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.

Trypanosoma vivax, T. congolense "forest type" and T. simiae: prevalence in domestic animals of sleeping sickness foci of Cameroon.

In order to better understand the epidemiology of Human and Animal trypanosomiasis that occur together in sleeping sickness foci, a study of prevalences of animal parasites (Trypanosoma vivax, T. congolense "forest type", and T. simiae) infections was conducted on domestic animals to complete the previous work carried on T. brucei gambiense prevalence using the same animal sample. 875 domestic animals, including 307 pigs, 264 goats, 267 sheep and 37 dogs were sampled in the sleeping sickness foci of Bipindi, Campo, Doumé and Fontem in Cameroon. The polymerase chain reaction (PCR) based method was used to identify these trypanosome species. A total of 237 (27.08%) domestic animals were infected by at least one trypanosome species. The prevalence of T. vivax, T. congolense "forest type" and T. simiae were 20.91%, 11.42% and 0.34% respectively. The prevalences of 7 vivax and T. congolense "forest type" differed significantly between the animal species and between the foci (p < 0.0001); however, these two trypanosomes were found in all animal species as well as in all the foci subjected to the study. The high prevalences of 7 vivax and T congolense "forest type" in Bipindi and Fontem-Center indicate their intense transmission in these foci.

The influence of self-fertilization and pairing on life-history traits in the freshwater snail Bulinus forskalii (Gastropoda, planorbidae).

This study analyses the basic reproductive biology of the schistosonie-vector snail Bulinus forskalli. This hermaphroditic species can self-fertilize. Variation of the mating system was analysed in two populations from Cameroon in a three-step experiment. (i) The fecundity (number of eggs and of egg capsules) of isolated virgin individuals from both populations was followed over a month, as well as the survival of offspring from these individuals. Estimates of these parameters were intermediate. Our results also indicate that inbreeding depression has a maximum value of about one-half in both populations. No difference was observed between populations for both the overall fecundity of adults and survival of offspring. (ii) Individuals were then paired either within, or between populations over three successive days, and the number of copulations recorded. Copulations were observed in all situations, including between-population pairs. No significant dlifference in the number of copulations per pair was detected. However, this value decreased with time, and reciprocal copulations were significantly less numerous between than within populations. (iii) Individuals that successfully copulated as female in (ii) were isolated, and their fecundity was followed over 3 weeks. The fecundity of individuals that had been maintained isolated over the whole experiment served as a control. No significant difference was on the whole detected. Our results collectively suggest that the individuals from the two populations studied are rather selfers, confirming that B. forskalii is a preferentially selfing species. There is also some pre-zygotic isolation against crosses between populations.

Morphogenesis of Bipartition in two species of the genus Ptychostomum (Ciliophora, Hysterocinetidae).

Two new species of Hysterocinetidae ciliates living parasitically in the alimentary tract of Alma nilotica are described along with the process of stomatogenesis in these species. The buccal primordium of the opisthe originates from the multiplication of kinetosomes of some somatic kineties forming a disorganized field. One row of kinetosomes, a second and a third are successively isolated from this field. As soon as this stage with three rows of kinetosomes is reached, the peristomial infraciliature of the old buccal infraciliature progressively migrates to the equator of the cell. There, it gives rise to the buccal infraciliature of the proter through resorption and reorganization of the old buccal infraciliature. The description of this morphological development is related to both nuclear division stages and the organization of the sucker cytoskeleton of the ciliates. Our findings contradict the well admitted idea concerning the autonomous characteristic of the stomatogenesis within the Hysterocinetidae. We propose that these ciliates should be considered as a separate group within the Oligohymenophora.

Toxic effects of some major polyaromatic hydrocarbons found in crude oil and aquatic sediments on Scenedesmus subspicatus.

The green alga, Scenedesmus subspicatus was exposed for 7 days to a series of PAHs (polyaromatic hydrocarbons) of increased molecular weight from two to five rings [naphthalene (Nap), anthracene (Ant), phenanthrene (Phe), pyrene (Pyr) and benzo(a)pyrene (BaP)]. The toxicity measured as population growth inhibition by individual PAH to the S. subspicatus followed the order: BaP>Pyr>Ant>Phe>Nap. These results confirmed that the toxicity potential of PAHs seems to be strongly influenced by their physico-chemical properties (aqueous solubility, K(ow), coefficient of volatilization, etc.) and the conditions of algae culture (light, presence of nitrate ions, etc.). Consequently, Nap, Phe and Ant having low k(ow) values and low coefficient of volatilization values were less toxic than BaP with the highest k(ow) value, indicating for example why Nap with the lowest EC(50) value was nearly 2 x 10(5) times lower than that of BaP. Moreover, nitrate ions seemed to act directly on the degree of hydroxylated radical reactivity of PAHs, since BaP always remained the most toxic of the compounds tested. The results were also agreed with the QSAR model for toxicity prediction of PAHs to many aquatic organisms.

