Overexpression of cytochrome P450 and esterase genes involved in permethrin resistance in larvae and adults of Culex quinquefasciatus.

Mosquitoes are important vectors of several arthropod-borne diseases, which remain a priority for epidemiological research. Mosquito vector control strategies have traditionally relied on chemical insecticides such as synthetic pyrethroids. However, the indiscriminate use of pesticides has resulted in the development of resistance in many mosquito species. In insects, resistance evolves primarily through the overexpression of one or more gene products from the cytochrome P450, carboxylesterase, and glutathione superfamilies. The current study examined the expression of cytochrome P450 CYP6M2, CYP6AA7, CYP6Z2, CYP9J34, α-Esterase, Esterase B1, and neuroactin genes in larvae and adults of a permethrin-resistant (PerRes) and susceptible (Sus) Culex quinquefasciatus strains. The results showed that the CYP6AA7 gene was overexpressed (10-fold) in larvae and adults with PerRes (p < 0.01) followed by CYPJ34 (9.0-fold) and CYP6Z2 (5.0-fold) compared to the Sus, whereas fewer changes in CYP6M gene expression were observed in PerRes adults (p < 0.05), and no expression was found in larvae. The esterase gene was overexpressed in PerRes larvae (9.0-fold) followed by adults (2.5-fold) compared to the susceptible strain. Based on data, the present study suggests that cytochrome P450, CYP6AA7, CYP6Z2, CYP9J34, α-Esterase, Esterase B1, and neuroactin genes were involved in permethrin resistance in larval and adult Cx. quinquefasciatus.

Impact of MiraNet® long-lasting insecticidal net against Anopheles arabiensis wild population of Northern Tanzania.

Despite high levels of pyrethroid resistance reported in malaria vectors, long-lasting insecticidal nets (LNs) still play a key role in controlling malaria transmission. This study tested the efficacy of MiraNet®, a pyrethroid-based LN against a wild population of Anopheles arabiensis in northern Tanzania. DuraNet® was used as a positive control in this evaluation. Standard WHO laboratory bioefficacy evaluations of MiraNet and DuraNet that were unwashed or had been washed 20 times indicated optimal knockdown and mortality for both net types against a susceptible strain of Anopheles gambiae s.s. Standard experimental hut evaluations were conducted to evaluate the efficacy of both nets against a wild population of An. arabiensis. The killing effect of MiraNet was 54.5% for unwashed and 50% for 20 times washed while DuraNet achieved 44.4% mortality for unwashed and 47.4% for 20 times washed against wild An. arabiensis. Both DuraNet and MiraNet exhibited significantly higher killing effects (> 44.4%). There was no significant difference in deterrence or induced exophily detected between the treatment arms for either net. Additionally, there were no adverse effects reported among hut sleepers. The results of this study indicate that the pyrethroid net MiraNet can be used effectively against wild populations of An. gambiae s.l. of low to moderate resistant levels from Northern Tanzania.

Anti-mosquito properties of Pelargonium roseum (Geraniaceae) and Juniperus virginiana (Cupressaceae) essential oils against dominant malaria vectors in Africa.

BACKGROUND: More than 90% of malaria cases occur in Africa where the disease is transmitted by Anopheles gambiae and Anopheles arabiensis. This study evaluated the anti-mosquito properties of Juniperus virginiana (JVO) and Pelargonium roseum (PRO) essential oils (EOs) against larvae and adults of An. gambiae sensu lato (s.l.) from East Africa in laboratory and semi-field conditions. METHODS: EOs was extracted from the aerial green parts of Asian herbs by hydrodistillation. Their constituents were characterized by gas chromatography-mass spectrometry (GC-MS). Larvicidal activities of JVO, PRO, and PRO components [citronellol (CO), linalool (LO), and geraniol (GO)] were investigated against An. gambiae sensu stricto (s.s.). The percentage of knockdown effects and mortality rates of all oils were also evaluated in the adults of susceptible An. gambiae s.s. and permethrin-resistant An. arabiensis. RESULTS: GC-MS analyses identified major constituents of JVO (sabinene, dl-limonene, ß-myrcene, bornyl acetate, and terpinen-4-ol) and PRO (citronellol, citronellyl formate, L-menthone, linalool, and geraniol). Oils showed higher larvicidal activity in the laboratory than semi-field trials. The LC50 values for JVO/PRO were computed as 10.82-2.89/7.13-0.9 ppm and 10.75-9.06/13.63-8.98 ppm in laboratory and semi-field environments, respectively at exposure time of 24-72 h. The percentage of knockdown effects of the oils were also greater in An. gambiae s.s. than in An. arabiensis. Filter papers impregnated with JVO (100 ppm) and PRO (25 ppm) displayed 100% mortality rates for An. gambiae s.s. and 3.75% and 90% mortality rates, for An. arabiensis populations, respectively. Each component of CO, LO, and GO exhibited 98.13%, 97.81%, and 87.5%, respectively, and a mixture of the PRO components indicated 94.69% adult mortality to permethrin-resistant An. arabiensis. CONCLUSIONS: The findings of this study show that PRO and its main constituents, compared to JVO, have higher anti-mosquito properties in terms of larvicidal, knockdown, and mortality when applied against susceptible laboratory and resistant wild populations of An. gambiae s.l. Consequently, these oils have the potential for the development of new, efficient, safe, and affordable agents for mosquito control.

