Florence A. Oyieke

Mechanical transmission of Trypanosoma evansi and T. congolense by Stomoxys niger and S. taeniatus in a laboratory mouse model.

Abstract.Mechanical transmission of Trypanosoma evansi (South American origin) and T. congolense of Kilifi DNA type (Kenyan origin) was studied in laboratory mice using the African stable flies Stomoxys niger niger and S. taeniatus. Altogether, 355 flies were interrupted after feeding on infected blood and then transferred immediately to an uninfected mouse to complete feeding. Microscopy and subinoculation of triturated flies into uninfected mice demonstrated the survival of T. congolense in Stomoxys for up to 210 min and T. evansi for up to 480 min. Parasites survived for much longer periods in the digestive tract than inside or on the mouthparts. Trypanosoma congolense was transmitted only by S. n. niger, and only at low rates of 3, 8 and 10% using flies of different feeding histories: fed on blood the previous day, freshly caught, and teneral. Trypanosoma evansi was transmitted by both Stomoxys species at higher rates: S. taeniatus range 13–18%;S. n. niger range 17–35%. The highest transmission rate occurred with the combination of teneral S. n. niger and T. evansi.

Electric nets and sticky materials for analysing oviposition behaviour of gravid malaria vectors

BackgroundLittle is known about how malaria mosquitoes locate oviposition sites in nature. Such knowledge is important to help devise monitoring and control measures that could be used to target gravid females. This study set out to develop a suite of tools that can be used to study the attraction of gravid Anopheles gambiae s.s. towards visual or olfactory cues associated with aquatic habitats.MethodsFirstly, the study developed and assessed methods for using electrocuting nets to analyse the orientation of gravid females towards an aquatic habitat. Electric nets (1m high × 0.5m wide) were powered by a 12V battery via a spark box. High and low energy settings were compared for mosquito electrocution and a collection device developed to retain electrocuted mosquitoes when falling to the ground. Secondly, a range of sticky materials and a detergent were tested to quantify if and where gravid females land to lay their eggs, by treating the edge of the ponds and the water surface. A randomized complete block design was used for all experiments with 200 mosquitoes released each day. Experiments were conducted in screened semi-field systems using insectary-reared An. gambiae s.s. Data were analysed by generalized estimating equations.ResultsAn electric net operated at the highest spark box energy of a 400 volt direct current made the net spark, creating a crackling sound, a burst of light and a burning smell. This setting caught 64% less mosquitoes than a net powered by reduced voltage output that could neither be heard nor seen (odds ratio (OR) 0.46; 95% confidence interval (CI) 0.40-0.53, p < 0.001). Three sticky boards (transparent film, glue coated black fly-screen and yellow film) were evaluated as catching devices under electric nets and the transparent and shiny black surfaces were found highly attractive (OR 41.6, 95% CI 19.8 – 87.3, p < 0.001 and OR 28.8, 95% CI 14.5 – 56.8, p < 0.001, respectively) for gravid mosquitoes to land on compared to a yellow sticky film board and therefore unsuitable as collection device under the e-nets. With a square of four e-nets around a pond combined with yellow sticky boards on average 33% (95% CI 28-38%) of mosquitoes released were collected. Sticky materials and detergent in the water worked well in collecting mosquitoes when landing on the edge of the pond or on the water surface. Over 80% of collected females were found on the water surface (mean 103, 95% CI 93–115) as compared to the edge of the artificial pond (mean 24, 95% CI 20–28).ConclusionA square of four e-nets with yellow sticky boards as a collection device can be used for quantifying the numbers of mosquitoes approaching a small oviposition site. Shiny sticky surfaces attract gravid females possibly because they are visually mistaken as aquatic habitats. These materials might be developed further as gravid traps. Anopheles gambiae s.s. primarily land on the water surface for oviposition. This behaviour can be exploited for the development of new trapping and control strategies.

Development of a Gravid Trap for Collecting Live Malaria Vectors Anopheles gambiae s.l.

Background Effective malaria vector control targeting indoor host-seeking mosquitoes has resulted in fewer vectors entering houses in many areas of sub-Saharan Africa, with the proportion of vectors outdoors becoming more important in the transmission of this disease. This study aimed to develop a gravid trap for the outdoor collection of the malaria vector Anopheles gambiae s.l. based on evaluation and modification of commercially available gravid traps. Methods Experiments were implemented in an 80 m2 semi-field system where 200 gravid Anopheles gambiae s.s. were released nightly. The efficacy of the Box, CDC and Frommer updraft gravid traps was compared. The Box gravid trap was tested to determine if the presence of the trap over water and the trap’s sound affected catch size. Mosquitoes approaching the treatment were evaluated using electrocuting nets or detergents added to the water in the trap. Based on the results, a new gravid trap (OviART trap) that provided an open, unobstructed oviposition site was developed and evaluated. Results Box and CDC gravid traps collected similar numbers (relative rate (RR) 0.8, 95% confidence interval (CI) 0.6–1.2; p = 0.284), whereas the Frommer trap caught 70% fewer mosquitoes (RR 0.3, 95% CI 0.2–0.5; p < 0.001). The number of mosquitoes approaching the Box trap was significantly reduced when the trap was positioned over a water-filled basin compared to an open pond (RR 0.7 95% CI 0.6–0.7; p < 0.001). This effect was not due to the sound of the trap. Catch size increased by 60% (RR 1.6, 1.2–2.2; p = 0.001) with the new OviART trap. Conclusion Gravid An. Gambiae s.s. females were visually deterred by the presence of the trapping device directly over the oviposition medium. Based on these investigations, an effective gravid trap was developed that provides open landing space for egg-laying Anopheles .

