This paper reports the experimental transmission of a bird parasite into jirds. Infective larvae of Cardiofilaria nilesi obtained from laboratory colonized Coquillettidia crassipes mosquitoes which had fed on an infected chicken were inoculated subcutaneously into jirds. The number of larvae per jird varied from 10 to 228. Microfilaraemia appeared 22 to 89 days after inoculation of the infective larvae. Experiments were carried out with 24 jirds through six generations extending over a period of 22 months and 17 produced patent infections. At present 8 infected jirds are being maintained in the laboratory; their patent periods ranging from 6 to 13 months. However, the longest patent period observed was about thirteen months. The percentage of adults recovered in autopsied jirds ranged from 0 to 40 with an average of 16. The chicken showed a microfilarial periodicity with the peak microfilarial density around 2200 hours. However, in jirds there was a change in sub-periodicity. This model in the jird may be very useful for the screening of filaricides and in immunological work.
R. sabanus and R. muelleri are very common in the lowland forests of Malaysia. In nature they are infected with Breinlia sp. and D. ramachandrani. In an attempt to determine whether they are also susceptible to subperiodic B. malayi and thereby being potential reservoirs of infection of the disease, 24 R. muelleri and 17 R. sabanus were experimentally infected with the parasite. Results show that although they can support the full development of the parasite, they are poor hosts. This confirms the observation that in Malaysia natural infection of Rattus spp. with the parasite has not been seen. These rats therefore are probably not important in the zoonotic transmission of subperiodic B. malayi in Malaysia.
Surveillance methods for Coquillettidia crassipes were studied in an open housing estate near Kuala Lumpur using three types of traps Trinidad 10 trap, modified Lard can trap and IMR trap, each baited with chicken or pigeon. All traps attracted Cq. crassipes. There was no significant difference in the catches in the three traps. There was also no significant difference between chicken and pigeon as bait. Catches at heights of 1.5, 3, 4.5 and 6 m did not show any significant difference in density. Cq. crassipes was active at night with an early peak during the first hour of the night and a minor peak between 0100 and 0200 hours. The activity of the parous and nulliparous sections of the population was similar, except that a higher proportion of the parous females was active during the second peak compared with the nulliparous females. The parous rate was 22.3%, and the probability of survival through one day for two gonotrophic cycles was 0.711 and 0.650. The infection rate for Cardiofilaria was 29 out of 1052 (2.76%) and the infective rate (L3 larvae) was 13 out of 1052 (1.24%). 48.3% of the infected Cq. crassipes had a worm burden of more than ten larvae. One of the chickens in the traps was positive for microfilariae of Cardiofilaria four weeks after exposure as bait. Laboratory bred Cq. crassipes fed on this chicken produced infective larvae in ten days, and these were inoculated into clean chickens and pigeons. Microfilariae appeared in the chickens but not in pigeons. The adult worms recovered await identification.
The leaf-monkeys, Presbytis cristata and Presbytis melalophos, experimentally infected with subperiodic Brugia malayi, have been used for studies on the pathoimmunology of the infection and the screening of potential filaricides during the last 6-8 years, and considerable information on the pattern of microfilaraemia and adult worm recoveries have been obtained. The prepatent periods in 97 P. cristata and 45 P. melalophos, each infected with about 200 infective larvae, were similar, these being approximately 70 and 68 days respectively. Although all infected animals became microfilaraemic, the peak geometric mean count was much higher in P. cristata than in P. melalophos, this being 182.0 and 65.8 per ml blood respectively. Mean adult worm recovery expressed as the percentage of the infective dose was 4.7% and 2.5%, respectively. Most worms were recovered from the sacral nodes/thoracic duct or inguinal lymph nodes in these animals. In view of the higher worm recovery and the higher peak microfilaraemia attained, it is concluded that P. cristata is a better model for the infection than P. melalophos.
