Natural Rabies Infection in a Domestic Fowl (Gallus domesticus): A Report from India

Background

Rabies is a fatal encephalitis caused by viruses belonging to the genus Lyssavirus of the family Rhabdoviridae. It is a viral disease primarily affecting mammals, though all warm blooded animals are susceptible. Experimental rabies virus infection in birds has been reported, but naturally occurring infection of birds has been documented very rarely.

Principal Findings

The carcass of a domestic fowl (Gallus domesticus), which had been bitten by a stray dog one month back, was brought to the rabies diagnostic laboratory. A necropsy was performed and the brain tissue obtained was subjected to laboratory tests for rabies. The brain tissue was positive for rabies viral antigens by fluorescent antibody test (FAT) confirming a diagnosis of rabies. Phylogenetic analysis based on nucleoprotein gene sequencing revealed that the rabies virus strain from the domestic fowl belonged to a distinct and relatively rare Indian subcontinent lineage.

Significance

This case of naturally acquired rabies infection in a bird species, Gallus domesticus, being reported for the first time in India, was identified from an area which has a significant stray dog population and is highly endemic for canine rabies. It indicates that spill over of infection even to an unusual host is possible in highly endemic areas. Lack of any clinical signs, and fewer opportunities for diagnostic laboratory testing of suspected rabies in birds, may be the reason for disease in these species being undiagnosed and probably under-reported. Butchering and handling of rabies virus- infected poultry may pose a potential exposure risk.     

The Cdk5 Kinase Downregulates ATP-Gated Ionotropic P2X<Subscript>3</Subscript> Receptor Function Via Serine Phosphorylation

Cdk5 is an endogenous kinase activated by the neuronal-specific protein p35 and implicated in multiple neuronal functions, including modulation of certain pain responses. We investigated whether Cdk5 could regulate ATP-gated P2X₃ receptors that are members of the family of membrane proteins expressed by sensory neurons to transduce nociception in baseline and chronic pain. To study the potential P2X₃ receptor modulation by Cdk5, we co-transfected rat P2X₃ receptors and Cdk5 into HEK cells and observed increased P2X₃ receptor serine phosphorylation together with downregulation of receptor currents only when these genes were transfected together with the gene of the Cdk5 activator p35. The changes in receptor responses were limited to depressed current amplitude as desensitization and recovery were not altered. Transfection of p35 with P2X₃ similarly downregulated receptor responses, suggesting that this phenomenon could be observed even with constitutive Cdk5. The present data indicate a novel target to express the action of Cdk5 on membrane proteins involved in pain perception.     

Variability of United States Isolates of Macrophomina phaseolina Based on Simple Sequence Repeats and Cross Genus Transferability to Related Genera Within Botryosphaeriaceae

Twelve simple sequence repeat (SSRs) loci were used to evaluate genetic diversity of 109 isolates of Macrophomina phaseolina collected from different geographical regions and host species throughout the United States (US). Genetic diversity was assessed using Nei’s minimum genetic distance, and the usefulness of each locus was determined by calculating the polymorphism information content (PIC). A total of 98 alleles were detected and of these 31 were unique to individual genotypes. Eight of twelve loci were highly informative with PIC values greater than 0.50. The majority of pairwise comparisons of genetic distance were greater than 0.60 indicating moderate to high genetic diversity. Dendrograms based on the genetic dissimilarities were created for the 109 isolates of which 79 were from soybean. Some clustering by host and geography was noted, but, the dendrograms generally grouped isolates independent of host or geography. Additionally, sequencing of the internal transcribed spacer region (ITS) for 10 isolates revealed that all of these isolates were 99% similar. Three SSR loci from M. phaseolina were cross amplified in other genera in the Botryosphaeriaceae. This was the first study of genotyping and assessing genetic diversity of M. phaseolina isolates collected from a widespread host and geographic range across the US with SSRs. With an additional 34 loci publically available for M. phaseolina, the results indicate that previously developed SSRs from one species can be used in future population, ecological, and genetic studies of M. phaseolina and other genera within the Botryosphaeriaceae.     

