Skeletal pathology and bone mineral density changes in wild muskrats (Ondatra zibethicus) and red squirrels (Tamiasciurus hudsonicus) inhabiting arsenic polluted areas of Yellowknife, Northwest Territories (Canada): A radiographic densitometry study.

The City of Yellowknife is a known hotspot of arsenic contamination and there is a growing body of evidence suggesting that local wildlife in the vicinity of the abandoned Giant Mine site may be at risk of decreased bone mineralization and various bone disorders. The purpose of this study was to preliminarily measure bone mineral density (BMD) changes and investigate the incidence, pattern, and severity of bone lesions in wild muskrats and red squirrels breeding in three (3) catchment areas at different distances from the Giant Mine Site in Yellowknife, Northwest Territories (Canada): ~2 km (location 1), ~18 km (location 2), and ~40-100 km (location 3). Full femoral bones of 15 muskrats and 15 red squirrels were collected from the three sampling locations (5 from each location) and subjected to radiographic analysis and densitometric measurements. The patterns and severities of bone lesions, including changes in bone mineral density, were evaluated and compared between groups. As levels were significantly higher in the bones of muskrats caught from location 1 and 2, relative to location 3. Further, As and Cd levels were significantly higher in the bones of squirrels caught from locations 1 and 2 relative to squirrels caught from location 3. The preliminary results from bones revealed that radiographic abnormalities such as bone rarefaction, osteopenia, and thinning of the femoral shafts with significant ossific cystic lesions and bowing were the most common skeletal pathologies found in bones of red squirrels from the three locations. Radiographic appearances of massive sclerosis and dysplasia, including severe osteocondensation and osteopathia striata-like abnormalities, were found in the bones of muskrats from all the sampling locations. Densitometric evaluation showed no significant differences between the three locations in the bone parameters measured. However, there was a statistically significant correlation between As content in the bones of muskrats and percent fat content in the femur samples, which suggests that accumulation of As could have been a causal factor for a change in percent fat in femurs of muskrats.

Comparative study of arsenic toxicosis and ocular pathology in wild muskrats (Ondatra zibethicus) and red squirrels (Tamiasciurus hudsonicus) breeding in arsenic contaminated areas of Yellowknife, Northwest Territories (Canada).

The Giant Mine is an abandoned gold mine in Yellowknife, Northwest Territories, Canada. Throughout its operation from 1948 to 2004, the Giant Mine released heavy amounts of arsenic trioxide into the environment, thus contaminating the soil and surface water within and around the vicinity of the mine site. Chronic arsenic (As) poisoning negatively impacts wildlife health and can induce multi-organ damages including neurodegeneration and visual dysfunction depending on concentration and duration of exposure. The aim of the current study was to comparatively assess retina layer changes and prevalence of ocular lesions in wild rodent populations (i.e. muskrats and red squirrels) breeding in arsenic endemic areas of Yellowknife, near the vicinity of the abandoned Giant mine site (∼2 km radius), at an intermediate location (approximately 20 km from the mine area) as well as a reference location (spanning 52-105 km from the city of Yellowknife, Canada). Eye globes were removed from euthanized muskrats and squirrels from the three sampling locations with increasing distance from the Giant mine area. Optical Coherence Tomography (OCT) was used to attempt a pan-retinal layer assessment, and histologic examination was utilized for assessment and confirmation of ocular lesions. The retinal layers were measured and statistically compared between the groups based on sampling locations to enhance the scope of histologic evaluations. The preliminary results revealed that thicknesses of ganglion cell layer (GCL), retina nerve fibre layer (NFL), and inner retina layer (IR) were statistically reduced in the muskrats from arsenic endemic area, particularly near the vicinity of the Giant mine compared to the control group. Generalized ocular pathology was histologically confirmed in all the muskrats from the arsenic endemic areas with the manifestation of moderate to severe lymphocytic plasmacytic uveitis (LPU), keratitis and subcapsular cataracts. Inner retinal degeneration was also observed in all the muskrats from the arsenic endemic areas, while muskrats from the control group were predominantly normal. Three muskrats from the control group were noted to have a mild LPU and keratitis. Significant histopathologic changes were not detected in the squirrel eyes from the three groups except for incidental mild cornea scars from all the locations. In general, these preliminary findings confirm the presence of ocular lesions and retina abnormalities in wild muskrats in the Yellowknife area and provide the first evidence of visual dysfunction and impairment in wildlife inhabiting arsenic endemic areas of Canada.