[Hygiene and microbiologic quality of ice creams produced in Cameroon. A public health problem]. / Hygiène et qualité microbiologique des crèmes glacées produites au Cameroun. Un problème de santé publique.

Three hundred samples of ice cream produced in two main towns in Cameroon (Douala and Yaoundé) are evaluated for their hygienic quality. The microbiologic examinations show that many of them are contaminated with bacteria of faecal origin, pathogenic Staphylococcus and Salmonella respectively in 71.3, 49.6 and 5% of the products examinations in the study. The use of non potable water and the disrespect of the hygienic rules during the production are the main causes of contamination. Recommendations are made to preserve the public health in the developing country.

[Experimental study of the compatibility between Schistosoma haematobium and two species of Bulinus in Cameroon]. / Etude expérimentale de la compatibilité entre Schistosoma haematobium et deux espèces de bulins au Cameroun.

A study on the compatibility between Schistosoma haematobium from three remote localities (Mourtourwa, Gounougou and Kékem) and four populations of Bulinus truncatus (Gounougou, Ngaoundéré, Bertoua and Kékem) and four populations of B. globosus (Mourtourwa, Ouroudoukoudje, Bafia and Yaoundé) was undertaken in order to estimate the risk of extension of urinary schistosomiasis in Cameroon. First generation of offspring from wild Bulinus was exposed to miracidia liberated by schistosome eggs extracted from patient urine. Between the 25th and the 60th day post-infestation the number of snails still alive, the number emitting cercariae and the prepatent period duration were noted. Results showed that all B. truncatus samples were susceptible to the three strains of parasite whereas only B. globosus of Mourtourwa and Ouroudoukoudje were susceptible to S. haematobium from Mourtourwa. The schistosome infection rate was then significantly higher in B. truncatus and the prepatent period significantly lower than in B. globosus. The compatibility characterised by a high infection rate and a low prepatent period was significantly better in homopatric couples than in allopatric combinations. The results suggested that B. truncatus might be potentially more implicated than B. globosus to the extension of the urinary bilharziasis in Cameroon.

[Evaluation of bacterial and polluting loads of effluent from activated sludge wastewater treatment plants in Yaounde, Cameroon]. / Evaluation de la charge polluante et de la charge bactérienne des rejets des stations d'épuration a boues activées Yaoundé (Cameroun).

Activated sludge wastewater treatment plants in Yaounde are plagued by failures in their operation. A microbiological and physico-chemical study was carried out on wastewater effluent of an hospital, university campus, and two residential areas made up of more than 2,000 apartments connected to two collective sewase systems. Wastewater sampling was performed in the form of daily cycles during which composite samples were made every two hours with samples collected every half hour. Variables measured included incoming flow rates, suspended solids, pH, electrical conductivity, ammonia nitrogen, total Kjeldahl nitrogen (combination of ammonia and organic nitrogen) and total reactive phosphorus, BOD5 and COD. Microorganisms were isolated on selective culture media using membrane filter procedures. The results obtained show that effluent discharged in receiving waters are, in general, not treated. The suspended solids contents of raw sewage range from 120 to 2,600 mg/l. In General Hospital a mean value of suspended solids of 31.8 mg/l was obtained for treated wastewater. pH values of both raw and treated sewage range from 6 to 8.5 units. Mean values of the conductivity of treated and raw wastewater range from 449.7 to 1,038.3 microseconds/cm. The TKN contents of untreated wastewater are comprised between 36 and 259.2 mg/l. These values range from 5 to 20 mg/l for treated effluent in the General Hospital plant. Large amounts of total hydrolysable phosphorus varying from 3.8 to 27.9 mg/l are present in wastewater from the plants. Influent BOD5 of Grand Messa, Cite Verte and university campus wastewater range from 110 to 530 mgO2/l. Values recorded indicate an important input of organic matter to receiving waters. In the case of biotic components, test germs densities registered in the influent of Grand Messa and those of Cite Verte are comprised between 9 x 106 and 88 x 107 CFU/100 ml for fecal coliforms, and between 9 x 105 and 74 x 107 CFU/100 ml for fecal streptococci. The densities of Aeromonas hydrophila in Grand Messa and Cite Verte influent fluctuate between 30 x 106 and 65 x 108 CFU/100 ml. Influent of university campus harbour 10 x 105. Densities of Pseudomonas aeruginosa remain below 70 x 105 CFU/100 ml. Treated effluent BOD5 values range from 48 to 150 mgO2/l. Such values indicate an inefficient treatment. The decrease in bacterial densities is limited to 2 logarithmic units. Treated effluent still harbour 10 x 103 to 10 x 105 CFU/100 ml for fecal coliforms, and 20 x 102 to 12 x 104 CFU/100 ml for fecal streptococci. These values range from 10 x 104 to 50 x 105 CFU/100 ml for Aeromonas hydrophila. Pseudomonas aeruginosa densities remain in general below 40 x 104 CFU/100 ml. Discharges from these wastewater treatment plants are responsible for the permanent fecal contamination of the environment and a lack of proper hygiene. Furthermore, they produce overfertilization of receiving waters.

