Residual malaria transmission: Magnitude and drivers of persistent Plasmodium infections despite high coverage of control interventions in Burkina Faso, West Africa.

This study collected baseline data on malaria vectors to characterize the drivers and the factors of persistent malaria transmission in two villages in the western part of Burkina Faso. Mosquitoes were collected in each village using the Human landing catch and pyrethrum spray catch and identified using the morphological keys. Molecular analyses were performed for the identification of An. gambiae complex species, the detection of Plasmodium infection and kdr-995F mutation. Anopheles mosquito larvae were also collected in the same villages, reared to adult's stage for the WHO tube and cone tests performing. The physical integrity of the LLINs already used by people in each village was assessed using the proportional hole index (pHI). An. gambiae s.l. was the main malaria vector accounting for 79.82% (5560/6965) of all collected mosquitoes. The biting pattern of An. gambiae s.l. was almost constant during the survey with an early aggressiveness before 8 p.m. and later biting activity after 6 a.m. The EIR varied from 0.13 to 2.55 infected bites per human per night (average: 1.03 infected bites per human per night). An. gambiae s.l. populations were full susceptible to Chlorpyrifos-methyl (0.4%) and Malathion (5%) with high kdr-995F mutation frequencies (>0.8). The physical integrity assessment showed high proportion of good nets in Santidougou compared to those collected in Kimidougou. This study highlighted a persistence of malaria transmission despite the intense use of vector control tools as LLINs and IRS by correlating mosquito biting time and human behavior. It provided a baseline guide for the monitoring of the residual malaria transmission in sub-Saharan Africa and encouraging the development of new alternative strategies to support the current malaria control tools.

Epidemiological evaluation of onchocerciasis along Ogun River System, southwest Nigeria.

BACKGROUND & OBJECTIVE: Epidemiological studies were carried out to assess the prevalence and community microfilarial load (CMFL) of onchocerciasis after repeated annual treatment with ivermectin along Ogun river System, southwest Nigeria. METHOD: Skin snips were taken from consented participants in 11 selected communities along the River system. The microfilarial load of the community was estimated. RESULTS: The prevalence and CMFL varied significantly in the communities (p <0.05). The prevalence of onchocerciasis ranged from 19.1 to 45.6%, while the CMFL ranged from 0.11 to 1.03 microfilariae per skin snip. The CMFL recorded was <5 microfilariae per skin snip, i.e. recognized by WHO as threshold value in certifying the communities to be free of onchocerciasis as public health problem, thus, signifying the possibility of onchocerciasis elimination in the study area. CONCLUSION: Efforts should therefore be intensified to achieve improved ivermectin coverage and compliance in annual ivermectin treatment in order to completely eliminate onchocerciasis as a public health problem in the studied communities.

[Effectiveness of a polymerase chain reaction using seminested and multiplex strategy for the identification of the three main bacteria responsible for meningitis in Burkina Faso]. / Efficacité d'une technique de réaction de polymérisation en chaîne (seminested et multiplex) pour l'identification des trois principales espèces bactériennes responsables de méningites au Burkina Faso.

A prospective study (from August 2006 to April 2007) was carried out with 214 cerebrospinal fluid samples with suspicion of bacterial meningitis. The aim of the study was to assess the effectiveness of the simultaneous detection of Neisseria meningitidis, Streptococcus sp. and Haemophilus influenzae using seminested polymerase chain reaction strategy. Among the 214 samples tested by both PCR and culture, the overall confirmation rate was 64% for PCR and 40.1% for culture (P = 2 x 10⁻6). Taking culture method as the standard reference, the overall sensitivity of PCR was 98.8% and specificity, 59.4%. The sensitivity of PCR was 100, 97.3 and 100% respectively for N. meningitidis, Streptococcus sp. and H. influenzae with respective specificities of 70, 93.2 and 97.2%. In conclusion, the seminested PCR strategy is a sensitive method and it can be implemented in the reference public health laboratories for an exhaustive microbiological surveillance of bacterial meningitis.

[Main characteristics of the street food sector in Bobo-Dioulasso, Burkina Faso]. / Caractéistiques de l'alimentation de rue dans la ville de Bobo-Dioulasso, Burkina Faso.