Removal of Arsenic(III) from Waste Water Using Lactobacillus acidophilus

Lactobacillus acidophilus was used for the removal of As(III) from 50–2000 ppb As(III)-containing water solution. Biosorption of As(III) by L. acidophilus was dependent on concentration (50 to 2000 ppb) and time (0 to 3 h).L. acidophilus(1 mg dry wt/ml) was able to remove 30, 60, 300, 420, 600 ppb As(III) from 50, 100, 500, 1000, and 2000 ppb of As(III)-containing water solution, respectively, within 3 h at pH 7. Moreover, by increasing the biomass of L. acidophilus(2 mg dry wt/ml) removal of As(III) was enhanced 1.66, 1.33, 1.16, 1.42, and 1.33 times, respectively. Fourier transform infrared (FTIR) and electron spectroscopy for chemical analysis (ESCA) spectrum of As(III)-loaded biomass was also investigated. An FTIR sample spectrum of L. acidophilus fresh biomass and As(III)-loaded biomass showed band stretching of fresh and As(III)-loaded biomass for O-H, 3423.43 to 3385.04 cm^?1, and for C-O, 1742.82 to 1731.14 cm^?1, and signified that –OH and –CO groups were also involved in the removal of As(III) from As(III)-containing water solution.     

BRIT1/MCPH1 Is Essential for Mitotic and Meiotic Recombination DNA Repair and Maintaining Genomic Stability in Mice

BRIT1 protein (also known as MCPH1) contains 3 BRCT domains which are conserved in BRCA1, BRCA2, and other important molecules involved in DNA damage signaling, DNA repair, and tumor suppression. BRIT1 mutations or aberrant expression are found in primary microcephaly patients as well as in cancer patients. Recent in vitro studies suggest that BRIT1/MCPH1 functions as a novel key regulator in the DNA damage response pathways. To investigate its physiological role and dissect the underlying mechanisms, we generated BRIT1 ^−/− mice and identified its essential roles in mitotic and meiotic recombination DNA repair and in maintaining genomic stability. Both BRIT1 ^−/− mice and mouse embryonic fibroblasts (MEFs) were hypersensitive to γ-irradiation. BRIT1 ^−/− MEFs and T lymphocytes exhibited severe chromatid breaks and reduced RAD51 foci formation after irradiation. Notably, BRIT1 ^−/− mice were infertile and meiotic homologous recombination was impaired. BRIT1-deficient spermatocytes exhibited a failure of chromosomal synapsis, and meiosis was arrested at late zygotene of prophase I accompanied by apoptosis. In mutant spermatocytes, DNA double-strand breaks (DSBs) were formed, but localization of RAD51 or BRCA2 to meiotic chromosomes was severely impaired. In addition, we found that BRIT1 could bind to RAD51/BRCA2 complexes and that, in the absence of BRIT1, recruitment of RAD51 and BRCA2 to chromatin was reduced while their protein levels were not altered, indicating that BRIT1 is involved in mediating recruitment of RAD51/BRCA2 to the damage site. Collectively, our BRIT1-null mouse model demonstrates that BRIT1 is essential for maintaining genomic stability in vivo to protect the hosts from both programmed and irradiation-induced DNA damages, and its depletion causes a failure in both mitotic and meiotic recombination DNA repair via impairing RAD51/BRCA2''s function and as a result leads to infertility and genomic instability in mice.     

Prediction of wheat response to an application of phosphorus under field conditions using diffusive gradients in thin-films (DGT) and extraction methods

The ability of the Diffusive Gradients in Thin Films (DGT) technique and two other established testing methods (Colwell, resin) to predict wheat responsiveness to applied P from 35 field trials across southern Australia was investigated. Regression analysis of relative early dry matter production and grain yield responses demonstrated that the DGT method predicted plant responsiveness to applied P more accurately than Colwell P and resin P at sites where maximum yields were reached with P rates used (20 out of 35). The measured concentration in soils at the DGT surface, C_DGT, explained 74% of the variation in response for both early dry matter and grain, compared to 7% for early dry matter and 35% for grain using the resin P method. No significant relationships could be obtained for Colwell P although modifying the Colwell test data using Phosphorus Buffering Index resulted in a correct response prediction for 11 of the 20 field sites compared to 18 for DGT and 14 for resin P. These observations suggest that the DGT technique can assess plant available P in soils with significantly greater accuracy than traditional soil P testing methods. The critical P threshold, expressed as C_DGT, was 255 μg L^?1 for early dry matter and 66 μg L^?1 for grain.     

