Optimization of breeding output for larval stage of Anopheles gambiae (Diptera: Culicidae): prospects for the creation and maintenance of laboratory colony from wild isolates.

Domesticating anopheline species from wild isolates provides an important laboratory tool but requires detailed knowledge of their natural biology and ecology, especially the natural breeding habitats of immature stages. The aim of this study was to determine the optimal values of some parameters of Anopheles gambiae larval development, so as to design a standard rearing protocol of highland isolates, which would ensure: the biggest fourth instars, the highest pupae productivity, the shortest duration of the larval stage and the best synchronization of pupation. The density of larvae, the size of breeding water and the quantity of food supplied were tested for their effect on larval growth. Moreover, three cheap foodstuffs were selected and tested for their capability to improve the breeding yield versus TetraMin® as the standard control. The larval density was a very sensitive parameter. Its optimal value, which was found to be ≈1 cm-2 surface area, yielded a daily pupation peak of 38.7% on day 8 post-oviposition, and a global pupae productivity of 78.7% over a duration range of three days. Anopheles gambiae's larval growth, survival and developmental synchronization were density-dependent, and this species responded to overcrowding by producing smaller fourth instars and fewer pupae, over elongated immature lifetime and duration range of pupae occurrence, as a consequence of intraspecific competition. While shallow breeding waters (<3 cm) produced a higher number of pupae than deeper ones, no effect of the breeding habitat's absolute surface area on larval development was observed. Increasing the daily food supply improved the pupae productivity but also boosted the water pollution level (which was assessed by the biological oxygen demand (BOD) and the chemical oxygen demand (COD)) up to a limit depending on the food quality, above which a rapid increase in larval mortality was recorded. The food quality that could substitute the manufactured baby fish food was obtained with weighed mixture of 1 wheat+1 shrimp+2 fish. On establishing an anopheline mosquito colony in the laboratory, special care should be taken to design and maintain the appropriate optimal values of larval density, water depth, daily diet quantity and nutritional quality.

In vitro nematicidal activity of extracts of Canthium mannii (Rubiaceae), on different life-cycle stages of Heligmosomoides polygyrus (Nematoda, Heligmosomatidae).

The increasing prevalence of anthelmintic-resistant strains of helminths, drug residues in animal products and high cost of conventional anthelmintics has created an interest in studying medicinal plants as an alternative source of anthelmintic. The potential nematicidal activities of four extracts from the bark of Canthium mannii (Rubiaceae) stem were investigated in vitro. Extracts were diluted in distilled water (DW) to obtain five different concentrations (1.5, 2.0, 2.5, 3.0 and 3.5 mg/ml) and put in contact with eggs and larvae of Heligmosomoides polygyrus. The different stages of the life cycle were also put in contact with the same concentration of mebendazole (MBZ, positive control). One millilitre of each extract at different concentrations and control were added to 1 ml solution containing 30-40 eggs or 10-15 larvae (L1, L2 and L3) and distributed in different Petri dishes. The eggs and larvae were incubated at 24 degrees C and exposure times were: 48 h for un-embryonated eggs, 6 h for embryonated eggs; 2, 4, 6 and 24 h for L1 and L2 larvae, 24-48 h for infective larvae (L3), and 5 days for the larval development test (from L1 to L3). DW and 1% dimethyl sulphoxide (DMSO) were used as placebo and DMSO control, respectively. Significant effects were obtained with three of the four extracts, and differences were observed depending on the parasite stage. Cold water extract (CWE), hot water extract (HWE) and ethanol extract (ETE) inhibited embryonic development (40, 45 and 10%) and hatching of embryonated eggs (40, 85 and 80%), respectively, at 3.5 mg/ml. Only ETE killed L1 (97.18%) and L2 (92.68%) larvae of H. polygyrus after 24 h at 3.5 mg/ml and drastically reduced the production rate (6% at 3.0 and 3.5 mg/ml) of infective larvae (L3) after 5 days of incubation compared to other extracts (P < 0.05). However, the infective larvae of H. polygyrus were resistant to the effect of each of the tested products (extracts and mebendazole). These in vitro results suggested that extracts of C. mannii, used by traditional healers in Dschang, Western Region of Cameroon (Central Africa) to cure intestinal helminthiasis and abdominal pains of their patients, possess nematicidal properties. The active principles responsible for the activity could be secondary metabolites such as alkaloids and saponins present in the extracts. It is suggested that further experiments incorporating in vivo purification of extracts and toxicological investigations should be carried out.

The gastro-intestinal helminth infections of domestic fowl in Dschang, western Cameroon.

Three hundred and fifty one chickens purchased from the Dschang animal market were examined for gastro-intestinal helminths. Ten species were found with the following prevalences: Heterakis brevispiculum (59.3%), Ascaridia galli (51.6%), Hymenolepis carioca (48.4%), Dispharynx spiralis (20.8%), Tetrameres americana (17.1%), Amoebotaenia cuneata (15.1%), Raillietina tetragona (14.5%), Syngamus trachea (13.7%), Hymenolepis cantaniana (5.7%) and Capillaria contorta (2.0%). Infections were predominantly mixed (93.5%). The infection rates were not influenced by host sex except for A. galli which was more prevalent in cocks. Older chickens showed some resistance to A. cuneata and S. trachea. Parasite prevalence and/or worm burdens were generally higher during the rainy season (April to October).

Periodic release of Eimeria species oocysts from chicken during daytime hours in a tropical environment.

Every two hours from 6 to 18 hrs, feces were collected from 62 native chickens in Dschang, Western Cameroon. Fecal Eimeria oocyst counts were made using a McMaster cell. Oocysts were most abundant at 16 and 18 hrs when air humidity was high. This particular timing of oocyst release was therefore considered adaptive for survival and transmission of coccidia and ideal for diagnosis.