Plasmodium falciparum-induced kidney and liver dysfunction in malaria patients in Freetown; Sierra Leone

This study was undertaken to investigate the effect of Plasmodium falciparum infection on kidney and liver function parameters in malaria patients in Freetown; Sierra Leone. Blood samples taken from 64 malaria patients and 64 non-malaria volunteers at Abanita and Blue Shield Hospitals; Freetown Sierra Leone between January to April; 2009 were examined. Changes in serum biochemical parameters were analysed using normal range values as baseline. Serum bilirubin; alkaline phosphatase (ALP); alanine aminotransferase (ALT) and aspartate aminotransferase (AST) concentrations were significantly elevated in falciparum malaria patients compared to their non-malaria counterparts which is an indication of defective liver function. Most of patients with falciparum malaria also have significantly high serum concentrations of urea; creatinine; sodium and potassium showing alteration in kidney function. This study suggests that malaria parasites could be responsible for derangement of kidney and liver functions in patients and could therefore contribute to organ damage in affected individuals if not treated

Dynamics of knockdown pyrethroid insecticide resistance alleles in a field population of Anopheles gambiae s.s. in southwestern Nigeria.

BACKGROUND & OBJECTIVES: Pyrethroid insecticide resistance in the malaria vector Anopheles gambiae Giles is mainly associated with reduced target site sensitivity arising from a single point mutation in the sodium channel gene, often referred to as knockdown resistance (kdr). This resistance mechanism is widespread in West Africa and was reported for the first time in Nigeria in 2002. Here we present changes in the susceptibility/resistance status of the molecular 'M' and 'S' forms of An. gambiae and the frequency of the kdr alleles from 2002-05. METHODS: Adult anophelines were sampled quarterly inside human dwellings from January 2002 to December 2005 and adults reared from wild larvae were identified using morphological keys. Samples belonging to the An. gambiae complex were subjected to PCR assays for species identification and detection of molecular 'M' and 'S' forms. Insecticide susceptibility tests were carried out using standard WHO procedures and test kits only on 2-3 days old adult An. gambiae s.s. reared from larval collections. The kdr genotypes were determined in both live and dead specimens of An. gambiae s.s. using alleles-specific polymerase chain reaction diagnostic tests. RESULTS: The overall collection showed that the molecular 'S' form was predominant (> 60%) but the proportions of both forms in the mosquito populations from 2002-05 were not statistically different. Both forms also occurred throughout the period without apparent relationship to wet or dry season. Insecticide susceptibility tests did not show any significant increase in the resistance status recorded for either Permethrin or DDT from 2002-05, rather, an improvement in the susceptibility status of the mosquitoes to these insecticides was observed from 2004-05 relative to the tests performed in 2002-03. CONCLUSION: The proportion of the molecular 'M' and 'S' form of An. gambiae and the kdr frequencies have not increased significantly from 2002 when it was first reported in Nigeria. However, the findings on susceptible mosquitoes exhibiting the kdr gene need further investigation. Further monitoring of this may provide additional information on the ongoing debate on the possibility of restriction in gene flow and reproductive barriers in these sympatric taxa.

Behaviour and population dynamics of the major anopheline vectors in a malaria endemic area in southern Nigeria.

BACKGROUND & OBJECTIVES: Anopheline mosquitoes consist of a large number of species each of which differs from another in population, resting and feeding behaviour in relation to the prevailing conditions in the locality. A longitudinal study was carried out to investigate the population dynamics, resting and feeding behaviour of the major anopheline species found in a rain forest zone of Nigeria. METHODS: Mosquitoes resting and biting indoors were collected using WHO standard techniques and supplemented with outdoor-biting collections in the study areas between January and December 2004. Samples were sorted and identified microscopically for morphological features while molecular identification was carried out using polymerase chain reaction (PCR) techniques. RESULTS: PCR-based tests showed that both indoor and outdoor collections constitute three groups of Anopheles mosquitoes, An. gambiae s.l. Giles (68.6%), An. funestus Giles (30.7%) and An. moucheti Evans (0.7%). Of the 1342 female Anopheles mosquitoes collected indoors, 799 were caught resting and 543 were caught biting. The outdoor-biting population accounted for 28.8% of the total collections (n=1885). There was no significant difference (p > 0.05) in the biting activities (indoors and outdoors) of these species in four villages. However, An. arabiensis and An. moucheti were more exophagic with >60% of their biting occurring outdoors while An. gambiae and An. funestus were more endophagic with >55% of their biting occurring indoors. The human-vector contact with An. gambiae and An. funestus (indoors) was about 73.3 and 66.7%, respectively as against 27.7% in An. arabiensis and 25.3% in An. moucheti. This gave the corresponding man-biting rates (MBR) of 17.5 bites/man/night for An. gambiae, 14.6 bites/man/night for An. funestus, 6.7 bites/man/night for An. arabiensis and 4.3 bites/man/night for An. moucheti. Moreover, the number of An. gambiae and An. funestus caught resting indoors was significantly higher than the other two species (p < 0.05). The wet season collections showed thatAn. gambiae caught were more than 67% of the total catch while An. arabiensis was predominant in the dry season (chi2 = 75.44, df = 3, p < 0.01). INTERPRETATION & CONCLUSION: The present study highlights some aspects of the behaviour of anopheline mosquitoes in southern Nigeria which is an important component of epidemiological study of malaria. This information provides basis to the understanding of the role played by the identified anopheline species in malaria transmission and a baseline to formulate malaria control programme.

