Study on the prevalence of intestinal parasitic infections and the assessment of the efficacy of albendazole in soil-transmitted helminths in school-going children in East Sikkim.

BACKGROUND: The problem of intestinal parasitic infections (IPIs) in children is one of the most worrisome problems worldwide. The latest estimates indicate that more than 880 million children are in the need of treatment for these parasites. OBJECTIVE: The aim of this study was to measure the prevalence of IPIs in school-going children in East Sikkim, India, and to assess the efficacy of single-dose albendazole (ALB) in children infected with soil-transmitted helminths (STHs). SUBJECTS AND METHODS: A total of 300 stool samples were collected from the schoolchildren of government schools of East Sikkim. Samples were processed for the identification of IPIs by direct microscopy and formalin-ether concentration method. Fecal egg counting was carried out for STH by Stoll's egg counting technique, pre- and posttreatment with single-dose ALB. The second stool samples were collected 10-14 days posttreatment of ALB. Cure rate (CR) and the fecal egg reduction rate (ERR), the two most widely used indicators for assessing the efficacy of an anthelmintic, were used in this study. The data were analyzed and the results were interpreted statistically. RESULTS: The overall prevalence of the IPIs was 33.9%. Helminthic infection was 4.6% and protozoan infection was 29.3%. Among helminthes Ascaris lumbricoides and among protozoans Entamoeba spp. were the dominant intestinal parasites. For drug efficacy, A. lumbricoides had CR 55.5% and ERR 81.4%. Moreover, for Trichuris trichiura, CR and ERR was 100%. The study has shown less efficacy against A. lumbricoides infections compared to T. trichiura. CONCLUSION: The study provides useful insight into the current prevalence of IPIs in school-going children in government schools in East Sikkim region. Keeping in view of less efficacy of ALB, it is necessary to keep the monitoring of development of drug resistance simultaneously.

Molecular characterization and phylogenetic analysis of Explanatum explanatum in India based on nucleotide sequences of ribosomal ITS2 and the mitochondrial gene nad1.

The aim of this study was to analyze the phylogenetic relationship between Explanatum explanatum populations in India and other countries of the Indian subcontinent. Seventy liver amphistomes collected from four localities in India were identified as E. explanatum based on the nucleotide sequences of ribosomal ITS2. The flukes were then analyzed phylogenetically based on the nucleotide sequence of the mitochondrial gene nad1 in comparison with flukes from Bangladesh and Nepal. In the resulting phylogenetic tree, the nad1 haplotypes from India were divided into four clades, and the flukes showing the haplotypes of clades A and C were predominant in India. The haplotypes of the clades A and C have also been detected in Bangladesh and Nepal, and therefore, it seems they occur commonly throughout the Indian subcontinent. The results of AMOVA suggested that gene flow was likely to occur between E. explanatum populations in these countries. These countries are geographically close and have been historically and culturally connected to each other, and therefore, the movements of host ruminants among these countries might have been involved in the migration of the flukes and their gene flow.

Application of water quality index for the assessment of suitability of natural sources of water for drinking in rural areas of east Sikkim, India.

In Sikkim, especially in the rural areas where there is no supply of treated water for drinking and other domestic uses, natural surface water is the only source. The objective was to assess the water quality of natural sources of water in the rural areas of East Sikkim using a water quality index (WQI) for different seasons. A total of 225 samples, that is, 75 in winter, 75 in summer, and 75 in monsoon were collected from different sources for physicochemical analysis, and a WQI was calculated. The water quality values ranged 32.01-96.71. The results showed that most of the water samples were in poor condition (85.3%) and very few of them were in good condition (2.6%). The water quality of the natural sources indicated that the water is poor-quality and not totally safe for human consumption, and that it needs treatment before consumption.

Molecular phylogenetic analysis of Fasciola flukes from eastern India.

