Early biting rhythm in the Afro-tropical vector of malaria, Anopheles arabiensis, and challenges for its control in Ethiopia

The biting cycle of the malaria vector Anopheles arabiensis Patton (Diptera: Culicidae) was assessed by hourly light trap collections in three villages in Tigray, northern Ethiopia. Hourly catches were conducted in two houses in each village, for four consecutive nights. Light traps were set from 18.00 hours to 07.00 hours in houses in which people slept under untreated bednets. Anopheles arabiensis showed early biting activities, which peaked between 19.00 hours and 20.00 hours in the three villages; over 70% of biting activity occurred before 22.00 hours, when people typically retire to bed. This early biting activity may have a negative impact on the efficiency of bednets to control malaria.     

Efficacy of permethrin-impregnated curtains for malaria vector control

Preliminary results obtained by the use of permethrin-impregnated curtains against the Afrotropical malaria vectors Anopheles gambiae Giles s.l. and An.funestus Giles are reported and discussed. Field trials were carried out in villages near Ouagadougou, Burkina Faso. Houses were provided with curtains made from 100% cotton netting, impregnated with permethrin at the dose of 1 g a.i./m2, to cover the doorway, the window(s) and the space under the eaves. Entomological data collected during the period 1985-86 showed residual permethrin activity for about a year, and almost complete prevention of indoor-resting mosquitoes. Increased exit-rate and mortality-rate of house-entering malaria vectors were also obtained. Utilization of this malaria vector control method in primary health care programmes is advocated.     

Longitudinal follow‐up of malaria transmission dynamics in two villages in a Sahelian area of Niger during a nationwide insecticide‐treated bednet distribution programme

Malaria transmission was monitored in two villages in the Sahel zone of Niger over 4 years. During this period, a nationwide vector control programme was carried out in which insecticide‐treated bednets were distributed free to mothers of children aged <5 years. Anopheles gambiae and Anopheles arabiensis (Diptera: Culicidae) were found to be the major malaria vectors. The dynamics of An. gambiae s.l. did not vary dramatically over the study period although the proportion of female mosquitoes found resting indoors decreased in both villages and, in one village, the parity rate and sporozoite index were significantly reduced after bednet distribution. By contrast with An. gambiae, the dynamics of Anopheles funestus altered greatly after the bednet distribution period, when adult density, endophagous rate and sporozoite rates decreased dramatically. Our observations highlight the importance of quantifying and monitoring the dynamics and infections of malaria vectors during large‐scale vector control interventions.     

Development of a genetic sexing strain of Anopheles arabiensis for KwaZulu‐Natal,South Africa

An efficient sexing system is important for the release of sterile males for any control programme using the sterile insect technique. This study describes the development and characterization of a new genetic sexing strain from South Africa (GMK), needed for the planned implementation of such a programme in northern KwaZulu‐Natal Province. The base colony used was a locally modified laboratory strain of Anopheles arabiensis containing a sex‐linked gene conferring dieldrin resistance to male mosquitoes. Female A. arabiensis mosquitoes from northern KwaZulu‐Natal were mated with these males and backcrossed to introduce the dieldrin resistance gene to the Y chromosome. The resulting strain therefore had an overall genotype representing the local population but with the Y chromosome containing the dieldrin resistance gene. Life‐history characteristics, stability of the sex‐linked resistance marker, and reduction in dieldrin waste were investigated. The strain showed semi‐sterility exhibited by low egg hatch rates, faster development in the immature stages and longer adult survivorship compared with the parental strains. While the GMK strain carrying the dieldrin‐resistant gene was successfully established, the stability of the gene is limited, requiring periodic purification. Dieldrin waste can be limited by treating many more eggs than currently recommended.     

Blood-feeding behaviour of the malarial mosquito Anopheles arabiensis: implications for vector control

