Browsing by Author "Meenambigai, TV"
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ArticleItem Open Access Assessment of nano selenium effect in developing zebra fish embryos(2019) Vaishnavi, AS; Mangala Gowri, A; Valli, C; Meenambigai, TV; Baskaran, D; TANUVASNano particles explicitly need to be non toxic not only for biomedical applications but for other areas that ends up in environmental exposures. Various platforms are being used from in vitro cell culture to higher vertebrate models. However, small efficient, more informative but cost effective well established Zebrafish models are the choice for preclinical studies due to the advantage of sharing not less than 85 percent homology with human genome biology. Systematic vision gained by toxicity assessment of food using various models from in vitro cell-based assays, biochemical assays, in vivo animal to clinical settings paved way for a better food safety. The present study was designed to find out the comparison and toxicity between Nano Selenium (NaSe) and Inorganic Selenium (Inorg Se) on the embryonic development of Zebrafish. Different concentration of dietary Se was used on developing Zebra fish embryos. The results indicated that with the increased NaSe and Inorg Se concentration, different observable deformities such as growth retardation, shrinkage of chorion, yolk sac edema, lack of pigmentation; tail deformities and scoliosis in developing embryos were observed. The NaSe at 0.1mg/L concentration showed the highest hatchability and healthy embryo development. This study is conducted to identify the optimal NaSe concentration that could be used for food fortification through supplementation in developing zebra fish embryos.ArticleItem Open Access Classical swine fever virus vaccines: An updated review(2021) Rathnapraba, S.; Ramesh, A.; Meenambigai, TV; TANUVASClassical swine fever (CSF) is one of the most devastating viral infectious disease affecting the members of Suidae family, which causes a severe impact on the global economy. The CSF aetiological agent is the Classical Swine Fever Virus (CSFV) an enveloped, single, positive stranded RNA virus belonging to Pestivirus genus and Flaviviridae family. The disease is notifiable to the World Organization for Animal Health (OIE) due to its enormous consequences on porcine health and the pig industry. Response policy to notification of an outbreak differs among different countries. In CSF endemic countries a mass vaccination approach is adopted for the disease control. Further, the negative aspects related with mass stamping out policies, including elevated costs and ethical issues, point out vaccination as the main control measure against future outbreaks. Thus, the current review enlightens information to acquire a clear understanding about the status quo regarding the vaccination against the disease in endemic areas.ArticleItem Open Access Cloning and expression of Vp2 protein of infectious bursal disease virus(2020) Kumari, C Sworna; Meenambigai, TV; Mangalagowri, A.; Palanisamy, A.; Sarathchandra, G.; TANUVASThe VP2 protein of highly virulent infectious bursal disease virus (hvIBDV) was cloned and expressed in pRSET A vector with high level of protein expression. VP2 is the host protective protein with major neutralizing epitopes was cloned and expressed in E. coli expression system BL21- DE3. The protein expression was evaluated by induction studies with IPTg and characterized by SDS PAGE and western blotting analysis. Thus, the present study focusing on expression of the immunodominant VP2 protein that may be used as a protein boost in DNA vaccine production strategy for enhanced immunity against IBDV infection in chickens in the near future.ArticleItem Open Access COMPARATIVE ANALYSIS OF MICROSCOPIC AGGLUTINATION TEST AND IgM ELISA IN THE SERODIAGNOSIS OF LEPTOSPIROSIS(2005) Meenambigai, TV; Govindarajan, R; Jayakumar, V; Koteeswaran, A; TANUVASMicroscopic agglutination test (MAT) and IgM enzyme linked immunosorbent assay (ELISA) are the two tests currently employed in the serological diagnosis of leptospirosis. Under certain instances the samples positive by IgM ELISA in other laboratories were found to be negative in our laboratory by MAT and vice-versa. This prompted us to conduct a comparative study. The IgM ELISA was compared with the standard MAT as a method for detecting antibodies in humans against pathogenic Leptospira interrogans serovars.ArticleItem Open Access COMPARATIVE EFFICACY OF COMPETITIVE ELISA OVER AGID FOR DETECTION OF BLUETONGUE VIRUS ANTIBODIES(2005-01) Prabhakar, TG; Meenambigai, TV; Govindarajan, R; Daniel Joy Chandran, N; Koteeswaran, A; TANUVASThe disease caused by bluetongue virus (BTV), a prototype of Orbivirus genus of the family Reoviridae, is endemic in certain pockets of southern districts of Tamil Nadu. It acts as a barrier