The impact of Anopheles gambiae egg storage for mass rearing and production success.

BACKGROUND: Mass rearing requires a large colony from which male individuals can be harvested for sterilization and release. Attention is needed when monitoring life parameters of the reared population, knowing that any variations within the target population would lead to mismatching between two populations. The aim of this study was to assess the impact of Anopheles gambiae sensu stricto (s.s.) egg storage on hatchability and life history traits. For each parameter, comparison was made between freshly laid and stored eggs in three densities (40, 80, 120 eggs). METHODS: Anopheles gambiae s.s. freshly laid eggs were collected from the Tropical Pesticide Research Institute (TPRI) insectary. Eggs to be stored were kept at - 20 °C for 10 min and then transferred to refrigerators at 4 °C for intervals of 5, 10, 15, 20, and 25 days. After respective storage days, the eggs were transferred from refrigerators to ambient temperature of (25 ± 2) °C for 24 h and then placed in incubators for 24 h. Thereafter eggs were hatched. The egg hatchability, emerged larvae development, larvae survival and emerged adult sex ratios were monitored. RESULTS: This study found that hatching rates decreased with increase in storage time. The difference was significant in eggs stored for 10 and 15 days (P < 0.05). There were no significant differences in hatching rates between An. gambiae eggs stored for 5 days and freshly hatched eggs (P > 0.05). Anopheles larvae development (L1 to pupae) was not significantly affected by storage time across all hatching densities. The study also found that larvae survival decreased with increase in egg storage time. However, there was no significant difference between larvae from freshly hatched eggs and those from eggs at 5 and 10 storage days (P > 0.05) but not for eggs stored for 15 days. Furthermore, there was a decrease in emerged adult males and increase in females relative to increased time of egg storage. The difference was significant (P < 0.05) at 15 storage days but not for eggs stored for 5 and 10 days (in triplicate densities). CONCLUSION: From this study it was concluded that storing An. gambiae eggs at 4 °C and 48 ± 2% relative humidity (RH) for 5 days is the optimal condition and time that did not affect egg hatching rates, larval development and survivorship and emerged adult mosquito sex ratio.

Malaria mosquito control in rice paddy farms using biolarvicide mixed with fertilizer in Tanzania: semi-field experiments.

BACKGROUND: The wide distribution of malaria mosquito breeding sites within tropical environments limits the mosquito larval source management efforts to control malaria. Rice farming contributes substantially in supporting malaria mosquito productivity in tropical countries. To overcome this challenge, this study was carried out to determine the effect of applying a mixture of biolarvicide and fertilizer on mosquito larvae density in rice farms under semi-field conditions in Tanzania. METHODS: A semi-field experiment was designed to determine the timing of application of a biolarvicide, Bacillus thuringiensis israelensis (Bti) and fertilizer (di-ammonium phosphate-DAP or urea) and assess their effect on mosquito larvae density and rice grain outputs. The experiment had five blocks (4 treatment arms and one control arm) and each had four replicates. Treatment arms had different intervals of days between treatments for mixtures of fertilizer and biolarvicides. The dosages used were 10 g of Bti/16 M2 and 160 g of DAP/Urea/16 m2. RESULTS: In overall, the intervention blocks (with biolarvicide) had lowest mean mosquito larvae abundance compared to control block (F = 22.42, P < 0.001). Similarly, the control arm maintained highest density of Anopheles gambiae sensu lato larvae compared to interventions blocks (F = 21.6, P < 0.001). The best determined timing for application of Bti was in 7 and in 10 days (F = 3.753, P < 0.001). There was neither significant different in mean rice grain harvest per ten panicle (F = 1.453, P = 0.27) nor mean difference in rice grain harvest (F = 1.479, P = 0.26) per intervention arms. CONCLUSION: The findings of this study have shown that application of a mixture of Bti and fertilizer have impact on both mosquito larvae density and maintaining yield rice harvest. Thus, application of a combination of biolarvicide and fertilizer can be an alternative approach in malaria mosquito intervention among rice farming communities of rural Tanzania.