Feeding behaviour of the malaria vector Anopheles arabiensis Patton (Diptera: Culicidae) was monitored for 12 months (March 2003-February 2004) in the Konso District of southern Ethiopia (5 degrees 15'N, 37 degrees 28'E). More than 45 000 An. arabiensis females were collected by host-baited sampling methods (light-traps, human landing catches, cattle-baited traps) and from resting sites (huts and pit shelters). In the village of Fuchucha, where the ratio of cattle : humans was 0.6 : 1, 51% of outdoor-resting mosquitoes and 66% of those collected indoors had fed on humans, human baits outdoors caught > 2.5 times more mosquitoes than those indoors and the mean catch of mosquitoes from pit shelters was about five times that from huts. Overall, the vast majority of feeding and resting occurred outdoors. In the cattle camps of Konso, where humans slept outdoors close to their cattle, approximately 46% of resting mosquitoes collected outdoors had fed on humans despite the high cattle : human ratio (17 : 1). In both places, relatively high proportions of bloodmeals were mixed cow + human: 22-25% at Fuchucha and 37% in the cattle camps. Anthropophily was also gauged experimentally by comparing the numbers of mosquitoes caught in odour-baited entry traps baited with either human or cattle odour. The human-baited trap caught about five times as many mosquitoes as the cattle-baited one. Notwithstanding the potential pitfalls of using standard sampling devices to analyse mosquito behaviour, the results suggest that the An. arabiensis population is inherently anthropophagic, but this is counterbalanced by exophagic and postprandial exophilic tendencies. Consequently, the population feeds sufficiently on humans to transmit malaria (sporozoite rates: 0.3% for Plasmodium falciparum and 0.5% for P. vivax, by detection of circumsporozoite antigen) but also takes a high proportion of meals from non-human hosts, with 59-91% of resting mosquitoes containing blood from cattle. Hence, classical zooprophylaxis is unlikely to have a significant impact on the malaria vectorial capacity of An. arabiensis in Konso, whereas treating cattle with insecticide might do.     

Comparative evaluation of carbosulfan- and permethrin-impregnated curtains for preventing house-entry by the malaria vector Anopheles gambiae in Burkina Faso

Pyrethroid-impregnated bednets and curtains are widely employed to reduce the risk of malaria transmission, but pyrethroid-resistance is becoming more prevalent among malaria vector Anopheles mosquitoes (Diptera: Culicidae). As an alternative treatment for curtains, we assessed carbosulfan (a carbamate insecticide) in comparison with permethrin as the standard pyrethroid, against endophilic female mosquitoes of the Anopheles gambiae Giles complex in a village near Ouagadougou, Burkina Faso. The main criterion evaluated was the impact of curtains (hung inside windows, eaves and doorways) on the number of An. gambiae s.l. females active indoors at night. Light-traps were operated overnight (21.00-06.00 hours beside occupied untreated bednets) to sample mosquitoes in houses fitted with net curtains treated with carbosulfan 0.2 g ai/m2 or permethrin 1 g ai/m2 or untreated, compared with houses without curtains. The treated and untreated curtains significantly reduced the numbers of mosquitoes collected indoors, compared with houses without curtains. Carbosulfan-treated curtains had a highly significantly greater effect than permethrin-treated or untreated curtains, the scale of the difference being estimated as three-fold. However, there was no significant difference between the impact of untreated and permethrin-treated curtains on densities of An. gambiae s.l. trapped indoors. Samples of the An. gambiae complex comprised An. arabiensis Patton and both the S- and M-forms of An. gambiae Giles s.s. Susceptibility tests revealed some resistance to DDT and low frequencies of permethrin-resistance, insufficient to explain the poor performance of permethrin on curtains. Among survivors from the diagnostic dosage of permethrin were some specimens of all three members of the An. gambiae complex, but the kdr resistance mechanism was detected only in the S-form of An. gambiae s.s. Questions arising for further investigation include clarification of resistance mechanisms in, and foraging behaviour of, each member of the An. gambiae complex in this situation and the need to decide whether carbosulfan-treated curtains are acceptably safe for use to reduce risks of malaria transmission.     

Development and evaluation of a novel nano‐scale vector for siRNA

To synthesize a lipid‐cationic polymer (LCP) containing brassidic acid side chain and to investigate its transfection efficiency and characteristics as a siRNA gene vector. The LCP was chemically synthesized and its nucleic acid binding capacity was determined by gel electrophoresis. HeLa‐EGFP and TH1080‐EGFP cell lines were transfected with siRNA against enhanced green fluorescent protein (EGFP) gene using a LCP to investigate the transfection efficiency. An MTT assay was performed to evaluate the cellular toxicity of the LCP vector. Its degradability and stability under acidic conditions were also investigated. The LCP vector possessed high DNA binding capacity. More than 73% of the cellular fluorescence was inhibited by the LCP‐mediated transfection of siRNA against EGFP gene, indicating that vector had high transfection efficiency. Cellular viability was about 95% at the optimum transfection efficiency of LCP, suggesting that the cellular toxicity of LCP was very low. The LCP was also observed to be degradable; moreover, it could be easily stored at normal temperature. A gene vector used for the transfection of siRNA was successfully fabricated from synthesized LCP. Its numerous excellent properties entitle values for further scientific research. J. Cell. Biochem. 111: 881–888, 2010. © 2010 Wiley‐Liss, Inc.     

