Epidemiology and characterization of leptospirosis at an urban and provincial site in Thailand.

Patients with FUOs at the Children's Hospital in Bangkok and the Chao Phya Abhai Bhu Bejhr Hospital in Prachinburi were screened for leptospirosis by blood and urine culture in addition to microagglutination testing of their serum. Animal populations in urban and periurban areas of Bangkok were surveyed for evidence of leptospira infection. Three rural sites near the Prachinburi Provincial Hospital were also surveyed. The rodents' and domestic animals' blood, urine, and/or kidney cell samples were cultured for leptospira. Sera from these animals were also tested for leptospira antibody. The bataviae serovar was the most commonly detected leptospiral agent in both man and animals. Presenting symptoms varied with age with children showing primarily fever, vomiting, headache, abdominal and generalized muscle pain and diarrhea whereas adults had fever, headache, anorexia, muscle pain and constipation. Blood samples from patients suspected of having leptospirosis were tested for antibody by the MAT and cultured in EMJH media. The following serogroups were identified: bataviae, autumanalis, javanica, hebdomadis, and pyrogens. Leptospirosis incidence in humans was much higher in the rainy/flooding year of 1983 compared to the relatively dry year of 1984. Results of our animal surveillance studies indicate that in addition to rats, which have previously been mentioned, dogs, bandicoots, cattle and pigs could be the source of human leptospirosis infection in both urban and provincial locations in Thailand.

Anti-rabies virus IgM in serum and cerebrospinal fluid from rabid dogs.

An anti-rabies IgM antibody capture radio immunoassay was used to test serum and cerebrospinal fluid from 37 dogs held in quarantine for suspicion of rabies. Rabies was confirmed in dogs that died by mouse inoculation and subsequent examination of mouse brains by fluorescent antibody technique to detect rabies antigen. The mean counts per minute (CPM) of iodinated anti-rabies gamma globulin coupled IgM rabies antibody in CSF and serum from rabid dogs were significantly higher than in CSF and serum from non-rabid dogs. Mean CPM from rabid dogs was greater in CSF than in sera, in contrast with non-rabid dogs, from which mean cpm was higher in sera than CSF, suggesting that antibody may have been synthesized in the CSF. To evaluate this test further, a dog was infected by rabies virus, and serial serum and CSF specimens were collected until the time of death. IgM anti-rabies antibody developed in the CSF and serum 29 days following infection, and rose just before the dog died of rabies on day 34. The rabies MAC RIA is potentially useful as a diagnostic method in quarantined dogs with rabies-like illness. Perhaps more importantly, it may be applied to better understand the immunopathogenicity of rabies.

The neurovirulence of flaviviruses in crab-eating monkeys (Macaca fascicularis).

The neurovirulent properties of attenuated dengue-2 and yellow fever (YF) vaccines, dengue-2 (DEN-2) and Japanese encephalitis (JE) viruses were studied in crab-eating monkeys (Macaca fascicularis). Number of central nervous system sites (as proportion affected) with neurovirulence (NV) lesions were compared. The results indicate that these monkeys reliably developed NV-lesion when inoculated with either JE or YF vaccine viruses (87%). NV-lesions occurred in a minority when inoculated with DEN-2 vaccine virus, were of minimal severity (9%), were probably biologically insignificant, and were of equal or less severity than lesions produced by its parental virus (10%).

Serologic evidence of Hantaan-like virus in rodents and man in Thailand.

A serologic survey was conducted to determine the presence of antibody to Hantavirus in rodents in Thailand. Sera from over 300 rodents were tested by an immunofluorescent antibody method. Bandicota indica, a field rodent, was found to have a high incidence of infection (20-24%) in 2 locations. A Hantavirus was isolated from lung samples of B. indica. When sera were tested from humans living in Kanchanaburi and several locations in Bangkok, those people living in close proximity to the infected B. indica had greater than 30% prevalence of positive antibody titers.

Intense transmission of Japanese encephalitis virus to pigs in a region free of epidemic encephalitis.

Epidemic Japanese encephalitis recurs annually in the northern provinces of Thailand, but in the southern provinces cases of human encephalitis are rare. We investigated transmission of Japanese encephalitis virus (JEV) to pigs in southern Thailand. Blood specimens from one hundred young pigs at abattoirs in three southern provinces were tested for JEV hemagglutination inhibiting (HAI) antibodies. Seventy-four percent were positive. Ten seronegative sentinel pigs were placed at five locations in one southern province. Seven of the ten pigs developed JEV HAI and JEV IgM ELISA antibodies within two weeks of placement. JEV was isolated from all seven seroconverting sentinel pigs from blood specimens collected 3 to 11 days after placement. Fifteen light-trap mosquito collections at the five locations all included known JEV vectors, some in large numbers. We conclude that there is intense transmission of JEV to pigs in southern Thailand despite the rare occurrence of human encephalitis in the same region.

Doxycycline prophylaxis for experimental leptospira infection in non-human primates and hamsters.

Doxycycline was effective as a chemoprophylactic agent for experimental Leptospira infection in non-human primates and hamsters. Monkeys injected intraperitoneally with Leptospira bataviae, and receiving only diluent as treatment developed a leptospiremia during the first week and later leptospires were cultured from the cerebrospinal fluid and urine. Monkeys treated daily with oral doxycycline for 10 days beginning one day before infection had a shortened period of detectable leptospiremia, and organisms were never detected in the cerebrospinal fluid or urine. Even an oral dose of doxycycline 2 hours before infection and on day 7 prevented the later infection of the cerebrospinal fluid and urine. In hamsters, doxycycline treatment prevented deaths from acute Leptospira infection and when hamsters were treated daily for 4 or more days, renal infection was prevented. The results of animal studies, the susceptibility of LC0475 and the five other isolates to doxycycline in vitro, and lack of evidence for antibiotic resistance in culture suggests this antibiotic may be useful as a prophylactic drug for high risk groups and an effective treatment for leptospirosis in Thailand.

Sporozoite-induced Plasmodium cynomolgi infections in captive born Macaca fascicularis.

Capability of captive born cynomolgus monkeys to substitute for rhesus in the Plasmodium cynomolgi radical curative antimalarial drug development model was examined. Eighteen monkeys divided into 3 groups were given standard or high doses of sporozoites intravenously. One group of 4 received 0.8 - 1.6 X 10(6) and a second group of 8 received 0.3 - 1.0 X 10(7) sporozoites. The third group of 6 was splenectomized and then received 3.0 - 4.0 X 10(6). The 2 groups of intact monkeys developed a persistent low level parasitemia; however, gametocyte production was poor. The splenectomized group developed a persistent parasitemia with a higher mean, which more closely resembled rhesus parasitemias. A high, post-patent leukocytosis consisting primarily of lymphocytes was observed in this group. Good gametocyte production resulted in the splenectomized group and oocysts were produced from all lots of Anopheles dirus which fed on them. Following clearance of blood forms, relapse potential was demonstrated in the 2 splenectomized monkeys tested. In this study the splenectomized captive born cynomolgus appeared to be capable of supplementing rhesus as an antimalarial drug testing model.