to the movement of sheep and goats within the districts of Tamil Nadu and also to other neighbouring states. The most widely used assay for the detection of BTV infection is agar gel immuno diffusion (AGID) test (Jochim and Chow, 1969; Stott and Osburn, 1983; Abu Elzein, 1984).ArticleItem Open Access Detection of Bluetongue Virus in Ovines by Nested Polymerase Chain Reaction(2017-07) Meenambigai, TV; Sworna Kumari, C; Balakrishnan, G; Shoba, K; Ganesan, PI; TANUVASBluetongue is a vector borne viral disease and its causative agent is Blue tongue virus (BTV). Detection of bluetongue virus by nested polymerase chain reaction (PCR) from the clinical samples provides a valuable tool to study the epidemiology of BTV infection in susceptible livestock. The present study reports detection of bluetongue virus in blood samples of sheep from different parts of Tamil Nadu assayed by nested polymerase chain reaction using two pairs of oligonucleotide primers designed from non structural protein 1 -NS1 Gene.ArticleItem Open Access DIAGNOSIS OF BOVINE LEPTOSPIROSIS BY 16s rRNA BASED POLYMERASE CHAIN REACTION(2014) Balakrishnan, G; Meenambigai, TV; Roy, Parimal; TANUVASThe present study was undertaken to diagnose acute case of bovine leptospirosis by PCR. A total of 115 serum samples collected from clinically suspected (33 cattle and 32 buffaloes) and apparently healthy (50) buffaloes were subjected to PCR. Among the clinically suspected animals, the PCR detected leptospiral DNA in 21 (32.31 per cent) samples. The clinical signs in these positive cases included abortion, repeat breeders, jaundice and haemorrhagic mastitis. Apparently healthy animals also gave positive results in PCR (6.00 per cent) which might be due to subclinical leptospirosis.ArticleItem Open Access DIFFERENTIAL METHODS FOR ANALYZING THE OUTER MEMBRANE PROTEINS OF PATHOGENIC SEROGROUPS OF LEPTOSPIRA(2015) Meenambigai, TV; Ravikumar, G; Balakrishnan, G; Monashree, R; TANUVASThe leptospiral outer membrane has a relatively complex protein profile Pathogenic mechanisms of Leptospira interrogans, the causal agent of leptospirosis, remain largely unknown. The outer membrane of pathogenic Leptospira species grown in culture media contains lipopolysaccharide (LPS), a porin (OMPL), and several lipoproteins, including LipL36 and LipL41. Loa22 was shown to be a protein having a C-terminal OmpA consensus domain. Loa22 was detected among pathogenic leptospires but not among nonpathogenic leptospires, suggesting the possible involvement of this protein in virulence. Loa22 is located in the outer membrane and a small portionis exposed on the cell surface. Thus, Loa22 may be a candidate for a novel vaccine against infection with pathogenic leptospires. The purpose of the study was to analyze the outer membrane proteins of pathogenic serogroups of leptospira by different methods whole cell solubilization, insoluble method and detergent method to identify the best method for extraction of proteins. Results of this study suggest that whole cell solubilization method was found promising for outer membrane protein extraction.ArticleItem Open Access Dissolving microneedles in drug delivery: An outlook(2020) Meenambigai, TV; Kumari, C Sworna; Sarathchandra, G.; Reddy, YKM; TANUVASDissolving microneedle (DMN) are micron sized needle that delivers the drug with painless penetration, excellent therapeutic efficacy, and with relative safety. It has an innovative transdermal delivery system with attractive scientific and industrial interests as oligonucleotide delivery, vaccine delivery and insulin delivery. Dissolving microneedles have a variety of representative biomedical applications such as disease diagnosis and treatment, immunobiological administration, and cosmetology. It holds a great promise for biomedical applications. The wearable device based dissolving microneedle patches will be desirable to integrate disease diagnosis and treatment in future.ArticleItem Open Access Evaluation of enriched canine hematopoietic progenitor cell colony forming potential(2018) Nandhini, S.; Mangalagowri, A.; Indhumathi, S.; Vaishnavi, AS; Meenambigai, TV; Bharanidharan, GR; TANUVAS; NandhiniHematopoietic stem cells (HSCs) are the well characterized adult stem cell that has been accepted for autologous cell therapy after minimal manipulation. The clinical use of HSCs to treat a variety of human and animal disorders has made them a key building block in the foundation of regenerative medicine. They represent a potential source of adult stem cells for transplantation in Veterinary Medicine. In the present study, Canine peripheral blood from apparently healthy animals were subjected for progenitor cells separation using an enrichment protocol and subjected for colony formation cell assay. The standardized simple protocol for progenitor cell enrichment and CFC assay