Mapping of schistosomiasis and soil-transmitted helminthiasis in the regions of Littoral, North-West, South and South-West Cameroon and recommendations for treatment.

BACKGROUND: The previous nationwide mapping of schistosomiasis and soil-transmitted helminthiasis (STH) in Cameroon was conducted 25 years ago. Based on its results, mass drug administration (MDA) of praziquantel was limited to the three northern regions and few health districts in the southern part of Cameroon. In 2010, we started the process of updating the disease distribution in order to improve the control strategies. Three of the ten regions of Cameroon were mapped in 2010 and the data were published. In 2011, surveys were conducted in four additional regions, i.e. Littoral, North-West, South and South-West. METHODS: Parasitological surveys were conducted in March 2011 in selected schools in all 65 health districts of the four targeted regions, using appropriate research methodologies, i.e. Kato-Katz and urine filtration. RESULTS: The results showed significant variation of schistosomiasis and STH prevalence between schools, villages, districts and regions. Schistosoma haematobium was the most prevalent schistosome species, with an overall prevalence of 3.2%, followed by S. mansoni (3%) and S. guineensis (1.2%). The overall prevalence of schistosomiasis across the four regions was 7.4% (95% CI: 6.7-8.3%). The prevalence for Ascaris lumbricoides was 19.5% (95% CI: 18.3-20.7%), Trichuris trichiura 18.9% (95% CI: 17.7-20.1%) and hookworms 7.6% (95% CI: 6.8-8.4%), with an overall STH prevalence of 32.5% (95% CI: 31.1-34.0%) across the four regions. STH was more prevalent in the South region (52.8%; 95% CI: 48.0-57.3%), followed by the South-West (46.2%; 95% CI: 43.2-49.3%), the North-West (35.9%; 95% CI: 33.1-38.7%) and the Littoral (13.0%; 95% CI: 11.3-14.9%) regions. CONCLUSIONS: In comparison to previous data in 1985-87, the results showed an increase of schistosomiasis transmission in several health districts, whereas there was a significant decline of STH infections. Based on the prevalence data, the continuation of annual or bi-annual MDA for STH is recommended, as well as an extension of praziquantel in identified moderate and high risk communities for schistosomiasis.

Four new species of Hysterocinetida (Protozoa: Ciliophora) from the digestive tract of earthworms collected to the lower Nyong estuary (South Coast, Cameroon).

Hysterocinetida are a large assemblage of ciliates characterized by sucker and buccal apparatus. They have a wide distribution in the digestive tract of oligochaete annelids of the genus Alma that inhabit hydromorphic parts of non-anthropic hydrosystems. On the basis of morphological, morphometric and infraciliature characteristics revealed by ammoniacal silver carbonate and 4',6-Diamidino-2-Phenyl Indole (DAPI) staining techniques, four forms were identified as new members belonging to three genera: Preptychostomum, Proptychostomum and Thurstonia. Preptychostomum donendaensis n. sp. is ovoid with a slightly narrowed anterior part and is commensal to the digestive tract of Alma emini and A. nilotica. There are 20-23 kineties on the lower side and 42-45 on the upper side. Proptychostomum gigas n. sp. is pear-shaped, with a narrowed anterior part and a wide posterior part (125-138 µm × 80-85 µm). There are 40 to 44 kineties, equally distributed on both sides of the ciliate. Its sucker is circular and measures 31-34 µm in diameter. The peristome follows the contour of the cell and the infundibulum has a turn of spire. Thurstonia emini n. sp. is identified in the digestive tract of A. emini. The cell is elongate, with the anterior pole narrower than the truncate posterior pole (90-115 µm × 40-55 µm). The kineties are equally distributed on both sides of the ciliate. The sucker is in the shape of an inverted U. Thurtonia nilotica n. sp. is commensal of the digestive tract of A. nilotica. The cell is pear-shaped, with the anterior part narrowed and the posterior part swollen (110-114 µm × 85-92 µm). 46 to 50 kineties are unevenly distributed on both sides of the ciliate. The sucker is shaped like an inverted lunar crescent.

Population genetic structure of the malaria vector Anopheles nili in sub-Saharan Africa.

