhipicephalus microplus tick isolates were subjected to Adult Immersion Test (AIT) and Larval Packet Test (LPT) using deltamethrin. The mean % of ixodid tick infested cattle population was 63.2%Ā±10.9. Positive correlation was found between the tick burden on household cattle and Resistance Factor (R=0.66). Further, 44% variance in the tick burden on cattle is explained by LPT based resistance factor suggesting deltamethrin resistance level is one of the contributing factors to the tick load on cattle. Analysis of DNA sequence of sodium channel gene domain II-S4-5 linker region of all deltamethrin resistant R. microplus tick isolates, confirmed the absence of point mutation at the 190th nucleotide which suggests the possible mutation in other regions of sodium channel gene of R. microplus tick isolates of south India and/or alternate resistance mechanisms.ArticleItem Open Access Biosafety Concerns During the Collection, Transportation, and Processing of COVID-19 Samples for Diagnosis(2020) Karthik, K.; Aravindh Babu, RP; Dhama, K.; Chitra, M. Ananda; Kalaiselvi, G.; Senthilkumar, TMA; Raj, G. Dhinakar; TANUVASThe coronavirus disease 2019 (COVID-19) pandemic, which started in China, has created a panic among the general public and health care/laboratory workers. Thus far, there is no medication or vaccine to prevent and control the spread of COVID-19. As the virus is airborne and transmitted through droplets, there has been significant demand for face masks and other personal protective equipment to prevent the spread of infection. Health care and laboratory workers who come in close contact with infected people or material are at a high risk of infection. Therefore, robust biosafety measures are required at hospitals and laboratories to prevent the spread of COVID-19. Various diagnostic platforms including of serological, molecular and other advanced tools and techniques have been designed and developed for rapid detection of SARS-CoV-2 and each has its own merits and demerits. Molecular assays such as real-time reverse transcriptase polymerase chain reaction (rRT-PCR) has been used worldwide for diagnosis of COVID-19. Samples such as nasal swabs or oropharyngeal swabs are used for rRT-PCR. Laboratory acquired infection has been a significant problem worldwide, which has gained importance during the current pandemic as the samples for rRT-PCR may contain intact virus with serious threat. COVID-19 can spread to workers during the sampling, transportation, processing, and disposal of tested samples. Here, we present an overview on advances in diagnosis of COVID-19 and details the issues associated with biosafety procedures and potential safety precautions to be followed during collection, transportation, and processing of COVID-19 samples for laboratory diagnosis so as to avoid virus infection.ArticleItem Open Access Genome Sequencing and Comparative Genomics of Indian Isolates of Brucella melitensis(2021) Karthik, K.; Anbazhagan, S.; Thomas, P.; Chitra, M. Ananda; Senthilkumar, T.M.A.; Sridhar, R.; Raj, G. Dhinakar; TANUVASBrucella melitensis causes small ruminant brucellosis and a zoonotic pathogen prevalent worldwide. Whole genome phylogeny of all available B. melitensis genomes (n = 355) revealed that all Indian isolates (n = 16) clustered in the East Mediterranean lineage except the ADMAS-GI strain. Pangenome analysis indicated the presence of limited accessory genomes with few clades showing specific gene presence/absence pattern. A total of 43 virulence genes were predicted in all the Indian strains of B. melitensis except 2007BM-1 (ricA and wbkA are absent). Multilocus sequence typing (MLST) analysis indicated all except one Indian strain (ADMAS-GI) falling into sequence type (ST 8). In comparison with MLST, core genome phylogeny indicated two major clusters (>70% bootstrap support values) among Indian strains. Clusters with <70% bootstrap support values represent strains with diverse evolutionary origins present among animal and human hosts. Genetic relatedness among animal (sheep and goats) and human strains with 100% bootstrap values shows its zoonotic transfer potentiality. SNP-based analysis indicated similar clustering to that of core genome phylogeny. Among the Indian strains, the highest number of unique SNPs (112 SNPs) were shared by a node that involved three strains from Tamil Nadu. The node SNPs involved several peptidase genes like U32, M16 inactive domain protein, clp protease family protein, and M23 family protein and mostly represented non-synonymous (NS) substitutions. Vaccination has been followed in several parts of the world to prevent small ruminant brucellosis but not in India. Comparison of Indian strains with vaccine strains showed that M5 is genetically closer to most of the Indian strains than Rev.1 strain. The presence of most of the virulence genes among all Indian strains and conserved core genome compositions suggest the use of any circulating strain/genotypes for the development of a vaccine candidate for small ruminant brucellosis in India.ArticleItem Open Access Immunochemical Localization Of Cross-Reactive Proteins Of Bladderworms with Special Reference to Detection of Specific Antigen in Cystic Echinococcosis of Sheep(TANUVAS, Chennai, 2021-03) Jeyathilakan, N.; Basith, S. Abdul; John, Lalitha; Chandran, N. Daniel Joy; Raj, G. Dhinakar; TANUVAS; ChappaImmunochemical localization of antigenic fractions of ovine bladder worms viz., hydatid cyst, Coenurus cerebralis and Cysticercus tenuicollis is important to identify cross reactive as well as specific proteins. SDS PAGE and western blot technique were carried out to analyze various portions of three ovine bladderworms and to identify the specific protein of cystic echinococcosis. SDS PAGE analysis of fluid antigens revealed 9,6 and 9 bands for hydatid cyst(HCFA),Coenurus cerebralis(CCFA)and Cysticercus tenuicollis(CTFA) respectively and 24, 38 and 68 kDa protein bands were common in HCFA and CCFA. The common protein band between HCFA and CTFA was identified as 28kDa.24 kDa protein which was common between CCFA and CTFA. Scolex antigen revealed 3,6 and 9 bands respectively for HPSA, CCSA and CTSA. The common protein bands between HPSA and CTSA were 29, 72 and 98 kDa. Protein bands 12, 42,98 and 112 kDa were common between CCSA and CTSA. Membrane antigen revealed 4, 4 and 6 bands respectively for HGMA, CCMA and CTMA. The common protein band between CCMA and CTMA was 16kDa only. Western blot analysis revealed that the low molecular weight protein 8 kDa from HCFA was specific for cystic echinococcosis. Cross reaction was noticed between HPSA and CTMA as well as between HGMA and CTMA.ArticleItem Open Access Establishment of Embryonic Stem Cell like Cell Colonies from In-vitro Produced Buffalo Compact Morulae(TANUVAS, Chennai, 2019-05) Satheshkumar, S.; Brindha, K.; ShaliniPriya, M.; Parthiban, M.; Palanisammi, A.; Raj, G. Dhinakar; TANUVASThe study was aimed at establishing the embryonic stem cells (ESC) like cell colonies from in-vitro produced buļ¬alo (Bubalus bubalis) compact morula (CM). A total of 33 CM were subjected for zonalysis using pronase and embryonic cells were derived by three different methods viz., T 1: Intact CM; T 2: Disaggregation of CM by gentle pipetting and T 3: Slicing of CM using Bard Parker blade.ArticleItem Open Access A PILOT STUDY ON CRYOPRESERVATION OF CANINE PLATELETS(TANUVAS, Chennai, 2017-09) Kumar, M. Ranjith; Selvaraj, P.; Prathaban, S.; Raj, G. Dhinakar; Maroudam, V.; TANUVASIn veterinary medicine, though the need for platelet transfusions are very high, given to the thrombocytopenic crisis occurring in many blood parasitic diseases and immune mediated diseases of dogs.