Techno‐economic evaluation of a two‐step biological process for hydrogen production

An integrated biological process for the production of hydrogen based on thermophilic and photo‐heterotrophic fermentation was evaluated from a technical and economic standpoint. Besides the two fermentation steps the process also includes pretreatment of the raw material (potato steam peels) and purification of hydrogen using amine absorption. The study aimed neither at determining the absolute cost of biohydrogen nor at an economic optimization of the production process, but rather at studying the effects of different parameters on the production costs of biohydrogen as a guideline for future improvements. The effect of the key parameters, hydrogen productivity and yield and substrate concentration in the two fermentations on the cost of the hydrogen produced was studied. The selection of the process conditions was based mainly on laboratory data. The process was simulated by use of the software Aspen Plus and the capital costs were estimated using the program Aspen Icarus Process Evaluator. The study shows that the photo‐fermentation is the main contributor to the hydrogen production cost mainly because of the cost of plastic tubing, for the photo‐fermentors, which represents 40.5% of the hydrogen production cost. The costs of the capital investment and chemicals were also notable contributors to the hydrogen production cost. Major economic improvements could be achieved by increasing the productivity of the two fermentation steps on a medium‐term to long‐term scale. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2010     

Fuzzy rule‐based decision support system for evaluation of long‐established forest restoration projects

Although the importance of monitoring and evaluation of restoration actions is increasingly acknowledged, availability of accurate, quantitative monitoring data is very rare for most restoration areas, particularly for long‐established restoration projects. We propose using fuzzy rule‐based expert systems to evaluate the degree of success of restoration actions when available information on project results and impacts largely relies on expert‐based qualitative assessments and rough estimates of quantitative values. These systems use fuzzy logic to manage the uncertainty present in the data and to integrate qualitative and quantitative information. To illustrate and demonstrate the potential of fuzzy rule‐based systems for restoration evaluation, we applied this approach to seven forest restoration projects implemented in Spain between 1897 and 1952, using information compiled in the REACTION database on Mediterranean forest restoration projects. The information available includes both quantitative and expert‐based qualitative data, and covers a wide variety of indicators grouped into technical, structural, functional, and socioeconomic criteria. The fuzzy rule‐based system translates expert knowledge of restoration specialists and forest managers into a set of simple logic rules that integrate information on individual indicators into more general evaluation criteria. The rule‐based approach proposed here can be readily applicable to any kind of restoration project, provided that some information, even if vague and uncertain, is available for a variety of assessment indicators. The evaluation of long‐established forest restoration projects implemented in Spain revealed important asymmetries in the degree of restoration success between technical, structural, functional, and socioeconomic criteria.     

Efficiency of fungus‐impregnated black cloths combined with Imidacloprid for the control of adult Aedes aegypti (Diptera: Culicidae)

Entomopathogenic fungi are potential candidates for use in integrated vector management. However, efficient delivery systems for these fungi need to be investigated. It is known that adult mosquitoes are attracted to dark surfaces, and therefore, black cotton cloths impregnated with Metarhizium anisopliae alone or in combination with the insecticide imidacloprid (IMI) were tested under laboratory conditions. Black cloths impregnated with fungus were also tested in large‐cage trials under natural extradomicile conditions. Blood‐fed Rockefeller and wild‐type strain Aedes aegypti had higher levels of survival when compared with sucrose‐fed counterparts following exposure to fungus‐impregnated cloths. However, when blood‐fed A. aegypti were exposed to a combination of M. anisopliae + IMI, the survival rates were statistically equal to those of sucrose‐fed females. Large‐cage trials showed significant decreases in A. aegypti survival following a minimum 12 h exposure of the mosquitoes to fungus‐impregnated cloths. Increased exposure times results in further reductions in survival. The synergism between M. anisopliae and IMI resulted in reduced survival rates independent of feeding regime under laboratory conditions. Fungus‐impregnated cloths tested under simulated field conditions, considered to be unfavourable for fungal infection, resulted in significant reductions in adult A. aegypti survival. We are currently testing the combined use of fungi and insecticides against blood‐fed insects under simulated field conditions.

Significance and Impact of the Study

The use of fungus‐impregnated cotton cloths is a promising point source application method for the control of adult Aedes aegypti, and this strategy could be incorporated into an integrated vector management programme aiming to reduce the incidence of dengue fever.     

