A mark-release-recapture experiment with Mansonia mosquitos in Malaysia.

An experiment was carried out with Mansonia mosquitos in an area endemic for subperiodic Brugia malayi to assess the applicability of the mark-release-recapture method to these mosquitos. An estimated 17,880 individuals of six species of Mansonia were marked with fluorescent dust and released: 453 Ma. annulata, 305 Ma. annulifera, 6,200 Ma. bonneae, 516 Ma. dives, 3,998 Ma. indiana and 6,408 Ma. uniformis. Twenty-three marked individuals were recaptured. Most recaptures were made one or two nights after their release, but one Ma. annulifera was recaptured five nights later and one Ma. bonneae had been marked and released 6-11 nights previously. The recaptured mosquitos were collected between 0.5 and 2.4 km from their release points.

[Clostridium bifermentans serovar malaysia, a new anaerobic bacterium pathogen to mosquito and blackfly larvae]. / Clostridium bifermentans serovar malaysia, une nouvelle bactérie anaérobie pathogène des larves de moustiques et de simulies.

A strain of Clostridium bifermentans individualized as serovar malaysia (C.b.m.) according to its specific H antigen is toxic to mosquito and blackfly larvae when given orally. The toxicity occurs in sporulated cells which contain, in addition to spores, proteinic parasporal inclusion bodies and feather-like appendages; the amino acid content of the inclusion bodies is similar to that of Bacillus thuringiensis serovar israelensis (B.t.i.) and B. sphaericus crystals. The toxicity to Anopheles stephensi is as high as that of B.t.i. and the best strains of B. sphaericus. Culex pipiens is somewhat less susceptible, and Aedes aegypti much less. Pure parasporal inclusion bodies, isolated by ultracentrifugation on sucrose gradients, are highly toxic to mosquito larvae. The larvicidal power is destroyed by heating at 80 degrees C or by treatment with 50 mM NaOH. It is preserved by freeze-drying. The innocuity to mice of the sporulated cells is shown by different routes of administration: force-feeding, percutaneous, subcutaneous, intraperitoneal or intravenous injections. The potential for the biological control of mosquito and blackfly larvae is suggested.

The transmission of an avian filarioid Cardiofilaria nilesi to the jird, Meriones unguiculatus.

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.

Laboratory colonization and life cycle of Coquillettidia crassipes in Malaysia.

Methods are described for the laboratory colonization of Coquillettidia crassipes. The highest rate of insemination occurred in 60 x 60 x 120 cm cages and better insemination in laboratory adapted F15 generation. Embryonation and hatchability of eggs ranged from 69.6 to 97.9% and 63.3 to 94.3% respectively. Gravid females laid egg rafts on water in 500 ml breakers with small leaves of Salvinia for resting. Newly hatched larvae were set up in a basal medium of guinea pig dung and water or liver powder, yeast powder and water. Larvae attached to aquatic plants or 'Keaykolour' ruffia snow white paper. The cultures with paper gave better yields. At present 21 generations of Cq. crassipes have been reared in the laboratory.

Identification of Brugia malayi in vectors with a species-specific DNA probe.

We evaluated the potential value of a cloned sequence of genomic DNA of Brugia malayi as a species-specific probe. Clone pBm 15 reacted with all stages of 8 different geographic isolates of B. malayi and cross-hybridized with microfilariae of B. timori. It did not hybridize with Wuchereria bancrofti or with B. pahangi, W. kalimantani, Dirofilaria repens, Breinlia booliati or Cardiofilaria species, animal filariids that can be sympatric with B. malayi. P32-labeled clone pBm 15 correctly identified mosquitoes infected even with 1 infective larva of B. malayi. This specific DNA probe should be an invaluable tool to monitor control programs of Brugian filariasis.

Field studies on the surveillance of Coquillettidia crassipes (Van der Wulp) and the isolation of a strain of Cardiofilaria in peninsular 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.

Field and laboratory observations on Coquillettidia crassipes in relation to transmission of Brugia malayi in Peninsular Malaysia.

Field observations were made on Coquillettidia crassipes during a study of Mansonia in a swamp forest ecotype in Tanjong Karang. There was an increase in abundance in July consistent with the increase in abundance of Mansonia and an increase in rainfall. The biting cycle showed a dramatic early peak during the period 1900-2000 hours. The probability of daily survival through one day for the first three gonotrophic cycles was 0.770, 0.722 and 0.759. Two of the 54 Cq. crassipes dissected were infective, with two and 25 L3 larvae of Brugia. Both subperiodic B. malayi and B. pahangi developed into L3 larvae in laboratory bred Cq. crassipes. The index of experimental infection was higher for B. pahangi. Mansonia bonneae and Ma. uniformis showed higher indices of experimental infection than Cq. crassipes for subperiodic B. malayi. It is concluded that in an endemic area with a high density of Cq. crassipes it could act as a secondary vector of Brugian filariasis.

Evaluation of the effectiveness of permethrin and DDT impregnated bed-nets against Anopheles maculatus.

A laboratory evaluation of cotton and nylon bed-nets treated with permethrin and DDT was carried out by releasing blood-fed An. maculatus for 30 minutes in the treated net. Results show almost 100% mortality up to one year after treatment. A ten minute exposure on the outside of the net also gave similar results. Even after washing the nets continued to give very good results. A method of treating the nets is described.

