Insecticidal effects of some selected plant extracts against Anopheles stephensi (Culicidae: Diptera).

BACKGROUND: The use of synthetic insecticides against mosquitoes may lead to resistance development and potential health hazards in humans and the environment. Consequently, a paradigm needs to shift towards the alternative use of botanical insecticides that could strengthen an insecticide resistance management programme. This study aimed to assess the insecticidal effects aqueous, hexane, and methanol crude leaf extracts of Calpurnia aurea, Momordica foetida, and Zehneria scabra on an insectary colony of Anopheles stephensi larvae and adults. METHODS: Fresh leaves of C. aurea, M. foetida and Z. scabra were collected and dried, then separately ground to powder. Powdered leaves of test plants were extracted using sonication with aqueous, hexane, and methanol solvents. The extracts were concentrated, and a stock solution was prepared. For comparison, Temephos (Abate®) and control solutions (a mixture of water and emulsifier) were used as the positive and negative controls, respectively. Different test concentrations for the larvae and the adults were prepared and tested according to WHO (2005) and CDC (2010) guidelines to determine lethal concentration (LC) values. Mortality was observed after 24 h exposure. The statistical analyses were performed using Statistical Package for the Social Sciences (SPSS) software (Kruskal-Wallis test) and R software (a generalized linear model was used to determine LC50 and LC90 values of the extracts). RESULTS: The lowest LC50 values were observed in aqueous extracts of M. foetida followed by Z. scabra extract and C. aurea leaves at 34.61, 35.85, and 38.69 ppm, respectively, against the larvae. Larval mortality was not observed from the hexane extracts and negative control, while the standard larvicide (temephos) achieved 100% mortality. Further, the adulticidal efficacy was greatest for aqueous extract of Z. scabra with LC50 = 176.20 ppm followed by aqueous extract of C. aurea (LC50 = 297.75 ppm). CONCLUSION: The results suggest that the leaf extracts of the three test plants have the potential of being used for the control of vector An. stephensi larvae and adult instead of synthetic mosquitocides. Further studies need to be conducted to identify the active ingredients and their mode of action.

An update on the distribution, bionomics, and insecticide susceptibility of Anopheles stephensi in Ethiopia, 2018-2020.

BACKGROUND: Anopheles stephensi, an invasive malaria vector, was first detected in Africa nearly 10 years ago. After the initial finding in Djibouti, it has subsequently been found in Ethiopia, Sudan and Somalia. To better inform policies and vector control decisions, it is important to understand the distribution, bionomics, insecticide susceptibility, and transmission potential of An. stephensi. These aspects were studied as part of routine entomological monitoring in Ethiopia between 2018 and 2020. METHODS: Adult mosquitoes were collected using human landing collections, pyrethrum spray catches, CDC light traps, animal-baited tent traps, resting boxes, and manual aspiration from animal shelters. Larvae were collected using hand-held dippers. The source of blood in blood-fed mosquitoes and the presence of sporozoites was assessed through enzyme-linked immunosorbent assays (ELISA). Insecticide susceptibility was assessed for pyrethroids, organophosphates and carbamates. RESULTS: Adult An. stephensi were collected with aspiration, black resting boxes, and animal-baited traps collecting the highest numbers of mosquitoes. Although sampling efforts were geographically widespread, An. stephensi larvae were collected in urban and rural sites in eastern Ethiopia, but An. stephensi larvae were not found in western Ethiopian sites. Blood-meal analysis revealed a high proportion of blood meals that were taken from goats, and only a small proportion from humans. Plasmodium vivax was detected in wild-collected An. stephensi. High levels of insecticide resistance were detected to pyrethroids, carbamates and organophosphates. Pre-exposure to piperonyl butoxide increased susceptibility to pyrethroids. Larvae were found to be susceptible to temephos. CONCLUSIONS: Understanding the bionomics, insecticide susceptibility and distribution of An. stephensi will improve the quality of a national response in Ethiopia and provide additional information on populations of this invasive species in Africa. Further work is needed to understand the role that An. stephensi will have in Plasmodium transmission and malaria case incidence. While additional data are being collected, national programmes can use the available data to formulate and operationalize national strategies against the threat of An. stephensi.

Exposure to DDT and HCH congeners and associated potential health risks through khat (Catha edulis) consumption among adults in South Wollo, Ethiopia.

