Pathogenic and Genotypic Characterization of a Japanese Encephalitis Virus Isolate Associated with Reproductive Failure in an Indian Pig Herd

Background

India is endemic to Japanese encephalitis virus (JEV) and recurrent outbreaks occur mainly in rice growing areas. Pigs are considered to be the amplifying host for JEV and infection in gestating pigs results in reproductive failure. Most studies conducted on JEV infection in Indian pigs have been serological surveys and very little is known about JEV genotypes circulating in pigs. So the potential risk posed by pigs in JEV transmission and the genetic relationship between viruses circulating in pigs, mosquitoes and humans is poorly understood.

Methodology/Principal Findings

This study was conducted in pigs with a history of reproductive failure characterized by stillborn piglets with neuropathological lesions. Japanese encephalitis (JE) suspected brain specimens inoculated intracerebrally into mice and Vero cells resulted in successful isolation of JEV/SW/IVRI/395A/2014. Clinicopathological observations in infected mice, demonstration of JEV antigen in brain, and analysis of the envelope protein identified the swine isolate as being neurovirulent. Phylogenetic analysis based on prM and E gene sequences showed that it belonged to genotype III. This swine isolate was closely related to JEV associated with the 2005 outbreak in India and JaoArS982 from Japan. Phylogenetic analysis of JEV strains collected between 1956 and 2014 in India categorized the GIII viruses into different clades blurring their spatial distribution, which has been discernible in the previous century.

Conclusions/Significance

Isolation of JEV from stillborn piglets and its close genetic relationship with viruses detected at least three decades ago in humans and mosquitoes in Japan suggests that the virus may have been circulating among Indian pigs for several decades. The close similarity between the present swine isolate and those detected in humans affected in the 2005 outbreak in Uttar Pradesh, India, suggests the need for more intensive surveillance of pigs and implementation of suitable strategies to control JE in India.     

The mycoflora and incidence of aflatoxin,zearalenone and sterigmatocystin in dairy feed and forage samples from Eastern India and Bangladesh

The mycoflora of a range of mixed ruminant feed and forage samples collected in Eastern India and Bangladesh were determined. Most fungi isolated from the samples were those associated with low moisture contents. Several potentially mycotoxigenic fungi and thermotolerant species were present particularly in the rice straw. The most frequently isolated fungi were: Aspergillus versicolor (Vuill.) Tiraboschi, Penicillium citrinum Thorn, Eurotium species, Wallemia sebi (Fr.) v. Arx, Aspergillus penicilloides Speg. and yeasts. Fusarium moniliforme Sheldon and Cladosporium species were prevalent in straw but not other feeds. Levels of certain individual fungi, particularly thermotolerant types, were higher in Bangladesh rice straw although overall counts were similar to those of the Indian samples. Mycotoxin analyses for aflatoxins, zearalenone and sterigmatocystin were carried out. Aflatoxins were detected in significant amounts in some feed samples from Eastern India but not in rice straw from India and Bangladesh. Zearalenone occurred in a Paspalum straw sample and three other feed samples from India and sterigmatocystin was detected in three rice straw samples from Bangladesh.     

Classical autophagy proteins LC3B and ATG4B facilitate melanosome movement on cytoskeletal tracks

Macroautophagy/autophagy is a dynamic and inducible catabolic process that responds to a variety of hormonal and environmental cues. Recent studies highlight the interplay of this central pathway in a variety of pathophysiological diseases. Although defective autophagy is implicated in melanocyte proliferation and pigmentary disorders, the mechanistic relationship between the 2 pathways has not been elucidated. In this study, we show that autophagic proteins LC3B and ATG4B mediate melanosome trafficking on cytoskeletal tracks. While studying melanogenesis, we observed spatial segregation of LC3B-labeled melanosomes with preferential absence at the dendritic ends of melanocytes. This LC3B labeling of melanosomes did not impact the steady-state levels of these organelles but instead facilitated their intracellular positioning. Melanosomes primarily traverse on microtubule and actin cytoskeletal tracks and our studies reveal that LC3B enables the assembly of microtubule translocon complex. At the microtubule-actin crossover junction, ATG4B detaches LC3B from melanosomal membranes by enzymatic delipidation. Further, by live-imaging we show that melanosomes transferred to keratinocytes lack melanocyte-specific LC3B. Our study thus elucidates a new role for autophagy proteins in directing melanosome movement and reveal the unconventional use of these proteins in cellular trafficking pathways. Such crosstalk between the central cellular function and housekeeping pathway may be a crucial mechanism to balance melanocyte bioenergetics and homeostasis.     

