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Biomedical subjects

S C Pal

Publications and source records attributed to S C Pal.

At least 19 recordsLinked to original sources

Campylobacter species as a cause of diarrhoea in children in Calcutta.

From 1985 to 1988, 857 children (aged between 1 day and 60 months) admitted to hospital with diarrhoea and 241 controls (aged between 5 days and 60 months) were examined for campylobacters and other enteric pathogens by means of conventional methods. The difference between the isolation rates of campylobacters in those cases in which no other enteric pathogen was found (4.8%) and controls (6.2%) was not significant (P greater than 0.05). Strains of Campylobacter jejuni/coli were isolated throughout the year with higher isolation rates during the summer and monsoon months. Mixed infections were very common. Watery diarrhoea (97.6% cases) was the most common clinical presentation of patients found to be infected solely by C. jejuni/coli. Most patients infected with campylobacters were mildly to moderately dehydrated. Biotype I of C. jejuni and C. coli was the dominant biotype associated with cases and controls. All strains of C. jejuni/coli, regardless of their source, were found to be sensitive to erythromycin. From this study, it appears that enteric infections with campylobacters among children in Calcutta are common but often asymptomatic.

Bacterial Typing Techniques

Virulence traits of Aeromonas strains in relation to species and source of isolation.

The virulence traits of 39 well-defined clinical (29 strains) and environmental (10 strains) isolates of Aeromonas (16 A. hydrophila, 12 A. sobria and 11 A. caviae) were examined by a variety of assays to delineate differences, if any, in the enteropathogenic potential in relation to species and the source of isolation. The distribution of enterotoxin (ent), cytotoxin (cyt) and haemolysin (hae) producing strains of Aeromonas did not correlate to species and source of isolation. The extracellular virulence phenotype Ent+ Cyt+ Hae+ was the most common one among all the three species although unique phenotypes associated prominently with either A. hydrophila or A. sobria were also discernible. None of the cytotoxin or haemolysin producing strains hybridized with the vt1/vt2 or the tdh/trh gene probes, respectively, indicating that these two factors of Aeromonas were distinct. Haemagglutination of human O group erythrocytes was not related to the source of isolation or production of enterotoxin, cytotoxin or haemolysin but appeared to be related to species. The strains which did not exhibit cell-associated haemagglutination belonged to either A. hydrophila or A. caviae. Haemagglutination unaffected by fucose, mannose and galactose was the dominant inhibition pattern exhibited mainly by the clinical haemagglutinating strains of the three species. Only one clinical strain of A. caviae showed a diffuse pattern of adherence to HeLa cells. Expression of the 5 bacterial enzymes by strains of Aeromonas did not fall into a readily discernible pattern in relation to species or source of isolation. From this study, it is clear that the mechanism of the pathogenesis of Aeromonas is a multifactorial one.

Aeromonas

Entero-adherent Escherichia coli is an important diarrhoeagenic agent in infants aged below 6 months in Calcutta, India.

Escherichia coli adherent to HEp-2 and HeLa cells were isolated from the faeces of 43 (19.7%) of 218 hospitalised infants aged below 6 months with acute diarrhoea. No conventional virulence factors, including enterotoxin production--heat-labile (LT) or heat-stable (ST), the verotoxin (VT) or shiga-like toxin (SLT)--or the invasive phenotype (determined by the Sereny test) could be detected among these isolates. Out of the 43 isolates, 16 (37.2%) were of the known enteropathogenic O:K serogroups--enteropathogenic E. coli (EPEC). The remaining 27 (62.8%) isolates showed different types of adherence to HEp-2 and HeLa cells which was diffuse (40.7%), localised (37.0%), or both (22.3%); they were identified as entero-adherent E. coli (EAEC). The EAEC isolates adhered to HEp-2 and HeLa cells in the presence of mannose, lactose, fucose, galactose, and fetuin, indicating that adhesion was not specific for these sugars or glycoprotein. Haemagglutination and the salt aggregation test (SAT) did not correlate with patterns of adherence. The results of this study indicate that LA-EAEC is an important aetiological agent of acute diarrhoea in infants aged below 6 months in Calcutta.