Fasciola flukes from eastern India were characterized on the basis of spermatogenesis status and nuclear ITS1. Both Fasciola gigantica and aspermic Fasciola flukes were detected in Imphal, Kohima, and Gantoku districts. The sequences of mitochondrial nad1 were analyzed to infer their phylogenetical relationship with neighboring countries. The haplotypes of aspermic Fasciola flukes were identical or showed a single nucleotide substitution compared to those from populations in the neighboring countries, corroborating the previous reports that categorized them in the same lineage. However, the prevalence of aspermic Fasciola flukes in eastern India was lower than those in the neighboring countries, suggesting that they have not dispersed throughout eastern India. In contrast, F. gigantica was predominant and well diversified, and the species was thought to be distributed in the area for a longer time than the aspermic Fasciola flukes. Fasciola gigantica populations from eastern India were categorized into two distinct haplogroups A and B. The level of their genetic diversity suggests that populations belonging to haplogroup A have dispersed from the west side of the Indian subcontinent to eastern India with the artificial movement of domestic cattle, Bos indicus, whereas populations belonging to haplogroup B might have spread from Myanmar to eastern India with domestic buffaloes, Bubalus bubalis.

Antibiotic susceptibility profile of bacteria isolated from natural sources of water from rural areas of East sikkim.

BACKGROUND: Contamination of water, food, and environment with antibiotic-resistant bacteria poses a serious public health issue. OBJECTIVE: The objective was to study the bacterial pollution of the natural sources of water in east Sikkim and to determine the antimicrobial profile of the bacterial isolates. MATERIALS AND METHODS: A total of 225 samples, 75 each during winter, summer, and monsoon season were collected from the same source in every season for bacteriological analysis by membrane filtration method. Antibiotic susceptibility test was performed using standard disc diffusion method. RESULTS: A total of 19 bacterial species of the genera Escherichia, Klebsiella, Proteus, Salmonella, Shigella, Enterobacter, Citrobacter, Morganella, Pseudomonas, Acinetobacter, Flavobacterium, and Serratia were isolated and their antimicrobial sensitivity tested. Generally, most bacterial isolates except Salmonella and Shigella species were found resistant to commonly used antibiotics such as ampicillin (57.5%), trimethoprim/sulfamethoxaole (39.1%), amoxicillin/clavulanic acid (37.4%), cefixime (34.5%), tetracycline (29.1%), ceftazidime (26.3%), ofloxacin (25.9%), amikacin (8.7%), and gentamicin (2.7%) but sensitive to imipenem and piperacillin/tazobactam. CONCLUSION: Natural sources of water in east Sikkim are grossly contaminated with bacteria including enteropathogens. The consumption of untreated water from these sources might pose health risk to consumers.

Paragonimus & paragonimiasis in India.

Ever since the discovery of the first indigenous case in 1981, paragonimiasis has gained recognition as a significant food borne parasitic zoonosis in India. The data available on the occurrence of paragonimiasis, until today, may be just the tip of an iceberg as the study areas covered were restricted to Northeast Indian States. Nevertheless, the results of research on paragonimiasis in India have revealed valuable information in epidemiology, life cycle, pathobiology and speciation of Indian Paragonimus. Potamiscus manipurensis, Alcomon superciliosum and Maydelliathelphusa lugubris were identified as the crab hosts of Paragonimus. Paragonimus miyazakii manipurinus n. sub sp., P. hueit'ungensis, P. skrjabini, P. heterotremus, P. compactus, and P. westermani have been described from India. P. heterotremus was found as the causative agent of human paragonimiasis. Ingestion of undercooked crabs and raw crab extract was the major mode of infection. Pulmonary paragonimiasis was the commonest clinical manifestation while pleural effusion and subcutaneous nodules were the common extra-pulmonary forms. Clinico-radiological features of pulmonary paragonimiasis simulated pulmonary tuberculosis. Intradermal test, ELISA and Dot-immunogold filtration assay (DIGFA) were used for diagnosis and epidemiological survey of paragonimiasis. Phylogenitically, Indian Paragonimus species, although nested within the respective clade were distantly related to others within the clade.