would be applicable in clinical autologous cell therapy and can be used to analyze effects of drugs and biomolecules that induce erythropoiesis in canines as a model applicable in analysis of cell based clinical therapy.ThesisItem Restricted EXPRESSING THE INFLUENCE OF EPIDERMAL GROWTH FACTOR AND LEUKEMIA INHIBITORY FACTOR IN THE DEVELOPMENT OF BOVINE PREIMPLANTATION EMBRYOS(2018) Vinayak, Sadekar Gautami; Meenambigai, TV; Mangalagowri, A; Reena, D; TANUVASAn array of growth factors and cytokines are associated in the development of embryos under in vivo and in vitro conditions. These growth factors and cytokines have autocrine and paracrine cell-cell signaling actions that contribute to yielding healthier embryos. Their actions in the embryo include modulation of cell gene expression and metabolic function that in turn influence cell survival and differentiation, ultimately impacting embryo implantation competence and post-implantation development. The effect of Epidermal Growth Factor (EGF) and Leukemia Inhibitory Factor (LIF) in the development of IVM/IVF bovine oocytes /embryos and their gene expression analysis were investigated in this study.ArticleItem Open Access EXTRACTION AND AMPLIFICATION OF LEPTOSPIRAL DNA FROM KIDNEY TISSUES OF RAT(Indian Veterinary Association, 2007) Meenambigai, TV; Meena, S; Govindarajan, R; Jayakumar, Vajiravelu; Samuel, RM; Koteeswaran, A; TANUVASMost human outbreaks of leptospirosis are confined to developing tropical and subtropical countries (Faine et al. 1999). Reservoirs of infection include chronically infected wild and domesticated carrier animals. Transmission is effected directly from contaminated urine or indirectly by entry from environments that permit leptospiral survival. Rodents contributes for 60% of transmission of the disease (Ratnam et aI., 1987). Maintenance of leptospires in chronically infected carrier animals makes the control of the disease difficult and results in significant economic losses in cattle, pigs and horses (Donahue, 2001). isolation of leptospira is a long term process. For early diagnosis, in the present study the usefulness of PCR in amplification of leptospiral DNA from kidney tissues of rat employing two methods of DNA extraction is reported.ArticleItem Open Access GROWTH AND ADAPTATION OF LEPTOSPIRES IN LOW PROTEIN MEDIUM(2015) Balakrishnan, G; Meenambigai, TV; Roy, Parimal; TANUVASThe present study was undertaken to grow leptospires from high protein (20 per cent BSA) to protein free media (without BSA),which revealed growth at the concentration of 20 per cent to one per cent BSA on the 4th day of incubation. No growth could be observed below one per cent level of BSA. Hence, the leptospires were adapted to grow from high protein (10 per cent) to low protein (upto 0.2 per cent) concentration of BSA. In this adaptation process, growth in low protein medium containing 0.2 per cent BSA was observed to a concentration of >1.4 x 109 organisms / ml by day 10 and 11. Hence, the low protein medium containing 0.2 per cent BSA can be used for growing leptospires for vaccine preparation so as to minimize the adverse effects of protein containing media.ThesisItem Restricted HEMATOPOIETIC PROGENITOR CELL CHARACTERIZATION IN CANINES(2018) Nandhini, S; Mangala Gowri, A; Meenambigai, TV; Baranidharan, GR; TANUVASBlood cells are responsible for constant maintenance and immune protection of every cell type of the body and this relentless and brutal work requires cells that have the greatest powers of self-renewal and are designated as Hematopoietic progenitor cells (HPCs). Peripheral blood stem cells in circulation have become the most common source of hematopoietic stem cells intended for transplantation after minimal manipulation. In vitro and in vivo animal studies have shown that the enriched CD34+marrow cells of different species can give rise to the multiple blood cell lineages and may provide long-term hematopoiesis, suggesting that CD34 could be regarded as a cell surface marker of primitive hematopoietic stem cells. This has made it possible to enrich HSCs from mice and human beings for molecular characterization and transplantation purposes. Homeobox (Hox), sonic hedgehog (SHH), and Wingless-type MMTV integration site family (Wnt) are known to modulate the self-renewal and expansion of hematopoietic progenitor/stem cells in humans and mice. Unlike cytokines, Hox, SHH, and Wnt are highly conserved among species from flies to humans but studies regarding the self-renewal and expansion of the HSC are extremely limited in dogs. Hence in the present research, a rapid CD34+population enrichment and immuno separation protocol has been standardized in canine blood. The expression of signaling molecules for self renewal and colony forming potential of the isolated CD34+ population of cells from Canine blood showed that