An outbreak of El Tor cholera associated with a tribal funeral in Irian Jaya, Indonesia.

An outbreak of El Tor biotype cholera occurring in a rural village in Irian Jaya, Indonesia was evaluated for risk factors associated with death from cholera. Among those dying in the village during the epidemic, a significant association between membership in one of the five tribal groups in the village complex was associated with an elevated risk of suffering a cholera death (odds ratio = 5.9). Interviews with members of the decedents' families revealed a very strong association (odds ratio = 11.6) between risk of cholera death and having attended the two day funeral of a woman who died of a cholera-like illness a few days prior to an outbreak of cholera-like diarrheal disease in the village complex. Recent flooding may have contributed to the creation of an environment conducive to cholera transmission.

Enterotoxigenic Escherichia coli and other causes of infectious pediatric diarrheas in Jakarta, Indonesia.

A hospital stool survey of Indonesian children less than 5 years of age determined the prevalence of diarrhea caused by enterotoxigenic Escherichia coli (ETEC) and other bacterial enteropathogens, compared to non-diarrheic control patients. ETEC were the second most frequent cause of diarrhea, isolated from 16 of 194 (8.2%) of patient's stools compared to 2 of 97 (2.1%) of control stools. The highest prevalence was in infants 12 to 23 months of age (17.9%).

The use of a coagglutination test to presumptively identify Salmonella typhi in bone marrow-oxgall medium cultures from typhoid fever patients.

A study was conducted to test a coagglutination procedure for detection of Salmonella typhi in bone marrow cultures from suspected typhoid patients admitted to Friendship Hospital, Jakarta, Indonesia. The results of the coagglutination tests were compared to the results from standard cultural isolation and identification. Bone marrow aspirates (356) were cultured in oxgall medium and aliquots subcultured daily for 7 days while simultaneously testing for the presence of Salmonella group D and Vi antigens using coagglutination (COAG). S. typhi was isolated from 220 (62%) of the cultures and the D- and Vi-COAG tests were positive for those same cultures. The COAG test was also negative for 6 cultures containing S. paratyphi A. The COAG results were available within 10 minutes after 18 to 24 hours incubation of the primary cultures whereas the isolation and confirmed identification took 2 to 3 days longer. The COAG test is valuable as an aid to rapidly identify S. typhi in bone marrow-oxgall cultures.

A penicillinase test incorporated in the rapid carbohydrate fermentation method for rapid detection of beta-lactamase producing Neisseria gonorrhoeae.

A modification of the acidometric (phenol red) test for penicillinase producing N. gonorrhoeae was incorporated into the rapid fermentation method for rapid screening and identification of PPNG strains. Two hundred and twenty-four non-penicillinase-producing N. gonorrhoeae, 55 penicillinase-producing N. gonorrhoeae, 87 N. meningitidis and 89 N. lactamica were included in this study. Results of the modified test were comparable with the iodometric and penicillin disk diffusion susceptibility and were obtainable within 1 to 5 minutes.

Isolation of Salmonella typhi from standard whole-blood culture vs blood-clot cultures.

The use of 10% oxgall and bile broth medium, both supplemented with freshly prepared 100 u/ml streptokinase, for isolating Salmonella typhi by clot culture technique was evaluated and compared against whole blood culture systems (3 ml blood in 9 ml media and 8 ml blood in 72 ml media). These gave a 1:4 and 1:10 blood to medium ratio, respectively. Clot cultures in 10% oxgall (CLOX) gave a 57% positive isolation rate for S. typhi. A similar result was obtained from clot cultures in bile broth medium (CLBB). A total of 184 samples identified as positive for S. typhi were tested. There was no significant difference between the use of 10% oxgall or bile broth medium when used for clot culture. The whole blood culture systems still showed a significantly better rate of isolation than the clot culture methods.

A CAMP phenomenon between Vibrio cholerae biotype El Tor and staphylococcal B-hemolysin.

A CAMP phenomenon was demonstrated by Vibrio cholerae biotype El Tor and B-lysin producing Staphylococcus aureus in 5% sheep red blood cells-tryptic soy agar medium. All 394 El Tor vibrio strains tested, all showed a crescent-shaped hemolysis (positive CAMP) when the cultures were incubated in a candle jar whereas 67% were CAMP positive when incubated aerobically. Only 9% of the isolates produced detectable hemolysin in a standard tube test using heart infusion broth and 72% in a tube test using heart infusion broth containing 1% glycerol. Seven classical V. cholerae tested were CAMP negative. The CAMP reaction is easy to perform and may be useful for routine use in the differentiation of V. cholerae biotype El Tor from classical V. cholerae.

An evaluation of alkaline peptone water for enrichment of Vibrio cholerae in feces.