Laboratory colonization of Mansonia uniformis, Ma. indiana and Ma. bonneae in Malaysia.

Methods are described for the laboratory colonization of Mansonia uniformis, Ma. indiana and Ma. bonneae in Malaysia. Gravid females oviposited in 500 ml beakers with a layer of water covered with small leaves of Salvinia. Newly hatched larvae were set up in a basal medium of guinea pig dung and water or liver powder, yeast powder and water. Larvae attached to aquatic plants or 'Keaykolour' ruffia snow white paper. The cultures with paper gave better yields than those with plants. Production of Ma. uniformis was higher than the other two species. Twelve generations of Ma. uniformis and 11 generations of Ma. indiana and Ma. bonneae were monitored in the laboratory.

Anopheles flavirostris incriminated as a vector of malaria and Bancroftian filariasis in Banggi Island, Sabah, Malaysia.

Seven villages in Banggi Island, Sabah, Malaysia, were surveyed four times to evaluate the roles of local mosquitoes as vectors of malaria and Bancroftian filariasis. 11 species of Anopheles were found biting man. 53.9% of the anophelines caught were An. flavirostris, 27.1% An. balabacensis, 6% An. donaldi and 4.2% An. subpictus. Infective malaria sporozoites, probably of human origin, were found in two of 336 An. flavirostris and 12 of 308 An. balabacensis. Sporozoites, probably of a non-human Plasmodium, were found in An. umbrosus. Nine of 1001 An. flavirostris and four of 365 An. balabacensis harboured L2 or L3 filarial larvae identified as those of Wuchereria bancrofti. This is the first record of An. flavirostris as a natural vector of malaria and W. bancrofti in Sabah.

Enzyme polymorphism in Mansonia uniformis, a mosquito vector of brugian filariasis.

Three temporal samples of a wild population of Mansonia uniformis were analysed for genetic variation at six gene-enzyme systems. Adenylate kinase, hexokinase (3 loci) and cathodal malate dehydrogenase were monomorphic. Phosphoglucomutase, glucose phosphate isomerase, isocitrate dehydrogenase and anodal malate dehydrogenase were polymorphic. Each of the polymorphic loci was represented by three alleles. The average heterozygosity or gene diversity was 0.0437.

Filariasis in Bengkoka Peninsula, Sabah, Malaysia: bionomics of Mansonia sp.

Studies on age groups within activity cycles, age composition and survivorship in natural populations of Mansonia in Kampung Pantai, Bengkoka Peninsula of Sabah state have been described. Early activity of 3-5 parous Ma. bonneae during the first hour after sunset was noted. Age composition of Ma. bonneae at forest shade, indoor and outdoor of house, comparative buffalo vs human bait outdoor in Kampung Pantai showed all round high parous rates ranging from 66.7 to 75.4%. Population 3-parous and older ranged from 18.8 to 26.7%. Nine of the 14 infective Ma. bonneae were 3-parous and this segment of the population was engaged in active transmission. High parous rates were observed in Ma. dives and Ma. uniformis taken in small numbers. Parous rates of Ma. bonneae taken in Kampung Delima and Kampung Taradas were also high. Estimates of daily survivorship of Ma. bonneae and Ma. dives determined by two methods were very high.

Filariasis in Bengkoka Peninsula, Sabah, Malaysia: vector studies in relation to the transmission of 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.

Mosquito fauna of the Bengkoka Peninsula, Sabah, Malaysia.

A total of 37 species of mosquitoes from seven genera were collected in six villages in the Bengkoka Peninsula, Sabah State, during two visits in 1981 in connection with studies on malaria and filariasis. Fifty-five per cent of the total mosquitoes collected were Mansonia. An. collessi constituted a new record of the species from Sabah. An. balabacensis was found to be naturally infected with sporozoites. Ma. bonneae was found to be naturally infected with Brugia, probably B. malayi. Parous rates of An. balabacensis and Ma. bonneae were very high with consequent high probability of survival ideally suiting transmission of malaria and filariasis.

A small scale field trial with Bacillus thuringiensis against culicine mosquitoes, Kelang, Malaysia.

Bacillus thuringiensis israeliensis (BTI) against culicine mosquitoes was tried out in cement sullage drains in Kelang municipal area at a dosage of 0.15 ppm. and 0.6 ppm. The results of the trial showed that at 0.15 ppm. the BTI was not effective, but at 0.6 ppm. it was effective giving about 95% kill. There was no residual effect and treatment had to be repeated weekly.

Multiple alleles and low variability of glycerol-3-phosphate dehydrogenase in Aedes albopictus (Diptera: Culicidae) from Peninsular Malaysia.

1. A total of 8 samples from three natural populations and a laboratory strain of Aedes albopictus were analysed for glycerol-3-phosphate dehydrogenase phenotypes by means of horizontal starch-gel electrophoresis. 2. The electrophoretic phenotypes were governed by three codominant Gpd alleles. 3. There was low variability, with the heterozygosity in the variable samples ranging from 0.02 to 0.12. 4. The commonest allele in all the population samples was GpdB which encoded an electrophoretic band with intermediate mobility. 5. There was no temporal or spatial variation.