Isolation, characterization and genomic analysis of vB-AhyM-AP1, a lytic bacteriophage infecting Aeromonas hydrophila.

AIMS: Aeromonas hydrophila is a zoonotic pathogen displaying resistance to multiple antibiotics. Here, we aim to develop a candidate biocontrol agent against A. hydrophila. METHODS AND RESULTS: In this study, we isolated and characterized the phage vB-AhyM-AP1 from sewage. It showed lytic activity against A. hydrophila strains. One-step growth curve revealed that the latent period lasted for 40 min. The burst size of one lytic cycle was 1413 PFU per infected cell. Temperature stability studies showed that the phage vB-AhyM-AP1 was active over temperatures ranging from 4 to 45°C for 1 h. pH stability studies indicated that the phage remained active within a pH range of 5-10 after 24 h of incubation. Stability tests in salt solutions showed that the phage was stable at salinities ranging from 0·1 to 2%. The phage also showed stabilities in organic solvents when incubated for 10 min. The Illumina Hiseq sequencing of its genome indicated that the phage vB-AhyM-AP1was a jumbo phage with a genome size of 2, 54 490 bp and GC content of 40·3%. The phylogenetic analysis of the terminase large subunit and major capsid protein indicated that the phage closely clustered with other Tevenvirinae phages. The genome encoded 455 ORFs and 22 tRNAs. The phage resulted in a reduction of 0·8 log units of viable A. hydrophila cells in biofilms grown on PVC coupons maintained in a low nutrient medium for 10 days. CONCLUSIONS: The phage showed lytic activity against planktonic and biofilm cells of A. hydrophila. Genome-based prediction showed it to be a strictly lytic phage without any virulence or antibiotic resistance genes indicating safety for environmental and clinical applications. SIGNIFICANCE AND IMPACT OF THE STUDY: The multidrug-resistant strains of A. hydrophila pose a significant health risk to both cultured fishes and consumers leaving few options for treatment. Phage vB-AhyM-AP1 may be used as a candidate biocontrol agent against A. hydrophila strains.

Draft Genome Sequence of Commercial Textile Dye-Decolorizing and -Degrading Bacillus subtilis Strain C3 Isolated in India.

Bacillus subtilis C3, a commercial textile dye-decolorizing and -degrading bacterium, was isolated from the common effluent treatment plant (CEPT) of the Jetpur textile dyeing and printing industrial sector situated in the district of Rajkot, Gujarat, India. Here, we present the annotated 4.18-Mb draft genome sequence of B. subtilis C3, providing information about the metabolic pathways involved in decolorization and degradation of several commercial textile azo dyes. Thus, we confirm B. subtilis C3 as a potential candidate for bioremediation of textile effluents.

Prevalence of adhesive genes among uropathogenic Escherichia coli strains isolated from patients with urinary tract infection in Mangalore.

The study was carried out to detect the adhesive genes pap (pyelonephritis associated pili), sfa (S fimbrial adhesin) and afa (afimbrial adhesin) from Escherichia coli strains isolated in patients diagnosed with urinary tract infection (UTI). A total of 23% of the isolates were positive for pap, sfa and afa genes with a prevalence of 60.87% (14/23), 39.1% (9/23) and 39.1% (9/23), respectively. Prevalence of multiple adhesive genes was 8.7% (2/23) for pap and afa, 30.43% (7/23) for pap and sfa. Significant numbers of isolates were positive for Congo red binding (80%) and haemolysin production 60%. The prevalence of multiple adhesive genes indicate the potential to adhere and subsequently cause a systemic infection among UTI patients.

Presence of T3SS2ß genes in trh⁺ Vibrio parahaemolyticus isolated from seafood harvested along Mangalore coast, India.

