Geographical Assessment of Rickettsioses in Indonesia.

To expand the documentation of rickettsioses in Indonesia, we conducted an ectoparasite and small mammal investigation involving four major islands: Java, Sumatra, Sulawesi, and Kalimantan. Coastal and highland regions on each island surveyed were chosen to represent different ecologies in Indonesia. Indication of the presence of Rickettsia spp. was evident in all areas sampled. Typhus group rickettsiae-specific antibodies had significantly higher prevalence among small mammals captured in Java compared to the other islands surveyed (78% in coastal and 50% in highland regions) and the prevalence of spotted fever group rickettsiae-specific antibodies was significantly higher in Kalimantan than the other islands investigated. Hosts and vectors were restricted by Rickettsia spp. but not by coastal or highland regions. Our findings expand the range in which rickettsial pathogens have been documented within the Indonesian archipelago and point to a significant risk to human health.

Survey of Anopheles mosquitoes (Diptera:Culicidae) in West Sumba District, Indonesia.

In August 2007, mosquitoes were collected using four different collection methods at 2 upland interior and 2 coastal villages in West Sumba District, East Nusa Tenggara Province, Indonesia. Methods included human-baited and unbaited tent and malaise traps, human-landing collections (HLC), and unbaited CDC light traps. Mosquitoes were identified to species by morphological characters and all anophelines were tested for malaria circumsporozoite protein (CSP) using an enzyme-linked immunosorbent assay (ELISA). During six trap nights, 4,174 Anopheles mosquitoes belonging to 13 species were captured and identified: An. aconitus, An. annularis, An. barbirostris, An. flavirostris, Hyrcanus Group species, An. indefinitus, An. kochi, An. leucosphyrus group, An. maculatus s.l., An. subpictus s.l., An. sundaicus s.l., An. tessellatus, and An. vagus. Of potential disease vectors, An. annularis, An. subpictus, and An. vagus were the most frequently collected species in the upland interior sites, whereas An. sundaicus, An. subpictus, and An. vagus were most commonly found along the coast. The predominant species from evening human-landing collections (mosquitoes per human) were An. subpictus and An. vagus in the upland interior and An. sundaicus along the coast. All mosquitoes were non-reactive for Plasmodium CSP. One specimen of the An. leucosphyrus group was captured from indoor HLC in Tenateke Village, an upland interior location. This finding appears to represent a new collection record for Sumba Island.

Rickettsial infections of fleas collected from small mammals on four islands in Indonesia.

Ectoparasites were sampled from small mammals collected in West Java, West Sumatra, North Sulawesi, and East Kalimantan, Indonesia, in 2007-2008 and were screened for evidence of infection from bacteria in the Rickettsaceae family. During eight trap nights at eight sites, 208 fleas were collected from 96 of 507 small mammals trapped from four orders (379 Rodentia; 123 Soricomorpha; two Carnivora; three Scandentia). Two species of fleas were collected: Xenopsylla cheopis (n = 204) and Nosopsyllus spp. (n = 4). Among the 208 fleas collected, 171 X. cheopis were removed from rats (Rattus spp.) and 33 X. cheopis from shrews (Suncus murinus). X. cheopis were pooled and tested for DNA from rickettsial agents Rickettsia typhi, Rickettsia felis, and spotted fever group rickettsiae. R. typhi, the agent of murine typhus, was detected in X. cheopis collected from small mammals in West Java and East Kalimantan. R. felis was detected in X. cheopis collected from small mammals in Manado, North Sulawesi. R. felis and spotted fever group rickettsiae were detected in a pool of X. cheopis collected from an animal in East Kalimantan. Sixteen percent of the X. cheopis pools were found positive for Rickettsia spp.; four (10.8%) R. typhi, one (2.7%) R. felis, and one (2.7%) codetection of R. felis and a spotted fever group rickettsia. These data suggest that rickettsial infections remain a threat to human health across Indonesia.

Laboratory and field testing of bednet traps for mosquito (Diptera: Culicidae) sampling in West Java, Indonesia.

Surveillance of medically important mosquitoes is critical to determine the risk of mosquito-borne disease transmission. The purpose of this research was to test self-supporting, exposure-free bednet traps to survey mosquitoes. In the laboratory we tested human-baited and unbaited CDC light trap/cot bednet (CDCBN) combinations against three types of traps: the Mbita Trap (MIBITA), a Tent Trap (TENT), and a modified Townes style Malaise trap (TSM). In the laboratory, 16 runs comparing MBITA, TSM, and TENT to the CDCBN were conducted for a total of 48 runs of the experiment using 13,600 mosquitoes. The TENT trap collected significantly more mosquitoes than the CDCBN. The CDCBN collected significantly more than the MBITA and there was no difference between the TSM and the CDCBN. Two field trials were conducted in Cibuntu, Sukabumi, West Java, Indonesia. The first test compared human-baited and unbaited CDCBN, TENT, and TSM traps during six nights over two consecutive weeks per month from January, 2007 to September, 2007 for a total of 54 trapnights. A total of 8,474 mosquitoes representing 33 species were collected using the six trapping methods. The TENT-baited trap collected significantly more mosquitoes than both the CDCBN and the TSM. The second field trial was a comparison of the baited and unbaited TENT and CDCBN traps and Human Landing Collections (HLCs). The trial was carried out from January, 2008 to May, 2008 for a total of 30 trap nights. A total of 11,923 mosquitoes were collected representing 24 species. Human Landing Collections captured significantly more mosquitoes than either the TENT or the CDCBN. The baited and unbaited TENT collected significantly more mosquitoes than the CDCBN. The TENT trap was found to be an effective, light-weight substitute for the CDC light-trap, bednet combination in the field and should be considered for use in surveys of mosquito-borne diseases such as malaria, arboviruses, and filariasis.