Mechanical transmission ofTrypanosoma evansiandT. congolensebyStomoxys nigerandS. taeniatusin a laboratory mouse model

Abstract.Mechanical transmission of Trypanosoma evansi (South American origin) and T. congolense of Kilifi DNA type (Kenyan origin) was studied in laboratory mice using the African stable flies Stomoxys niger niger and S. taeniatus. Altogether, 355 flies were interrupted after feeding on infected blood and then transferred immediately to an uninfected mouse to complete feeding. Microscopy and subinoculation of triturated flies into uninfected mice demonstrated the survival of T. congolense in Stomoxys for up to 210 min and T. evansi for up to 480 min. Parasites survived for much longer periods in the digestive tract than inside or on the mouthparts. Trypanosoma congolense was transmitted only by S. n. niger, and only at low rates of 3, 8 and 10% using flies of different feeding histories: fed on blood the previous day, freshly caught, and teneral. Trypanosoma evansi was transmitted by both Stomoxys species at higher rates: S. taeniatus range 13–18%;S. n. niger range 17–35%. The highest transmission rate occurred with the combination of teneral S. n. niger and T. evansi.

Habitat preferences and activity patterns of glossina swynnertoni Austen (diptera: glossinidae) in Aitong, Masai Mara, Kenya

The habitat preferences and activity patterns of Glossina swynnertoni Austen were studied using Siamese traps, electrified screens and vehicle patrols in Aitong, southwestern Kenya in three habitats: large thicket, wooded grassland and an Acacia community. Except in the Acacia community, G. swynnertoni occurred in association with Glossina pallidipes Austen. Apparent density varied significantly among the three habitats, being highest in the Acacia community, intermediate in wooded grassland and lowest in large thickets. Density also varied seasonally, with higher densities occurring in the rainy season than in the dry season in all three areas. Vehicle patrol was a more effective and rapid method for sampling G. swynnertoni than either Siamese traps or electrified screens. However, vehicle patrol catches were strongly biased in favour of males (4:1). Diurnal activity in the Acacia community was unimodal for both sexes, with peaks of activity occurring at 1100–1200 h for males and at 1400–1500 h for females. Both sexes remained active in the afternoon but activity declined rapidly towards dusk (1700–1800 h).RésuméLes préférences d’habitat et les rythmes d’activité de Glossina swynnertoni Austen ont été étudiés à l’aide de pièges de Siamese, d’écrans electrifies et de récoltes (patrouilles) en voiture, à Aitong, dans le Sud-ouest du Kenya dans trois habitats des grands bosquets, de la savane arborée et une forêt d’acacias. A l’exception de la forêt d’acacias, G. swynnertoni est rencontrée en association avec G. pallidipes Austen. La densité apparente varie significativement entre les trois habitats; elles est la plus forte dans la forêt d’acacias, intermédiaire en savane arborée et plus faible dans les grands bosquets. La densité varie également avec la saison, avec de plus fortes densités observées en saison des pluies par rapport à la saison sèche dans les trois habitats. Les captures réalisées lors des patrouilles en voiture sont plus efficaces et rapides pour l’échantillonnage de G. swynnertoni que les pièges de Siamese, et que les écrans électrifiés; ces captures sont cependant fortement biaisées en faveur des mâles (4:1). L’activité diurne dans la forêt d’acacias est unimodale pour les deux sexes, avec des pics d’activité entre 1100–1200 h pour les mâles et 1400–1500 pour les femelles. Les deux sexes restent actifs dans l’après-midi mais l’activité décline rapidement vers le crépuscule (1700–1800 h).

Malaria vector abundance is associated with house structures in Baringo County, Kenya

Malaria, a major cause of morbidity and mortality, is the most prevalent vector borne disease in Baringo County; a region which has varied house designs in arid and semi-arid areas. This study investigated the association between house structures and indoor-malaria vector abundance in Baringo County. The density of malaria vectors in houses with open eaves was higher than that for houses with closed eaves. Grass thatched roof houses had higher density of malaria vectors than corrugated iron sheet roofs. Similarly, mud walled houses had higher vector density than other wall types. Houses in the riverine zone were significantly associated with malaria vector abundance (p<0.000) possibly due to more varied house structures. In Kamnarok village within riverine zone, a house made of grass thatched roof and mud wall but raised on stilts with domestic animals (sheep/goats) kept at the lower level had lower mosquito density (5.8 per collection) than ordinary houses made of same materials but at ground level (30.5 mosquitoes per collection), suggestive of a change in behavior of mosquito feeding and resting. House modifications such as screening of eaves, improvement of construction material and building stilted houses can be incorporated in the integrated vector management (IVM) strategy to complement insecticide treated bed nets and indoor residual spray to reduce indoor malaria vector density.

