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The populations of scrub typhus vector chiggers were compared in two developing oil palm areas, one 5 years old and the other 7 years old at the inception of the study. Both areas were located within the same oil palm scheme in central Peninsular Malaysia. Leptotrombidium (L.) deliense, a principal vector of scrub typhus in Malaysia, was found in reduced numbers in the older oil palm habitat. This reduction is attributed to changes in the microhabitat, specifically the elimination of grasses between the oil palm trees due to canopy shading and to cultural practices.

The strains of Rickettsia tsutsugamushi found in naturally infected, laboratory-reared Leptotrombidium (Leptotrombidium) arenicola and L. (L.) fletcheri chiggers were characterized by direct immunofluorescence (FA) and by mouse and monkey virulence tests. The strains existing in the L. (L.) arenicola chiggers consisted of different combinations of TA716, TA763, TA686, Karp, and Kato. In addition to these five strains, Gilliam was found in the L. (L.) fletcheri chiggers. Results indicate that individual chiggers can be simultaneously infected with several antigenic strains of R. tsutsugamushi. Although these antigens appear to remain stable within familial lines when several generations were viewed, the antigenic patterns observed in two succeeding generations did not always correlate. This variable expression of antigens was considered to be due to a quantitative fluctuation from one generation to the next in the strains of rickettsiae combined with a lack of sensitivity of the direct FA test in detecting small numbers of antigenically different rickettsiae. Phenotypic variation was considered to be a less probable explanation. Morbidity and mortality were minimal in ICR mice fed upon by individual chiggers of either species, but infection rates were 85-99%. Tissue suspensions prepared from mice infected by L. (L.) arenicola produced higher mortality and longer duration of illness in mice than those prepared from L. (L.) fletcheri-infected mice. Silvered leaf and cynomolgus monkeys were fed upon by the two species of chiggers or inoculated with the mouse tissue suspensions. In both cases, minimal clinical responses were observed.

Fifty-one Rickettsia tsutsugamushi isolates from small mammals collected in central Peninsular Malaysia serologically characterized by direct immunofluorescence using eight prototype strains. Karp-related (TA763, TA686, TA716) antigens were found in 90.2% of the isolates.

A high prevalence of murine typhus was found in the human and rodent populations of Senaling, West Malaysia. In addition to 2 index cases which prompted the investigation, 45% of the humans and 35% of the rodents had IFA titers of 1/50 or greater. Serological studies provided evidence of recent infections in 3 human subjects. In a control group of Malaysian soldiers IFA titers of 1/50 or greater were found in only 13 (5%) of 265 sera tested.

R. tsutsugamushi strains found in chiggers collected from 3 different sites throughout Thailand were antigenically characterized. Monotypic infections were observed in 76.7% of the chiggers. Karp was the most predominant strain, followed by TA716, TA763, TA686 and Kato. This study represents the first confirmed report of R. tsutsugamushi infection in 8 different chigger species in Thailand.

An epidemiological study in a mature oil palm estate in Peninsular Malaysia has demonstrated a low prevalence of R. tsutsugamushi infection in small mammals. The direct fluorescent antibody technique for assaying infections in chiggers proved more sensitive than mouse inoculation. Most infections in both chiggers and rodents were caused by the Karp strain.

The life cycle of 5 generations of Leptotrombidium (L.) fletcheri infected with Rickettsia tsutsugamushi and reared under ambient temperatures in Malaysia was presented and compared with a colony reared at a constant 27 degrees C (Neal and Barnett, 1961). In general our colony had a longer generation time (average of 54 days from engorged larvae to adult compared with 37 days) and produced fewer eggs (average of 127.9 compared with 900.0) than the comparison colony. Possible factors causing these differences are discussed.

Laboratory-reared, Rickettsia tsutsugamushi-infected Leptotrombidium arenicola and L. fletcheri chiggers were fed on 1 and 2 human volunteers respectively. All subjects developed typical clinical signs and symptoms of scrub typhus beginning days 8-10 post chigger attachment (PCA); these included fever, severe headache, myalgia, regional lymphadenopathy, and eschar. The two L. fletcheri subjects developed a transient generalized rash on days 3-4 after the onset of fever, and these two individuals also appeared to suffer a more severe clinical disease. Rickettsemias were detected in all three volunteers beginning on day 7 PCA, 1-3 days before the onset of clinical disease. Rises in indirect fluorescent antibody titers occurred starting on days 13-19 PCA (day 4-11 post fever) and in Weil-Felix OXK titers starting on days 26-22 PCA (days 7-14 post fever). These results strongly suggest that the use of laboratory-reared chiggers is a reliable means of transmitting scrub typhus infections to volunteers.

Rickettsia tsutsugamushi organisms were detected and typed antigenically by direct immunofluorescence in mites from laboratory-maintained infected colonies of Leptotrombidium (Leptotrombidium) fletcheri and L. (L.) arenicola. Rickettsiae were identified most readily in unengorged larvae, but were also discernable in engorged larvae and all post-larval stages of the vectors.

Leptotrombidium (Leptotrombidium) umbricola, described here as a new species, is a member of the L. (L.) deliense group and most closely resembles L. (L.) vivericola. L. (L.) umbricola was collected from the ground surface and from animal hosts, in similar habitats to the scrub typhus vector, L. (L.) deliense. The host and habitat distribution records and the Rickettsia tsutsugamushi infection rates within unengorged specimens suggest that L. (L.) umbricola may be an important vector of scrub typhus in Peninsular Malaysia.

Over 1,200 isolations of R. tsutsugamushi were obtained over 7 generations of L. (L.) arenicola and no reproducible differences in virulence or antigenic markers could be detected among these isolations. The F8 generation yielded rickettsia with the same antigenic makeup and virulence properties as the original female parent mite. When these isolates were inoculated into mice they elicited the same predominant antibody specificities, i.e. they all produced antibody which reacted with the TA 763 and Karp strains. These were the same as the major antigens detected by direct fluorescent antibody staining of the isolates.

Leptotrombidium (Leptotrombidium) arenicola Traub, a vector of scrub typhus, had previously been found to occur in the coastal vegetation behind the edge of open sand along the beaches of Peninsular Malaysia. Surveys of the west coast beaches of Sabah and Sarawak were conducted to determine if this species occurs in similar habitat in East Malaysia on the island of Borneo. Leptotrombidium (L.) arenicola was not collected from the eighteen sites studied. Of the 11,982 mite larvae collected, 55 per cent were L.(L.) deliense (Walch), a well-known, widespread vector of scrub typhus.

L. (L.) deliense was the predominant vector of scrub typhus in a mature oil palm estate, but a small number of L. (L.) fletcheri (0.1% from rodents) and L. (L.) vivericola (0.02% from rodents and 8.0% from black plates) was also collected. Although good correlation between L. (L.) deliense collected from rodents and from black plates was not established, either method may serve as a general indicator of population fluctuations over a period of time. For the most part, the vectors of scrub typhus were limited to litter piles, and thus, the possibility of contracting scrub typhus within this type of habitat was minimal.