Application of vaginal tampon as an alternative to nasal swabs for higher recovery of DNA from sheep and goats for PCR based diagnosis of bovine tuberculosis.

A study was conducted to find the applicability of vaginal tampons as an alternative to regular cotton swabs as a nasal secretion collection tool for the higher recovery of DNA. Nasal secretions were collected from sheep and goats using regular cotton swab and tampon swab. The mean yield and purity of the DNA extracted from tampon were significantly higher than that of the DNA extracted from cotton swab. The tampon swabs resulted higher DNA recovery than the cotton swabs after they were allowed to absorb M. bovis culture. The tampon swab was also found to be more sensitive in detecting M. bovis by PCR. This study concluded that vaginal tampons are having a higher absorption capacity with more DNA yield and can be used as a nasal swab in the diagnosis of bovine tuberculosis.

Temporal pattern and risk factors for occurrence of Canine Rabies in Chennai.

Rabies is one of the most important zoonoses resulting in a high case fatality rate in humans. Most of the human Rabies cases are due to dog bites which can be prevented by effective vaccination in dogs. Globally, epidemiological studies on understanding the seasonality and risk factors for occurrence in canines are limited. The present study aimed to understand the temporal pattern of Rabies occurrence in Chennai city of Tamil Nadu, India, and address the suggestive clinical signs for better clinical ante-mortem rabies diagnosis. Data of 598 suspected canine hippocampus brain smear samples with Seller's staining and/or FAT percent positivity of 71.57% (428/598) from March 2010 to February 2019 were included in this study. Cross-correlation between rabies cases and meteorological factors showed that maximum temperature (lag 15), morning relative humidity (lag 0 and lag 5) and evening relative humidity (lag 4) were significantly associated with rabies cases. Auto-Regressive Integrated Moving Average (ARIMA) model with exogenous variables (significant lags of meteorological variables) was used to fit the time series of canine Rabies in Chennai. In logistic regression analysis, the following risk factors were found to be playing a significant role in Rabies positivity viz., behavioural changes in dogs (P < 0.001), free-roaming, unprovoked biting, hyper salivation (P < 0.05), dog bite history and drop jaw (P < 0.01). Hence, the study results highlight the need for continuous surveillance of canine Rabies for devising and implementing future preventive strategies and is helpful to establish the above-identified risk factors as a criterion to help in clinical rabies diagnosis.

A Study on Insulin Levels and the Expression of Glut 4 in Streptozotocin (STZ) Induced Diabetic Rats Treated with Mustard Oil Diet.

The study was undertaken to evaluate the therapeutic effect of mustard oil incorporated diet in streptozotocin (STZ)-induced type 1 diabetic rats. Dietary composition has shown to play a significant role in improving insulin sensitivity. Various authors have reported the hypoglycemic effect of mustard oil in experimentally induced diabetic rats. In the present study, reverse transcriptase polymerase chain reaction (RT-PCR) was done to analyze the Glut 4 expression in STZ induced diabetic rats as it is a key player in glucose homeostasis. The effect of mustard oil on serum biochemical parameter and insulin levels was also studied. Twenty-four male Wistar rats were randomly divided into three different groups with each containing eight animals. The first, second and third groups were control, diabetic control and treatment group with mustard oil respectively. All the rats in respective groups were fed for 60 days with iso-caloric mash diet containing 8% lipid. Diabetes was induced by intra-peritoneal administration of STZ (40 mg/kg body weight). A highly significant reduction in blood glucose level, with an increase in insulin activity was observed in mustard oil-treated diabetic rats when compared to control group indicating anti-hyperglycemic activity of mustard oil. Mustard oil-treated diabetic rats showed increased expression of  Glut 4 in muscle tissue when compared to diabetic control. A significant reduction in the levels of triacylglycerols, total cholesterol, VLDL and LDL and raised plasma HDL were noticed in mustard oil-treated diabetic rats when compared to diabetic control rats. Histopathological studies revealed a mild regeneration of ß cells of pancreas in mustard oil-treated diabetic rats. The results from our investigation suggest that mustard oil elicits hypoglycemic effect by increased insulin activity and up-regulation of  Glut 4 gene expression in muscle tissue of STZ-induced diabetic rats.