To investigate the sector of food sold in the streets of Bobo-Dioulasso and identify relevant information for action, a survey on knowledge and practices of street food vendors and consumers was conducted in June 2005. Data have been collected in 928 street food selling posts. Structured questionnaires were used to collect data from 874 street vendors and 2474 consumers. Street food sites are concentrated in places where administration and trade activities are usually running. The street food seller is a married and illiterate woman of 32 years old. Cereals (48.5%), meat (33.9%), milk (9.6%) and fruits (4.4%) are the basic consumables. The street food consumer is a non married man, 27 years old working in profit-making activity. Consumers use many criteria to choose the place to eat, at times or permanently. The street food sector represents a source of income and induces change in household eating habits. Street food in Bobo-Dioulasso needs to be better organised, by using an holistic approach that involves all the actors.

Year to year and seasonal variations in vector bionomics and malaria transmission in a humid savannah village in west Burkina Faso.

A longitudinal entomological study was carried out from 1999 to 2001 in Lena, a humid savannah village in the western region of Burkina Faso in order to establish malaria vector bionomics and the dynamics of malaria transmission. In the first year, malaria transmission was mainly due to An. gambiae s.s., but during the two later years was due to An. funestus, which were observed in high frequency towards the end of the rainy season. PCR identification of samples of An. gambiae s.1. showed 93% to be An. gambiae s.s. and 7% An. arabiensis. An. funestus constituting more than 60% of the vectors were identified in PCR as An. funestus s.s. The persistence of intense vectorial activity in this village was probably due to the road building in a swampy area creating a semi-permanent swamp that provided large sites for larval mosquitoes. These swampy sites seemed to be more favorable for An. funestus than for An. gambiae s.s. Thus, land development must be monitored and subjected to planning to minimize vector proliferation. Such a system of planning could lead to the restriction or even elimination of the swamp that is the source of larvae developing in the heart of the village.

[Influence of social perceptions and practices on the use of bednets in the malaria control programme in Ivory Coast (West Africa)]. / Representations sociales et pratiques liées à l'utilisation des moustiquaires dans la lutte contre le paludisme en Côte d'Ivoire (Afrique de l'Ouest).

The National Malaria Programme in Ivory Coast has encountered difficulty in winning public acceptance of insecticide-treated bednets. We speculate that resistance to the use of bednets could be rooted in social perceptions, beliefs and practices in the communities. The purpose of this study was to identify sociocultural and environmental factors that could be used to support promotion strategies and acceptance of impregnated bednets in Ivory Coast. Survey findings confirmed that bednets were not in widespread use among the population (25%). The most widely used methods were burning mosquito coils (50%) and indoor spraying (31%). Use of impregnated bednets was low (6%). Most survey respondents (73%) indicated initial appreciation for the effectiveness of bednets in protecting against mosquitoes as a nuisance. However only 9% of respondents thought that impregnated bednets provided protection against malaria although they did not necessarily use them. Design was a determinant factor for the use, and even acceptance, of bednets. The population want rectangular, permanently impregnated bednets large enough to accommodate at least 2 persons. Cost was a major obstacle to wider use by the population. According to our data the best price for the population would be between 2000 and 2500 FCFA as compared to the current price of 3500 FCFA in Ivory Coast.

Design of Onchocerca DNA probes based upon analysis of a repeated sequence family.

Repeated DNA sequences have been instrumental in the development of DNA probes for many different parasites. Isolation of such DNA probes has generally been accomplished by differential screening of genomic libraries with total genomic DNA preparations. In the current work, a rational design strategy is presented for the development of oligonucleotide probes based upon repeated sequence families. A repeated sequence family present in the genome of Onchocerca parasites, designated O-150, has been amplified from various samples of genomic DNA using PCR. DNA sequence analysis of the resulting PCR products demonstrated that the sequences may be arranged into clusters within which the individual sequences are identical or nearly identical. Differences among the cluster consensus sequences have been exploited to explain the specificities of previously isolated O-150 based probes and to develop two new oligonucleotide probes. One of these probes hybridizes specifically to Onchocerca volvulus O-150 PCR products, while the second hybridizes specifically to O-150 PCR products from the closely related bovine parasite O. ochengi. These oligonucleotide probes have been used to characterize Onchocerca infective larvae isolated from wild caught infected flies in West Africa. Because repeated sequence families are a common feature of most genomes, including those of parasites, this method should be applicable to the rational design of oligonucleotide probes for other parasitic infections.

Mitochondrial alleles of Simulium damnosum sensu lato infected with Onchocerca volvulus.

Onchocerca volvulus infected Simulium damnosum s.l. were analysed by directed heteroduplex analysis. Of 73 infected flies, 68 produced heteroduplex products identical to those previously identified. All 6 major sibling species, except S. leonense, were present in this group. In the 5 remaining flies, 2 new heteroduplex patterns were noted. Molecular phylogenetic analysis of these samples suggested that they belonged to the S. squamosum/S. yahense subcomplex. The ability to reliably genotype adult flies will permit studies of the vectorial capacity of the sibling species of S. damnosum s.l. for the blinding and non-blinding strains of O. volvulus.