UNLABELLED: Vibrio parahaemolyticus is a seafood-borne pathogen autochthonous to the marine and estuarine ecosystem, responsible for gastroenteritis when contaminated raw seafood is consumed. The pathogenicity has been associated with thermostable direct haemolysin (TDH) and TDH-related haemolysin (TRH). Of late, the presence of T3SS2α and T3SS2ß gene clusters has been well documented in clinical isolates of Vibrio parahaemolyticus and known to play an essential role in pathogenesis. However, reports on the presence of T3SSß genes in V. parahaemolyticus isolated from the seafood and/or environmental samples are scanty. In this study, we have identified and analysed the distribution of the T3SS2ß genes in V. parahaemolyticus isolated from seafood harvested along southwest coast of India. Results showed that T3SS2ß genes are solely associated with trh⁺ and tdh⁺ /trh⁺ strains of V. parahaemolyticus. Reverse transcriptase PCR (RT-PCR) showed that the T3SS2ß genes identified in trh⁺ V. parahaemolyticus were transcriptionally active. To our knowledge, this study appears to be the first description on the presence of T3SS2ß-positive V. parahaemolyticus isolated from seafood in India. The study of T3SS2 along with other virulence factors will help in better understanding of the risk of seafood-borne illness due to V. parahaemolyticus. SIGNIFICANCE AND IMPACT OF THE STUDY: T3SSs (α or ß) are the important virulence factors of Vibrio parahaemolyticus that contribute to their pathogenicity in humans. This study demonstrated the presence of T3SS2ß genes in V. parahaemolyticus isolated from the seafood harvested along Mangalore coast. RT-PCR showed that the T3SS2ß genes identified in seafood isolates of V. parahaemolyticus were found to be functional. To the best of our knowledge, this is the first description of T3SS2ß genes in trh⁺ V. parahaemolyticus isolated from seafood in India. The presence of T3SS2 along with other virulence factors such as TDH and/or TRH highlights a potential health risk for seafood consumers.

A comparative evaluation of impact strength of conventionally heat cured and high impact heat cured polymethyl methacrylate denture base resins: an in vitro study.

UNLABELLED: The denture bases made by using polymethyl methacrylates of Acrylic resin family have excellent physical properties, simple to process and easy to reline, rebase and repair. One of the inherent disadvantages of this material is the liability to break during function. The strength assessment of acrylic resins have been made generally by transverse defection tests. AIM: To evaluate the impact strength valves of certain brands of commercially available denture base resins and suggest their suitability. MATERIALS AND METHODS: The denture bases we made using polymethyl methacrylates of acrylic resin family because they have excellent physical properties, simple to process and easy to reline and rebase. Six commercial brands of polymethyl meth- acrylate, namely Stellon (DPI-India), Acralyn-H (Asian Acrylate, India), Trevalon (Dentsply-England), Lucitone 199 (Dentsply/ York division), Acralyn-H (Super Unbreakable), Trevalon HI (Dentsply, Detray division, England) were tested by breaking them using Analog Pendulum (ASTM D 256). RESULTS: From the entire study the maximum impact strength was reported for Acralyn-H super unbreakable (Asian Acrylates, India) 62.19 joules. CONCLUSION: All the analysis led to conclusion that there is basic change in material composition within and among the different groups of denture base resins. CLINICAL IMPLICATIONS: The complete dentures made using denture base resins with high impact strength valves (e.g. Acralyn-H super unbreakable) will be more durable and can be used by the patient for considerable period of time, i.e. beyond 4 to 5 years.

Presence of typical and atypical virulence genes in vibrio isolates belonging to the Harveyi clade.

AIMS: The study was aimed at investigating the presence of typical and atypical virulence genes in isolates belonging to the Harveyi clade (Vibrio harveyi and Vibrio campbellii). METHODS AND RESULTS: Forty-eight vibrio isolates belonging to the Harveyi clade were screened for the presence of virulence genes that are typical for these bacteria and those found in human pathogenic vibrios such as Vibrio cholerae, Vibrio parahaemolyticus and Vibrio vulnificus and aquatic pathogenic Vibrio anguillarum. The virulence genes were amplified by PCR with specific primers, and the presence further confirmed by dot blot hybridization. The virulence genes vhh, chiA, vhpA, toxR(Vh), luxR and serine protease, typical of Harveyi clade were detected in all the isolates. The haemolysin gene hlyA and the virulence regulator gene toxR(Vc) specific to V. cholerae and the V. anguillarum-specific flagellum gene (flaC) were present in some of the isolates. Challenge tests with gnotobiotic Artemia nauplii did not show any correlation between the presence of the virulence genes and virulence of the isolates. CONCLUSION: From our results, there appears a remote possibility that vibrios belonging to the Harveyi clade might acquire virulence genes from other vibrios in the aquatic environment through horizontal gene transfer. SIGNIFICANCE AND IMPACT OF THE STUDY: Vibrios belonging to the Harveyi clade may be an important reservoir of virulence genes of other (human pathogenic) Vibrio species in the aquatic environment. The acquisition of virulence genes by horizontal transfer might increase the ability of Harveyi clade vibrios to infect aquatic organisms by increasing their virulence to a specific host by broadening their host range. The detection of such genes may forewarn the hatchery operators about a potentially virulent pathogen and thus help to develop management measures to handle the problem of vibriosis.

Mysore childhood obesity study.