An evaluation of methods for assessing the impacts of Bt‐maize MON810 cultivation and pyrethroid insecticide use on Auchenorrhyncha (planthoppers and leafhoppers)

1 Auchenorrhyncha (Planthoppers and Leafhoppers) are not only pests of many crops, but they are also nontarget organisms with respect to Bt‐protein expressing genetically modified plants. As herbivorous arthropods, planthoppers and leafhoppers ingest Cry proteins depending on their feeding behaviour. Consequently, they are directly exposed to these entomotoxic proteins and can also serve as a source of Cry protein exposure to predatory arthropods. Therefore, it is reasonable to use Auchenorrhyncha in the risk assessment of genetically modified crops. 2 During a 2‐year field study, we evaluated four different methods in terms of their feasibility to assess the impacts of plant‐incorporated protectants from Bt‐maize and of insecticide use on this group of arthropods. Visual assessment of plants, sweep netting, yellow traps and custom made sticky traps were utilized in field plots of Bt‐maize MON810, untreated near‐isogenic maize and insecticide‐treated near‐isogenic maize and were compared with respect to their capability to reflect the diversity and abundance of Auchenorrhyncha species. 3 Zyginidia scutellaris (Herrich‐Schäffer) (Cicadomorpha: Cicadellidae) represented more than 94% of all captured individuals in both years. The analysis of Z. scutellaris data showed no consistent differences between Bt‐maize MON810 and the untreated near isogenic hybrid, demonstrating no negative impact of MON810 on this species. Insecticide treatment, on the other hand, was not equivalent to the isogenic maize in terms of Z. scutellaris densities. Based on the collected data and on practical considerations, we recommend the combined use of transect‐wise sweep netting and sticky traps for the sampling of Auchenorrhyncha in maize.     

Semi‐field evaluation of a granulovirus and Bacillus thuringiensis ssp. kurstaki for season‐long control of the potato tuber moth,Phthorimaea operculella

There are few insecticidal options for potato tuber moth (PTM), Phthorimaea operculella Zeller (Lepidoptera: Gelechiidae), control late in the growing season. We evaluated the PTM granulovirus (PoGV) and Bacillus thuringiensis Berliner ssp. kurstaki (Btk) for season‐long control of PTM on potato foliage in 2006 and 2007. Compared to untreated controls, 10 weekly applications of PoGV (10¹³ occlusion bodies/ha) reduced PTM populations in replicated 1‐m³ field cages by 86–96% on pre‐harvest foliage and 90–97% on tubers added to cages shortly before harvest. Infection rates of 82–95% of L4 larvae by PoGV were noted within individual larval cohorts. Equivalently timed Btk treatments (1.12 kg product/ha) were significantly less effective at population suppression, with a 36–76% reduction in larvae recovered from tubers added to cages. A PoGV/Btk alternation was significantly more effective than Btk alone and as effective as PoGV in 2007, but not in 2006. There was some evidence that reduced rate PoGV treatments (10% rate or 50% application frequency) were less effective than the standard program. There were no treatment effects on percentage of tubers growing in the ground that were infested at harvest, which remained comparatively low at ≤8.1%. Bioassays were conducted to evaluate the residual activities of foliar deposits. Early‐season applications were highly effective for the first 24 h (≥93% mortality) with a steady decline in activity over 10 days. A second application, applied later in the season, showed similar patterns, although in this case Btk was less persistent than PoGV, whereas both agents provided significant larval mortality compared with controls over 14 days. Both PoGV and Btk provide alternatives to manage field infestations of PTM prior to harvest, thus reducing the risk of tuber infestations in storage.     

A host species‐informative internal control for molecular assessment of African swine fever virus infection rates in the African sylvatic cycle Ornithodoros vector

African swine fever virus (ASFV) infection in adult Ornithodoros porcinus (Murry 1877, sensu Walton 1979 ) ticks collected from warthog burrows in southern and East Africa was assessed using a duplex genomic amplification approach that is informative with respect to the invertebrate host species and infecting sylvatic cycle virus. DNA extracted from individual ticks was used as template for the simultaneous amplification of a C‐terminal 478‐bp ASFV p72 gene region and a ～313‐bp fragment of the tick mitochondrial 16S rRNA gene, under optimized reaction conditions. Within‐warthog burrow infection rates ranged from 0% to 43% using this approach, and phylogenetic analysis of 16S gene sequences revealed the presence of three geographically discrete O. porcinus lineages, but no support for subspecies recognition. False negatives are precluded by the inclusion of host species‐informative primers that ensure the DNA integrity of cytoplasmically located genome extracts. In addition, infection rate estimates are further improved as false positives arising from carry‐over contamination when performing a two‐step nested polymerase chain reaction are negated by the one‐step approach. Phylogenetic comparison of full‐length virus gene sequences with the partial C‐terminal p72 gene target confirmed the epidemiological utility of the latter in a sylvatic setting. The method is therefore of particular value in studies assessing the prevalence and diversity of ASFV in relation to the African sylvatic tick vector and holds potential for investigating the role of alternative tick species in virus maintenance and transmission.