Efficacy of the mermithid nematode, Romanomermis iyengari, for the biocontrol of Anopheles gambiae, the major malaria vector in sub-Saharan Africa.

BACKGROUND: The intensive use of chemical insecticides against mosquitoes has led to the development of widespread insecticide resistance. Control of Anopheles mosquitoes in malaria endemic areas of sub-Saharan Africa has become increasingly difficult. There is an urgent need for malaria control programmes to adopt more integrated mosquito management approaches that include sustainable, nonchemical solutions. The mermithid nematode Romanomermis iyengari is one of several natural control alternatives to synthetic pesticides for mosquito suppression. This study evaluated the effectiveness of the nematode R. iyengari for control of Anopheles gambiae. METHODS: The nematode R. iyengari was mass-produced, and pre-parasitic stage (J2) were used for laboratory and field experiments. In laboratory experiments, two concentrations of pre-parasitics (5 and 10 J2 per larva) were tested against first- (L1), second- (L2) and third-instar (L3) larvae of An. gambiae. Infected larvae were observed daily to determine their mortality rate and the number of post-parasitic nematodes emerging from dead larvae. In field experiments, 3500, 4000 and 5000 J2/m2 were sprayed in separate natural Anopheles breeding sites. After treatment, the larval mosquito density in the breeding sites was assessed every 5-7 days. RESULTS: Laboratory results showed that larval An. gambiae is susceptible to nematode infection: 100% L1 larvae died within 24 hours post-treatment, and 100% of both L2 and L3 larvae died within 7 days, regardless of nematode concentrations. The average number of post-parasitic nematodes emerging per larva increased with increasing nematode concentration. In field experiments, the monthly applications of 3500 to 5000 pre-parasitic nematodes per m2 eliminated larval mosquito development in Anopheles- and mixed breeding sites. Larval mosquito density dramatically decreased five days after the first treatment in all treated sites and was maintained at a very low level during the whole experimental period. Basically, only early instar larva were detected in treated sites throughout the test period. The average number of post-parasitic nematodes emerging per larva collected in treated sites was 1.45, 2, and 5.7 respectively for sites treated with 3500, 4000, and 5000 J2/m2. CONCLUSIONS: Malaria mosquito larvae is susceptible to R. iyengari infection in West Africa. Parasitism intensity depends on tested nematode concentrations. Monthly application of 3500 J2/ m2 was enough to control effectively larval An. gambiae in wetlands and floodable locations in West Africa.

Large-scale production of the malaria vector biocontrol agent Romanomermis iyengari (Nematoda: Mermithidae) in Benin, West Africa.

Background: The mermithid nematode Romanomermis iyengari is one of several natural control alternatives to synthetic pesticides for mosquito suppression. The commonly used mass rearing procedure of R. iyengari involves the use of coarse sand as a substrate for nematode maturation and oviposition. The coarse sand technique gives excellent nematode productivity in North America. However, under West African climatic conditions, this technique generates relatively lesser amounts of infectious worms. We evaluated coconut coir fibres as a replacement for coarse sand to improve yields in large-scale production of R. iyengari in Benin, West Africa. Materials and Methods: Culex quinquefasciatus was the host for the nematodes, and mosquitoes were blood-fed on chickens. Four days after blood feeding, egg rafts were collected and transferred into trays, each containing 2 l of water. The mosquito larvae were fed with fish food. When the mosquito larvae reached the second instar, preparasites (J2) were added (3 J2/larva) to the incubation trays. Eight days after infection, post-parasitic juveniles were separated from the water containing dead mosquito larvae and other debris using sieves and needles; 2 g of them were deposited in containers with coarse sand or coconut coir fibres and water. Three hours later, the water was drained, the jars covered and stored for eight weeks, after which J2 abundance was determined, using a total of 320 containers for each substrate. The abundance of J2 preparasites was also assessed 3-5 months after storage to determine the impact of long-term storage on the J2 yield. Results: After 2 months storage, 2 g of post-parasites (~457 females and 583 males) yielded an average of 559,300±6094 J2 and 155,818±4427 J2 per container for coconut fibres and for coarse sand, respectively. During long-term storage, yields of J2 on coconut fibres substrate slowly decreased from 442,180±9322 J2 (3 months storage) to 163,632±12,416 J2 per container (5 months storage). On coarse sand substrate, the yield was relatively low and decreased from 49,812±1200 J2 at 3 months storage to 3046±229 J2 at 5 months storage. Conclusion: Under West African climatic conditions, coconut coir fibres gave significantly higher preparasitic nematode yields than the coarse sand technique.

Potential of the bush mint, Hyptis suaveolens essential oil for personal protection against mosquito biting.

We studied the potential of the essential oil extract from the bush mint, Hyptis suaveolens, for use against mosquito biting under both laboratory and field conditions. In the laboratory, the repellency of various concentrations (1-6%) of the essential oil was assessed against Anopheles gambiae, based on a 15-min landing and biting on treated forearms of volunteers. In the laboratory, the percentage of mosquitoes landing on the forearm was 42, 33, 23, 23, 9, and 2 for 1%, 2%, 3%, 4%, 5%, and 6% essential oil concentration, respectively; and 92 and 91 for the solvent (isopropanol) and untreated control, respectively. The percentage of mosquitoes taking a blood meal was 22, 12, 13, 12, 5, and 3 for 1%, 2%, 3%, 4%, 5%, and 6% essential oil, respectively; and 52 and 51 for the solvent and control, respectively. In the field, the 6% essential oil repelled all mosquitoes immediately postapplication; this activity declined to 75% after 5 h. The repellent action of the 8% essential oil concentration was higher, 97% after 5 h. Based on these data, the essential oil of H. suaveolens appears to be a good candidate for use in the integrated management of mosquito vectors of disease.