Surface modified zeolite, a novel carrier material for Azotobacter chroococcum

A new immobilization matrix based on zeolite has been developed to immobilize Azotobacter chroococcum, for fixing nitrogen, with an intention to hold the cells in the root zone of the plants and to protect them under stressful conditions. The matrix has been developed by modifying the surface of the zeolite with surfactant. This enhances the hydrophobicity of the material and also modifies the surface charge, which in turn enhances the immobilization. Surface modified zeolite-A (SMZ-A) has been compared with commercial zeolite-A (CZA) for immobilization efficiency. CZA is non-toxic for A. chroococcum but is inefficient to adsorb the cells whereas SMZ-A showed 100% adsorption of the microbial cells wherein it was observed that for 1 l of broth culture with total viable count of 10⁸ cfu ml⁻¹ cells of A. chroococcum, a minimum dose of 0.7 g SMZ-A and minimum contact time of 10 h is required to achieve 100% adsorption. Adsorption was confirmed by the cell count and light as well as scanning electron microscopy. Most importantly, the cells adsorbed on SMZ-A could fix the atmospheric nitrogen up to 13 mg g⁻¹ sucrose consumed, which was comparable with the control (unadsorbed cells), which confirms the survival and nitrogen fixation activity of the bacteria. Responsible Editor: Euan K. James.     

Scavenging effect of Indian grape polyphenols on 2,2'-diphenyl-1-picrylhydrazyl(DPPH) radical by electron spin resonance spectrometry

Antioxidant potency of Indian grape cultivars varying in their skin color, seed and polyphenol content (Bangalore blue, Pandhari sahebi, Sharad seedless and Thompson seedless) and their components (whole grapes, pulp with skin and seeds) was examined as 2,2'-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity using electron spin resonance spectrometry. The total polyphenols in Indian grapes ranged between 3-51%. Extracted polyphenols caused a concentration dependent and significant loss in DPPH radical signal, similar to known antioxidants-Vitamin C, catechin and procyanidin B3 used as references. Among seedless cultivars, polyphenols from Sharad was more potent as antioxidant than Thompson, showing IC50 values of 1250 +/- 30 and 2650 +/- 125 microg/ml, respectively. The inhibitory effect of polyphenols from seedless grape cultivars was as effective as that of seeded variety. The results indicate that polyphenols extracted from Indian grapes/ components (with /without seeds) exhibited free radical scavenging activity and their chemopreventive properties need to be exploited by in vivo model system.     

Downregulation of NADPH oxidase, antioxidant enzymes, and inflammatory markers in the heart of streptozotocin-induced diabetic rats by N-acetyl-L-cysteine

We investigated the effect of N-acetyl-l-cysteine (NAC) on the expression of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, antioxidant enzymes, and inflammatory markers in diabetic rat hearts. Metabolic parameters, free 15-F(2t)-isoprostane level, protein expression of NADPH oxidase, superoxide dismutase (SOD), heme oxygenase (HO-1), interleukin-6 (IL-6), and cyclooxygenase-2 (COX-2) were analyzed in control and streptozotocin-induced diabetic rats treated with or without NAC in drinking water for 8 wk. The cardiac protein expression of p67(phox) and p22(phox) was increased in diabetic rats, accompanied by increased NADPH-dependent superoxide production. As a compensatory response to the increased NADPH oxidase, the protein expression of Cu-Zn-SOD and HO-1 and the total SOD activity were also increased in diabetic rat hearts. Consequently, cardiac free 15-F(2t)-isoprostane, an index of oxidative stress, was increased in diabetic rats, indicating that the production of reactive oxygen species becomes excessive in diabetic rat hearts. Cardiac inflammatory markers IL-6 and COX-2 were also increased in diabetic rats. NAC treatment prevented the increased expression of p22(phox) and translocation of p67(phox) to the membrane in diabetic rat hearts. Subsequently, the levels of cardiac free 15-F(2t)-isoprostane, HO-1, Cu-Zn-SOD, total SOD, IL-6, and COX-2 in diabetic rats were decreased by NAC. Consequently, cardiac hypertrophy was attenuated in diabetic rats treated with NAC. The protective effects of NAC on diabetic rat hearts may be attributable to its protection of hearts against oxidative damage induced by the increased NADPH oxidase and to its reduction in cardiac inflammatory mediators IL-6 and COX-2.