A comparison was made to determine the sensitivity of direct inoculation of thiosulfate citrate bile salts agar (TCBS) and alkaline peptone water (APW) enrichment prior to direct inoculation of TCBS to culture Vibrio cholerae from feces of patients with gastroenteritis. V. cholerae was isolated from 611 feces specimens. Of those, 535 were isolated in TCBS and APW-TCBS, 15 in only TCBS and 61 in only APW-TCBS. V. parahemolyticus (21) and non-agglutinating vibrios (11) were also isolated but more often in direct inoculated TCBS than APW-TCBS cultures. Maximum isolation sensitivity of V. cholerae and V. parahemolyticus from feces is obtained by both direct inoculation of TCBS and enrichment in APW prior to TCBS inoculation.

Improved method, using staphylococcal beta-hemolysin, for detection of hemolysin(s) produced by Vibrio cholerae biotype El Tor.

A tube test using brain heart infusion broth and staphylococcal B-lysin (HIBL) was devised to improve the detection of Vibrio cholerae El Tor hemolysin. Fifty six (100%) strains of V. cholerae serotypes Ogawa (28) and Inaba (28) were positive by the hemolysin test whereas 4 Inaba and 2 Ogawa were positive by a standard tube test using heart infusion broth (HIB) and 20 Ogawa and 18 Inaba were positive by another tube test using HIB containing glycerol (HIBG). Seven classical V. cholerae strains tested were negative by the 3 methods. The HIBL tube test was faster and more sensitive than the other 2 methods and showed that hemolysin was present that would otherwise have gone undetected by the other 2 methods using HIB or HIBG.

Slide coagglutination for Salmonella typhi antigens in broths inoculated with feces from typhoid fever patients.

Salmonella typhi antigens D, Vi and d were readily detected, by slide coagglutination, in mannitol selenite (MSB) and dulcitol selenite (DSB), Salmonella enrichment broths 4 hours after inoculation with feces from 60 patients with bacteriologically confirmed typhoid fever. Positive coagglutination also occurred using MSB and DSB inoculated with fecal specimens obtained from 16 patients from whom S. typhi was not cultured. Twelve of these later seroconverted to Salmonella O antigen. None of the MSB or DSB inoculated with feces from 50 healthy control subjects, gave a positive coagglutination test. The coagglutination method appears to have potential as a rapid test for the detection of antigens of S. typhi in MSB and DSB broths inoculated with feces from patients with suspected typhoid fever.

Detection of Salmonella C1, D and V1 antigens, by coagglutination, in blood cultures from patients with Salmonella infections.

Protein A-containing Staphylococcus aureus was coupled to Salmonella C1, D and Vi monovalent antisera to produce C1-, D- and Vi-COAG reagents. The reagents were used to detect their homologous Salmonella antigens in blood cultures (BC). The D and Vi antigens were detected in 79 of 239 BC from patients with suspected typhoid fever and Salmonella typhi was later isolated from the same 79 BC. The C1 antigen was detected in 8 BC from which only S. oranienburg was later isolated. The COAG test was generally positive at the same time the BC became culture positive. However, because of subculture and biochemical identification requirements the COAG test could be interpreted 1-2 days before culture results were available. The COAG test can be used to presumptively identify Salmonella typhi and Salmonella group C1 in blood cultures before the culture results are available.

Chloramphenicol resistant strains in salmonellosis in Jakarta.

During an observation period of 20 months (from January 1978 to September 1979) 123 children with clinically suspected salmonellosis were admitted to the Department of Pediatrics, 70 males and 53 females varying in age from 17 days to 14 years. S. typhi or S. enteriditis was isolated from the stool, urine, blood or cerebrospinal fluid in 85% (105/123) of the cases. The results of the microbiologic examination showed that 28 out of the 105 cases (27%) were resistant to chloramphenicol of which the S. sero-group C1 was predominant. It appeared that 28 cases conformed to the clinical data. In all these cases chloramphenicol was replaced by other antibiotics according to the sensitivity test. The mortality rate was 7% (9/123). Four of the fetal cases were from the chloramphenicol resistant group.

A coagglutination method for presumptive identification of Salmonella typhi.

Salmonella group D, Vi and d antisera were used to sensitize A-containing staphylococcal cells. The coagglutination (COAG) reagents thus obtained, termed D-COAG, Vi-COAG and d-COAG were used to test growth taken from Kligler Iron Agar slants following 8-10 hours incubation. In 188 recently isolated strains of S. typhi, the D antigen was detected in 134 (71%) by direct reaction with the D-COAG reagent and 51 (27%) others following a simple treatment with 50% ethanol. Group D antigen was detected in the remaining 3 only following boiling of a suspension of each culture. The Vi-COAG reagent reacted with 184 (98%) of the cultures. The 4 remaining cultures were also negative for Vi antigen by the conventional slide agglutination test but D-COAG reagent was specific for other group D Salmonella but with none of the other groups tested, while the Vi-COAG reagent also reacted with a strain of Vi-containing Citrobacter furendii. The d-COAG reagent agglutinated with all S. typhi strains but none of the other non-S. typhi isolates tested.