Studies on Escherichia coli as a cause of acute diarrhoea in Calcutta.

The prevalence of different types of diarrhoea-producing Escherichia coli among 240 patients with acute diarrhoea in hospital was investigated. The 25 patients (10.4% of the total) from whose faeces we isolated enteropathogenic E. coli (EPEC) were all less than 5 years old but the 29 (12.1%) from whom we isolated enterotoxigenic E. coli (ETEC) were of various ages, most of them greater than 12 years old. No enteroinvasive E. coli (EIEC) strains were isolated. ETEC strains that produced heat-labile toxin (LT) were encountered more often than those that produced either heat-stable toxin (ST) alone or both LT and ST. The ETEC isolates were distributed among eight different serotypes, the commonest being O148:H28 (38%). Correlations between enterotoxin production, serotype pattern and possession of colonisation factor antigens I and II were observed.

Role of rotavirus as the cause of acute paediatric diarrhoea in Calcutta.

Of the 245 acute paediatric diarrhoea cases admitted to the Infectious Diseases Hospital, Calcutta between July, 1979 and June, 1981, rotavirus was detected in the faeces of 55 (22.4%) patients either as the sole aetiological agent or in association with other enteropathogens. Children aged six months to two years were most commonly infected. The virus was detected throughout the year with higher incidence during the winter months of both years. The frequency of detection of major enteropathogens other than rotavirus was Vibrio cholerae biotype El Tor (31.0%), enteropathogenic Escherichia coli (8.2%) and enterotoxigenic E. coli (6.5%).

Kanagawa phenomenon and serotypic pattern of Vibrio parahaemolyticus strains isolated from various sources in Calcutta.

A total of 135 strains of Vibrio parahaemolyticus isolated during the period 1976-77 from human gastroenteritis cases and various categories of environmental sources, e.g., crustaceans, fish and different water samples (pond, stored, open well and tap water) were tested for their Kanagawa phenomenon and serotypic pattern. Amongst the 80 human isolates tested, 69 strains (86.3%) were Kanagawa phenomenon positive, and 60 strains (75.0%) were serologically typable--the dominant serotype being 05:K15. Of the 25 isolates from various water sources 10(40.0%) were Kanagawa phenomenon positive, and 17(68.0%) were serologically typable and 4 (23.5%) of them belonged to serotype 05:K15. All the 30 strains isolated from crustaceans and fish were Kanagawa phenomenon negative and 22 (73.3%) of them were serologically typable, belonging to heterogeneous serotypes. The results of this study also indicate the possible role of water in the transmission of V. parahaemolyticus infection in Calcutta slums.

Aetiological studies on hospital in-patients with acute diarrhoea in Calcutta.

Bacterial enteropathogens and rotavirus were sought in 356 cases with acute diarrhoea admitted to the Infectious Diseases Hospital, Calcutta. One or more pathogens were isolated from 74.7% of the cases. Single enteropathogens could be detected from 66% and multiple enteropathogens from 8.7% of the patients. Vibrio cholerae biotype El Tor, rotavirus, V. parahaemolyticus, and enteropathogenic and enterotoxigenic Escherichia coli were the major pathogens detected. Rotavirus was detected from 7.6% of the cases. A higher rate of detection of rotavirus was seen in children younger than two years. Campylobacter jejuni could be isolated from the faeces of six (15%) of 40 cases either as a single pathogen or in association with V. cholerae biotype El Tor.

An epidemic of rotavirus diarrhoea in Manipur, India.

Recurrent epidemics of acute diarrhoeal disease in young children have occurred during the winter months in Manipur (India) for many years. During November 1979, children admitted to hospital for acute diarrhoea were investigated and, in addition to pathogenic Escherichia coli and Shigella rotovirus was detected by ELISA in a number of stool samples, especially of those less than one year old. The importance of this finding and the need for further study are discussed.

Occurrence and significance of Cryptosporidium infection in Calcutta.

During a 2-year study, Cryptosporidium oocysts were detected in 32 (5.6%) of 566 hospitalized paediatric diarrhoea cases and 2 (1.2%) of 167 non-diarrhoeic individuals. Cryptosporidium was the sole pathogen detected in 17 (3.0%) of the 32 positive cases; in the other 15 it occurred in combination with one or more other established enteropathogen(s). The frequency of detection of the parasite was highest in the 0-6 months age group; no sex-specific difference was discernible. The detection rate of the parasite was highest during the monsoon and post-monsoon months. Most of the patients had watery stools with a mild to moderate degree of dehydration, with the diarrhoea lasting for less than 7 d.