Enhanced Bioavailability of Boswellic Acid by Piper longum: A Computational and Pharmacokinetic Study.

Chronic inflammation is a key culprit factor in the onset and progression of several diseases. Novel and pharmacologically effective therapeutic approaches are needed for new treatment remedy or improved pharmacokinetics and pharmacodynamics for existing synthetic drugs, in particular natural products. Boswellic acids are well-known natural products, with capacity to effectively retard inflammation without severe adverse effects. However, the therapeutic use of Boswellic acids are greatly hindered by its poor pharmacokinetic properties. Co-administration strategies that facilitate the oral absorption and distribution of Boswellic acids should lead to a safe and more effective use of this product prophylactically and therapeutically in inflammatory disorders. In this study, we examined the effect of Piper longum extract on the absorption and bioavailability of Boswellic acid in rabbits. In addition, we further explored computational pharmacodynamic interactions between Piper longum and Boswellic acid. Piper longum extract at 2.5 and 10 mg/kg, increased the bioavailability of Boswellic acid (p < 0.05). Based on our drug-based computational modeling, cytochrome P450 (CYP450)-mediated mechanism was involved in increased bioavailability. These findings confirmed that Piper longum with Boswellic acid may be administered orally together for effective therapeutic efficacy. Thus, our studies support the application of Piper longum with Boswellic acid as a novel therapeutic avenue in diseases associated with inflammation.

Avian parvovirus: classification, phylogeny, pathogenesis and diagnosis.

Poultry parvoviruses identified during the early 1980s are found worldwide in intestines from young birds with enteric disease syndromes as well as healthy birds. The chicken parvovirus (ChPV) and turkey parvovirus (TuPV) belong to the Aveparvovirus genus within the subfamily Parvovirinae. Poultry parvoviruses are small, non-enveloped, single-stranded DNA viruses consisting of three open reading frames, the first two encoding the non-structural protein (NS) and nuclear phosphoprotein (NP) and the third encoding the viral capsid proteins 1 (VP1 and VP2). In contrast to other parvoviruses, the VP1-unique region does not contain the phospholipase A2 sequence motif. Recent experimental studies suggested the parvoviruses to be the candidate pathogens in cases of enteric disease syndrome. Current diagnostic methods for poultry parvovirus detection include PCR, real-time PCR, enzyme linked immunosorbent assay using recombinant VP2 or VP1 capsid proteins. Moreover, sequence-independent amplification techniques combined with next-generation sequencing platforms have allowed rapid and simultaneous detection of the parvovirus from affected and healthy birds. There is no commercial vaccine; hence, the development of an effective vaccine to control the spread of infection should be of primary importance. This review presents the current knowledge on poultry parvoviruses with emphasis on taxonomy, phylogenetic relationship, genomic analysis, epidemiology, pathogenesis and diagnostic methods.

Molecular Characterization of Thermostable Newcastle disease virus Isolated from Pigeon.

The HN and L gene sequences of an Indian isolate of Newcastle disease virus was analyzed prior to and after exposure to 56 °C at tenth passage and fifteenth passage to study the variations at molecular level. In the HN gene sequence of progenitor and thermostable strain, substitution of K373I, F374L, M516R, D517V were considered to contribute to the increase in the stability of the protein. In the L gene of the thermostable strain, variations were observed at many positions and among these the substitutions at position P675H K677R, K893D, R1132K, had charged amino acids, and at L656A, F657V, F869L, T886I, M899I, G1131V, V1675L, had hydrophobic amino acids that could be related to increased stability of L protein at high temperatures. The changes in amino acid sequence in HN and L gene of the thermostable strain might render structural variations that might have contributed to the stability of the strain at higher temperature.

Analysis of the Fusion Protein Cleavage Site of Newcastle disease virus Isolates from India Reveals Preliminary Evidence for the Existence of II, VI and VII Genotypes.

