Immune response and intestinal permeability in children with acute gastroenteritis treated with Lactobacillus rhamnosus GG: a randomized, double-blind, placebo-controlled trial.

BACKGROUND: Probiotics have a possible role in the treatment of pediatric acute gastroenteritis. We report the effect of the probiotic Lactobacillus rhamnosus GG (LGG) on intestinal function, immune response, and clinical outcomes in Indian children with cryptosporidial or rotavirus diarrhea. METHODS: Children with gastroenteritis aged 6 months to 5 years, testing positive for either rotavirus or Cryptosporidium species in stool (coinfections were excluded), were randomized to LGG (ATCC 53103) or placebo, once daily for 4 weeks. Baseline demographic and clinical details were obtained. Sera were tested for immunoglobulin G (IgG) and immunoglobulin A (IgA) antibodies to Cryptosporidium and rotavirus, and the lactulose to mannitol ratio for intestinal permeability was determined at baseline and at the end of follow-up. RESULTS: Of the 124 children enrolled, 82 and 42 had rotavirus and cryptosporidial diarrhea, respectively. Median diarrheal duration was 4 days; one-third of the children had severe diarrhea. Baseline and clinical parameters were comparable between children receiving LGG and placebo. At the end of follow-up, fewer children with rotavirus diarrhea on LGG had repeated diarrheal episodes (25% vs 46%; P = .048) and impaired intestinal function (48% vs 72%; P = .027). Significant increase in IgG levels postintervention (456 vs 2215 EU; P = .003) was observed in children with rotavirus diarrhea receiving LGG. Among children with cryptosporidial diarrhea, those receiving LGG showed significant improvement in intestinal permeability. CONCLUSIONS: LGG has a positive immunomodulatory effect and may be useful in decreasing repeated episodes of rotavirus diarrhea. Improvement in intestinal function in children with rotavirus and cryptosporidial gastroenteritis emphasizes the role of probiotics in treating intestinal impairment after infection. CLINICAL TRIALS REGISTRATION: CTRI/2010/091/000339.

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.

Rotavirus shedding in symptomatic and asymptomatic children using reverse transcription-quantitative PCR.

Reverse transcription-real-time polymerase chain reaction (RT-qPCR) for the VP6 gene was used to study group A rotavirus shedding in children with symptomatic and asymptomatic rotavirus infection. Sequential stool samples (n = 345) from 10 children with rotavirus associated diarrhea and from five children (n = 161) with asymptomatic rotavirus infection were collected over a period of 2 months. A RT-qPCR assay on the samples using a rotavirus VP6 plasmid standard demonstrated high reproducibility, with an inter-assay coefficient of variation (CV) of 1.40-2.97% and an intra-assay CV of 0.03-3.03%. The median duration of shedding was longer in children with diarrhea compared to asymptomatic children (24 days vs. 18 days; P = 0.066). The median quantitation cycle (C(q)) at presentation in symptomatic children was 17.21 compared to 30.98 in asymptomatic children (P = 0.086). The temporal pattern in symptomatic children consisted of a high initial viral shedding coinciding with the duration of diarrhea, followed by a rapid fall, and then a small increase in secondary shedding 21 days later. Compared to children with rotavirus diarrhea, those with asymptomatic infection shed lower quantities of virus throughout the observation period. No difference was noted between the G and P genotypes of samples collected at onset of infection and during the shedding period. Shedding was intermittent in a subset of children in both groups. RT-qPCR is a useful method to characterize shedding patterns.

Rotavirus antigenemia in Indian children with rotavirus gastroenteritis and asymptomatic infections.