Neuropathological changes in wild muskrats (Ondatra zibethicus) and red squirrels (Tamiasciurus hudsonicus) breeding in arsenic endemic areas of Yellowknife, Northwest Territories (Canada): Arsenic and cadmium accumulation in the brain and biomarkers of oxidative stress.

The brain is one of the critical organs particularly susceptible to the damaging effects of chronic arsenic poisoning and there is a growing body of evidence that suggest that oxidative stress plays a key role in the pathogenesis of neurodegenerative disorders. The aim of this present work was to comparatively assess biomarkers of oxidative stress and status of antioxidant enzyme activities in the brains of muskrats and squirrels breeding in arsenic endemic areas, specifically near the vicinity of the abandoned Giant mine site (~2 km radius), and an intermediate location approximately 20 km from the mine area and in reference locations spanning 52-105 km from the city of Yellowknife, Northwest Territories (Canada). Analysis included measurement of total arsenic and cadmium concentration in the nails, brain, and stomach content of muskrats and squirrels, in addition to biochemical evaluation of lipid peroxidation levels and antioxidant enzymes defense: catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GPx) in the brain tissues. The results revealed that arsenic concentration in the nails of muskrats collected closest to the vicinity of the mine area was in the range of 11 to 35.1 times higher than those from the reference site. The maximum concentration of arsenic in the nails of muskrats from the intermediate location was 47.6 times higher than the maximum concentration observed in the reference muskrats. Cadmium was generally undetected in the nails of muskrats and squirrels from the three sampling locations. Arsenic in the gut contents of muskrats from the arsenic affected area was 4.5 to 49.1 times higher than those from the reference site. Cadmium levels in the guts of muskrats from the mine area almost doubled those from the reference site. Arsenic accumulated in the nails of squirrels from the areas closest to the mine but was undetected in the squirrel nails from the reference location. The maximum arsenic levels in the stomach content of squirrels from the mine area was ~40 times higher than those from the reference site. Arsenic did not accumulate in the brains of muskrats, but cadmium was detected in a few brains of muskrats. Brains of squirrels from the mine area and intermediate locations accumulated both arsenic and cadmium. The brains of squirrels and muskrats from the arsenic affected area showed no evidence of increased lipid peroxidation compared to the animals from the reference site. However, SOD, CAT and GPx activities in the brains of animals from the arsenic endemic areas tended to be higher compared to the control sites. This is the first study documenting evidence of oxidative stress and altered antioxidant enzyme activities in brains of wild rodent population in arsenic endemic areas of Canada.

Altered neurotransmission and neuroimaging biomarkers of chronic arsenic poisoning in wild muskrats (Ondatra zibethicus) and red squirrels (Tamiasciurus hudsonicus) breeding near the City of Yellowknife, Northwest Territories (Canada).