BACKGROUND: Anopheles nili is a widespread efficient vector of human malaria parasites in the humid savannas and forested areas of sub-Saharan Africa. Understanding An. nili population structure and gene flow patterns could be useful for the development of locally-adapted vector control measures. METHODS: Polymorphism at eleven recently developed microsatelitte markers, and sequence variation in four genes within the 28s rDNA subunit (ITS2 and D3) and mtDNA (COII and ND4) were assessed to explore the level of genetic variability and differentiation among nine populations of An. nili from Senegal, Ivory Coast, Burkina Faso, Nigeria, Cameroon and the Democratic Republic of Congo (DRC). RESULTS: All microsatellite loci successfully amplified in all populations, showing high and very similar levels of genetic diversity in populations from West Africa and Cameroon (mean Rs = 8.10-8.88, mean He = 0.805-0.849) and much lower diversity in the Kenge population from DRC (mean Rs = 5.43, mean He = 0.594). Bayesian clustering analysis of microsatellite allelic frequencies revealed two main genetic clusters in the dataset. The first one included only the Kenge population and the second grouped together all other populations. High Fst estimates based on microsatellites (Fst > 0.118, P < 0.001) were observed in all comparisons between Kenge and all other populations. By contrast, low Fst estimates (Fst < 0.022, P < 0.05) were observed between populations within the second cluster. The correlation between genetic and geographic distances was weak and possibly obscured by demographic instability. Sequence variation in mtDNA genes matched these results, whereas low polymorphism in rDNA genes prevented detection of any population substructure at this geographical scale. CONCLUSION: Overall, high genetic homogeneity of the An. nili gene pool was found across its distribution range in West and Central Africa, although demographic events probably resulted in a higher level of genetic isolation in the marginal population of Kenge (DRC). The role of the equatorial forest block as a barrier to gene flow and the implication of such findings for vector control are discussed.

Morphology and infraciliature of two new earthworm ciliates, Hoplitophrya polymorphus sp. nov. and Anoplophrya simplex sp. nov. (Ciliophora: Oligohymenophorea: Astomatia).

Morphological and infraciliature studies carried out using pyridinated ammoniacal silver carbonate and the 4',6-diamidino-2-phenylindole (DAPI) staining techniques, led to the identification of two new species of ciliates pertaining to the subclass Astomatia. The first species, Hoplitophrya polymorphus sp. nov., displays two main cellular forms: the elongated form (150-247 µm long and 40-87 µm width) and the stocky form (140-170 µm long and 70-98 µm width). The macronucleus is generally skinny in the elongated forms and ribbon-shaped in the stocky forms. The common feature of the two cellular shapes uniting them in the same species is the identical structure of their skeletal apparatus, a V-shaped element located in an apical depression and bearing skeletal fibres on its ventral face. The second species, Anoplophrya simplex sp. nov., is totally deprived of skeletal apparatus. The cell shape is fusiform and dorsoventrally flattened (105-180 µm long and 65-125 µm width). Six to 12 pulsatile vacuoles form two rows arranged symmetrically on either side of the macronucleus. These two newly identified species highlight the extremely rich diversity of ciliates inhabiting the digestive tract of tropical earthworms.

Population genetic structure of the malaria vector Anopheles moucheti in south Cameroon forest region.

We used recently developed microsatellite DNA markers to explore the population genetic structure of the malaria vector, Anopheles moucheti. Polymorphism at 10 loci was examined to assess level of genetic differentiation between four A. moucheti populations from South Cameroon situated 65-400 km apart. All microsatellite loci were highly polymorphic with a number of distinct alleles per locus ranging from 9 to 17. Fst estimates ranging from 0.0094 to 0.0275 (P < 0.001) were recorded. These results suggest a very low level of genetic differentiation between A. moucheti populations. The recently available microsatellite loci revealed useful markers to assess genetic differentiation between geographical populations of A. moucheti in Cameroon.

Malaria vectors and urbanization in the equatorial forest region of south Cameroon.

Entomological surveys were carried out in the town of Mbalmayo and in the nearby rural village of Olama, within the equatorial forest zone of Cameroon. Mosquitoes were captured when landing on human volunteers and by pyrethrum spray catches. Malaria vectors captured were Anopheles gambiae Giles (M and S forms) and A. moucheti Evans in both areas, together with A. funestus Giles in Mbalmayo. One A. marshallii (Theobald) specimen infected by Plasmodium falciparum was found in Olama. Anopheles moucheti was the most abundant anopheline species caught in Olama, while A. gambiae was the most abundant in Mbalmayo. All these vectors were highly anthropophilic as indicated by the fact that only 5 of 201 blood meals analysed had been taken from non-human hosts. Plasmodium falciparum was the only malaria parasite species found in Mbalmayo, while P. malariae was also found in Olama. The annual entomological inoculation rate was estimated at 129 infective bites/person/year in Mbalmayo and 322 in Olama. Comparison with data published in 1955 from Mbalmayo, before expansion of the town, showed the impact of urbanization on the composition of the vector system and malaria transmission dynamics. Such changes should be considered when implementing sustainable control measures.