Newcastle disease virus (NDV) has been a threat to poultry industry in most of the developing countries with a wide variety of avian species being susceptible, coupled with the presence of mobile wild bird reservoirs contributing not only to the vast genomic diversity of this virus but also to the diagnostic failures. NDV of multiple genotypes (I-XI) is known to be prevalent and reported worldwide. However, there is a paucity of information on the circulating genotypes of NDV in India. This study utilized the fusion protein cleavage site (FPCS) sequence to determine the different genotypes of NDV circulating in India. Our results indicate that majority of NDV isolates from southern states of India namely, Tamil Nadu, Kerala and Karnataka were found to belong to genotype II. However, some of the strains from Tamil Nadu and most from Uttar Pradesh belong to genotype groups VI and VII. Interestingly, three isolates recovered from Tamil Nadu grouped with genotype IV viruses (namely Herts/33) which had not been hitherto reported to GenBank since 1989. This preliminary information points to the existence of multiple genotypes and also the need for efficacy studies with vaccines incorporating multiple genotypes in controlling virulent NDV (vNDV) outbreaks in India.

Pathotyping of a Newcastle disease virus isolated from peacock (Pavo cristatus).

This report describes Newcastle disease in peacock and the isolation and characterization of the virus. The virus had an intracerbral pathogenicity index of 1.71 and mean death time of 47 h. The isolate had multiple basic amino acids at the fusion protein cleavage site sequence ((110)GGRRQRRFIG(119)) with a phenylalanine at residue 117. Biological and molecular characterization revealed that the virus is velogenic. Phylogenetic analysis placed the isolate in genotype II.

Pathotyping of Newcastle disease virus isolates from pet birds.

Four Newcastle disease virus (NDV) isolates obtained from a pigeon, lory, parrot, and love bird were subjected to biological and molecular characterization. All the isolates were identified as velogenic with intracerebral pathogenicity indices (ICPI) of 1.9-2.0. All the isolates had a 112RRQKRF117 motif in the fusion protein cleavage site (FPCS), typical for pathogenic NDV. Phylogenetic analysis placed the isolates along with a velogenic Indian isolate of Cl group recovered during 1987.

Biological and molecular characterization of Indian isolates of Newcastle disease virus from pigeons.

Five Newcastle disease virus (NDV) isolates from pigeons were characterized by biological and molecular methods. Four of the five isolates were found to be velogenic with high intracerebral pathogenicity indices (ICPI). The fusion protein cleavage site (FPCS) sequences of these isolates had multiple basic amino acids RRQKRF at positions 112-116 and a phenyl alanine at position 117 characteristic of velogenic isolates. Three of these velogenic isolates were phylogenetically related to mesogenic vaccine virus strain and the fourth one to a few exotic velogenic isolates. The lentogenic isolate obtained in this study was identical with the LaSota strain.

ANTIMICROBIAL SCREENING OF LEAVES OF Memecylon umbellatum.

The present study includes antimicrobial activity of Memecylon umbellatum Alcoholic extract showed maximum antibacterial activity Staphylococcus aureus. (gram positive) and it also showed antibacterial activity against gram negative bacteria and also alcoholic extract alone showed slight antifungal activity.

Characterisation of Newcastle disease viruses isolated in India.

Eleven Newcastle disease virus (NDV) isolates obtained from outbreaks of disease in chickens (9) and Japanese quail (2) in Tamil Nadu, India were characterised in pathogenicity tests, antigenically, using mouse monoclonal antibodies (MAbs), and other established tests devised to distinguish between different strains. All 11 isolates were shown to be highly virulent for chickens. In indirect immunoperoxidase tests used to assess the ability of a panel of 28 MAbs to bind to infected cell cultures, 10 of the isolates showed an identical reaction pattern, the other isolate (No. 4) failed to react with one MAb which bound to cells infected with the other isolates. Isolates 9 was unstable at pH 3 while the other 10 were stable. All other properties were shared by the 11 isolates.