Synergistic growth of Bacillus and Pseudomonas and its degradation potential on pretreated polypropylene

This study investigated biodegradation of physically pretreated polypropylene (PP) by using two different combinations of microorganisms, namely, Bacillus flexus + Pseudomonas azotoformans(B1) and B. flexus + B. subtilis(B2), for a period of 12 months. The growth rate of (B1) was found to be high throughout the study period, and reached a maximum of 1 × 10¹⁴ colony-forming units (CFU)/mL. At the end of the experiment, the polymers become hydrophilic. Carbonyl indices showed that ultraviolet (UV)-treated polymers started degrading faster than the thermally treated PP. The thermogravimetric analysis also revealed that UV-treated PP exposed to the B. flexus + P. azotoformans combination for 1 year exhibited maximum degradation (22.7%). The gravimetric weight loss method showed 1.95% weight loss followed by 1.45% with B. flexus + B. subtilis. The changes in the carbonyl indices of the polymer through Fourier-transform infrared (FTIR) analysis also support the degradation.     

Association between Milk and Milk Product Consumption and Anthropometric Measures in Adult Men and Women in India: A Cross-Sectional Study

Background

The nutritional aetiology of obesity remains unclear, especially with regard to the role of dairy products in developing countries.

Objective

To examine whether milk/milk product consumption is associated with obesity and high waist circumference among adult Indians.

Methods

Information on plain milk, tea, curd and buttermilk/lassi consumption assessed using a Food Frequency Questionnaire was obtained from the cross-sectional sib-pair designed Indian Migration Study (3698 men and 2659 women), conducted at four factory locations across north, central and south India. The anthropometric measures included were Body Mass Index (BMI) and Waist Circumference (WC). Mixed-effect logistic regression models were conducted to accommodate sib-pair design and adjust for potential confounders.

Results

After controlling for potential confounders, the risk of being obese (BMI≥25 kg/m²) was lower among women (OR = 0.57;95%CI:0.43−0.76;p≤0.0001) and men (OR = 0.67;95%CI: 0.51−0.87;p = 0.005), and the risk of a high WC (men: >90 cm; women: >80 cm) was lower among men (OR = 0.71;95%CI:0.54−0.93;p = 0.005) and women (OR = 0.79;95%CI:0.59−1.05;p>0.05) who consume ≥1 portions of plain milk daily than those who do not consume any milk. The inverse association between daily plain milk consumption and obesity was also confirmed in sibling-pair analyses. Daily tea consumption of ≥1 portion was associated with obesity (OR = 1.51;95%CI:1.00−2.25;p>0.050) and high WC (OR = 1.65;95%CI:1.08−2.51;p>0.019) among men but not among women but there was no strong evidence of association of curd and buttermilk/lassi consumption with obesity and high waist circumference among both men and women.

Conclusions

The independent, inverse association of daily plain milk consumption with the risk of being obese suggests that high plain milk intake may lower the risk of obesity in adult Indians. However, this is an observational finding and uncontrolled confounding cannot be excluded as an explanation for the association. Therefore, confirmatory studies are needed to clarify this relationship.     

Molecular Characterization of Brinjal (Solanum melongena L) Cultivars using RAPD and ISSR Markers

Molecular characterization of 19 advanced cultivars and landraces of brinjal was carried out using RAPD and ISSR markers. Twenty-nine RAPD primers generated a total of 240 amplified fragments, while 23 anchored and non-anchored ISSR primers produced 299 fragments. Of these, 66 (27.5%) RAPD and 56 (18.73%) ISSR fragments were polymorphic. All the cultivars could be distinguished based on RAPD and/or ISSR profiles. A set of two RAPD primers, OPW 11 and OPX 07, was adequate to distinguish all the 19 cultivars. On the other hand, a minimum of ten ISSR primers were required to achieve the same result. Eleven cultivars could be identified by the unique presence or absence of one to four markers. The correlation between primer Rp and the number of cultivars distinguished by RAPD was r = 0.873, while that for ISSR it was r = 0.327. The correlation between PIC of primer and the number of cultivars distinguished was r = 0.324 for RAPD, while for ISSR primers it was r = ? 0.066. The probability of chance identity between two cultivars for RAPD and ISSR markers was calculated as 8.94×10^?4 and 2.25×10^?2, respectively. The average Jaccard’s similarity coefficient between cultivars based on combined RAPD and ISSR data was estimated to be 0.919. The UPGMA analysis grouped the cultivars into three main clusters with significant bootstrap support. While the cultivars bred at Indian Agricultural Research Institute, New Delhi formed one sub-cluster; others did not show a prominent region-based clustering.