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Background: The effectiveness of Long-Lasting Insecticidal Nets and Indoor Residual Spraying in malaria vector control is threatened by vector resistance to insecticides. Knowledge of mosquito habitats and patterns of insecticide resistance would facilitate the development of appropriate vector control strategies. Therefore, we investigated An. coluzzii larval habitats and resistance to insecticides in the Manoka rural island area compared with the Youpwe suburban inland area, in Douala VI and II districts respectively., Methods: Anopheline larvae and pupae were collected from open water bodies in December 2013 and April 2014 and reared until adult emergence. Two to four day old emerging females were morphologically identified as belonging to the An. gambiae complex and used for WHO susceptibility tests with 4 % DDT, 0.75 % permethrin, and 0.05 % deltamethrin, with or without piperonyl butoxide (PBO) synergist. Control and surviving specimens were identified down to the species using a PCR-RFLP method. Survivors were genotyped for kdr L1014 mutations using Hot Oligonucleotide Ligation Assay., Results: In both study sites, ponds, residual puddles, boats, and drains were identified as the major An. gambiae s.l. larval habitats. A total of 1397 females, including 784 specimens from Manoka and 613 from Youpwe, were used for resistance testing. The two mosquito populations displayed resistance to DDT, permethrin and deltamethrin, with variable mortality rates from 1 % to 90 %. The knock-down times were also significantly increased (at least 2.8 fold). Pre-exposure of mosquitoes to PBO did not impact on their mortality to DDT, conversely the mortality rates to permethrin and deltamethrin were significantly increased (7.56 ≤ X(2) ≤ 48.63, df = 1, p < 0.01), suggesting involvement of P450 oxidases in pyrethroid resistance. A subsample of 400 An. gambiae s.l. specimens including 280 control and 120 survivors from bioassays were all found to be An. coluzzii species. Only the kdr 1014 F mutation was found in survivors, with 88.5 % (N = 76) and 75 % (N = 44) frequencies in Youpwe and Manoka respectively., Conclusion: This is the first report of An. coluzzii resistance to insecticides in an insular area in Cameroon. Since permanent larval habitats have been identified, larval source management strategies may be trialed in this area as complementary vector control interventions.

A trial of permethrin-treated nets (PTNs) versus untreated nets (UTNs) was conducted in Pitoa (north Cameroon), where the main malaria vectors, Anopheles gambiae s.l. and Anopheles arabiensis, show metabolic-based permethrin resistance. The deterrent effect of permethrin greatly reduced A. gambiae biting rate inside rooms where PTNs were installed. After 3 months of net use, malaria reinfection rate was significantly lower in children sleeping under a PTN, but no such effect was observed after 6 months. Parasitaemia was not significantly different between the two arms. These findings suggest good, although transitory, personal protection against malaria conferred by PTNs in an area of metabolic-based permethrin resistance.

Some populations of Anopheles gambiae s.l. from Cameroon were reported to develop resistance to DDT or pyrethroids but were free of the kdr mutation "Leucine-Phenylalanine" (Leu-Phe). This study reports on the metabolic activity of non-specific esterases (NSEs), mixed function oxidases (MFOs), and glutathione S-transferases (GSTs), three enzyme systems commonly involved in insecticide resistance. Biochemical assays were performed in DDT or pyrethroid-resistant populations of An. gambiae s.l. from Douala, Mbalmayo, Pitoa, and Simatou neighborhoods. Enzyme activity was compared to the Kisumu-susceptible reference strain using the Mann-Whitney test. Most of the tested samples had elevated NSE activity (P < 0.02). The Douala sample evenly displayed elevated GST activity (P < 0.001), while high MFO level was recorded in the Pitoa sample (P < 0.001). MFO or GST levels were sometimes lower or similar to that of the Kisumu strain. These results suggest metabolic detoxification is a major DDT or pyrethroid resistance mechanism and emphasize the need for further investigations on An. gambiae s.l. resistance mechanisms in Cameroon.

Background: Malaria is a major public health problem in Cameroon. Unlike in the southern forested areas where the epidemiology of malaria has been better studied prior to the implementation of control activities, little is known about the distribution and role of anophelines in malaria transmission in the coastal areas., Methods: A 12-month longitudinal entomological survey was conducted in Tiko, Limbe and Idenau from August 2001 to July 2002. Mosquitoes captured indoors on human volunteers were identified morphologically. Species of the Anopheles gambiae complex were identified using the polymerase chain reaction (PCR). Mosquito infectivity was detected by the enzyme-linked immunosorbent assay and PCR. Malariometric indices (plasmodic index, gametocytic index, parasite species prevalence) were determined in three age groups (<5 yrs, 5-15 yrs, >15 yrs) and followed-up once every three months., Results: In all, 2,773 malaria vectors comprising Anopheles gambiae (78.2%), Anopheles funestus (17.4%) and Anopheles nili (7.4%) were captured. Anopheles melas was not anthropophagic. Anopheles gambiae had the highest infection rates. There were 287, 160 and 149 infective bites/person/year in Tiko, Limbe and Idenau, respectively. Anopheles gambiae accounted for 72.7%, An. funestus for 23% and An. nili for 4.3% of the transmission. The prevalence of malaria parasitaemia was 41.5% in children <5 years of age, 31.5% in those 5-15 years and 10.5% in those >15 years, and Plasmodium falciparum was the predominant parasite species., Conclusion: Malaria transmission is perennial, rainfall dependent and An. melas does not contribute to transmission. These findings are important in the planning and implementation of malaria control activities in coastal Cameroon and West Africa.