Accurate identification of filarial parasites in mosquitoes poses a major problem for the coordination of filariasis control programs. Traditional methods are tedious, and some are not specific enough to give satisfactory results. Amplification of specific gene sequences by primer-directed polymerase chain reaction (PCR) has been increasingly utilized as a diagnostic tool. However, current protocols for the extraction of parasite DNA from mosquito samples are tedious and could lead to failure of PCR amplification. We demonstrate that the use of Chelex is an efficient method for DNA extraction from mosquitoes and the parasite and that PCR amplification with primers specific for Brugia malayi yields a band of the expected size. The PCR products were transferred to a nylon membrane with Southern blotting, and a B. malayi-specific digoxigenin-labeled probe confirmed the sequence similarity of the PCR-amplified fragment and increased the sensitivity of the PCR assay. Use of this probe enabled us to detect PCR-amplified product from B. malayi even when a product was not visible on an ethidium bromide-stained agarose gel. This increased sensitivity allowed us to detect the parasite in the heads of mosquitoes.
The possible depression of cell-mediated immunity by long-term Brugia malayi infection in jirds (Meriones unguiculatus) was investigated. Different groups of infected jirds were sensitized with dinitrofluorobenzene, sheep red blood cells, Dirofilaria immitis adult antigens and B. malayi adult antigens. The 24-hour delayed type hypersensitivity skin response to testing with antigen was measured as an in vivo correlate of cell-mediated immunity. The delayed-type hypersensitivity responses to dinitrofluorobenzene, sheep red blood cells and D. immitis antigens were normal but the response to B. malayi antigens was significantly depressed, confirming that long-term B. malayi infection depresses cell-mediated immunity and that this depression is specific to B. malayi antigens.
Four Presbytis cristata were treated with oral ivermectin at the same time as the subcutaneous inoculation of 100 infective larvae monthly for three months. Two animals given 0.2 mg/kg monthly and two others given 0.3 mg/kg monthly as well as three control animals became patent for microfilaraemia. However, only 1% of the infective dose was recovered as adult worms from animals in the higher drug dosage group compared to 8.2% and 6.2% in the lower dosage and control groups respectively.
Adult worms of the rural strain of Wuchereria bancrofti in Peninsular Malaysia obtained from a successful experimental transmission in an immunosuppressed Macaca fascicularis are described for the first time. Although the worms, especially females, were slightly smaller, they were similar in morphology to those of the periodic and non-periodic W. bancrofti previously described.
Comparative studies of vector efficiency were done with the Liverpool and Malaysian strains of Aedes (Finlaya) togoi for subperiodic Brugia malayi and Brugia pahangi. The Malaysian strain of A. togoi was found to take in fewer microfilariae under the same experimental conditions than the Liverpool strain. Also, for various microfilarial densities in the host's peripheral blood, the Malaysian strain had less mean infective larvae per fed mosquito than the Liverpool strain. The microfilarial intake of A. togoi was not affected by the site of feeding on the host affected by the site of feeding on the host. Most of the mosquitoes took in fewer microfilariae than expected. It is concluded from these studies that the Malaysian strain of A. togoi is a susceptible and reasonably good vector for subperiodic B. malayi and B. pahangi. Further field studies should be carried out to determine its importance as a natural vector of Brugian filariasis.
Using seven methods of surveillance, 58 species of mosquitoes from nine genera were in Pantai and the two neighbouring villages during two visits in 1982. Ma. bonneae was the most prevalent species attracted to man. In the forest shade Ma. bonneae and Ma. dives showed activity throughout the 24 hours with peak biting during 1900-2100 hours. An. balabacensis exhibited peak activity shortly after midnight. Inside and outside house, Ma. bonneae showed similar activity except that it ceased during the day. Mansonia was only mildly zoophilic. CDC light traps gave poor yields of mosquitoes. Pyrethrum spray catch inside houses early morning did not include any Mansonia. Outdoor day resting catch included Ma. bonneae fed on man. Transmission of Brugia, probably human filariasis, by Ma. bonneae occurred in Pantai and in the two neighbouring villages. One infection in Ma. dives was found in Pantai. The monthly infective biting rate and monthly transmission potential for Ma. bonneae were estimated at the forest shade and outside the house in Pantai.