Bacterial Adhesion

Detection of cholera toxin by a highly sensitive bead-enzyme linked immunosorbent assay.

A bead-enzyme linked immunosorbent assay (bead-ELISA) for detection and quantification of cholera toxin (CT) in broth cultures of Vibrio cholerae O1 has been developed. Under optimal buffer and pH conditions the bead-ELISA could consistently detect 40 pg/ml of CT. None of the ingredients of commonly used media for in vitro culture of V. cholerae O1 hindered the performance of the bead-ELISA. Evaluation of the sensitivity and specificity of the bead-ELISA against the commonly used reversed passive latex agglutination (RPLA) test for detection of CT was performed using a collection of 239 strains of V. cholerae O1 (including both biotypes and serotypes) which were examined by a gene probe encoding for the A1 subunit of CT. Although both the assays were highly specific, the bead-ELISA was more sensitive than the RPLA. Quantification of CT by the bead-ELISA revealed that the concentration of CT produced by the strains of V. cholerae O1 which were negative by the RPLA was lower than 1 ng/ml and therefore below the minimum detection ability of the RPLA. The bead-ELISA is a simple, specific and highly sensitive assay for routine detection of CT and is recommended for routine use in clinical microbiology laboratories.

Antigen-Antibody Reactions

Reassessment of the prevalence of heat-stable enterotoxin (NAG-ST) among environmental Vibrio cholerae non-O1 strains isolated from Calcutta, India, by using a NAG-ST DNA probe.

A collection of 521 environmental isolates of Vibrio cholerae which were previously examined by the suckling mouse assay and found to be negative for the heat-stable enterotoxin NAG-ST were reassessed by a recently developed DNA probe for NAG-ST. A total of 12 (2.3%) of the isolates hybridized with the NAG-ST probe. By using a cholera toxin (CT) DNA probe, the CT gene was detected in six of the strains in the collection, although none of the isolates of V. cholerae non-O1 hybridized with both of the toxin probes. All of the NAG-ST and CT probe-positive strains were hemolysin positive. Thirty-fold-concentrated supernatants of the three representative NAG-ST DNA probe-positive V. cholerae non-O1 strains gave positive fluid accumulation ratios in the suckling mouse assay even after heating (100 degrees C for 5 min) and also inhibited the binding of a NAG-ST monoclonal antibody to the bound NAG-ST in a competitive enzyme-linked immunosorbent assay (ELISA). Likewise, all six CT probe-positive V. cholerae non-O1 strains produced in vitro CT when examined by the CT bead ELISA. HindIII digest patterns of chromosomal DNA from the representative NAG-ST gene-positive strains were visually indistinguishable. Between the groups of NAG-ST probe-positive strains examined, there was a variation in the hybridizable fragments, with one group of strains exhibiting a hybridizable fragment similar to that of the NRT 36 reference strain; a smaller HindIII fragment hybridized with the NAG-ST probe in the other group of strains.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Taxonomical implications of the emergence of high frequency of occurrence of 2,4-diamino-6,7-diisopropylpteridine-resistant strains of Vibrio cholerae from clinical cases of cholera in Calcutta, India.

Of the 110 consecutive isolates of Vibrio cholerae recovered from cholera patients admitted to the Infectious Diseases Hospital, Calcutta, India, between July 1989 and October 1990, 90 and 82.7% were resistant to 10 and 150 micrograms of 2,4-diamino-6,7-diisopropylpteridine (O/129), respectively. Additionally, all O/129-resistant strains of V. cholerae were multiply resistant to antimicrobial agents. Except in the cases of four strains, resistance to O/129 was invariably linked with resistance to co-trimoxazole. Although O/129 susceptibility is still a useful test for Vibrio identification, resistance of V. cholerae to this compound in local areas might occasionally pose a problem.

Cholera

Evaluation of the bead enzyme-linked immunosorbent assay for detection of cholera toxin directly from stool specimens.