Rice farmers' perceptions and acceptability in the use of a combination of biolarvicide (Bacillus thuringiensis var. israeliensis) and fertilizers application for malaria control and increase rice productivity in a rural district of central Tanzania.

BACKGROUND: The use of larval source management as a supplementary intervention for malaria control has not been widely used in rural Africa due to perceived high costs and complex logistics. To reduce the cost of larviciding in rice farming communities, concurrent application of biolarvicides and fertilizer in rice fields was introduced to control malaria vectors larvae and improve rice grain yields. The present study determined rice farmers' perceptions and acceptability in the use of a combination of biolarvicide and fertilizers in farming practices. METHODS: This was a qualitative study conducted among rice farmers at Kilangali village, south-central Tanzania. Semi-structured interviews and three focus group discussions (FGDs) were conducted with men and women who participated in the biolarvicide and fertilizer application project. The interviews and discussion focused on knowledge, attitudes and perceptions of participants on the use of the innovation in their farming practices and their willingness to pay for the innovation. RESULTS: A total of 40 (mean age = 38.8 ± 10.12 years) rice farmers were involved in the study. Overall, all farmers agreed that it was possible to apply the two products concurrently with minimal challenges. The trust on the safety of biolarvicides on both human and paddy health was high. Respondents reported no challenge in preparation and applying the product in their rice fields. Over half (56.6%) of the participants reported an average decrease in mosquito density in their households and a quarter (26.6%) of them reported a decrease in mosquito population in their farms. Similarly, 93.3% of the participants reported that the intervention had reduced malaria risk in their households. In general, all participants expressed willingness to contribute to a biolarvicide and fertilizer programme and to use the approach in their farming practices. CONCLUSION: Community-based concurrent application of biolarvicides and fertilizer in rice fields was feasible and led to a perceived reduction in mosquito density. Willingness to pay for the larviciding/fertilizer approach was expressed by participants and they accepted to use the approach in their future farming practices. However, the impact of this approach on malaria transmission and rice grain harvest need to be evaluated in a large-scale programme.

Bio-efficacy and wash resistance of MAGNet long-lasting insecticidal net against wild populations of Anopheles funestus in experimental huts in Muheza, Tanzania.

BACKGROUND: The decline in malaria cases and vectors is major milestone in fighting against malaria. The efficacy of MAGNet long-lasting insecticidal nets (MAGNet LLIN), an alpha-cypermethrin incorporated long-lasting net, with the target dose ± 25% of 5.8 g active ingredient (AI)/kg (4.35-7.25 g AI/kg) was evaluated in six veranda-trap experimental huts in Muheza, Tanzania against freely flying wild population of Anopheles funestus. METHODS: MAGNet LLINs were tested against wild, free-flying, host-seeking An. funestus mosquitoes over a period of 6 weeks (total of 36 nights in the huts). MAGNet LLIN efficacy was determined in terms of mosquito mortality, blood-feeding inhibition, deterrence, induced exiting, personal protection, and insecticidal killing over 20 washes according to WHO standardized procedures. Efficacy was compared with reference to a WHOPES recommended approved LLINs (DuraNet) and to a net conventionally treated (CTN) treated with alpha-cypermethrin at WHO-recommended dose and washed to just before cut-off point. The efficacy of MAGNet was evaluated in experimental huts against wild, free-flying, pyrethroid-resistant An. funestus. The WHO-susceptibility method was used to detect resistance in wild Anopheles exposed to 0.75% permethrin. Mosquito mortality, blood-feeding inhibition and personal protection were compared between untreated nets and standard LLINs. Blood-feeding rates were recorded and compared between the 20 times washed; blood-feeding rates between 20 times washed MAGNet LLIN and 20 times washed WHOPES-approved piperonyl butoxide (PBO)/pyrethroid were not statistically different (p > 0.05). RESULTS: The results have evidently shown that MAGNet LLIN provides similar blood-feeding inhibition, exophily, mortality, and deterrence to the standard approved LLIN, thus meeting the WHOPES criteria for blood feeding. The significantly high feeding inhibition and personal protection over pyrethroid-resistant An. funestus recorded by both unwashed and 20 times washed MAGNet compared to the unwashed DuraNet, the WHOPES-approved standard pyrethroid-only LLIN provides proof of MAGNet meeting Phase II WHOPES criteria for a LLIN. CONCLUSION: Based on this study, MAGNet has been shown to have a promising impact on protection when 20 times washed against a highly resistant population of An. funestus.