Operational vector-borne disease surveillance and control: closing the capabilities gap through research at overseas military laboratories.

Malaria, dengue fever, chikungunya virus, leishmaniasis, and a myriad of other vector-borne diseases pose significant threats to the warfighter and to the overall combat effectiveness of units. Military preventive medicine (PM) assets must accurately evaluate the vector-borne disease threat and then implement and/or advise the commander on countermeasures to reduce a particular threat. The success of these measures is contingent upon the biology of the disease vector and on the tools or methods used to conduct vector/pathogen surveillance and vector control. There is a significant gap between the tools available and those required for operational PM assets to provide real-time, effective surveillance and control. A network of US Army and US Navy overseas laboratories is focused on closing the current capabilities gap. Their mission is to develop and field test tools and methods to enhance the combatant commander's ability to identify and mitigate the threat posed by these vector-borne diseases.

Bionomics of Anopheles spp. (Diptera: Culicidae) in a malaria endemic region of Sukabumi, West Java, Indonesia.

A 15-month bionomic study of Anopheles species was conducted in two ecologically distinct villages (coastal and upland) of Sukabumi District, West Java, Indonesia from June 2006 to September 2007. Mosquitoes were captured using human-landing collections at both sites. During the study, a total of 17,100 Anopheles mosquitoes comprising 13 Anopheles species were caught: 9,151 at the coastal site and 7,949 at the upland site. Anopheles barbirostris, Anopheles maculatus, and Anopheles vagus were the predominant species caught at the coastal site, and Anopheles aconitus, Anopheles barbirostris, and An. maculatus predominated in the upland site. Overall, species were exophagic at both sites, but there was variation between species. Anopheles aconitus was endophagic at the coastal site, exophagic at the upland site, collected most often in April 2007 and had a peak landing time between 22:00 and 23:00. Anopheles sundaicus was only collected at the coastal site, exophagic, collected most often in October 2006, and had a peak landing time between 19:00 and 20:00. Potential malaria vector species such An. aconitus, An. maculatus, and An. sundaicus were present throughout the year. None of the 7,770 Anopheles tested using CSP-ELISA were positive for malaria, although the risk for malaria outbreaks in Sukabumi district remains high.

Integration of insect parasitic nematodes (Rhabditida steinernematidae) with insecticides for control of pest mole crickets (Orthoptera: Gryllotalpidae: Scapteriscus spp.).

To develop a successful integrated pest management program for pest mole crickets (Orthoptera: Gryllotalpidae: Scapteriscus spp.), it is important to ascertain the compatibility of infective juveniles of insect parasitic nematodes and chemical insecticides. Aqueous solutions of five pesticides (acephate, bifenthrin, deltamethrin, fipronil, and imidacloprid) used in turfgrass to control mole crickets were tested for compatibility with Steinernema scapterisci Nguyen & Smart (Rhabditida: Steinernematidae) in the laboratory. Survival of S. scapterisci was >95% in solutions of acephate, bifenthrin, and imidacloprid. Infectivity of S. scapterisci in adult Scapteriscus vicinus Scudder was >60% in acephate and bifenthrin; however, infectivity was <40% in imidacloprid. The entomopathogenic nematode was compatible with most insecticides tested without significantly reduced survival or infectivity.

Development of a toxic bait for control of eastern lubber grasshopper (Orthoptera: Acrididae).

This study assessed baits for eastern lubber grasshopper, Romalea guttata (Houttuyn). When offered a choice among several grain-based baits (rolled oats, wheat bran, oat bran, yeast, corn meal, cornflakes) and vegetable oils (canola, corn, peanut, soybean), eastern lubber grasshopper adults preferred bait consisting of wheat bran carrier with corn oil as an added phagostimulant. Other carriers were accepted but consumed less frequently. Discrimination by eastern lubber grasshoppers among oils was poor. Similarly, addition of flavorings (peppermint, anise, lemon, banana) resulted in few significant effects. The carbaryl, wheat bran, and oil bait developed in this study was effective at causing eastern lubber grasshopper mortality in field-cage studies. Significant mortality occurred even though grasshoppers had to locate dishes of bait in a large cage, and could feed on daylilies, or grass growing through the bottom of the cage, rather than on the bran flakes. Consumption of as little as a single carbaryl-treated bran flake could induce mortality, although individuals varied greatly in their susceptibility. The bait matrix developed in this study was readily consumed when in the presence of some plant species. We expect that wheat bran and corn oil bait would be most effective as protection for less preferred plants (tomato, pepper, eggplant, leek, parsley, fennel, daylily, lily of the Nile, and canna lily) because baits were readily consumed in the presence of these plants. Plants that are readily consumed in the presence of bait (preferred plants) included butter crunch lettuce, carrot, yellow squash, cauliflower, collards, green onion, chive, cucumber, cabbage, cantalope, endive, red leaf lettuce, society garlic, caladium, and amaryllis. Baits are likely to be less effective in the presence of such plants. On average, vegetables in Solanaceae (i.e., tomato, pepper, and eggplant) and Apiaceae (i.e., fennel and parsley) elicited high levels of bait-feeding activity, indicating that these vegetables were not highly preferred. The plants tested from Liliaceae, Cucurbitaceae, Asteraceae, and Brassicaceae elicited an intermediate-to-low level of bait feeding.