We conducted this study to document the prevalence of obesity, overweight and underweight in the school children aged 5 to 16 years from Mysore. 5 Principal Investigators and 13 Co-Investigators trained the teachers of 139 schools (Private--111, Govt--28) to record the vital statistics of the children studying in their schools. A total of 43,152 school children (23,527 boys and 19,625 girls) were surveyed. 36,354 children were from private schools and 6798 children were from Government (Govt) schools. Indian Academy of Pediatrics growth charts were used as reference. The prevalence of obesity, overweight and underweight were 3.4%, 8.5% and 17.2%, respectively. The prevalence of obesity was maximum in the age group of 5-7 years and in those from private schools.

PLA2 mediated arachidonate free radicals: PLA2 inhibition and neutralization of free radicals by anti-oxidants--a new role as anti-inflammatory molecule.

PLA2 enzyme catalyses the hydrolysis of cellular phospholipids at the sn-2 position to liberate arachidonic acid and lysophospholipid to generate a family of pro-inflammatory eicosanoids and platelet activating factor. The generation of pro-inflammatory eicosanoids involves a series of free radical intermediates with simultaneous release of reactive oxygen species (superoxide and hydroxyl radicals). Reactive oxygen species formed during arachidonic acid metabolism generates lipid peroxides and the cytotoxic products such as 4-hydroxy nonenal and acrolein, which induces cellular damage. Thus PLA2 catalyzes the rate-limiting step in the production of pro-inflammatory eicosanoids and free radicals. These peroxides and reactive oxygen species in turn activates PLA2 enzyme and further attenuates the inflammatory process. Therefore scavenging these free radicals and inhibition of PLA2 enzyme simultaneously by a single molecule such as antioxidants is of great therapeutic relevance for the development of anti-inflammatory molecules. PLA2 enzymes have been classified into calcium dependent cPLA2 and sPLA2 and calcium independent iPLA2 forms. In several inflammatory diseases sPLA2 group IIA is the most abundant isoform identified. This isoform is therefore targeted for the development of anti-inflammatory molecules. Many secondary metabolites from plants and marine sponges exhibit both anti-inflammatory and antioxidant properties. Some of them include flavonoids, terpenes and alkaloids. But in terms of PLA2 inhibition and antioxidant activity, the structural aspects of flavonoids are well studied rather than terpenes and alkaloids. In this line, molecules having both anti-oxidant and PLA2 inhibitions are reviewed. A single molecule with dual activities may prove to be a powerful anti-inflammatory drug.

Cytochrome P450 3A: genetic polymorphisms and inter-ethnic differences.

Ethnicity is a demographic variable that plays an important role in interindividual variability of drug metabolism and response. The genetic variations of drug-metabolizing enzymes exhibiting interindividual differences of drug metabolism also show differences between populations. The reason for this is that the frequency of a polymorphism is found to differ between populations. The other reason is that different variants are seen in different populations. Most drugs are biotransformed in the body by cytochrome P450. The CYP3A isozymes are responsible for the metabolism of 50-60% of all currently prescribed drugs. Studies have shown that there is variability in CYP3A activity and also inter-ethnic differences in CYP3A-mediated drug metabolism. The purpose of this review is to focus on the genetic polymorphism and ethnic variations in CYP3A-mediated oxidative drug metabolism.

Fine needle aspiration cytology of chondroblastoma and chondromyxoid fibroma. A report of two cases.

The fine needle aspiration cytology in two cases of chondroblastoma and chondromyxoid fibroma are described. The diagnosis of chondroblastoma was made on a recurrent tumor of which the histopathology was known, whereas chondromyxoid fibroma was diagnosed initially on fine needle aspiration cytology. The radiologic appearances and differential diagnoses of these chondroid neoplasms are discussed.

Use of positive-negative deviant analyses to improve programme targeting and services: example from the TamilNadu Integrated Nutrition Project.

This paper describes the characteristics of southern rural Indian children who grow best (positive deviants) and worst (negative deviants) as compared to median growers. A 100 each of positive and negative deviants and 120 median growers were selected after analysing the 12-month growth patterns (weight-for-age) of 2954 children enrolled in the TamilNadu Integrated Nutrition Project (TINP), a major health and nutrition intervention project covering nearly one million children in rural south India. The determinants of poor growth that have been left unaddressed by 6 years of TINP exposure are delineated to address the question of what more needs to be done. Further, the rationale for differential targeting of services to negative deviants and to median growers is discussed, as are the implications for programme evaluation. Data indicate that the next generation of projects targeted at the most needy (negative deviants) should address the issues of gender discrimination in childcare, of breastfeeding, of diarrhoeal disease, and of maternal empowerment. Such interventions will, however, not improve the growth of median growers in the direction of positive deviance. Instead, programmes targeted at the median growers need to support the hygienic use of nonbreast milk supplements. Improving family wealth will also improve the nutritional status of the median growers, but less so than for the negative deviants.