Khat (Catha edulis) chewing is widespread in the region of East Africa. Even low levels of organochlorine pesticides (OCPs) in khat could induce public health concern. In a market-based study, from five popular khat varieties, a total of 35 composite khat samples were analyzed for dichlorodiphenyltrichloroethane (DDT) and its main transformation products, and four hexachlorocyclohexane (HCH) isomers. Extraction was carried out by quick, easy, cheap, effective, rugged and safe method (QuEChERS). OCP concentrations were determined by head space solid phase microextraction coupled to gas chromatography-mass spectrometry (HS-SPME-GC-MS). Every sample contained ß-HCH above the maximum residue limit set by the European Commission. For total DDT, this was the case for 25.7% of the samples. The ratios of (p,p'-DDD + p,p'-DDE) to p,p'-DDT were less than one for 85% of khat samples, demonstrating recent use of DDT in khat farmlands. Conversely, the ratio of ß-HCH to total HCH varied from 0.56 to 0.96, implying historical input of technical HCH. Assuming a daily chewable portion of 100 g, dietary intakes of p,p'-DDT, total DDT and total HCH by adults ranged from 3.12 to 57.9, 6.49 to 80.2 and 39.2 to 51.9 ng (kg body weight)-1 day-1, respectively. These levels are below acceptable levels suggested by international organizations. Chewing khat showed lower non-cancer health risk, but showed relatively higher cancer risk in terms of OCPs. Because khat is chewed without being subjected to any treatment, uncertainties associated with estimated intakes and health risks should be low. Therefore, this practice is of great concern.

Ecology and Infection Status of Sand Flies in Rural and Urban Cutaneous Leishmaniasis Endemic Areas in Northwest Ethiopia.

Cutaneous leishmaniasis (CL) caused by Leishmania aethiopica is transmitted by Phlebotomus longipes in northern Ethiopia. No studies have been conducted to investigate the transmission dynamics of CL, despite its high endemicity in both rural and urban settings. Evidence on the ecology and behavior of the vector from this area are required to develop integrated disease control strategies. Sand flies were collected in the dry and wet seasons in 2021 in CL-endemic rural Gindmeteaye and urban Addis-Alem in northwest Ethiopia. Trapping was performed with sticky and Centers for Disease Control and Prevention (CDC) light traps in three habitats, including inside patients' houses, peridomestic areasand in caves/rocky areas. Sand flies were morphologically identified to species level. Female Phlebotomus species were categorized according to blood feeding status and tested by spliced-leader (SL-) ribonucleic acid (RNA) polymerase chain reaction (PCR) to screen for Leishmania infection. Of 1161 sand flies, the majority (77%) were P. longipes, six (0.5%) were P. orientalis and the remaining were Sergentomyia. The abundance of the 430 female P. longipes was significantly linked to seasonality (p < 0.001), with the majority in the dry season occurring in the outdoor rocky (37%) and peridomestic (34%) sites, while, in the wet season, most (62%) were captured indoors. This seasonality was more pronounced in rural Gindmeteaye, where housing construction is poor. The number of blood-fed and gravid P. longipes was significantly higher in the wet (31%; 22%), compared to the dry season (13%; 8%), and their proportion was highest indoors. Eighteen (4%) female P. longipes were Leishmania positive, with highest infection prevalence in caves (7% compared to 3% indoors, p = 0.022), and in the dry season (6%, p < 0.001). Phlebotomus orientalis specimens were all captured in May in rural Gindmeteaye, five indoors and one in a peridomestic site. Further research should be conducted to investigate the absolute contribution of humans and indoor transmission to the transmission cycle of CL. Inhabitants of endemic villages should be made aware that evening outdoor activities near caves may increase their exposure to infectious sand flies. Whether P. orientalis can breed and become infected at high altitudes should be further studied.

Phlebotomine sandflies (Diptera: Psychodidae) of Ethiopia.

Phlebotomine sandflies have a long history of association with humans, which makes them the only proven natural vectors of Leishmania species, the parasitic protozoans that cause leishmaniases in humans and animals. In Ethiopia, the three forms of leishmaniases, viz., visceral, dermal and mucocutaneous are endemic in different parts of the country. Since the first report of phlebotomine sandflies in Ethiopia in 1936, the distribution of different species and their role in the transmission of leishmaniases have been extensively studied. The objective of this review was to summarize the patchy information and give an updated list of phlebotomine sandfly species in Ethiopia and their known geographical distribution in the country. Peer-reviewed literature search was conducted using online databases. All articles published which focus on distribution and medical importance of Phlebotomus and Sergentomyia species of Ethiopia starting from 1936 up to 2022 were reviewed. Until July 2022, 65 phlebotomine sandfly species have been reported, belonging to the genus Phlebotomus and Sergentomyia. The genus Phlebotomus in Ethiopia is represented by six subgenera such as Adlerius, Anaphlebotomus, Larroussius, Paraphlebotomus Phlebotomus and Synphlebotomus, whereas the genus Sergentomyia is represented by six subgenera, namely Grassomyia, Parrotomyia, Parvidens, Rondanomyia, Sergentomyia, and Sintonius.

Cutaneous leishmaniasis in Kutaber District, Ethiopia: Prevalence, sand fly fauna and community knowledge, attitude and practices.