The influence of age on insecticide susceptibility of Anopheles arabiensis during dry and rainy seasons in rice irrigation schemes of Northern Tanzania.

BACKGROUND: Insecticide resistance is the major emerging challenge facing the malaria vector control programmes in Tanzania. Proper monitoring and detection is of paramount importance guiding the vector control programmes. This paper presents the effect of mosquito aging on insecticide resistance status in Anopheles arabiensis populations in dry and rainy seasons in northern Tanzania. METHODS: Anopheles gambiae s.l. larvae were sampled from rice fields in both dry and rainy seasons and reared in the insectary to adults. The emerged females in batches of 2, 3, 5, and 10 days old were exposed to six insecticides (deltamethrin, permethrin, lambda-cyhalothrin, DDT, bendiocarb and pirimiphos-methyl) to see the effects of age on insecticide resistance. Mosquitoes were exposed to insecticides using WHO standard susceptibility test kits. Knockdown was recorded during the 1-h exposure, while mortality and resistance ratio were recorded 24 h later. Mosquito specimens were identified to species level using the polymerase chain reaction (PCR) method. RESULTS: Among the 326 specimens processed by PCR, 323 (99.1%) were identified as Anopheles arabiensis. There was reduced mortality (ranging from 61 to 97.7%) when adults reared from larvae were exposed to all pyrethroids and bendiocarb in both dry and rainy seasons, while they were fully susceptible to DDT and pirimiphos-methyl. There was a significant increase in mortality rate with increase in mosquito's age in both dry and rainy seasons following exposure to pyrethroids (DF = 1, P < 0.05). Mosquitoes showed significantly higher mortality rates in the rainy season than in the dry season after being exposed to pyrethroids (DF = 1, P < 0.05). Higher mortality rates (94.0-99.8%) were observed in all ages and seasons when mosquitoes were exposed to bendiocarb compared with pyrethroids. Pirimiphos-methyl was only tested in the rainy season so no comparison with dry season mosquitoes could be made. CONCLUSIONS: Results showed that An. arabiensis were resistant to pyrethroids in both seasons and that the young age groups exhibited higher levels of resistance compared with the older age groups. Mosquitoes were full susceptible to DDT and pirimiphos-methyl irrespective of the season and age.

Why some sites are responding better to anti-malarial interventions? A case study from western Kenya.

BACKGROUND: In sub-Saharan Africa, malaria interventions over the last decades have been successful in reducing both mortality and morbidity. In western Kenya however some areas experience contrasting outcomes of the ongoing interventions while the causes for this observation remains not yet clearly known. METHODS: The WHO insecticide (deltamethrin) susceptibility test of the common malaria vectors was studied. Multiple surveys on household use and hospital prescriptions of antimalarial drugs from 2003 to 2015 were done. Along with this, cross sectional surveys on their availability in the local drug dispensing outlets were also done in 2015. Monthly precipitations and air temperature data was collected along with systematic review on abundance and composition of common malaria vectors in the study area before and during interventions. The above factors were used to explain the possible causes of contrasting outcome of malaria interventions between the three study sites. RESULTS: Areas with malaria resurgence or sustained high transmission (Kombewa and Marani) showed higher composition of Anopheles funestus sensu lato (s.l.) than the previously abundant Anopheles gambiae sensu stricto (s.s.) and the later had higher composition to an area with a sustained infection decline (Iguhu). Anopheles gambiae s.l. from Kombewa showed highest resistance (50% mortality) upon exposure to WHO deltamethrin discriminating dosage of 0.75% while those from Marani and Iguhu had reduced resistance status (both had a mean mortality of 91%). Sampled An. funestus s.l. from Marani were also highly resistant to deltamethrin as 57% of the exposed vectors survived. An increasing of mean air temperature by 2 °C was noted for Marani and Iguhu from 2013 to 2015 and was accompanied by an increased rainfall at Marani. Community drug use and availability in selling outlets along with prescription in hospitals were not linked to the struggling control of the disease. CONCLUSIONS: The malaria vector species composition shift, insecticide resistance and climatic warming were the likely cause of the contrasting outcome of malaria intervention in western Kenya. Surveillance of malaria parasite and vector dynamics along with insecticide resistance and vector biting behaviour monitoring are highly recommended in these areas.

