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

Abstract

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.

Figure 1.

Nematode production room

Figure 2.

Post-parasitic R. iyengari (20× magnification)

Figure 3.

Post-parasitic nematodes on coconut coir fibres (containers on the left) and on coarse sand (containers on the right)

Figure 4.

Average (± SE) abundance of infective preparasites and eggs obtained per container after 2 months storage of post-parasitic nematodes. (n=320 for each of the substrates)

Figure 5.

Impact of long-term storage on the yield (average ± SE) of infectious preparasites. (n=40 for each column)