Status of insecticide resistance in malaria vectors in Kwale County, Coastal Kenya

BackgroundThe strategy for malaria vector control in the context of reducing malaria morbidity and mortality has been the scale-up of long-lasting insecticidal nets to universal coverage and indoor residual spraying. This has led to significant decline in malaria transmission. However, these vector control strategies rely on insecticides which are threatened by insecticide resistance. In this study the status of pyrethroid resistance in malaria vectors and it’s implication in malaria transmission at the Kenyan Coast was investigated.ResultsUsing World Health Organization diagnostic bioassay, levels of phenotypic resistance to permethrin and deltamethrin was determined. Anopheles arabiensis showed high resistance to pyrethroids while Anopheles gambiae sensu stricto (s.s.) and Anopheles funestus showed low resistance and susceptibility, respectively. Anopheles gambiae sensu lato (s.l.) mosquitoes were further genotyped for L1014S and L1014F kdr mutation by real time PCR. An allele frequency of 1.33% for L1014S with no L1014F was detected. To evaluate the implication of pyrethroid resistance on malaria transmission, Plasmodium falciparum infection rates in field collected adult mosquitoes was determined using enzyme linked immunosorbent assay and further, the behaviour of the vectors was assessed by comparing indoor and outdoor proportions of mosquitoes collected. Sporozoite infection rate was observed at 4.94 and 2.60% in An. funestus s.l. and An. gambiae s.l., respectively. A higher density of malaria vectors was collected outdoor and this also corresponded with high Plasmodium infection rates outdoor.ConclusionsThis study showed phenotypic resistance to pyrethroids and low frequency of L1014S kdr mutation in An. gambiae s.l. The occurrence of phenotypic resistance with low levels of kdr frequencies highlights the need to investigate other mechanisms of resistance. Despite being susceptible to pyrethroids An. funestus s.l. could be driving malaria infections in the area.

Secondary bacterial infections and antibiotic resistance among tungiasis patients in Western, Kenya

Tungiasis or jigger infestation is a parasitic disease caused by the female sand flea Tunga penetrans. Secondary infection of the lesions caused by this flea is common in endemic communities. This study sought to shed light on the bacterial pathogens causing secondary infections in tungiasis lesions and their susceptibility profiles to commonly prescribed antibiotics. Participants were recruited with the help of Community Health Workers. Swabs were taken from lesions which showed signs of secondary infection. Identification of suspected bacteria colonies was done by colony morphology, Gram staining, and biochemical tests. The Kirby Bauer disc diffusion test was used to determine the drug susceptibility profiles. Out of 37 participants, from whom swabs were collected, specimen were positive in 29 and 8 had no growth. From these, 10 different strains of bacteria were isolated. Two were Gram positive bacteria and they were, Staphylococcus epidermidis (38.3%) and Staphylococcus aureus (21.3%). Eight were Gram negative namely Enterobacter cloacae (8.5%), Proteus species (8.5%), Klebsiellla species (6.4%), Aeromonas sobria (4.3%), Citrobacter species (4.3%), Proteus mirabillis(4.3%), Enterobacter amnigenus (2.1%) and Klebsiella pneumoniae (2.1%). The methicillin resistant S. aureus (MRSA) isolated were also resistant to clindamycin, kanamycin, erythromycin, nalidixic acid, trimethorprim sulfamethoxazole and tetracycline. All the Gram negative and Gram positive bacteria isolates were sensitive to gentamicin and norfloxacin drugs. Results from this study confirms the presence of resistant bacteria in tungiasis lesions hence highlighting the significance of secondary infection of the lesions in endemic communties. This therefore suggests that antimicrobial susceptibility testing may be considered to guide in identification of appropriate antibiotics and treatment therapy among tungiasis patients.

Mechanical transmission ofTrypanosoma evansiandT. congolensebyStomoxys nigerandS. taeniatusin a laboratory mouse model: Mechanical transmission by Stomoxys

Abstract.Mechanical transmission of Trypanosoma evansi (South American origin) and T. congolense of Kilifi DNA type (Kenyan origin) was studied in laboratory mice using the African stable flies Stomoxys niger niger and S. taeniatus. Altogether, 355 flies were interrupted after feeding on infected blood and then transferred immediately to an uninfected mouse to complete feeding. Microscopy and subinoculation of triturated flies into uninfected mice demonstrated the survival of T. congolense in Stomoxys for up to 210 min and T. evansi for up to 480 min. Parasites survived for much longer periods in the digestive tract than inside or on the mouthparts. Trypanosoma congolense was transmitted only by S. n. niger, and only at low rates of 3, 8 and 10% using flies of different feeding histories: fed on blood the previous day, freshly caught, and teneral. Trypanosoma evansi was transmitted by both Stomoxys species at higher rates: S. taeniatus range 13–18%;S. n. niger range 17–35%. The highest transmission rate occurred with the combination of teneral S. n. niger and T. evansi.