Dirofilaria hongkongensis - A first report of potential zoonotic dirofilariosis infection in dogs from Tamil Nadu.

Dirofilariosisis, is a mosquito borne anthropo-zoonotic disease caused by the filarial nematodes of the genus Dirofilaria. The current paper reports a species of Dirofilaria, which has not been recorded in Tamil Nadu, in addition to differentiating it from the existing species of Dirofilaria spp. through morphological and molecular techniques. Micrometry of the microfilaria detected by Knott's technique as well as in Giemsa stained blood smears revealed that the length was on an average 280 µm to 310 µm whereas the width was 5 to 8 µm. The microfilariae were unsheathed with blunt heads and tapering tails. For molecular confirmation, the blood sample was subjected to genomic DNA isolation. PCR amplification using specific primers for D. hongkongensis was carried out using thermal cycler with the isolated genomic DNA as template. Molecular analyses revealed the band size of about 230 bp which confirmed the presence of D. hongkongensis. Phylogenetic analysis was done using UPGMA and MLM method. Dirofilaria is recognized as a zoonotic entity throughout the world, and dirofilariasis should be considered an emerging zoonosis in southern India. The control of ubiquitous mosquito population is essential to prevent the transmission of microfilaria from animals to arthropods and from arthropods to humans.

Cat fur mite Lynxacarus radovskyi in India.

A Persian cat with the complaint of lustreless dry coat, salt and pepper appearance was brought to the dermatology unit of Madras Veterinary College, Chennai. Microscopical examination of the tape impression and hair coat brushings revealed Lynxacarus radovskyi mites with eggs adhering to the hair shaft. The cat was treated with Ivermectin @ 300 µg/kg body weight subcutaneously once a week for 5 weeks after getting owner's consent. The cat was free from the parasites 5 weeks post treatment. Fomite and close contact might aid the spread of infestation. Isolation coupled with full course of treatment favours complete recovery from the infestation.

Serum Anti-Cryptosporidial gp15 Antibodies in Mothers and Children Less than 2 Years of Age in India.

Little is known about the type and longevity of the humoral response to cryptosporidial infections in developing countries. We evaluated serum antibody response to Cryptosporidium gp15 in 150 sets of maternal, preweaning and postinfection/end-of-follow-up sera from children followed up to 2 years of age to determine the influence of maternal and preweaning serological status on childhood cryptosporidiosis. Fifty two percent (N = 78) of mothers and 20% (N = 30) of children were seropositive preweaning. However, most positive preweaning samples from children were collected early in life indicating transplacental transfer and subsequent rapid waning of antibodies. Although 62% (N = 94) of children had a parasitologically confirmed cryptosporidial infection (detected by stool polymerase chain reaction) during the follow-up, only 54% (N = 51) of children were seropositive postinfection. Given there were striking differences in seropositivity depending on when the sample was collected, even though Cryptosporidium was detected in the stool of the majority of the children, this study indicates that antibodies wane rapidly. During follow-up, the acquisition or severity of cryptosporidial infections was not influenced by maternal (P = 0.331 and 0.720, respectively) as well as the preweaning serological status of the child (P = 0.076 and 0.196, respectively).

Detection and sequence analysis of accessory gene regulator genes of Staphylococcus pseudintermedius isolates.