A study of the titrimetric potassium silicofluoride method for determination of silica in aluminosilicate materials.

A critical study of the titrimetric potassium silicofluoride method for determination of silica reveals that quantitative precipitation of the sificofluoride is possible in 3.5-7.5N acid containing 1% sodium fluoride and >/ 12% potassium chloride. An aqueous wash solution (pH approximately 5.3) containing 0.1% sodium fluoride and 12% potassium chloride has been found ideal for washing the precipitate without significant hydrolysis even at 35 degrees C. The interference of large amounts of aluminium (up to 160 mg of Al(2)O(3)) is eliminated by precipitating the silicofluoride in 6-7.5N acid solution, the aluminium then forming practically no fluoro-complex, because of extensive protonation of fluoride. Interference by large amounts of boron (up to 160 mg of B(2)O(3)) is eliminated by precipitating silicofluoride with the minimum permissible potassium chloride concentration (12%) and washing with the aqueous wash solution at room temperature. Nitrazene Yellow has been used as a more sensitive indicator.

A study of the titrimetric potassium silicofluoride method for determination of silica in aluminosilicate materials.

A critical study of the titrimetric potassium silicofluoride method for determination of silica reveals that quantitative precipitation of the sificofluoride is possible in 3.5-7.5N acid containing 1% sodium fluoride and > trg 12% potassium chloride. An aqueous wash solution (pH approximately 5.3) containing 0.1% sodium fluoride and 12% potassium chloride has been found ideal for washing the precipitate without significant hydrolysis even at 35 degrees C. The interference of large amounts of aluminium (up to 160 mg of Al(2)O(3)) is eliminated by precipitating the silicofluoride in 6-7.5N acid solution, the aluminium then forming practically no fluoro-complex, because of extensive protonation of fluoride. Interference by large amounts of boron (up to 160 mg of B(2)O(3)) is eliminated by precipitating silicofluoride with the minimum permissible potassium chloride concentration (12%) and washing with the aqueous wash solution at room temperature. Nitrazene Yellow has been used as a more sensitive indicator.

A prospective study of phage types & biotypes of Salmonella enterica serotype Typhi isolated from hospitalized children in Kolkata, India.

BACKGROUND & OBJECTIVES: Kolkata and its suburbs in eastern India faced an epidemic of typhoid fever in 1990. A prospective, hospital and laboratory based study over a period of 12 yr (1990-2001), on the phage typing and biotyping pattern of Salmonella enterica serotype Typhi was carried out, to see if there has been a change. METHODS: A total of 338 S. enterica serotype Typhi isolates from 1491 blood samples were phage typed and biotyped. The mean age of isolation was calculated. RESULTS: The age distribution of subjects (neonates to 12 yr) has been analysed. Of the 338 (22.7%) isolates obtained, eight different S. enterica serotype Typhi phage types were detected. Biotype I (95.8%) was more prevalent as compared to biotype II (4.1%). Phage type E1 was the commonest phage type in Kolkata and its suburbs. INTERPRETATION & CONCLUSION: The mean age at isolation was found to be 6.7 +/- 3.3 yr. Biotype I was predominant and it was of interest that all strains of phage type E1 belonged to biotype I.

Validity of new phage typing scheme against Vibrio cholerae 01 biotype ElTor strains.

A total of 538 strains of V. cholerae 01 biotype ElTor were phage typed by the conventional Basu and Mukerjee and also the new typing scheme developed at the National Institute of Cholera and Enteric Diseases, Calcutta. The strains could be clustered into seven types by the new scheme as against only two by the conventional method. The results provide conclusive evidence on the validity of the new scheme for phage typing of V. cholerae strains.

Value of a single Widal test in the diagnosis of typhoid fever.

Results of a single Widal test in patients with bacteriologically confirmed typhoid fever (116), clinically suggestive but culture negative fever (170) and non-typhoidal febrile illness (98) and in normal control children (54) were analysed. Positive Widal test (antibody titre against S. typhi O antigen of 1:160) was recorded in 61.2 per cent of patients with bacteriologically confirmed typhoid fever and in 58.8 per cent with culture negative but clinically suggestive typhoid fever. In contrast, the same titre was observed in 10.2 per cent patients with other febrile illnesses of known etiology and in 1.8 per cent of normal children. Differences in the positivity of Widal test in patients with bacteriologically confirmed typhoid fever and clinically suggestive but culture negative fever were highly significant (P < 0.000001) when compared to that of patients with non-typhoidal febrile illnesses and normal controls. High specificity and positive predictive value in 1:160 dilution makes the Widal test acceptable as a diagnostic tool.