In vivo effect of Commiphora swynnertonii ethanolic extracts on Trypanosoma congolense and selected immunological components in mice.

BACKGROUND: The search for alternative trypanocidal compounds which can be available at affordable price is of paramount importance for control of trypanosomosis in human and animals. The current study evaluates the in vivo activity of ethanolic stem bark extracts on Trypanosoma congolense and selected immunological components in an inbred Swiss albino mouse model. METHODS: Groups of mice infected with T. congolense were treated with the stem bark extracts at a rate of 1000 mg/kg, 1500 mg/kg, and 2000 mg/kg, twice a day in one set and thrice a day in another setting for three days consecutively. Negative (infected and untreated) and positive (infected treated with diminazene diaceturate at 3.5 mg/kg) control groups were used. Levels of parasitaemia were monitored daily for the first 10 days and thereafter 2-3 times per week to the end of experiment. In the other setting, uninfected mice, randomized in groups were treated with the extract but categorized as: thorough mixed extract (TME) and supernatant extract (SE) each at 500 mg/kg and 1500 mg/kg, in 8 hourly intervals respectively for three days consecutively. Control group was administered with phosphate buffered saline with glucose at 0.1 ml/10 g in a similar manner as for the extract. Whole blood and spleen were taken 24 h after the last treatment for hematological and histopathological analysis. RESULTS: The groups that received the extracts at 8 hourly intervals drastically reduced the parasitaemia. The higher dose of SE significantly reduced the percentage of lymphocytes (P < 0.05). Both high and low dose of TME significantly reduced lymphocytes percent (P < 0.05) while percent of neutrophils and monocytes increased significantly (P < 0.05). Histopathological changes of the spleen in the mice treated with higher concentrations of the extract of C. swynnertonii were suggestive of lymphocytes toxicity. CONCLUSION: The current study has provided evidence that, in vivo trypanocidal activity of ethanolic bark extracts of C. swynnertonii is probably affected by its negative effect on humoral mediated immune response. Further studies are recommended to determine its potential as an alternative source of lead compounds for trypanocidal drug discovery.

Advancements in bait technology to control Glossina swynnertoni Austen, the species of limited distribution in Kenya and Tanzania border: A review.

Glossina swynnertoni is a savannah tsetse that is largely confined to the Serengeti-Mara [a very small part of East Africa covering northern Tanzania (Arusha and Manyara regions and parts of Shinyanga and Mara regions) extending Maasai Mara ecosystem in southwestern Kenya]. Nevertheless, it is of great concern to human and animal health and is one of the top target tsetse species for eradication. To achieve this eradication objective, it is important to know about its behaviour so that the appropriate tools/measures especially the right traps can be applied against it. In this paper G. swynnertoni is reviewed in terms of its behaviour, and development of traps for its survey and control. Glossina swynnertoni control is of paramount importance in Tanzania tourism industry and country's income. Since, G. swynnertoni is also distributed in national parks, control is vital as it might reduce tourists excursion/movement, by transmitting the African trypanosomiasis among travelers. Different literature search engines such as Google Scholar and PubMed were deployed for literature search. It was found that the behaviour of G. swynnertoni is relatively similar but unique from other tsetse flies. Its feeding cycle is 2½-3 days as opposed to 3-4 days observed in other tsetse species. The flight activity pattern varied between sex, with male having their peak at 1100-1200 hrs and females 1400-1600 hrs. The activity in both sexes decline rapidly towards the dusk (1700-1800 hrs). It was further that host odours, relatively smaller and vertically oriented devices, as well as host movement are the main attractive factors to this tsetse species, which can be exploited to design efficient artificial devices for control of G. swynnertoni . Therefore, due to its restricted distribution and threat it poses on tourism industry, deliberate efforts need to be made against G. swynnertoni as a next candidate to be eradicated using artificial bait technology.