Chronic arsenic poisoning has been shown to be a risk factor for the development of intellectual disability. Numerous human and animal studies have also confirmed that low-level arsenic exposure has deleterious effects on neurotransmission and brain structures which have been further linked to neurobehavioral disorders. The aim of this present work was to comparatively assess structural brain volume changes and alteration of two (2) neurotransmitters, specifically dopamine (DA) and serotonin (5-HT) in the brains of wild muskrats and squirrels breeding in arsenic endemic areas, near the vicinity of the abandoned Giant mine site in Yellowknife and in reference locations between 52 and 105 km from the city of Yellowknife. The levels of DA and 5-HT were measured in the brain tissues, and Magnetic Resonance Imaging (MRI) was used to attempt brain volume measurements. The results revealed that the concentrations of DA and 5-HT were slightly increased in the brains of squirrels from the arsenic endemic areas compared to the reference site. Further, DA and 5-HT were slightly reduced in the brains of muskrats from the arsenic endemic areas compared to the reference location. In general, no statistically significant neurotransmission changes and differences were observed in the brain tissues of muskrats and squirrels from both arsenic endemic areas and non-endemic sites. Although MRI results showed that the brain volumes of squirrels and muskrats were not statistically different between sites after multiple comparison correction; it was noted that core brain regions were substantially affected in muskrats, in particular the hippocampal memory circuit, striatum and thalamus. Squirrel brains showed more extensive neuroanatomical changes, likely due to their relatively smaller body mass, with extensive shrinkage of the core brain structures, and the cortex, even after accounting for differences in overall brain size. The results of this present study constitute the first observation of neuroanatomical changes in wild small mammal species breeding in arsenic endemic areas of Canada.

Chronic arsenicosis and cadmium exposure in wild snowshoe hares (Lepus americanus) breeding near Yellowknife, Northwest Territories (Canada), part 2: Manifestation of bone abnormalities and osteoporosis.

Various bone abnormalities, including osteoporosis, have been associated with chronic arsenic and cadmium exposure in experimental animal models, but information regarding the bone pathology of wild population of small mammals breeding in contaminated environment is limited. This present study was conducted to comparatively assess the prevalence and pattern of skeletal abnormalities in free ranging snowshoe hares inhabiting an area heavily contaminated by arsenic and other trace metals, near the vicinity of the abandoned Giant mine, and in a reference location approximately 20km from the city of Yellowknife, Northwest Territories, Canada. The femur and vertebrae of snowshoe hares from the mine area and reference location were subjected to bone densitometry examination and biomechanical testing using dual energy X-ray absorptiometry (DXA) and 3-point bending test. t-test results indicated that femoral densitometry parameters such as bone mineral density (BMD) (p=0.5), bone mineral content (BMC) (p=0.675), bone area (BA) (p=0.978) and tissue area (TA) (p=0.549) were not significantly different between locations. All densitometry parameters of the vertebrae (BMD, BA and TA) differed between locations (p<0.05), except for BMC (p=0.951) which showed no significant difference between the two locations. Vertebrae from the mine area also showed relatively lower BA and TA compared to the reference location. A constellation of skeletal abnormalities were also observed along the axial and appendicular bones respectively. Specifically, growth defects, osteoporosis, cortical fractures, sclerosis, and cyst like changes were commonly observed in the femurs and vertebrae of hares from both locations. With respect to biomechanical properties, only bone stiffness and peak load tended to be relatively reduced in specimens from the mine area, whereas work to failure was notably increased in specimens from the reference site compared to those from the mine area. Taken together, the results of this preliminary study suggest that chronic concomitant exposure to arsenic and cadmium may be involved in the etiology of various bone abnormalities, including osteoporosis in wild population of snowshoe hares from the Yellowknife area. The result presented in this study represent the first evaluation of osteological effects in free-ranging furbearers (snowshoe hares) diagnosed with arsenicosis, and concomitantly exposed to environmental levels of cadmium.

Chronic arsenicosis and cadmium exposure in wild snowshoe hares (Lepus americanus) breeding near Yellowknife, Northwest Territories (Canada), part 1: Evaluation of oxidative stress, antioxidant activities and hepatic damage.