A case of cutaneous paragonimiasis presented with minimal pleuritis.

Clinically, paragonimiasis is broadly classified into pulmonary, pleuropulmonary, and extrapulmonary forms. The common extrapulmonary forms are cerebral and cutaneous paragonimiasis. The cutaneous paragonimiasis is usually presented as a slowly migrating and painless subcutaneous nodule. The correct diagnosis is often difficult or delayed or remained undiagnosed until the nodule becomes enlarged and painful and the cause is investigated. We report here a case of cutaneous paragonimiasis in a male child who presented with mild respiratory symptoms. The diagnosis of paragonimiasis was based on a history of consumption of crabs, positive specific serological test, and blood eosinophilia. The swelling and respiratory symptoms subsided after a prescribed course of praziquantel therapy.

Evaluation of phenotypic with genotypic methods for species identification and detection of methicillin resistant in Staphylococcus aureus.

BACKGROUND: Phenotypic methods for the detection of methicillin resistance are inadequate, due to presence of hetero-resistant population and dependence of environmental factors that may affect the phenotypic expression of resistance. AIMS: Present study was conducted, to evaluate the efficacy of phenotypic methods for the identification of species and mec-A mediated resistance in S. aureus with polymerase chain reaction (PCR), and to assess the prevalence of the Panton-Valentine leukocidin (pvl) toxin in methicillin resistant S. aureus (MRSA) and overall S.aureus population. MATERIALS AND METHODS: A total of 200 clinical isolates of Staphylococci were subjected to phenotypic and genotypic methods for the species identification and detection of MRSA. RESULTS: The specificity and sensitivity of conventional methods in the detection of S.aureus, was found to be 100 and 97.59% respectively. However, the performance of phenotypic methods in the detection of MRSA were: Oxacillin disc diffusion (DD)-sensitivity 70.58%, specificity 75.75%; cefoxitin DD-sensitivity 86.27%, specificity 83.33%; and oxacillin agar dilution-sensitivity 92.15%, specificity 90.90%. PVL gene was detected in all mec-A positive isolates irrespective of their types. CONCLUSION: Phenotypic methods still preferred for the species identification, but for the reliable detection of MRSA an algorithm should include a combination of tests and apply a genotypic method for confirmation of resistance isolates showing discrepant results. Considering the high prevalence of PVL-MRSA, we recommend PCR as assay, as it has an advantage of simultaneous detection of mec-A and pvl genes by multiplex PCR.

Experimental infection with Paragonimus heterotremus metacercariae in laboratory animals in Manipur, India.

This study was aimed to find out the host-parasite relationship between Paragonimus heterotremus isolated as metacercariae from mountain crabs, Indochinamon manipurensis, in Manipur, India and laboratory animals such as puppies, albino rats, Swiss mice, guinea pigs, and rabbits, as experimental animals. The animals were fed with the metacercariae. Infected animals were sacrificed 35 to 430 days after feeding to recover worms, which were used to determine the developmental stages. Adult worms (n = 14) were recovered from 3 puppies > or = 70 days after feeding and immature worms (n = 25) were recovered from 2 other puppies 35 or 43 days after infection. The infection rate in puppies was 100%. Juvenile worms were recovered from 3 of 13 rats: 1 of 11 rats whose viscera and cavities were examined and both of two rats whose muscles were examined. Rats were not a suitable animal model for pulmonary infection with P. heterotremus. Mice, guinea pigs, and rabbits were also found to be insusceptible to pulmonary infection with P. heterotremus.

Cerebral paragonimiasis mimicking tuberculoma: First case report in India.