Roles and challenges of construction firms and public health entomologists in ending indoor malaria transmission in African setting.

Indoor malaria transmission reduction across sub-Saharan Africa has been attained through implementation of long-lasting insecticidal nets and indoor residual spray interventions with small-scale larval source management. Improvement of house structures in sub-Saharan Africa can lead to zero indoor malaria transmission with evidence from West Africa, East Africa and Middle East countries. Residual malaria transmission cannot be targeted well with LLINs and IRS alone, but with incorporation of house structures modifications it may be possible.

Trypanocidal activity of ethanolic extracts of Commiphora swynnertonii Burtt on Trypanosoma congolense.

BACKGROUND: African trypanosomosis is the disease caused by extracellular protozoan parasites of the genus Trypanosoma transmitted by tsetse flies. The current study has evaluated the trypanocidal activity of Commiphora swynnertonii extracts on Trypanosoma congolense. METHODS: The effect of ethanolic stem bark and resinous extracts on motility of T. congolense was evaluated by in vitro study at concentrations of 2 mg/ml and 4 mg/ml. Then, trypanocidal activity was evaluated by drug incubation infectivity test using mice at concentrations of 0.4 mg/ml and 2 mg/ml. In both studies negative (without drug) and positive (diminazene diaceturate) controls were used. RESULTS: The in vitro study showed that, ethanolic stem bark extract of C. swynnertonii at concentration of 4 mg/ml caused complete cessation of motility for T. congolense in 30 min. However, resinous ethanolic extract had delayed effect on cessation of motility of T. congolense observed at 90 and 100 min post-incubation at concentrations of 4 mg/ml and 2 mg/ml respectively. The drug incubation infectivity test study depicted that ethanolic stem bark extract at concentration of 2 mg/ml significantly (p = 0.000) reduced the infectivity of T. congolense in mice. However, it did not vary significantly (P =0.897) with group treated with diminazene diaceturate incubated mixture. CONCLUSION: The current study has provided evidence that, ethanolic stem bark extract of C. swynnertonii possess trypanocidal activity against T. congolense. Based on these findings, further studies are recommended to determine its potential as a lead to trypanocidal drug discovery.

Symptomatic malaria diagnosis overestimate malaria prevalence, but underestimate anaemia burdens in children: results of a follow up study in Kenya.

BACKGROUND: The commonly accepted gold standard diagnostic method for detecting malaria is a microscopic reading of Giemsa-stained blood films. However, symptomatic diagnosis remains the basis of therapeutic care for the majority of febrile patients in malaria endemic areas. This study aims to compare the discrepancy in malaria and anaemia burdens between symptomatic diagnosed patients with those diagnosed through the laboratory. METHODS: Data were collected from Western Kenya during a follow-up study of 887 children with suspected cases of malaria visiting the health facilities. In the laboratory, blood samples were analysed for malaria parasite and haemoglobin levels. Differences in malaria prevalence between symptomatic diagnosis and laboratory diagnosis were analysed by Chi-square test. Bayesian probabilities were used for the approximation of the malaria and anaemia burdens. Regression analysis was applied to: (1) determine the relationships between haemoglobin levels, and malaria parasite density and (2) relate the prevalence of anaemia and the prevalence of malaria. RESULTS: The prevalence of malaria and anaemia ranged from 10% to 34%, being highest during the rainy seasons. The predominant malaria parasite was P. falciparum (92.3%), which occurred in higher density in children aged 2‒5 years. Fever, high temperature, sweating, shivering, vomiting and severe headache symptoms were associated with malaria during presumptive diagnosis. After conducting laboratory diagnosis, lower malaria prevalence was reported among the presumptively diagnosed patients. Surprisingly, there were no attempts to detect anaemia in the same cohort. There was a significant negative correlation between Hb levels and parasite density. We also found a positive correlation between the prevalence of anaemia and the prevalence of malaria after laboratory diagnosis indicating possible co-occurrence of malaria and anaemia. CONCLUSION: Symptomatic diagnosis of malaria overestimates malaria prevalence, but underestimates the anaemia burden in children. Good clinical practice dictates that a laboratory should confirm the presence of parasites for all suspected cases of malaria.