Previous gold mining activities and arsenopyrite ore roasting activities at the Giant mine site (1948 to 2004) resulted in the release of high amounts of arsenic and trace metals into the terrestrial and aquatic ecosystems of Yellowknife, Northwest Territories, Canada. While elevated levels of arsenic has been consistently reported in surface soils and vegetation near the vicinity of the Giant mine area and in surrounding locations, systematic studies evaluating the overall health status of terrestrial small mammals endemic to the area are lacking. The purpose of this present study was to evaluate and comparatively assess the biochemical responses and histopathological effects induced by chronic arsenic and cadmium exposure in wild snowshoe hares breeding near the city of Yellowknife, specifically around the vicinity of the abandoned Giant mine site and in reference locations. Analysis included measurement of total arsenic and cadmium concentration in nails, livers, kidneys, bones, stomach content of hares, in addition to histopathological evaluation of hepatic and ocular lesions. Biochemical responses were determined through measurement of lipid peroxidation levels and antioxidant enzymes activities (catalase, superoxide dismutase, glutathione peroxidase, and glutathione disulfide). The results revealed that arsenic concentration was 17.8 to 48.9 times higher in the stomach content, and in the range of 4 to 23 times elevated in the nails of hares from the mine area compared to the reference location. Arsenic and cadmium levels were also noted to be increased in the bones, renal and hepatic tissues of hares captured near the mine area compared to the reference site. Specifically, hares from the mine area showed nail cadmium levels that was 2.3 to 17.6 times higher than those from the reference site. Histopathological examination of the eyes revealed no specific ocular lesions, such as lens opacity (cataracts) or conjunctivitis; however, hares from both locations exhibited hepatic steatosis (fatty liver change). Lipid peroxidation levels were relatively increased and accompanied with reduced antioxidant enzyme activities in hares from the mine area compared to the hares from the reference site. The results of this preliminary study suggest that the snowshoe hares breeding near the vicinity of Yellowknife, including near the Giant mine area have been chronically exposed to elevated levels of arsenic and cadmium, which consequently led to the increased levels of oxidative stress and perturbation of antioxidant defense system in exposed animals. The results of this present study constitute the first observation of chronic arsenicosis in wild small mammal species in Canada.

Heavy metal bioaccumulation and histopathological alterations in wild Arctic hares (Lepus arcticus) inhabiting a former lead-zinc mine in the Canadian high Arctic: A preliminary study.

A preliminary study was undertaken to determine post-mining baseline accumulation of selected trace metals, and histopathological alterations in free-living arctic hares (Lepus arcticus) inhabiting the vicinity of a former lead-zinc mine located on North Baffin Island in the Canadian High Arctic. Trace metal analysis included measurement of As, Cd, Fe, Pb and Zn in tissues, and histopathological assessment comprised of evaluation and scoring the severity of metal-induced hepatic and renal lesions. Metal contents in hepatic and renal tissues from hares from the mine area compared with the reference locations did not differ significantly suggesting that the animals are not uniformly exposed to background levels of metals in the environment. However, relatively higher accumulation pattern of Pb and Cd were noted in liver tissues of hare from the mine area compared to the background area, but did not induce increased lesions. Surface soils near the mine area contained relatively higher levels of trace metals (Zn>Mn>Pb>Cd>As) compared to reference soils, and with soil levels of Cd showing strong correlation with Cd accumulation in kidney tissues. Generally, both case and reference animals showed similar but varying severities of hepatic and renal lesions at the sublethal level, notably vascular congestion, occasional large hepatocyte nuclei, binucleate hepatocytes, yellow-brown pigmentation in the cytoplasm of hepatocytes and clustering of lymphocytes. Only hares with relatively higher accumulation of Pb from the mine area showed evidence of renal edema and hemorrhage of the capsular surface. This study constitutes the first assessment of metal induced histopathological alterations in arctic hares exposed to a historical mining area in the high arctic.

Genetic characteristics and changing antimicrobial resistance among Shigella spp. isolated from hospitalized diarrhoeal patients in Kolkata, India.