A highly sensitive bead enzyme-linked immunosorbent assay (bead ELISA) for detection of cholera toxin (CT) was evaluated for direct detection of CT from stool specimens of patients with acute secretory diarrhea. Of the 75 stool samples examined, 59 yielded biochemically, and serologically confirmed strains of Vibrio cholerae O1. The bead ELISA was positive for CT in stool supernatants in 50 (84.7%) of the 59 samples from which V. cholerae O1 was isolated. In addition, the bead ELISA was positive for three stool specimens which were negative by culture. The free CT present in 48 of the 50 stool samples positive by culture for V. cholerae O1 and for CT by bead ELISA was completely absorbed by anti-CT immunoglobulin G. All of the 59 strains of V. cholerae O1 biotype eltor isolated in this study produced in vitro CT. The concentration of CT present in the bead ELISA-positive stool samples ranged between 26 pg/ml and greater than 100 ng/ml. This evaluation study demonstrates that the bead ELISA is a sensitive and simple method for direct detection of CT in nonsterile stool samples, and we recommend routine use of this assay for detection of CT in stool samples and culture supernatants in clinical and reference laboratories.

Cholera Toxin

Influence of admission weight on neonatal mortality amongst hospitalised neonates in Calcutta.

The study was conducted on 785 neonates aged up to 28 days to evaluate the influence of admission weight on mortality. It was observed that there were 200 (25.5%) cases of septicaemia, 134(17.1%) of diarrhoea, 120(15.3%) each of prematurity related conditions and neonatal jaundice, 117(14.9%) of respiratory diseases and 94 (11.9%) cases of convulsion. There were total 182(23.18%) deaths comprising 70(38.5%) from prematurity related conditions, 40(22%) from diarrhoea, 35(19.2%) from respiratory diseases, 26(14.3%) from septicaemia, 8(4.4%) from neonatal jaundice and 3(1.6%) deaths from convulsion. The incidence of deaths among neonates weighing less than 2500 g on admission was 59.2% in diarrhoeal diseases, 53.4% in respiratory diseases and 44.6% in other conditions compared to those of 10%, 8.2% and 7.1% respectively in neonates having admission weight more than 2500 g. The findings are statistically significant. The results of the study indicate that low admission weight should be considered as a predictor of mortality among neonates.

Body Weight

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.

Anti-Bacterial Agents

Comparison of selective media for optimal recovery of Clostridium difficile from diarrhoeal stools.

Five selective media were compared for their efficacy in the recovery of C. difficile from stool specimens. Of 341 diarrhoeic stool samples, 38 (11%) yielded C. difficile. Eighty per cent of the isolates were detected on modified taurocholate cycloserine cefoxitin fructose agar (MTCCFA) and 73 per cent were detected on taurocholate cycloserine cefoxitin fructose agar (TCCFA). MTCCFA was also found superior to the other four media as it supported better growth of C. difficile colonies, by effectively suppressing the competing microflora. These results suggest that the recovery rate of C. difficile could be enhanced when routine media, incorporated with taurocholate and lower concentration of cycloserine and cefoxitin, is used for the isolation of C. difficile from diarrhoeic stool.

Clostridioides difficile

Efficacy of norfloxacin for shigellosis: a double-blind randomised clinical trial.

In a double-blind, randomised, clinical trial on 122 adults with acute Shigella dysentery, 60 patients were treated with norfloxacin and 62 with nalidixic acid. Of these, 32 patients in the norfloxacin group and 28 patients in the nalidixic acid group had Shigella in their stool. Patients of the two treatment groups were clinically comparable on admission. No significant differences in clinical responses were observed in the two groups among the Shigella-positive cases, Shigella-negative cases and among the total cases. All isolates of Shigella were susceptible to norfloxacin, whereas 13.8% of the strains were resistant to nalidixic acid.

Adult

Detection of specific anti-Giardia antibodies in the serodiagnosis of symptomatic giardiasis.