[Bacteriological quality of spring and well water in Yaounde (Cameroon)]. / Qualité bactériologique des eaux des sources et des puits de Yaoundé (Cameroun).

The drinking water supply in Yaounde, Cameroon, often fails to meet demand and this leads many people to seek water from springs and wells the microbiological quality of which is unknown. We carried out a microbiological survey of five spring water points and ten wells. The fifteen study sites were located in various residential districts and were selected according to geographical location and number of users. Microorganisms were isolated on specific culture media using the membrane filter technique. We found that these water supplies contained many types of bacteria including Pseudomonas aeruginosa, Aeromonas hydrophila and indicators of fecal contamination. Maximum annual concentrations differed at the various sites and over time, from 750 to 98,000 CFU/l for P. aeruginosa, 40 to 78,000 CFU/l for A. hydrophila and 300 to 58,000 CFU/l for fecal indicator bacteria. The density of fecal bacteria was strongly correlated (p < 0.01) with those of P. aeruginosa in spring water and A. hydrophila in well water. The degree of correlation between microbial concentrations differed between sites. The source of fecal contamination differed between sites and was animal, human or both. The densities of bacteria recorded were high. Therefore, consumers of water drawn from these sources are at short-term risk of contracting waterborne diseases.

Microbiological water quality of the Mfoundi River watershed at Yaoundé, Cameroon, as inferred from indicator bacteria of fecal contamination.

Using the membrane filtration technique to count total coliform (TC), fecal coliform (FC) and fecal streptococci (FS), the microbiological water quality of the Mfoundi River and four of its representative tributaries at Yaoundé, Cameroon, was assessed for human use and contact. Sampling was conducted so as to examine the potential origin of fecal contamination and how rainfall affects the measured concentrations of indicators organisms. Our results revealed that waters were not safe for human use or primary contact according to the standards for water quality established by the Word Health Organization (WHO). Indeed, these waters exhibited high concentrations of TC (Mean +/- SD = 5.6 x 10(8) +/- 2.5 x 10(6) CFU/100 ml), FC (Mean +/- SD = 6.8 x 10(5) +/- 2.4 x 10(3) CFU/100 ml) and FS (Mean +/- SD = 7.3 x 10(5) +/- 2.1 x 10(3) CFU/100 ml) that varied with the sampling sites and points. FC/FS ratio suggested that this contamination was more from warm-blooded animals than humans and correlation analysis points to the role of rainfall as a contributing factor, which enhanced the bacterial numbers detected. We conclude that there is a great potential risk of infection for users of waters from the Mfoundi River and its tributaries at Yaoundé.

Modeling growth for predicting the contamination level of guava nectar by Candida pelliculosa under different conditions of pH and storage temperature.

The combined effects of temperature (2-46 degrees C) and pH (1.55-6.25) on the growth of Candida pelliculosa isolated from guava nectar produced in Cameroon were studied using a turbidity method, ie measurement of optical density at 630 nm. A quadratic polynomial model was constructed to predict the effects and interactions of these two environmental conditions on the maximal optical density obtained (i2 = 0.97). The relation between optical density and population density of C. pelliculosa (CFU ml-1) was also established using an exponential regression (2 = 0.99). According to the model, maximal growth conditions were 37 degrees C and pH 6.25 for obtaining the maximal optical density of 1.25 corresponding to about 60 x 10(6) CFU ml-1. A good agreement of the model was found between the predicted values and the observed values of maximal optical density. The model was validated by the experimental values of maximal optical density obtained in the growth of C. pelliculosa in commercial guava nectar (pH 3.15).