Antimicrobial resistance of Gram-negative bacilli isolates from inpatients and outpatients at Yaounde Central Hospital, Cameroon.

OBJECTIVE: To determine and compare antimicrobial susceptibility patterns of pathogenic bacteria from inpatients and outpatients at a university teaching hospital in Yaounde, Cameroon. METHODS: Gram-negative bacilli isolates (n = 522), obtained from a wide range of clinical specimens (urine, pus and blood) from inpatients and outpatients at Yaounde Central Hospital between March 1995 and April 1998, were evaluated for resistance to antibiotics (amoxicillin, amoxicillin/clavulanate, piperacillin, cefazolin, cefoxitin, cefotaxime, ceftazidime, aztreonam, imipenem, gentamicin, tobramicin, ofloxacin and trimethoprim/sulfamethoxazole). RESULTS: Of the 522 isolates recorded, 80.3% were Enterobacteriaceae. A high incidence of resistance to amoxicillin (85%), piperacillin (75%) and trimethoprim/sulfamethoxazole (71%) was observed. The proportion of antimicrobial-resistant isolates from inpatients was significantly higher than that from outpatients (P < 0.05), except for piperacillin, tobramicin and trimethoprim/sulfamethoxazole. The combinations of antimicrobial and organism showed that the percentage of ceftazidime-resistant Pseudomonas aeruginosa and ceftazidime-resistant Enterobacter cloacae were 26.8% and 24% respectively. The rate of antimicrobial resistance in isolates from inpatients was not significantly higher than that in isolates from outpatients for all the antimicrobial/organism combinations, except for ceftazidime-resistant Escherichia coli, which was exclusively found in isolates from inpatients. Among Enterobacteriaceae, high and low level penicillinase (mostly in E. coli (13.6% and 11% respectively) and Klebsiella spp. (9% and 8% respectively) were the most important beta-lactam resistance phenotypes (31.2% and 23.6%, respectively). Wild type (exclusively observed in E. coli, Proteus mirabilis and Salmonella spp.) and low level penicillinase were higher in outpatient than inpatient isolates (wild type--17.9% vs 10.8% and low level penicillinase--29.4% vs 20.5%, respectively; P < 0.05). However, extended spectrum beta-lactamase strains (Klebsiella spp. (3.5%), E. coli (2.6%), Citrobacter spp. (0.7%), Enterobacter spp. (0.4%) and P. mirabilis (0.2%)) were exclusively recovered from inpatients. Penicillinase and high level cephalosporinase resistance phenotypes were frequently observed in non-fermenter Gram-negative bacilli (46.6% and 29.1% respectively). However, there were no significant differences in penicillinase and cephalosporinase resistance between inpatient and outpatient isolates. CONCLUSION: As the incidence of antimicrobial resistance is substantially higher in isolates from inpatient than outpatient pathogens, more resources should be allocated within the hospital to encourage good antibiotic practices and good hospital hygiene.

Mapping of schistosomiasis and soil-transmitted helminthiasis in the regions of centre, East and West Cameroon.

BACKGROUND: Schistosomiasis and soil-transmitted helminthiasis (STH) are widely distributed in Cameroon. Although mass drug administration (MDA) of mebendazole is implemented nationwide, treatment with praziquantel was so far limited to the three northern regions and few health districts in the southern part of Cameroon, based on previous mapping conducted 25 years ago. To update the disease distribution map and determine where treatment with praziquantel should be extended, mapping surveys were conducted in three of the seven southern regions of Cameroon, i.e. Centre, East and West. METHODOLOGY: Parasitological surveys were conducted in April-May 2010 in selected schools in all 63 health districts of the three targeted regions, using appropriate research methodologies, i.e. Kato-Katz and urine filtration. PRINCIPAL FINDINGS: The results showed significant variation of schistosomiasis and STH prevalence between schools, villages, districts and regions. Schistosoma mansoni was the most prevalent schistosome species, with an overall prevalence of 5.53%, followed by S. haematobium (1.72%) and S. guineensis (0.14%). The overall prevalence of schistosomiasis across the three regions was 7.31% (95% CI: 6.86-7.77%). The prevalence for Ascaris lumbricoides was 11.48 (95% CI: 10.93-12.04%), Trichuris trichiura 18.22% (95% CI: 17.56-18.90%) and hookworms 1.55% (95% CI: 1.35-1.78%), with an overall STH prevalence of 24.10% (95% CI: 23.36-24.85%) across the three regions. STH was more prevalent in the East region (46.57%; 95% CI: 44.41-48.75%) in comparison to the Centre (25.12; 95% CI: 24.10-26.17%) and West (10.49%; 95% CI: 9.57-11.51%) regions. CONCLUSIONS/SIGNIFICANCE: In comparison to previous data, the results showed an increase of schistosomiasis transmission in several health districts, whereas there was a significant decline of STH infections. Based on the prevalence data, the continuation of annual or bi-annual MDA for STH is recommended, as well as an extension of praziquantel in identified moderate and high risk communities for schistosomiasis.