BACKGROUND: Rotavirus gastroenteritis results in significant morbidity and mortality in Indian children. Although there are numerous studies on rotavirus diarrhea, there are few reports on antigenemia and extraintestinal presentations in these populations. METHODS: Following screening for rotavirus antigen of stool samples from children with and without acute gastroenteritis with a commercial enzyme immunoassay (EIA), a total of 199 stool and serum sample pairs were identified for additional testing. All EIA-positive stool samples were genotyped, and viral load estimated by real-time reverse-transcriptase polymerase chain reaction (RT-PCR). Serum samples were tested for rotavirus antigen by an in-house EIA, and antigen was quantified by optical density. Scoring of disease severity was performed for all hospitalized children. Data on extra-intestinal presentations were collected if available. RESULTS: Based on screening of stool samples by EIA, the study population could be divided into 3 groups, including 111 children with rotavirus diarrhea, 44 children with diarrhea and no rotavirus detected in stool specimens, and 44 children with asymptomatic rotavirus infection. Antigenemia was significantly higher among children with rotavirus diarrhea (50.4%) than among children with non-rotaviral diarrhea (16%) or asymptomatic infections (2.3%) (P < .001). Low copies of rotavirus were detected by RT-PCR in all 7 children with EIA-negative stool specimens and antigenemia. Presence and levels of rotavirus antigen in serum specimens correlated with stool viral load. Children with antigenemia had significantly more-severe disease but not more extraintestinal presentations than did children without antigenemia. CONCLUSIONS: Antigenemia occurs frequently in rotavirus infection and correlates with virus replication in the gut but not with extra-intestinal presentations.

Comparison of group counseling with individual counseling in the comprehension of informed consent: a randomized controlled trial.

BACKGROUND: Studies on different methods to supplement the traditional informed consent process have generated conflicting results. This study was designed to evaluate whether participants who received group counseling prior to administration of informed consent understood the key components of the study and the consent better than those who received individual counseling, based on the hypothesis that group counseling would foster discussion among potential participants and enhance their understanding of the informed consent. METHODS: Parents of children participating in a trial of nutritional supplementation were randomized to receive either group counseling or individual counseling prior to administration of the informed consent. To assess the participant's comprehension, a structured questionnaire was administered approximately 48-72 hours afterwards by interviewers who were blinded to the allocation group of the respondents. RESULTS: A total of 128 parents were recruited and follow up was established with 118 (90.2%) for the study. All respondents were aware of their child's participation in a research study and the details of sample collection. However, their understanding of study purpose, randomization and withdrawal was poor. There was no difference in comprehension of key elements of the informed consent between the intervention and control arm. CONCLUSIONS: The results suggest that the group counseling might not influence the overall comprehension of the informed consent process. Further research is required to devise better ways of improving participants' understanding of randomization in clinical trials.

Rotavirus infection in the neonatal nurseries of a tertiary care hospital in India.

BACKGROUND: The majority of neonatal rotavirus infections are believed to be asymptomatic, and protection from subsequent infection and disease has been reported in neonatally infected children. In this study, we present the results of a 4-year prospective surveillance in the neonatal nurseries of a tertiary care hospital in south India. METHODS: Stool samples from neonates admitted for >48 hours either with gastrointestinal (GI) symptoms or with nonenteric pathology were screened for rotavirus. Careful assessment of clinical data was carried out. G- and P-typing for all symptomatic rotavirus positive cases and equal number of asymptomatic controls from the same month was determined by reverse transcription polymerase chain reaction. RESULTS: Rotavirus was detected in 43.9% of 1411 neonates, including those with and without gastrointestinal disease. Rotavirus detection was significantly higher among neonates with GI disease (55.5%) than asymptomatic neonates (44.4%) (P < 0.001). Rotavirus was seen in association with diarrhea, vomiting, feed intolerance, necrotizing enterocolitis, hematochezia, gastroesophageal reflux, and abdominal distension. Diarrhea was significantly more frequent in neonates with rotavirus infection (P < 0.001) whereas uninfected neonates developed significantly more feeding intolerance (P < 0.001). Significantly greater proportion of term neonates with GI disease were positive for rotavirus than preterm neonates (P < 0.001). G10P[11] was the most common genotype associated with both symptomatic and asymptomatic infections. CONCLUSIONS: This study documents the high rates of rotavirus infection in the neonatal nurseries and the continuing detection of the G10P[11] strain associated with GI disease in Vellore.

Investigation of a hepatitis A outbreak in children in an urban slum in Vellore, Tamil Nadu, using geographic information systems.