The Laboratory and Semi-Field Larvicidal Effects of Essential Oil Extracted from Feronia limonia against Anopheles arabiensis Patton.

This study intended to evaluate the larvicidal activity of Feronia limonia leaf essential oil against the wild population of Anopheles arabiensis Patton larvae in laboratory and semi-field environments. Larvae mortality was observed after 12, 24, 48, and 72 hours of exposure. In laboratory condition, the essential oil showed good larvicidal activity against An. arabiensis (LC50 = 85.61 and LC95 = 138.03 ppm (after 12 hours); LC50 = 65.53 and LC95 = 117.95 ppm (after 24 hours); LC50 = 32.18 and LC95 = 84.59 ppm (after 48 hours); LC50 = 8.03 and LC95 = 60.45 ppm (after 72 hours), while in semi-field experiments, larvicidal activity was (LC50 = 91.89 and LC95 = 134.93 ppm (after 12 hours); LC50 = 83.34 and LC95 = 109.81 ppm (after 24 hours); LC50 = 66.78 and LC95 = 109.81 (after 28 hours); LC50 = 47.64 and 90.67 ppm (after 72 hours). These results give an insight on the future use of F. limonia essential oils for mosquitoes control.

Anopheles stephensi: a guest to watch in urban Africa.

Malaria vector control programs in Sub-Saharan Africa have invested many efforts and resources in the control of eight-sibling species of Anopheles gambiae complex and An. funestus group. The behaviour of sibling species of these vectors is well known and used for implementing the current intervention tools. The reports of An. stephensi in urban Africa with different habitats breeding behaviour is an alert on the success of malaria vector control efforts achieved so far. This communication intends to give an insight on what should be considered as a challenge for the management of An. stephensi in urban Africa to retain the achievement attained in malaria control.

Is it time for Africa to adopt primaquine in the era of malaria control and elimination?

Primaquine is a gametocytocidal drug known to significantly reduce malaria transmission. However, primaquine induces a dose-dependent acute hemolytic anemia (AHA) in individuals with glucose-6-phosphate dehydrogenase (G6PD) deficiency that has led to a limited use of the drug especially in Africa where the condition is common. The World Health Organization (WHO) now recommends a single low dose (SLD) of primaquine (0.25 mg/kg) as P. falciparum gametocytocidal without the need for prior screening of G6PD status. Adoption and implementation of SLD primaquine in Africa may probably reduce malaria transmission, a pre-requisite for malaria elimination. This review therefore, focused on the safety of primaquine for control of malaria in Africa. The literature search was performed using online database Google Scholar, PubMed, HINARI, and Science Direct. Search terms used were "malaria", "primaquine", "safety", "G6PD deficiency", "large scale" or "mass administration". Clinical trials in many African countries have shown SLD primaquine to be safe especially in a milder African G6PD A- variant. Likewise, large-scale primaquine administrations outside Africa involving hundreds of thousands to tenths of millions of participants and with severe variants of G6PD deficiency have also shown primaquine to be safe and well-tolerated. Fourteen deaths associated with primaquine have been reported globally over the past 6 decades, but none occurred following the administration of SLD primaquine. Available evidence shows that the WHO-recommended SLD primaquine dose added to effective schizonticides is safe and well-tolerated even in individuals with G6PD deficiency, and therefore, it can be safely used in the African population with the mildest G6PD A- variant.

Diet and Oviposition Deprivation Effects on Survivorship, Gonotrophic Dissociation, and Mortality of Anopheles gambiae s.s.

Diet quality is of paramount importance for egg batch size, longevity, and mortality of vector mosquitoes. Oviposition site presence and absence assumed to be dry season means a lot to the survivorship and mortality of most anthropophilic malaria vectors in sub-Saharan Africa. This study has assessed the effect of different diets and oviposition-site deprivation (OSD) on survivorship, longevity, and mortality of An. gambiae s.s. To determine the effect of diet and OSD on mortality, gonotrophic dissociation rates, longevity, and survivorship, six treatments were employed: Blood Fed with Oviposition (BFO), Blood Fed without oviposition (BF), Blood and Sugar Fed with Oviposition (BSFO), Blood and Sugar Fed without oviposition (BSF), Sugar Fed with Oviposition (SFO), and Sugar Fed without oviposition (SF). Mortality and gonotrophic dissociation were monitored daily. This study found that female mosquitoes offered blood meals with sugar solution and oviposition deprivation survived longer than those deprived of oviposition deprivation. Similarly, female mosquitoes fed on blood and provided with oviposition deprivation lived longer than those without oviposition deprivation. The gonotrophic dissociation rates were found to be lower in groups provided with oviposition deprivation. Our results show that OSD has a direct impact on the survivorship, gonotrophic dissociation rate, and longevity of the malaria anthropophilic vector, An. gambiae s.s., regardless of the diet.