Background: Insecticide-treated nets (ITNs) are an integral component of malaria control programmes in Africa. How much pyrethroid resistance in malaria vectors will impact on the efficacy of ITNs is controversial. The purpose of this study was to evaluate knockdown and killing effects of ITNs on a metabolic-based resistant or tolerant malaria vector strain., Methods: Bio-efficacy of 500 mg/m2 permethrin EC treated bednets was assessed on the OCEAC laboratory (OC-Lab) strain of Anopheles gambiae s.s.. This strain is resistant to DDT and tolerant to pyrethroids, with elevated mixed function oxidases. The Kisumu reference susceptible strain of A. gambiae s.s. was used as control. Nets were impregnated in February 1998 and used by households of the Ebogo village. Then they were collected monthly over six months for Bio-assays (WHO cone test). Knockdown and mortality rates were compared between the OC-Lab and the Kisumu strains, by means of the Mantel-Haenszel chi-square test., Results: During the whole trial, permethrin EC knockdown rates were impressive (mostly higher than 97%). No significant difference was observed between the two strains. However, the mortality rates were significantly decreased in the OC-Lab strain (40-80%) compared with that of the Kisumu strain (75-100%). The decrease of killing effect on the OC-Lab strain was attributed to permethrin EC tolerance, due to the high oxidase metabolic activity., Conclusion: These data suggested an impact of pyrethroid tolerance on the residual activity of ITNs. More attention should be given to early detection of resistance using biochemical or molecular assays for better resistance management.

The bioefficacy of cyfluthrin-impregnated bed-nets was evaluated in the agro-industrial town of Mbandjock (southern Cameroon). The objectives were to assess the knockdown and mortality rates, the protection against bloodfeeding mosquitoes, and the irritant effect of cyfluthrin (SOLFAC EW050)-impregnated bed-nets against a susceptible strain of Anopheles gambiae. Polyester bed-nets were impregnated and distributed to inhabitants of Mbandjock for use, then nets were retreated after 6 months. The uptake of active ingredient on nets ranged from 30.8 mg/m2 to 75.1 mg/m2 at the initial treatment and from 131.0 mg/m2 to 225.0 mg/m2 at retreatment. The susceptible Kisumu reference strain of Anopheles gambiae reared in our laboratory was used for bioassays. The knockdown rate on freshly treated nets (2 wk after treatment) ranged from 93 to 96% and the mortality rate ranged from 52 to 70%. During the 12-month trial, knockdown and mortality rates showed 2 peaks, respectively, in the 4th month (94-97% knockdown and 61-96% mortality) and 7th month (i.e., soon after retreatment; 89-98% knockdown and 86-100% mortality), separated by breakdowns on the 5th through 6th and 12th months (46-77% knockdown and 8-69% mortality). Knockdown and mortality rates decreased faster on the lower part of nets than on the top, suggesting that some external factors such as dirt and daily hand manipulation could impede the efficacy of treated nets. Exceedingly dirty nets were less effective than clean nets. The protective rate against bloodfeeding mosquitoes ranged from 60 to 100% during the 1st 4 months and decreased at 40-70% during the 5th and 6th months after the initial treatment. After retreatment, the protective rate ranged from 50 to 90% for the 1st 5 months and from 35 to 64% at the 6th month. More than 70% of mosquitoes that attempt to feed through treated or retreated nettings died within 24 h after contact. Cyfluthrin was found to be mildly irritant during the whole evaluation. This trial reveals that cyfluthrin EW050-treated nets were effective against a susceptible strain of An. gambiae.

A large-scale survey of Anopheles gambiae Giles, 1902 susceptibility to DDT, dieldrin, permethrin, and deltamethrin was conducted in the Republic of Cameroon. 15 field populations from various geographical areas were tested using World Health Organization test kits for adult mosquitoes. The laboratory Kisumu susceptible reference strain was tested as a control. Results showed that dieldrin and DDT resistance was still present in some populations, and indicated permethrin or deltamethrin resistance. Within the Anopheles gambiae complex, resistant individuals belonged to An. gambiae s.s. and An. arabiensis species. Both M and S molecular forms of An. gambiae s.s. were found resistant. In most of resistant populations, the knockdown times were 2-5-folds increased. However, none of the surviving mosquitoes was positive to the kdr "Leu-Phe" mutation using polymerase chain reaction (PCR) diagnostic test. These results likely suggested involvement of other resistance mechanism(s), such as enzyme detoxification or kdr "Leu-Ser" mutation. Researches on An. gambiae s.l. resistance should be promoted in Cameroon, to improve malaria vector control programs and to implement resistance management strategies.