Impact of urbanisation on bionomics and distribution of malaria vectors in Lagos, southwestern Nigeria.

BACKGROUND & OBJECTIVES: The patterns of annual, rainfall as well as average daily temperature have not changed drastically in the study area since 1960 when detailed baseline entomological surveys were carried out. However, the increase in human population from 1.2 to 10 million has resulted in both expansion of land and tremendous ecological and environmental change. This has led to drastic changes in vectors' densities as well as species' composition while the preferred larval habitat of malaria vectors has shifted to water reservoirs. A longitudinal study was carried out to investigate the impact of urbanisation on bionomics and distribution of malaria vectors in Lagos, a mega city in Nigeria. METHODS: Mosquitoes were collected indoors and outdoors using WHO standard techniques in the selected areas between January and December 2000. Specimens were identified using the morphological keys and PCR assays. ELISA tests were used for Plasmodium falciparum sporozoite infection. RESULTS: The Anopheles gambiae species-specific PCR identified 56% (435/777) of the An. gambiae s.1. as An. gambiae s.s. and 44% (342/777) as An. arabiensis. The molecular M and S forms represented 35.3 and 64.7% of the An. gambiae s.s. population, respectively. The An. funestus species-specific PCR identified 60% (239/401) of the An. funestus group as An. funestus s.s. and 40% (162/401) as An. rivulorum. The biting activity of An. funestus s.s. both indoors and outdoors attained a peak at 0200 and 2200 hrs, respectively, with a significant increase in the "pre-bed time" collections indoors (Chi2 = 6.15, df = 1, p < 0.05) and outdoors (Chi2 = 6.28, df = 1, p < 0.05). The overall outdoor collection was significantly higher (Chi2 = 28.23, df =3, p < 0.05) than that recorded indoors. The overall sporozoite rates for An. gambiae s.s., An. arabiensis and An. funestus were significantly different in both localities (Chi2 = 0.58, df = 2, p < 0.01). Infection rates in both indoor and outdoor collections were also different statistically (Chi2 = 0.67, df = 2, p < 0.01). INTERPRETATION & CONCLUSION: Large number and species of anopheline mosquitoes collected in the study area may be associated with the availability of aquatic breeding sites. A phenomenon leading to an increase in man-vector contact and a high level of risk to the crowded urban population is observed.

Distribution of the molecular forms of Anopheles gambiae and pyrethroid knock down resistance gene in Nigeria.

We investigated the distribution of the molecular M and S forms of Anopheles gambiae and the knock down resistance (kdr) gene associated with pyrethroid and DDT resistance in A. gambiae s.s. at 13 localities across Nigeria. Two-three days old adult female mosquito reared from larval collections were tested using standard WHO procedures, diagnostic test kits and impregnated papers to assess their pyrethroid resistance status. Specimens were identified by PCR assays and characterized for the kdr gene. DNA from adult A. gambiae s.s. collected from human dwellings were also tested for the presence of the kdr gene. The overall collection was a mix of the molecular M and S forms across the mangrove (63:37%), forest (56:44%), and transitional (36:64%) ecotypes, but almost a pure collection of the S form in the Guinea and Sudan-savanna. Results of insecticide susceptibility tests showed that mosquitoes sampled at seven localities were susceptible to permethrin, deltamethrin, and DDT, but populations of A. gambiae resistant to these insecticides were recorded at six other localities mainly in the transitional and Guinea-savanna ecotypes. The kdr gene was found only in the molecular S forms, including areas where both forms were sympatric. The overall kdr frequency was low: <47% in forest, 37-48% in the transitional, and 45-53% in Guinea-savanna. The data suggest that pyrethroid resistance in A. gambiae in Nigeria is not as widespread when compared to neighbouring West African countries.

Identification of three members of the Anopheles funestus (Diptera: Culicidae) group and their role in malaria transmission in two ecological zones in Nigeria.

The role of the Anopheles funestus group in malaria transmission was investigated in two ecological zones in Nigeria. Sampling was carried out at four sites each around Ibadan (forest) and Ilorin (savanna). Human landing catches were supplemented with indoor and outdoor resting collections. PCR was used to identify 1848 A. funestus group mosquitoes to species level (749 in the savanna, 1099 in the forest) and three species were identified. In the forest, A. funestus s.s. predominated (55.4%), followed by A. rivulorum (27.6%) and A. leesoni (17.0%). Anopheles funestus was found mostly indoors. Anopheles rivulorum and A. leesoni predominated in outdoor collections (P<0.001). Only Anopheles funestus s.s. was found in the savanna. ELISA analysis of 803 blood meal-positive specimens showed that over half of the blood meals were taken from humans in both ecotypes. The human blood index in A. funestus from the two study areas was similar. Anopheles funestus s.s. was the only species found positive for Plasmodium falciparum using ELISA, with overall infection rates of 2.3% and 1.0% in the forest and savanna respectively. The presence of three A. funestus species in Nigeria emphasizes the desirability of correct species identification within a malaria vector control programme.