BACKGROUND: Staphylococcus pseudintermedius (SP) is the major pathogenic species of dogs involved in a wide variety of skin and soft tissue infections. The accessory gene regulator (agr) locus of Staphylococcus aureus has been extensively studied, and it influences the expression of many virulence genes. It encodes a two-component signal transduction system that leads to down-regulation of surface proteins and up-regulation of secreted proteins during in vitro growth of S. aureus. The objective of this study was to detect and sequence analyzing the AgrA, B, and D of SP isolated from canine skin infections. MATERIALS AND METHODS: In this study, we have isolated and identified SP from canine pyoderma and otitis cases by polymerase chain reaction (PCR) and confirmed by PCR-restriction fragment length polymorphism. Primers for SP agrA and agrBD genes were designed using online primer designing software and BLAST searched for its specificity. Amplification of the agr genes was carried out for 53 isolates of SP by PCR and sequencing of agrA, B, and D were carried out for five isolates and analyzed using DNAstar and Mega5.2 software. RESULTS: A total of 53 (59%) SP isolates were obtained from 90 samples. 15 isolates (28%) were confirmed to be methicillin-resistant SP (MRSP) with the detection of the mecA gene. Accessory gene regulator A, B, and D genes were detected in all the SP isolates. Complete nucleotide sequences of the above three genes for five isolates were submitted to GenBank, and their accession numbers are from KJ133557 to KJ133571. AgrA amino acid sequence analysis showed that it is mainly made of alpha-helices and is hydrophilic in nature. AgrB is a transmembrane protein, and AgrD encodes the precursor of the autoinducing peptide (AIP). Sequencing of the agrD gene revealed that the 5 canine SP strains tested could be divided into three Agr specificity groups (RIPTSTGFF, KIPTSTGFF, and RIPISTGFF) based on the putative AIP produced by each strain. The AIP of SP contains serine and produce lactone ring structured AIP. CONCLUSION: Presence of AgrA, B, and D in all SP isolates implies the importance of this regulatory system in the virulence genes expression of the SP bacteria. SP isolates can be typed based on the AgrD auto-inducible protein sequences as it is being carried out for typing of S. aureus isolates. However, further studies are required to elucidate the mechanism of controlling of virulence genes by agr gene locus in the pathogenesis of soft tissue infection by SP.

Cryptosporidiosis among children in an endemic semiurban community in southern India: does a protected drinking water source decrease infection?

BACKGROUND: A quasi-experimental study was conducted to determine whether or not a protected water supply (bottled drinking water) could prevent or delay cryptosporidial infections among children residing in an endemic community. METHODS: A total of 176 children residing in a semiurban slum area in southern India were enrolled preweaning and received either bottled (n = 90) or municipal (n = 86) drinking water based on residence in specific streets. Weekly surveillance visits were conducted until children reached their second birthday. Stool samples were collected every month and during diarrheal episodes, and were tested for the presence of Cryptosporidium species by polymerase chain reaction. Differences in the incidence of cryptosporidiosis between bottled and municipal water groups were compared using Poisson survival models, and a propensity score model was developed to adjust for the effect of potential confounders. RESULTS: A total of 186 episodes of cryptosporidiosis, mostly asymptomatic, were observed in 118 (67%) children during the follow-up period at a rate of 0.59 episodes per child-year. Diarrhea associated with Cryptosporidium species tended to be longer in duration and more severe. Stunting at 6 months was associated with a higher risk of cryptosporidiosis (rate ratio [RR] = 1.40; 95% confidence interval [CI], 1.03-1.91). A higher gastrointestinal disease burden was also seen in children with cryptosporidiosis. Drinking bottled water was not associated with a reduced risk of cryptosporidiosis (adjusted RR = 0.86; 95% CI, .60-1.23). CONCLUSIONS: This study documented a high burden of cryptosporidiosis among children in an endemic Indian slum community. The lack of association between drinking bottled water and cryptosporidiosis suggests possible spread from asymptomatically infected individuals involving multiple transmission pathways.

Peptide aggregates: a novel model system to study self-assembly of peptides.

Ordered aggregates of Val-Leu-Pro-Phe, tetrapeptide 1, have been found in aqueous solutions. Evidence for the formation of aggregates for the above peptide was obtained by conductometric, pH metric, UV and fluorescence spectroscopic techniques. Values of critical micelle concentration (CMC) for the above peptide obtained by these methods are in good agreement with each other. The formation of organized aggregates of the peptide is favoured upon increasing the temperature (viz. the process of aggregation is endothermic). The aggregation number has been determined at different temperatures. Values of delta G0m, delta H0m, delta S0m and delta C0p have also been estimated. Binding studies with the 8-anilinonaphthyl sulfonic acid (ANS) and pyrene indicate that the interior of the aggregate is nonpolar. There are two processes with regard to the change of thermodynamical parameters like delta G0m, delta H0m, delta S0m, delta C0p and aggregation number (N). In the first process (from 5 degrees C to 40 degrees C) the driving force for aggregation seems to be the positive entropy because of water release due to intermolecular association of ionic moieties. The second process (from 40 degrees C and above) is due to intramolecular ionic interaction. The chemical shifts of the amide protons of the peptide have been presented in the light of inter- and intramolecular hydrogen-bond formation, and forces implicated in aggregation for both the first and second processes.