DNA probe-based classification of Simulium damnosum s. l.-borne and human-derived filarial parasites in the onchocerciasis control program area.

The development of polymerase chain reaction-based methods using strain- and species-specific DNA probes for Onchocerca volvulus has permitted classification of individual parasites from every stage of the parasite's life cycle. This technology has been applied on a large scale basis by Onchocerciasis Control Program (OCP) in West Africa. The primary objective of the OCP in using the DNA probes was to obtain accurate estimates of the annual transmission potential of the blinding strain of O. volvulus. The DNA probe classification of larvae collected throughout the OCP area demonstrated that larvae of less pathogenic strains of O. volvulus and other filarial parasites carried by Simulium damnosum s.l. have resulted in a significant overestimation of the annual transmission potential for blinding onchocerciasis. This effect is particularly pronounced along the southern border of the OCP, where the blinding and less pathogenic strains of O. volvulus coexist, and in the north of the control area, where animal parasites, particularly O. ochengi, may even predominate. A second objective of the OCP in applying the DNA probe technology was to determine the distribution of blinding and less pathogenic O. volvulus in infected individuals along the southern border of the control area. Results obtained from these studies have generally confirmed the distribution pattern established by previous epidemiologic studies. In addition, DNA probe classifications have demonstrated that in areas where the blinding and less pathogenic strains of O. volvulus coexist, a single individual may simultaneously be infected with both strains of the parasite.

Pool screen polymerase chain reaction for estimating the prevalence of Onchocerca volvulus infection in Simulium damnosum sensu lato: results of a field trial in an area subject to successful vector control.

Detection of infective parasites in the vector population can be an early indicator of recrudescence in areas freed of new cases of onchocerciasis. However, dissection of vector black flies is inefficient in areas subject to effective control. Recently, a polymerase chain reaction (PCR)-based assay has been used to detect a single Onchocerca volvulus-infected black fly in pools containing large numbers of uninfected flies. This method had not been validated on wild-caught black flies in an area subject to effective vector control. Here, we report a method of restricting the pool screen PCR assay to infectious parasites and the results of a field test in an area subject to long-term vector control. The prevalence of infection determined by dissection did not differ from that determined by pool screen PCR. The results suggest that the PCR assay may be a useful tool for epidemiologic surveillance for 0. volvulus infection.

Topical application of diethylcarbamazine to detect onchocerciasis recrudescence in west Africa.

The Onchocerciasis Control Programme in West Africa (OCP) has succeeded in eliminating blinding onchocerciasis as a public health problem throughout much of West Africa. The efforts of the OCP are now turning towards surveillance, with the goal of rapidly detecting and controlling outbreaks of infection in the onchocerciasis-free zone. With this goal in mind, cutaneous application of a solution of diethylcarbamazine (the DEC-patch test) was evaluated in 1996-99 as a method to detect patent Onchocerca volvulus infection in children and adolescents, a sentinel population for the detection of recrudescence. In an analysis of 1887 individuals in Côte d'Ivoire and Burkina Faso, the DEC-patch test produced prevalence estimates comparable to those obtained by skin snip. The sensitivity of the DEC-patch assay was marginally greater in children and adolescents than in adults, and was greater in individuals who had received prior Mectizan treatment. These data suggest that the DEC-patch test may be a useful tool for detecting recrudescence of O. volvulus infection in a sentinel population of children and young adults within the onchocerciasis-free zone created by the OCP.

[A case of cross infection by Onchocerca volvulus and Onchocerca ochengi in Simulium damnosum S.L]. / Un cas d'infectivite mixte par onchocerca volvulus et Onchocerca ochengi chez Simulium damnosum S.L.

During a routine entomological survey conducted within the framework of the Program to Control Onchocerciasis in West Africa, a female simulium forest fly was found to be contaminated by 13 Onchocerca volvulus larvae and 7 Onchocerca ochengi larvae. The two Onchocerca species were identified using specific DNA probes. We speculate that cross infection could be related either to behavioral factors, e.g. interruption of blood meals on two different hosts, or developmental factors, e.g. asynchronous development of parasites of the same species or specific differences in the duration of parasite cycles. Further study will be needed to determine the incidence and scope of cross infection in areas where accurate assessment of the impact of vector control on transmission of onchocerciasis in man is required.