Occurrence of multi-drug resistant Salmonella typhi in Calcutta.

Blood and faecal samples were collected from 122 hospitalised patients of Calcútta clinically suspected to have enteric fever, for isolation of S. typhi. It was isolated from 34.4, 4.9 and 4.1 per cent patients by blood culture, stool culture and by both respectively. The in vitro drug susceptibility testing showed that all the isolates were resistant to chloramphenicol, ampicillin and trimethoprim-sulphamethoxazole, but were uniformly susceptible to ciprofloxacin, norfloxacin and furazolidone. In view of the appearance of multi-drug resistant S. typhi in Calcutta, great care should be exercised in the use of newer quinolone derivatives.

Decreasing trend in the occurrence of Salmonella enterica serotype Typhi amongst hospitalised children in Kolkata, India during 1990-2000.

In a prospective hospital based surveillance, 1454 children clinically diagnosed as typhoid fever were enrolled during the period between 1990 to 2000. Of them 336 (23.1%) children were positive for Salmonella enterica serotype Typhi by blood culture. A declining trend of hospitalization and identification of the pathogen was observed from 1992 to 2000 as compared to 1990-1991. A declining trend of resistance to the commonly used anti-typhoid drugs was seen in the S. enterica serotype Typhi isolates. Recently in 2000, nine strains were detected as ciprofloxacin resistant. Misuse and overuse of ciprofloxacin for the treatment of typhoid fever influenced the development of ciprofloxacin resistant strains of S. enterica serotype Typhi in and around Kolkata.

Comparison between the multiplex PCR, sensitivity to biotype specific phages & polymyxin B for biotyping of Vibrio cholerae O1.

A total of 196 Vibrio cholerae O1 strains isolated between 1970 and 1996 were biotyped by multiplex PCR, susceptibility to polymyxin B and sensitivity to biotype specific phages. We modified the multiplex PCR by increasing the primer concentration of tcpA to improve the results. Comparison of the results of modified multiplex PCR and sensitivity to biotype specific phages and to polymyxin B showed that multiplex PCR was as efficient as phage typing for biotyping of V. cholerae O1. All the strains of V. cholerae O1 could be accurately distinguished based on polymyxin B sensitivity. Thus our results show that susceptibility of strains of V. cholerae O1 to polymyxin B is the easiest method to biotype V. cholerae O1 and is feasible in most laboratories when compared with multiplex PCR and sensitivity to biotype specific phages.

A longitudinal study of diarrhoea among children in Calcutta communities.

PIP: A longitudinal study was conducted to determine the profile of diarrheal diseases in a group of 383 children below 5 years of age in 2 typical slums of Calcutta (India). The overall annual incidence of diarrheal illness in these children during the 1st year of the study was found to be 1.1/child. However, the incidence was higher (1.9/child) in children below 2 years of age and declined progressively with advancing age. Most (99.5%) of the diarrheal episodes were mild in nature and not a single child required hospitalization; about 53.4% children had no diarrheal symptoms. Enteropathogens identified in diarrheal stool samples from 13.7% of children included enteropathogenic Escherichia coli (5.8%), rotaviruses (5.6%), shigellae (3.5%), Salmonella spp, (1.0%), Entamoebae histolytica (0.9%), Giardia lamblia (0.4%), Vibrio cholerae (0.6%), and V. parahemolyticus (0.3%).^ieng

Isolation of Salmonella enterica serotypes from children with diarrhoea in Calcutta, India.

The prevalence of Salmonella enterica serotypes, antimicrobial susceptibility, and phage typing of serovers were studied. Clinical presentations of the infected cases were also examined. The study was carried out during August 1993-September 1996 in and around Calcutta, India. In total, 1,025 faecal samples from hospitalized diarrhoeal children were screened for enteropathogens. Four S. enterica serotypes were identified in 157 (15.3%) cases as a single pathogen. S. enterica serotype Typhimurium was detected in 110 (70%) cases. S. Seftenberg, S. Infantis, and S. Virchow were detected in 28 (17.8%), 14 (8.9%), and 5 (3.2%) cases respectively. S. Typhimurium was isolated from 11 (3.2%) non-diarrhoeal control children. All of these children had acute watery diarrhoea, and 5% of them had severe dehydration, 40% had some dehydration, and 55% had no dehydration. Vomiting, fever, and diffused pain in abdomen were the associated presentations of these children. Most (95%) of them recovered with oral rehydration therapy only and without any antibiotics. In-vitro susceptibility testing showed that 120 of the 121 S. Typhimurium strains isolated from cases and controls were resistant to the commonly-used drugs. Thirteen of the 121 strains were phage-typeable and belonged to the phage type 193. However, no clinical or epidemiological significance could be established with these typeable strains. The findings of the study indicate that diarrhoeagenic Salmonella is one of the major pathogens causing diarrhoeal diseases in eastern India.