they were cells with proliferation and self renewal potential.PresentationItem Open Access In-vitro and in-vivo TLR-7 mRNA levels in response to lentogenic and velogenic pathotypes of Avian Avulavirus-1(2020-02) Chinju, C; Rajasekaran, Ranjani; Balakrishnan, G; Meenambigai, TV; John Kirubaharan, J; TANUVASTranscriptional studies in response to Avian Avulavirus-1 (AAv1) have gained importance in the recent past and have provided insight towards disease pathogenesis in chickens. The present study was focused towards elucidation of TLR-7 mRNA levels in response to D58 (lentogenic vaccine strain) and D162 (velogenic isolate) isolates of AAv1 in chicken embryo fibroblast cells (CEF) and chicken spleen at 1, 2, 3, 4 and 5dpi. The mRNA levels were determined by real-time PCR using SYBR-green chemistry. The TLR-7 mRNA levels were significantly up regulated to 18.90+0.03 in CEF cells and 12.33+0.02 in chicken spleen in response to lentogenic D58 at 1dpi. Similarly, in response to velogenic D162, TLR-7 mRNA levels were significantlyunregulated to 52.40+0.03 in CEF cells and 34.62+0.02 in chicken spleen. Later, in both CEF cells and chicken spleen, the mRNA levels of TLR-7 gradually declined from 2dpi in response to both lentogenic D58 and velogenic D162. TLR-7 has been reported to be a protein that plays specific role in detecting single stranded viral components and stimulating pro-inflammatory and anti-viral immune response. In the present study, the significantly up regulated levels of TLR-7 mRNA at 1dpi confirms the fact that TLR-7 levels were induced so as to stimulate necessary pro-inflammatory and anti-viral immune response against AAv-1. Further, it was observed that the virulence of AAv-1 pathotype also affected TLR-7 mRNA levels. This shows that TLR-7 mRNA levels vary with varying virulence of AAv1.ArticleItem Open Access Insights into veterinary vaccinology: Bygone and Future(2021) Meenambigai, TV; Anbukumar, K.; Madhanmohan, M.; TANUVASDevelopment of modern veterinary vaccines are challenging as against conventional vaccines. In the current era of genomics, intense progress has been made in the field of vaccinology and next generation vaccine approaches. Modern approaches in veterinary vaccinology greatly demands novel strategies for controlling emerging pathogens in livestock. This review corrugates a birds eye view of live attenuated vaccines, inactivated vaccines, DNA vaccines, subunit vaccines and genetically engineered vaccines. Reverse vaccinology approaches paves a new beginning for development of novel vaccines against infectious pathogens affecting the livestock and poultry sector. This conceptual framework for veterinary vaccinology leads to the rational development of next generation vaccines.ArticleItem Open Access Methodologies for cloning and expression of immunodominant protein LipL32 of pathogenic Leptospira(2020) Meenambigai, TV; Kumari, C Sworna; Balakrishnan, G.; TANUVASPathogenic serovars of Leptospira have wide antigenic diversity, which is mainly attributed to the lipopolysaccharide which is present in the outer membrane. The most common surface exposed immunodominant lipopolysaccharide protein is LipL32. In the present study the highly conserved surface exposed LipL32 protein was cloned and expressed in pRSET B vector. The His-tagged protein was purified and characterized by western blot analysis. The developed LiPL32 recombinant protein will serve as a novel diagnostic candidate for effective diagnosis of pathogenic forms of Leptospira in animals.ArticleItem Open Access Molecular Detection of Field Isolates of Infectious Bursal Disease Virus Adapted to African Green Monkey Kidney Cells(2017-11) Meenambigai, TV; Sworna Kumari, C; Balakrishnan, G; Shoba, K; Kumanan, K; TANUVASInfectious Bursal Disease (IBD) is an acute, highly infectious viral disease of young chicken. In the present study molecular detection of Infectious bursal disease virus was carried out from field samples (Bursa). A total 65 field samples were collected from different districts of Tamil Nadu and processed for detection by Reverse Transcriptase polymerase chain reaction (RT-PCR). The hypervariable region of VP2 gene encoding the neutralizing epitopes (633bp) was amplified. The very virulent samples were characterized by infecting in African green monkey kidney cells (Vero) upto 5 passages. The adaptation studies are carried out up to 120 hours to monitor the cytopathogenic effects (CPE) RT-PCR was carried out on cell culture adapted IBDV of every passage. The TCID50 per ml of the adapted virus at 5th passage was 1.58 x107.These findings concluded that the RT-PCR amplification of VP2 gene on bursal samples and characterization studies in African green monkey kidney cells would serve as a valuable tool for isolation and identification of IBDV for development of novel vaccines in future.