[Human onchocerciasis and "sowda" in the Republic of Yemen]. / Onchocercoses humaines et "sowda" en République du Yémen.

The geophysics of the north Yemen, associating a north-south directed mountainous fish bone (rising in more of 2,000 meters), to numerous rivers or "wadis" is convenient to the development of simulium shelters, main vectors for cutaneous filariasis to Onchocerca sp. Following several missions of bio-clinical and epidemiological evaluations in neighbouring villages of wadis, it has been possible to study different clinical aspects: one reminding the classical african onchocerciasis with generalized and diffused dermatitis, and, on an other hand, a hyperreactive dermatitis on one side of the body and associated with a collateral lymphatic ganglion. This disease is well known for local populations as "aswad" meaning "black" or "sowda". Clinically whatever the studied focus, coexists the two types of onchodermatitis (uni or bilateral). Yhe sowda patients are proportionally less numerous than those touched by the generalized type. Frequent eye lesions of the West African onchocerciasis are not found in sowda cases. In classical optical microscopy, microfilaria is morphologically indifferenciable between sowda and onchocerciasis clinical aspects. Skin snips were carried out on patients of both groups. Identification of microfilaria by molecular biology through the study of the DNA genome was done out of 5 skin snips. Microfilaria was kept dry between laminas and the DNA extracted from rehydrated microfilaria. DNA was intensified with specific primers of Onchocerca type (O150PCR). This phase was followed by hybridisation of amplification products by PCR to specific stains: OVS-2 for Onchocerca volvulus species, OCH for Onchocerca ochengi, PFS1 and PSS1-BT respectively for the forest strain and the savannah strain of Onchocerca volvulus as described previously. We can distinguish 2 kinds of answers based on the clinical origin of the snip-tests: the first one concern 3 patients with numerous dermal microfilariae but without any clinical sowda and corresponding to microfilaria O. volvulus type but different from the forest or savannah strains found in sub-Saharan Africa. The second one corresponds to 2 patients with less than 5 microfilaria in their snip-test. They show the typical clinical picture of sowda. They are identified as microfilaria type Onchocerca but they do not belong to species volvulus, or to species ochengi. It seems quite probable that the clinical picture of sowda be the result of developing onchocerciasis of animal origin and not identified as to day. The ivermectin, therapeutic of choice for African onchocerciasis in annual unique cure seems less effective in the coverage of sowda. In that case rehearsal of cures every 3 months would be necessary for mass campaigns to limit the transmission of this filariasis.

Epidemiological evaluation of onchocerciasis along Ogun River System, southwest Nigeria.

Background & objective: Epidemiological studies were carried out to assess the prevalence and community microfilarial load (CMFL) of onchocerciasis after repeated annual treatment with ivermectin along Ogun river System, southwest Nigeria. Method: Skin snips were taken from consented participants in 11 selected communities along the River system. The microfilarial load of the community was estimated. Results: The prevalence and CMFL varied significantly in the communities (p <0.05). The prevalence of onchocerciasis ranged from 19.1 to 45.6%, while the CMFL ranged from 0.11 to 1.03 microfilariae per skin snip. The CMFL recorded was <5 microfilariae per skin snip, i.e. recognized by WHO as threshold value in certifying the communities to be free of onchocerciasis as public health problem, thus, signifying the possibility of onchocerciasis elimination in the study area. Conclusion: Efforts should therefore be intensified to achieve improved ivermectin coverage and compliance in annual ivermectin treatment in order to completely eliminate onchocerciasis as a public health problem in the studied communities.

Intra-specific heterogeneity of the rDNA internal transcribed spacer in the Simulium damnosum (Diptera: Simuliidae) complex.

The internal transcribed spacer (ITS) of the rRNA gene cluster has been used as a model for the study of the action of concerted evolution and molecular drive on repeated sequence families. In contrast to this general finding, preliminary DNA sequence analysis of cloned representatives of the ITS from the West African black fly species complex Simulium damnosum s.1. demonstrated extensive intra-individual and intra-specific polymorphisms. Variability in the ITS was primarily confined to the ITS1 domain. The degree and type of intra-individual and intra-specific variability within the ITS was further characterized using gel electrophoresis, DNA hybridization, and heteroduplex analysis of the PCR products generated from the ITS1 domain. ITS1 copies from individual S. damnosum s.1. differed in length and sequence composition. These results, when taken together, demonstrate that a large degree of intra-individual and intra-specific heterogeneity exists in the ITS of S. damnosum s.1. The intra-individual heterogeneity was greater in the savanna-dwelling than forest-dwelling sibling species of S. damnosum s.1. This heterogeneity may be due in part to inter-breeding among sympatric sibling species, coupled with disturbance of S. damnosum s.1. populations resulting from intensive vector control efforts.