BACKGROUND & OBJECTIVE: An outbreak of symptomatic viral hepatitis in children less than 10 yr of age in Vellore, south India, was investigated and the disease pattern studied using serological and epidemiological methods, supplemented by geographic information systems (GIS) mapping. METHODS: Three cases of hepatitis A were identified during routine surveillance in a birth cohort House-to-house visits were undertaken to identify other symptomatic cases and samples collected for anti- HAV IgM, ELISA testing. All cases and controls were mapped and geo-referenced using Arc View GIS 3.3. Spatial clustering was investigated using SaTScan 7.0.1 software. Drinking water sources were tested for coliform counts with the most probable number technique. RESULTS: Of the 965 children surveyed, 26 (2.78%) had jaundice between February to July 2006. From the 26 patients, 11 (42.3%) blood samples were obtained and tested for anti-HAV IgM; 10 (90.9%) were found to be positive. Water analysis showed high coliform counts in all samples. No spatial clustering of cases could be detected. INTERPRETATION & CONCLUSION: The outbreak was identified because of the symptomatic presentation of the cases. Our study highlighted the increasing detection of symptomatic children with hepatitis A virus infection. Water sources in the area were contaminated and may have served as the source of infection. The lack of clustering in GIS analysis could be due to the common water source.

Severity of rotavirus gastroenteritis in an Indian population: report from a 3 year surveillance study.

This study investigated the severity of rotavirus gastroenteritis (RVGE) in hospitalized children less than 60 months of age and compared severity in the first five months of life to severity in children 6 to 23 months of age. Results from a 3 year surveillance study show an early peak of rotavirus disease, with 117 (31%) RVGE hospitalizations in children <6 months old. Higher incidence of severe dehydration, acidemia and acidosis at admission and prolonged hospitalization >7 days were seen in infants 0-5 months of age. The findings support the need for consideration of timely immunization or an accelerated immunization schedule with a birth dose to protect this vulnerable age.

Severity of rotavirus gastroenteritis in Indian children requiring hospitalization.

INTRODUCTION: The burden of rotavirus gastroenteritis is greatest in India and other developing countries. With the availability of two licensed vaccines and a number of additional vaccines in various stages of development and trial, analysis of detailed clinical information is essential for the development of a uniform method of severity assessment. METHODS: Diarrhoeal stool samples from 1001 children <5 years of age hospitalized with gastroenteritis were screened for rotavirus using a commercial enzyme immunoassay. Positive samples were confirmed by genotyping using hemi nested multiplex RT-PCR. Detailed clinical data was collected for gastroenteritis assessment for 934 children and extraintestinal presentations were analyzed in 470 children. Severity scoring was carried out for all children using the Vesikari score and in a subset by Clark's scoring system. RESULTS: Rotavirus was detected in 35.4% of samples tested between December 2005 and November 2008. Clark's and Vesikari scores showed moderate correlation but varied greatly in the categorization of severe disease. Using Clark's scoring, only 1.6% were categorized as presenting with severe disease in comparison to 66.1% by the Vesikari score. Association of extraintestinal symptoms with rotavirus gastroenteritis was not documented in this study. CONCLUSION: The assessment of disease severity using two common severity scoring systems highlights the difference in the categorization of "severe" disease. This underscores the need for a robust scoring system which is needed for vaccine trial and in post-licensure surveillance, because vaccine efficacy is estimated for protection against severe rotavirus gastroenteritis.

Disease and economic burden of rotavirus diarrhoea in India.

We used published and unpublished studies and national statistics to estimate the number of deaths, hospitalizations, and outpatient visits due to rotavirus diarrhoea and the associated national economic burden of disease in India. Annually in India, rotavirus diarrhoea causes an estimated 122,000-153,000 deaths, 457,000-884,000 hospitalizations, and 2 million outpatient visits in children <5 years of age. India spends Rs 2.0-3.4 billion (US$ 41-72 million) annually in medical costs to treat rotavirus diarrhoea. The use of specific interventions against rotavirus, such as newly available vaccines, would help prevent much of this large disease and economic burden.