The roles of circulating anti-Giardia IgM and IgG antibodies were assayed by the Enzyme linked immunosorbent assay (ELISA) using axenic Giardia lamblia whole trophozoites as antigen for routine serodiagnosis of Giardia infection. Detection of anti-Giardia IgM antibody was found to be useful in the early detection of symptomatic giardiasis, whereas IgG detection was not, since the latter could not discriminate between present infection and previous exposure. A mean optical density (OD) +/- 2 standard deviation (SD) value of 0.088 at 1:100 serum dilution was found optimal for the detection of anti-Giardia IgM antibodies as it completely separated out the acute Giardia cases from the Giardia-free controls. The same sera did not reveal any marked differences when tested for anti-Giardia IgG antibodies. Detection of IgM antibodies correlated with high sensitivity and specificity (96%) in an evaluation of a large number of sera collected from Giardia cases and controls.

Animals

Shigellosis in children: a prospective hospital based study.

From 1985 to 1988, fecal samples of 950 hospitalized children suffering from diarrhea or dysentery were screened for Shigella species using standard methods. Shigella species were isolated as sole pathogen from 192 (20.2%) cases and S. flexneri type 2 was the predominant serotype. Shigella infection was prevalent throughout the year with high isolation rate during the summer and early monsoon months. Shigella strains isolated during the period were resistant to most of the commonly used drugs for the treatment of shigellosis. Nearly 16% of the Shigella strains were also resistant to nalidixic acid. Presence of blood and mucus in stools (dysentery) was the common clinical presentation of shigellosis cases. Malnutrition was associated with longer duration of illness. High cases fatality rate (16.7%) was observed among hospitalized children infected with Shigella.

Child, Preschool

Electrophoretic mobility and immunoblot analysis of the outer membrane proteins of Aeromonas hydrophila, A. sobria and A. caviae.

The outer membrane profiles of three species of the genus Aeromonas were examined by means of SDS-PAGE and immunoblotting to identify species-specific polypeptides and antigens which could presumably be applied to differentiate Aeromonas spp. at the species or subspecies level. Profiles on an 11% discontinuous SDS-PAGE showed common band sharing at the 52 kD position. Species-specific bands for the three strains could also be detected. Immunoblots using heterologous LPS-adsorbed polyclonal antisera revealed demarcated common and uncommon antigens within the three species. Outer membrane preparations were immunoblotted against whole cell polyclonal antisera. The previously documented host pathogenicity of A. sobria correlated well with the immunoblots which showed antigenicity, especially due to the LPS, when compared with the other two species.

Aeromonas

Serovar, biotype, phage type, toxigenicity & antibiotic susceptibility patterns of Vibrio cholerae isolated during two consecutive cholera seasons (1989-90) in Calcutta.

Characteristics of V. cholerae isolated from patients of acute secretory diarrhoea admitted to the Infectious Diseases Hospital, Calcutta during two consecutive cholera seasons (1989 and 1990), with special emphasis on biotyping and toxigenicity, were investigated. The isolation rates of V. cholerae during 1989 and 1990 were 78 and 85.1 per cent respectively, with Inaba serotype dominating in 1989 and Ogawa in 1990. All the V. cholerae 01 strains isolated in this study belonged to biotype Eltor with phage type 4 dominating (48.8%). Most of the strains of V. cholerae were resistant to 10 and 150 micrograms/ml of 0/129 vibriostatic agent. Similarly, majority of the V. cholerae strains were resistant to furazolidone (95.7%), cotrimoxazole (83%) and tetracycline (63.1%) and several resistance patterns were encountered. All the V. cholerae 01 strains examined produced cholera toxin (CT) in amounts ranging between greater than 70 pg/ml and greater than 2.5 ng/ml. In contrast, all but one of the non-01 strains isolated in this study did not produce CT. Further studies are required to elucidate the mechanism involved in the pathogenesis of non-01 V. cholerae mediated diarrhoea.

Anti-Bacterial Agents

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.

Anti-Bacterial Agents