Characterization and Evaluation of Metarhizium spp. (Metsch.) Sorokin Isolates for Their Temperature Tolerance.

A field survey was done in teak (Tectona grandis F.) forests in South India to explore the entomopathogenic effect of Metarhizium anisopliae (Ascomycota: Sordariomycetes) against teak defoliator, Hyblaea puera (Lepidoptera: Hyblaeidae). About 300 soils and infected insect samples were collected during the survey and thirty-six fungal isolates were isolated from soil and insect samples and characterized. The fungi were cultured on PDAY with dodine and antibiotics. Generally, the EPF culture was incubated at 27 °C in darkness for 15 days. Virulence of the Entomopathogenic Fungi (EPF) ability to germinate under cold and heat temperatures was assessed in a culture impregnated with conidia. In the experiment, it was found that for the first time Metarhizium quizhouense, Metarhizium robertsii, and Metarhizium majus species caused significantly higher mortality to hosts. These isolates of M. anisopliae, M. robertsii, M. majus, and M. quizhouense were all considered to be effective virulent and environmentally adaptive. The Metarhizium isolates were recommended as effective bio-control agents through the field investigation of teak defoliator Hyblaea puera from South India forest. This study paves the way to utilize the indigenous isolates of EPF for the control of teak defoliator and to combat the pests thatare resistant to insecticide.

Cladophialophora bantiana metabolites are efficient in the larvicidal and ovicidal control of Aedes aegypti, and Culex quinquefasciatus and have low toxicity in zebrafish embryo.

Mosquitoes' current insecticide resistance status in available public health insecticides is a serious threat to mosquito control initiatives. Microbe-based control agents provide an alternative to conventional pesticides and insecticides, as they can be more targeted than synthetic insecticides. The present study was focused on identifying and investigating the mosquitocidal potential of Cladophialophora bantiana, an endophytic fungus isolated from Opuntia ficus-indica. The Cladophialophora species was identified through phylogenetic analysis of the rDNA sequence. The isolated fungus was first evaluated for its potential to produce metabolites against Aedes aegpti and Culex quinquefasciatus larvae in the 1-4th instar. The secondary metabolites of mycelium extract were assessed at various test doses (100, 200, 300, 400, and 500 µg/mL) in independent bioassays for each instar of selected mosquito larvae. After 48 h of exposure, A. aegypti expressed LC50 values of 13.069, 18.085, 9.554, and 11.717 µg/mL and LC90 = 25.702, 30.860, 17.275, and 19.601 µg/mL; followed by C. quinquefasciatus LC50 = 14.467, 11.766, 5.934, and 7.589 µg/mL, and LC90 = 29.529, 20.767, 11.192, and 13.296 µg/mL. The mean % of ovicidal bioassay was recorded 120 h after exposure. The hatchability (%) was proportional to mycelia metabolite concentration. The enzymatic level of acetylcholinesterase in fungal mycelial metabolite treated 4th instar larvae indicated a dose-dependent pattern. The GC-MS profile of C. bantiana extracts identified five of the most abundant compounds, namely cyclobutane, trans-3-undecene-1,5-diyne, 1-bromo-2-chloro, propane, 1,2,3-trichloro-2-methyl-, 5,5,10,10-tetrachlorotricyclo, and phenol, which had the killing effect in mosquitoes. Furthermore, the C. bantiana fungus ethyl acetate extracts had a strong larvicidal action on A. aegypti and C. quinquefasciatus. Finally, the toxicity test on zebrafish embryos revealed the induction of malformations only at concentrations above 1 mg/mL. Therefore, our study pioneered evidence that C. bantiana fungal metabolites effectively control A. aegypti and C. qunquefasciastus and show less lethality in zebrafish embryos at concentrations up to 500 µg/mL.