To study the prevalence pattern and trends in the phenotypic and genetic characteristics of shigellae, we tested 212 isolates isolated from diarrhoeal patients admitted to the Infectious Diseases Hospital, Kolkata, India, from November 2007 to October 2010. Prevalence of Shigella spp. was higher in the >5 years age group (69 %) than in children in the <5 years age group (31 %). Serotypes 2a, 3a and untypable isolates of Shigella flexneri were frequently detected. An increase in the isolation of Shigella sonnei (15 %) is a novel trend in this region. Fluoroquinolone resistance among S. flexneri serotypes 2a, 3a and other serogroups of shigellae is another evolving trend. The set gene was exclusively present in S. flexneri 2a, and the sen gene was detected in all serogroups. PFGE revealed the grouping of S. flexneri isolates according to their serotypes with approximately 80-100 % similarity, whilst Shigella dysenteriae type 2 and S. sonnei were clonal in nature. There was no demarcation in the prevalence of serotypes, antimicrobial resistance or clonality between the two age groups.

Role of probiotic in preventing acute diarrhoea in children: a community-based, randomized, double-blind placebo-controlled field trial in an urban slum.

Acute diarrhoea remains a major public health challenge in developing countries. We examined the role of a probiotic in the prevention of acute diarrhoea to discover if there was an effect directed towards a specific aetiology. A double-blind, randomized, controlled field trial involving 3758 children aged 1-5 years was conducted in an urban slum community in Kolkata, India. Participants were given either a probiotic drink containing Lactobacillus casei strain Shirota or a nutrient drink daily for 12 weeks. They were followed up for another 12 weeks. The primary outcome of this study was the occurrence of first episodes of diarrhoea. We assessed this during 12 weeks of intake of study agent and also for 12 weeks of follow-up. There were 608 subjects with diarrhoea in the probiotic group and 674 subjects in the nutrient group during the study period of 24 weeks. The level of protective efficacy for the probiotic was 14% (95% confidence interval 4-23, P<0·01 in adjusted model). The reduced occurrence of acute diarrhoea in the probiotic group compared to nutrient group was not associated with any specific aetiology. No adverse event was observed in children of either probiotic or nutrient groups. The study suggests that daily intake of a probiotic drink can play a role in prevention of acute diarrhoea in young children in a community setting of a developing country.

Multilocus variable-number tandem repeat analysis of Vibrio cholerae O1 El Tor strains harbouring classical toxin B.

Atypical Vibrio cholerae O1 strains - hybrid strains (strains that cannot be classified either as El Tor or classical biotype) and altered strains (El Tor biotype strains that produce classical cholera toxin) - are currently prevalent in Asia and Africa. A total of 74 hybrid and altered strains that harboured classical cholera toxin were investigated by multilocus variable-number tandem repeat analysis (MLVA). The results showed that the hybrid/altered strains could be categorized into three groups and that they were distant from the El Tor strain responsible for the seventh cholera pandemic. Hybrid/altered strains with a tandem repeat of the classical CTX prophage on the small chromosome were divided into two MLVA groups (group I: Mozambique/Bangladesh group; group III: Vietnam group), and altered strains with the RS1-CTX prophage containing the El Tor type rstR and classical ctxB on the large chromosome were placed in two MLVA groups (group II: India/Bangladesh group; group III: India/Vietnam group).

Classification of hybrid and altered Vibrio cholerae strains by CTX prophage and RS1 element structure.

Analysis of the CTX prophage and RS1 element in hybrid and altered Vibrio cholera O1 strains showed two classifiable groups. Group I strains contain a tandem repeat of classical CTX prophage on the small chromosome. Strains in this group either contain no element(s) or an additional CTX prophage or RS1 element(s) on the large chromosome. Group II strains harbor RS1 and CTX prophage, which has an E1 Tor type rstR and classical ctxB on the large chromosome.

Increasing antimicrobial resistance--an emerging problem in the treatment of shigellosis.