Genetic diversity and phylogenetic position of the subclass Astomatia (Ciliophora) based on a sampling of six genera from West African oligochaetes (Glossoscolecidae, Megascolecidae), including description of the new genus Paraclausilocola n. gen.

To more confidently assess phylogenetic relationships among astome ciliates, we obtained small subunit (SSU) rRNA sequences from nine species distributed in six genera and three families: Almophrya bivacuolata, Eudrilophrya complanata, Metaracoelophrya sp. 1, Metaracoelophrya sp. 2, Metaracoelophrya intermedia, Metaradiophrya sp., Njinella prolifera, Paraclausilocola constricta n. gen., n. sp., and Paraclausilocola elongata n. sp. The two new species in the proposed new clausilocolid genus Paraclausilocola n. gen. are astomes with no attachment apparatus, two files of contractile vacuoles, and an arc-like anterior suture that has differentiations of thigmotactic ciliature on the anterior ends of the left kineties of the upper surface. Phylogenetic analyses were undertaken using neighbor-joining, Bayesian inference, maximum likelihood, and maximum parsimony. The nine species of astomes formed a strongly supported clade, showing the subclass Astomatia to be monophyletic and a weakly supported sister clade to the scuticociliates. There were two strongly supported clades within the astomes. However, genera assigned to the same family were found in different clades, and genera assigned to the same order were found in both clades. Thus, astome taxa appear to be paraphyletic when morphology is used to assign species to genera.

The in-vitro antimicrobial activity of some medicinal plants against beta-lactam-resistant bacteria.

BACKGROUND: In effort to identify novel bacterial agents, this study was initiated to evaluate the antimicrobial properties of 17 crude extracts from 12 medicinal plants against beta-lactam-resistant bacteria. METHODOLOGY: The antimicrobial activities of plant extracts were evaluated against clinically proved beta-lactam-resistant bacteria (Klebsiella pneumoniae, Klebsiella oxytoca, Enterobacter cloacae, Serratia marcescens, Acinetobacter baumannii, Staphylococcus aureus and Enterococcus sp.) and reference strains of bacteria (Escherichia coli ATCC 35218, Enterobacter aerogenes ATCC 29751, E. aerogenes ATCC 13048, Pseudomonas aeruginosa ATCC 27853 and Enterococcus hirae ATCC 9790) by using disc-diffusion and agar-dilution assays. RESULTS: The crude plant extracts demonstrated broad spectrum activity against all bacteria tested with inhibition zones in the range of 8-30 mm. The minimal inhibitory concentration (MIC) values of different plant extracts against the tested bacteria were found to range from or= 10 mg ml(-1). The most active plant extracts were from Dortenia picta and Bridelia micrantha (MIC: 1.25-10 mg ml(-1)) on beta-lactam-resistant Gram-negative bacilli and the extracts from B. micrantha, Mallotus oppositifolius, Garcinia lucida, Garcinia. kola, Campylospermum densiflorum (leaves) and C. zenkeri (root) on beta-lactam-resistant Gram-positive cocci (MIC:

Screening of some medicinal plants from cameroon for beta-lactamase inhibitory activity.

In efforts to find new bioactive beta-lactamase inhibitors, this study investigated 16 Cameroonian plants belonging to 10 families which were evaluated for anti-beta-lactamase activity. The investigation showed that extracts 2, 6, 3 and 5 of the 16 plants investigated presented interesting in vitro beta-lactamase inhibition (over 90%), respectively, of the beta-lactamases TEM-1, OXA-10, IMP-1 and P99. These extracts were from Mammea africana (all beta-lactamases), Garcinia lucida, G. kola (OXA-10, IMP-1 and P99), Bridelia micrantha (OXA-10, P99), Ochna afzelii (OXA-10, P99), Prunus africana (IMP-1) and Adenia lobata (TEM-1). After elimination of tannins (according to the European Pharmacopoeia) the extracts from B. micrantha, G. lucida and M. africana were tested further for their anti-beta-lactamase activity. The extracts from B. micrantha and G. lucida exhibited potent inhibitory activity, respectively, of beta-lactamase OXA-10 (IC(50) = 0.02 mg/mL) and P99 (IC(50) = 0.01 mg/mL). The anti-beta-lactamase activity of M. africana extract was weak. The isolation and the structural elucidation of the active constituents of G. lucida and B. micrantha will provide useful leads in the development of beta-lactamase inhibitors.