An 8-year-old male child of Tuensang District, Nagaland, India, attended Civil Hospital, Tuensang, complaining of cough, fever, headache, and inability to move right arm since one month. On clinical suspicion of tubercular meningitis, anti-tubercular therapy was initiated and the patient was referred to the Naga Hospital Authority. A brain computed tomography scan revealed an isodense area with surrounding edema on the left parietal lobe, which was diagnosed as tuberculoma and the anti-tubercular therapy was continued. As there was no sign of clinical improvement on completion of the three-month-ATD regimen, the patient was investigated for paragonimiasis. Laboratory investigations revealed peripheral blood eosinophilia, raised ESR, Paragonimus egg-positive sputum, and positive Enzyme-linked immunosorbent assay (ELISA) and ID tests for paragonimiasis. The Bacillus Calmette-Guérin (BCG) test and Acid Fast Bacilli (AFB) sputum smears were negative. Chest roentgenogram showed no abnormal findings. A final diagnosis of pulmonary paragonimiasis associated with cerebral paragonimiasis was made. The patient responded to praziquantel therapy. Cerebral paragonimiasis is a serious extrapulmonary form of paragonimiasis, sometimes life-threatening, but curable with praziquantel. It should be included in the differential diagnosis of cerebral granulomatous and other space-occupying lesions.

Occurrence of the lung fluke, Paragonimus heterotremus in Manipur, India.

OBJECTIVES: To determine the natural crustacean host, the Paragonimus species, and to investigate the potential host-parasite relationship between Manipur Paragonimus and some of the laboratory animals. METHODS: The laboratory animals such as puppies, albino rats and Swiss mice were infected orally with metacercariae isolated from the fresh water crabs, Potamiscus manipurensis. The fecal specimens of the experimentally infected animals were examined microscopically for Paragonimus eggs at regular intervals. The animals were autopsied on days 35 to approximately 328 after infection and the isolated worms were flattened between glass slides and fixed in 70% alcohol. The worms were stained with carmine and mounted with Canada balsam for morphological studies. The eggs were collected in 5% formol saline solution for microscopy. The flukes were classified into 4 developmental stages. RESULTS: A total of 11 worms, 5 mature, 5 immature and 1 pre-adult were recovered. The morphological features of the metacercariae, worms and eggs were similar to those of Paragonimus heterotremus. CONCLUSION: Manipur is one of the rare areas in the world where Paragonimus heterotremus is prevalent and the puppies are ideal experimental animal host. This species may be one of the important causes of paragonimiasis in animals and humans in Manipur.

Occurrence of the lung fluke Paragonimus hueit'ungensis in Manipur, India.

BACKGROUND: Paragonimus has emerged as an important parasitic zoonosis in recent years in Manipur, India. However, no scientific data on the systematic and phylogenetic classification of lung fluke in India is available. Therefore, a study was conducted to determine the species of lung fluke prevalent in Manipur by infecting an experimental animal with Paragonimus metacercariae obtained from fresh water crabs. METHODS: Thirteen Paragonimus metacercariae isolated from the fresh water crabs, Potamiscus manipurensis, at Churachandpur, Manipur, India were used to infect a puppy orally using a Pasteur pipette. After the infection, fecal specimens were checked for Paragonimus eggs by microscopic examination. Autopsy was performed on day 121 post-infection. The worms recovered from the worm cysts were alcohol fixed, flattened and stained with carmine for morphological study. RESULTS: Four adult worms were recovered. The worms presented with singly spaced cuticular spines, and the ventral suckers were slightly larger than the oral suckers. The ovaries consisted of 4 to 5 lobed masses branched like a ginger stem, and the testes were irregularly lobed and elongated. One ofthe worms showed a different shape and size ofthe right testis. The morphological features of the worms are similar to the Chinese species, Paragonimus hueit'ungensis. CONCLUSION: The results of this study indicate the possibility of occurrence of P hueit'ungensis as 1 of the important Paragonimus species prevalent in Manipur, India after its initial discovery in China.