Shigellosis is a major cause of diarrhoea-related morbidity and mortality, especially in developing countries. Effective antibiotic treatment reduces the average duration of illness by reducing faecal excretion of the bacterium and preventing further transmission and potentially lethal complications. Treatment of shigellosis is currently limited by the high prevalence of multidrug-resistant strains of Shigella. Although fluoroquinolones are currently effective in treating adults, resistance to fluoroquinolones among Shigella spp. is emerging, and their use in children is subject to limitations. Azithromycin and third-generation cephalosporins are also effective in the treatment of shigellosis, but monitoring of Shigella isolates to detect the emergence of resistance is essential.

Phage typing, biotyping & antimicrobial resistance profile of Salmonella enterica serotype Typhi from Kolkata.

BACKGROUND & OBJECTIVE: Kolkata and its suburbs in eastern India are known to be endemic for typhoid fever. The objective of this study was to determine phage types, biotypes and antimicrobial resistance patterns of Salmonella enterica serotype Typhi isolated during the period 2003-2005 from a prospective surveillance for typhoid fever in two urban slums in Kolkata. METHODS: A total of 195 Salmonella enterica serotype Typhi isolated from blood cultures were phage typed, biotyped and tested for their antimicrobial susceptibility profile. RESULTS: Phage type E1 was the most common (60.3%) followed by phage type A among five phage types identified. Biotype I (95%) was predominant, 28 isolates were multidrug resistant (MDR) and most of the MDR strains belonged to phage type E1 and biotype I. INTERPRETATION & CONCLUSION: A single phage type and biotype were prevalent among the Salmonella enterica serotype Typhi isolates studied from Kolkata, India.

Molecular epidemiology of human astrovirus infections in Kolkata, India.

UNLABELLED: The study is aimed to determine the seasonal distribution and clinical characteristics of astroviruses associated with acute watery diarrhoea among children in Kolkata and characterize them at the molecular level. METHOD OF STUDY: Faecal specimens of acute watery diarrhoea cases (n=857) and non-diarrhoeic samples (n=211) from the hospitals and a nearby field community were screened with IDEIA Astrovirus detection kit; astrovirus co-infections with rotavirus and/or picobirnavirus were detected by RNA-PAGE and silver staining. Further RT-PCR was carried out using specific primers, viz. Mon340 (+) and Mon348 (-) targeting a highly conserved domain of ORF1a (289 bp) of human astroviruses. RESULTS: Astrovirus infection was detected in 50 cases (50/857); astroviruses were detected mostly in children aged 6-12 months (50%); all non-diarrhoeic samples (n=211) were negative for astrovirus. In 52% of astrovirus positive cases, the virus was detected as the sole agent; mixed infections were also detected with other diarrhoeic pathogens such as rotavirus (32%), picobirnavirus (2%), rotavirus and picobirnavirus (2%), picobirnavirus and Enterotoxigenic E. coli (ETEC) (2%), rotavirus and ETEC (2%), rotavirus and Enteroaggregative E. coli (EAEC) (2%), Enteropathogenic E. coli (EPEC) (2%), Shigella flexneri type 3a (2%) and Ascaris (2%). RT-PCR and sequencing of amplicons of astroviruses from Kolkata, with specific primers targeted to the conserved domain of ORF1a (289 bp) of the astrovirus genome, showed maximum homology to the astrovirus strain ("5-158") from Seoul (98%). RESULTS AND CONCLUSIONS: Clinical characteristics of the diarrhoeic children in Kolkata indicated that astrovirus infections were detected throughout the year and were associated with varying degree of dehydration and acute watery diarrhoea. In-depth molecular epidemiological surveillance of astroviruses in Kolkata is essential for better understanding of their overall genetic nature.

Epidemiological, microbiological & electron microscopic study of a cholera outbreak in a Kolkata slum community.