Occurrence of multi-resistant Salmonella typhimurium infection in a pediatric hospital at Calcutta.

Salmonella typhimurium was isolated from 55 (15.9%) of the 347 hospitalized diarrheal children and 14 (11.1%) of the 126 non-diarrheal controls. All the 98 asymptomatic children attending hospital outpatients were negative. Six (3.7%) of the 162 samples of different categories examined from the hospital were positive for S. typhimurium. Finger washing of one female food handler, feces of two cats of the wards, surface of wash basin, lavatory seat and shelf yielded positive isolations. In vitro antibiotic sensitivity testing showed majority of the strains were resistant against commonly used antimicrobial agents while they were uniformly sensitive to norfloxacin and ciprofloxacin. The present study points to cross-infection by multi-resistant S. typhimurium strains in the hospital wards.

Nosocomial rotavirus diarrhea in two medical wards of a pediatric hospital in Calcutta.

One hundred eighty nine children suffering from different medical problems were admitted in two wards of a pediatric hospital in Calcutta during the period between November 18, 1985 and February 10, 1986. Amongst them, 36 children developed nosocomial diarrhea and rotavirus was detected from 80.5% of the cases. The nosocomial rotavirus diarrhea cases had lesser frequency of stools and only mild dehydration but the course of illness was longer in comparison to that of the hospitalized rotavirus diarrhea cases. There is a possibility of spread of infection via fomites, environmental surfaces and most likely mothers.

Prospective study of nosocomial enteric infections in a pediatric hospital, Calcutta.

Hospital acquired enteric infections were investigated by studying 3138 children under 5 years of age who were admitted without diarrhea in nine medical words of a pediatric hospital, Calcutta during the period between March and September 1987. Three hundred and twenty (10.2%) children developed nosocomial diarrhea during their hospital stay. Fecal samples from 178 nosocomial diarrhea, 345 hospitalized diarrhea cases, 178 hospital controls and 200 outpatient controls were collected for detection of established enteropathogens. There were no statistically significant differences in the detection of most of the enteropathogens from fecal samples of nosocomial diarrhea, hospitalized diarrhea and hospital controls. Enteric pathogens were detected at a higher frequency (statistically significant) from fecal samples of nosocomial diarrhea cases as compared to outpatient controls. This study highlights the importance of most of the enteropathogens like Shigella, Salmonella, rotavirus, enteropathogenic E. coli as the cause of hospital cross infection. This study reinforces the importance of developing preventive measures in order to reduce the frequency of illness.

Multi-drug resistant typhoid fever with diarrhea.

OBJECTIVE: To provide information about the characteristics of diarrheal stool in multi-drug resistant typhoid fever and observe the clinical course after treatment with furazolidone or ciprofloxacin. SETTING: Hospital based. SUBJECTS AND METHODS: Twenty one male children who were positive for multi-drug resistant S. typhi by blood and stool cultures, having diarrhea at the time of hospitalization comprised the subjects. Serum and stool electrolytes were estimated. Stool samples were also processed to detect established enteropathogens, leukocytes and red blood cells. Children were treated either with furazolidone or ciprofloxacin and evaluated till recovery. RESULTS: Mean (+/- SD) pre-admission duration of fever and diarrhea of these cases were 19.1 (+/- 5.6) and 15.8 (+/- 4.6) days, respectively. Stool character in 81% of the patients was watery with mean (+/- SD) volume of stool 51.4 (+/- 25.1) ml per kg body weight in the first 24 hours of observation. Leukocyte count varied between 20-49 per high power field in 66.7% stool samples. Occult blood was present in only 19% cases. Fecal red blood cells in high power field were detected in 52.4% cases. Mean fecal electrolytes (mmol/liter) were as follows: sodium-53.8, potassium-51.4, chloride-41.6 and total CO2-24.3. Most of the children (71.4%) had no dehydration and had normal serum electrolytes. The isolated strains of S. typhi were multi-drug resistant. These children were treated successfully either with furazolidone or ciprofloxacin. CONCLUSION: The stools of multi-drug resistant typhoid fever patients were watery with little blood. Their electrolyte contents were more similar to the diarrheal stool seen in shigellosis rather than cholera. Uncontrolled observations revealed that children recovered with furazolidone or ciprofloxacin therapy.