Vector-parasite transmission complexes for onchocerciasis in West Africa.

BACKGROUND: In West Africa, there are two strains of the filarial parasite Onchocerca volvulus, which differ in their ability to induce ocular disease. Transmission studies have suggested that six sibling species of the parasite vector, the black fly Simulium damnosum sensu lato, allow development of the two strains of O volvulus with varying efficiency. We aimed to test the hypothesis of parasite-vector complexes, whereby the two parasite strains, known as forest and savanna, are preferentially transmitted by distinct groups of the species of S damnosum S l. METHODS: During 1993 and 1994, wild black flies were collected from 11 river basins within the area covered by the Onchocerciasis Control Programme (OCP). The flies were dissected and filarial larvae, ovaries, and malpighian tubules removed. Genomic DNA was extracted from larvae, and PCR amplification was used to classify O volvulus parasites as forest or savanna strains. PCR-amplified DNA from ovaries and malpighian tubules was used to distinguish sibling species of S damnosum s l. S yahense and S squamosum were distinguished by body colour. FINDINGS: 214 of 105105 flies dissected were infected with filarial larvae; 84 of these were infected with mature O volvulus parasites. Of the 35 savanna-dwelling infected flies. 17 carried forest-strain parasites and 18 savanna-strain parasites. Of the 45 infected flies identified as the forest dwelling sibling species. 20 carried savanna-strain parasites and 25 forest-strain parasites. No significant differences were found in the numbers of mature larvae of each strain carried by the forest-dwelling species of fly or in the number of forest and savanna larvae in savanna-dwelling vector species. INTERPRETATION: Vector-parasite transmission complexes do not currently play a part in the biology of O volvulus transmission in the area of the OCP in West Africa. This finding has important strategic implications for the future of efforts to control onchocerciasis in West Africa.

Determining the prevalence of Onchocerca volvulus infection in vector populations by polymerase chain reaction screening of pools of black flies.

An important variable in the epidemiology of arthropodborne diseases is the intensity of transmission, which is a function of host-vector contact and the prevalence of infection in the vector population. This latter value is often difficult to estimate. It is possible to envision the application of polymerase chain reaction (PCR) assays to this problem. To accomplish this, the assay must detect a single infected vector in a pool containing a large number of uninfected individuals. It must also be possible to calculate the prevalence of infection from the number of positive pools. A PCR assay for detecting Onchocerca volvulus in pools of vector black flies is described, and an algorithm is presented to calculate the prevalence of infection in the vector population, based upon the proportion of PCR-positive pools. This algorithm should be applicable to any disease for which a PCR assay is available.

The Simulium damnosum species complex: phylogenetic analysis and molecular identification based upon mitochondrially encoded gene sequences.

The DNA sequence of portions of the 16s rRNA and the NADH dehydrogenase subunit 4 (ND4) genes were used to determine phylogenetic relationships in the Simulium damnosum s.l. species complex. Results suggested that at least two major clades existed in the S. damnosum species complex, and that members of the S. damnosum s.l. species complex were not closely related to North American Simulium species. The sequence variability of the ND4 gene was exploited to develop a method to distinguish the sibling species of the S. damnosum s.l. species complex, based on directed heteroduplex analysis of PCR products derived from the ND4 gene. This method was capable of classifying the six sibling species into at least five groups.

Onchocerca volvulus: comparison of field collection methods for the preservation of parasite and vector samples for PCR analysis.

In recent years, methods for the identification of the filarial worm Onchocerca volvulus and its vector, blackflies of the Simulium damnosum complex (S. damnosum sensu lato (s.l.)), based on the amplification of parasite and vector DNA sequences with the polymerase chain reaction (PCR), have been developed. Routine application of these methods requires techniques for sample collection and preservation that are compatible with the limitations of field collection, yet preserve DNA in a form suitable for PCR. Two different methods for sample preservation were evaluated by the field collection teams and the DNA probe laboratory of the Onchocerciasis Control Programme in West Africa. The most successful involved the preservation of material from O. volvulus and its associated vectors in a dried state on microscope slides. Of over 1200 parasite samples preserved in this manner, more than 93% retained DNA yielding positive results in PCR analysis (1208/1291). Vector material (malpighian tubules and ovaries) preserved in the same manner on the same microscope slides also yielded DNA that was suitable for PCR.