BACKGROUND & OBJECTIVE: Epidemics of cholera caused by Vibrio cholerae O1 or O139 have been reported from different parts of India. Factors such as unsafe water supply, poor environmental sanitation, indiscriminate defaecation and lack of personal hygiene are mainly responsible for continued transmission of this disease. We report here epidemiological and microbiological findings of a localized outbreak of cholera, which occurred during March and April 2004 in the eastern part of Kolkata city. METHODS: The affected slum area has a population of 4409, predominantly muslims. Patients suffering from acute watery diarrhoea attended the health outposts organized by National Institute of Cholera and Enteric Diseases, Kolkata and International Vaccine Institute, South Korea as part of a routine surveillance programme at the locality as well as the emergency medical camp organized by the Kolkata Municipal Corporation. Stool and water samples were collected and tested for diarrhoeagenic pathogens in the laboratory. Bacteriophages specific for V. cholerae were isolates and studied electron microscopically for morphology. RESULTS: A total of 89 diarrhoea cases were reported giving an attack rate of 2 per cent. V. cholerae O1 biotype ElTor, serotype Ogawa was isolated as a sole pathogen from 15 (15.8%) of 89 stool samples screened. Water samples (2 from tube wells, 3 from municipal taps and 1 from well) showed presence of coliform bacilli with high MPN (Most Probable Number) count. Bacteriophages specific to V. cholerae were isolated from 2 of 6 water samples examined. A leakage was detected in the main pipeline supplying drinking water to that area. INTERPRETATION & CONCLUSION: The outbreak was caused by V. cholerae O1 (Ogawa) biotype ElTor. The presence of phages in the water samples was an additional indicator for V. cholerae contamination in this community. Occurrences of such outbreaks support vaccination against cholera as an alternative strategy.

Interaction between dietary calcium supplementation and chronic waterborne zinc exposure in juvenile rainbow trout, Oncorhynchus mykiss.

This study investigated the effects of dietary Ca2+ on branchial Ca2+ and Zn2+ uptake, new and total zinc accumulation in target tissues (gill, liver and kidney), calcium and zinc homeostasis, and acute tolerance to waterborne zinc in fish chronically exposed to waterborne zinc. Juvenile rainbow trout (Oncorhynchus mykiss) were maintained on a calcium-enriched diet [41.2 mg vs. 21.2 mg (control) calcium/g dry wt. of food] and chronic waterborne zinc exposure (2.3 micromol/L), both separately and in combination, for 28 days. Calcium-supplemented diet in the absence of waterborne zinc significantly reduced branchial Ca2+ and Zn2+ influx rates, and new and total zinc accumulations in target tissues relative to control. However it did not protect against the acute zinc challenge. In contrast, waterborne zinc exposure significantly increased branchial Ca2+ and Zn2+ influx rates, new and total zinc concentrations in target tissues, and acute zinc tolerance relative to control. Interestingly, no such changes in any of these parameters were recorded in fish treated simultaneously with elevated dietary Ca2+ and waterborne zinc, except acute zinc tolerance which was highest among all the treatments. Thus, we conclude that the interactions between elevated dietary Ca2+ and waterborne zinc can protect freshwater fish against waterborne zinc toxicity.

Food selection, growth and physiology in relation to dietary sodium chloride content in rainbow trout (Oncorhynchus mykiss) under chronic waterborne Cu exposure.