Antimicrobial activity against gram negative bacilli from Yaounde Central Hospital, Cameroon.

BACKGROUND: Antimicrobial resistance among bacteria pathogens is a world-wide issue. The antimicrobial susceptibility patterns of common pathogenic bacteria are essential to guide empirical and pathogen-specific therapy; unfortunately, these data are scarse in Cameroon. OBJECTIVE: To determine the antimicrobial susceptibility patterns of Gram-negative bacilli isolated in Yaounde Central Hospital Laboratory of Bacteriology. METHODS: Gram-negative bacilli isolates (n = 505), obtained from a wide range of clinical specimens (urine, pus and blood) in Yaoundé Central Hospital Laboratory of Bacteriology between March 1995 and April 1998, were evaluated for resistance to antibiotics (amoxicillin, amoxicillin/clavulanate, piperacillin, cefazolin, cefoxitin, cefotaxime, ceftazidime, aztreonam, imipenem, gentamicin, tobramicin, ofloxacin and trimethoprim/sulfamethoxazole) by the Kirby-Bauer disk diffusion method. RESULTS: High rates of resistance were found in most of the bacteria studied. Resistance to all isolates was mostly observed for amoxicillin (87%), piperacillin (74%) and trimethoprim/sulfamethoxazole (73%). Susceptibilities to third generation cephalosporins (cefotaxime, ceftazidime) and monobactame (aztreonam) were = 91% for Escherichia coli, = 71% for Klebsiella spp., = 98% for Proteus mirabilis, = 50% for Enterobacter spp. and Citrobacter spp. Pseudomonas aeruginosa was less susceptible to cefotaxime (2%) and aztreonam (33%), and highly susceptible to ceftazidime (72%) whereas Acinetobacter baumannii was highly resistant to aztreonam (100%), to cefotaxime (96%) and cetazidime (62%). Imipenem (98%) was the most active antibiotic followed by the ofloxacine (88%). Susceptibility of all isolates to gentamicin was 67%. CONCLUSION: These results indicate that surveillance to antimicrobial resistance in Cameroon is necessary to monitor microbial trends, antimicrobial resistance pattern, and provide information for choosing empirical or direct therapy to physicians.

Extended-spectrum-beta-lactamase-producing Enterobacteriaceae in Yaounde, Cameroon.

Organisms producing extended-spectrum beta-lactamases (ESBLs) have been reported in many countries, but there is no information on the prevalence of ESBL-producing members of the family Enterobacteriaceae in Cameroon. A total of 259 Enterobacteriaceae strains were isolated between 1995 and 1998 from patients at the Yaounde Central Hospital in Cameroon. Enterobacterial isolates resistant to extended-spectrum cephalosporin and monobactam were screened for ESBL production by the double-disk (DD) synergy test. Thirty-one (12%) of these Enterobacteriaceae strains were shown to be positive by the DD synergy test, suggesting the presence of ESBLs. Resistance to oxyimino-cephalosporins and monobactams of 12 (38.7%) of the 31 strains-i.e., 6 Klebsiella pneumoniae, 4 Escherichia coli, 1 Citrobacter freundii, and 1 Enterobacter cloacae strain-was transferred to E. coli HK-225 by conjugation. Resistance to gentamicin, gentamicin plus trimethoprim-sulfamethoxazole, or trimethoprim-sulfamethoxazole was cotransferred into 6, 2, and 1 of these transconjugants, respectively. All 12 transconjugants were resistant to amoxicillin, piperacillin, all of the cephalosporins, and aztreonam but remained susceptible to cefoxitin and imipenem. Crude extracts of beta-lactamase-producing transconjugants were able to reduce the diameters of inhibition zones around disks containing penicillins, narrow- to expanded-spectrum cephalosporins or monobactams when tested against a fully susceptible E. coli strain but had no effect on such zones around cefoxitin, imipenem, and amoxicillin-clavulanate disks. The beta-lactamases produced by the 12 tranconjugants turned out to be SHV-12 by DNA sequencing. Therefore, the ESBL SHV-12 is described for the first time in Cameroon.