Waterborne Cu is toxic to Na(+) and Cl(-) regulation in freshwater fish, and Cu is taken up, at least in part, via the Na(+)-transport pathway in the gills. Therefore, we hypothesized that freshwater fish may mitigate the toxic effects of waterborne Cu by selecting a NaCl-enriched diet over a normal diet. We tested this hypothesis in juvenile rainbow trout (Oncorhynchus mykiss) by offering them the choice between NaCl-enriched (1.9 mmol g(-1)Na(+)) and normal (0.2 mmol g(-1)Na(+)) diets under a chronic waterborne Cu exposure of 55 microg L(-1) for a period of 28 days. Contrary to expectation, trout exhibited a preference for NaCl-enriched diet under control conditions, while exposure to chronic waterborne Cu severely disrupted their normal feeding pattern with an accompanying loss of preference for the NaCl-enriched diet. Waterborne Cu exposure also severely affected appetite and growth. Both appetite and growth gradually recovered with time, but remained significantly impaired relative to Cu-unexposed fish until the end of the exposure. Waterborne Cu exposure also significantly increased Cu accumulations in target organs (gill, liver, and gut), plasma and whole body. However, Cu accumulation decreased substantially towards the end of the exposure in target organs and whole body as well as in plasma in Cu-exposed fish with dietary choice relative to Cu-exposed fish with normal diet. These adjustments were concurrent with the gradual recovery of appetite, which also led to increased ingestion of the NaCl-enriched diet. Interestingly, this elevated dietary uptake of NaCl produced significant stimulation of Na(+) efflux in Cu-exposed fish. Subsequently, it also led to significant elevation of Na(+) levels in target organs and whole body, and restored the decrease of plasma Na(+) and Cl(-) levels in Cu-exposed fish. The NaCl supplemented diet appeared to be beneficial in compensating Na(+) and Cl(-) losses from the body induced by waterborne Cu. Overall, these results demonstrate that a NaCl-enriched diet, although consumed in relatively reduced quantities due to the impairment of food selection and appetite, can help to protect freshwater fish against chronic waterborne Cu toxicity.

Epidemiology of HIV infection among brothel-based sex workers in Kolkata, India.

A cross-sectional study was carried out during January-June 2004 to assess the status of HIV infection among brothel-based sex workers in Kolkata city, India. Six hundred and twenty-two sex workers, selected from six brothels, were included in the study to assess their HIV status, and 362 sex workers, a subset of the above population, were interviewed to study their risk behaviour and practices. Blood samples were collected from each sex worker for testing HIV by an unlinked anonymous method. The prevalence of HIV infection was 9.6%, but was much higher among younger sex workers aged 20 years or less (27.7%) compared to the older age group (8.4%). This difference was statistically significant, indicating an association of younger ages with HIV infection [p = 0.006 and odds ratio (OR) = 4.18; (95% confidence interval (CI) 1.2-13.8)]. HIV was not associated with duration of sex work, average number of clients entertained per day, condom use, practice of sex during menstruation, pre-coital examination of penis for visible ulcer/discharge, suffering from sexually transmitted infections, and entertaining clients outside the brothel. There was a big gap between the reported and the evaluated condom use by sex workers. The results suggest that there is a need to develop suitable HIV intervention strategies, considering the socioeconomic and cultural aspects of city sex workers, with a provision for continuous monitoring and evaluation.

Species diversity and antimicrobial resistance of Shigella spp. isolated between 2001 and 2004 from hospitalized children with diarrhoea in Kolkata (Calcutta), India.

The incidence, phenotypic characteristics and antimicrobial resistance patterns of 193 Shigella strains isolated from 2489 hospitalized children with acute diarrhoea were studied during January 2001 to August 2004. S. flexneri (60%) was the most prevalent serogroup, followed by S. sonnei (23.8%), S. dysenteriae (9.8%) and S. boydii (5.7%). Since 2002, S. flexneri 2a was the most dominant serotype. Almost all S. flexneri strains exhibited resistance to ampicillin, co-trimoxazole, tetracycline, nalidixic acid and fluoroquinolones. After a lapse of almost 14 years, S. dysenteriae type 1 strains reemerged for the first time during 2002 and these strains were resistant to more than two antibiotics (multidrug resistance), including fluoroquinolones. An upsurge of similar resistance patterns was also noted among S. flexneri type 2a since December 2003. Resistance to fluoroquinolone increased year on year among S. dysenteriae type 1 and S. flexneri, but not in S. boydii or S. sonnei. Monitoring of antimicrobial susceptibility through a surveillance programme is recommended to select appropriate antibiotics for the effective treatment of shigellosis in this region.