Background: Cutaneous leishmaniasis (CL) affects 25% of the population living in the highlands of Ethiopia. CL intervention has not decreased the number of leishmaniasis patients. A cross-sectional study was conducted to determine CL prevalence, community's knowledge, attitude and practices (KAP), and the sand fly fauna in Kutaber district, northeast Ethiopia. Methods: A retrospective, community-based cross-sectional study was conducted in Boru Meda Hospital from December 2014-March 2021 to study CL prevalence of Kutaber district. A Pre-tested, well-structured questionnaire was used to collect data on the participants' socio-demographic characteristics, KAP towards CL and knowledge about sand fly vectors. Chi-square test and logistic regression analysis were used in the study, and data were analyzed using SPSS version 23 (p < 0.05). Results: A total of 10,002 (14.02%), of which 71,325 samples were confirmed as positive for CL. The infection rate of CL in females (7.1%) was a little bit higher than males (7.0%). More cases were recorded among 15-29 age category. The study also revealed that 77.1% of the respondents had poor knowledge about CL treatment, prevention, clinical presentation and disease transmission. Farmers tended to have poorer knowledge about sand flies than non-workers and students (32.7 vs. 35 and 44.1%; P = 0.049). Housewives had poorer knowledge about sand flies than farmers and workers (22.2 vs. 32.7 and 33.3%; P = 0.023). Phlebotomus longipes comprised the highest composition (80%) of the sand fly species identified in Kutaber district. Conclusions: The data showed that the community had poor knowledge about CL, vector, and transmission mode. CL preventive measures were prevalent, implying the need to raise CL awareness. Phlebotomus longipes was identified as the most dominant sand fly species which accounted for CL. The findings can be used in developing an effective control strategy to reduce CL transmission in the study area and elsewhere in Ethiopia.

Sero-prevalence of visceral leishmaniasis and its associated factors among asymptomatic individuals visiting Denan health center, southeastern Ethiopia.

BACKGROUND: In the Somali region of Ethiopia, visceral leishmaniasis (VL) is a public health concern. However, VL epidemiology and sand fly vectors have not been well studied in various areas of the regional state, including Denan district. Therefore, this study was conducted to determine the sero-prevalence, associated factors, and distribution of sand fly vectors of VL in Denan district, south-eastern Ethiopia. METHODS: A facility-based cross-sectional study was conducted from April to September 2021 among VL patients with classic signs and symptoms visiting Denan Health Center in south-eastern Ethiopia. Using a convenience sampling method, 187 blood samples were collected from individuals who visited Denan Health Center during the study period. Blood samples were subjected to Direct Agglutination Test for the detection of antibodies to VL. A pre-tested structured questionnaire was also used to gather information on risk factors and other characteristics of knowledge and attitude assessment. Sand flies were also collected from indoor, peri-domestic, mixed forest, and termite mounds using light and sticky traps to determine the fauna and abundance. RESULTS: The overall sero-prevalence rate was 9.63% (18/187). The sero-prevalence was significantly associated with outdoor sleeping (OR = 2.82), the presence of damp floors (OR = 7.76), and sleeping outdoor near animals (OR = 3.22). Around 53.48% of the study participants had previously heard about VL. Study participants practiced different VL control methods, including bed nets (42%), insecticide spraying (32%), smoking plant parts (14%), and environmental cleaning (8%). In total, 823 sand fly specimens, comprising 12 species in two genera (Phlebotomus and Sergentomyia), were trapped and identified. The most abundant species was Sergentomyia clydei (50.18%), followed by Phlebotomus orientalis (11.42%). Also, a higher proportion of P. orientalis was found in termite mounds (65.43%), followed by mixed forest (37.8%) and peri-domestic (20.83%) habitats. CONCLUSION: The study demonstrated a 9.63% sero-positivity of VL and a remarkable gap in knowledge, attitude, and practices towards VL. P. orientalis was also detected, which could be a probable vector in this area. Thus, public education should be prioritized to improve the community's awareness of VL and its public health impact. In addition, detailed epidemiological and entomological studies are recommended.

Laboratory Evaluation of Efficacy of the Larvicide Spinosad Against AnophelesStephensi in Jigjiga, Ethiopia.

We report the efficacy of a commercial formulation of the insecticide spinosad against larvae of Anopheles stephensi populations found in the city of Jigjiga, Somali Region, eastern Ethiopia. Batches of 25 larvae (late III to early IV instars) collected from large water storage reservoirs associated with construction sites (the primary An. stephensi larval site in the dry season) were tested under laboratory conditions against each insecticide at a dose recommended by the manufacturer (Natular® G30, 0.02 g/5 liter), following World Health Organization guidelines. Mortality at 24-48 h postexposure was 100%. Results show that spinosad is effective against An. stephensi larvae and suggest that it may be a useful tool as part of larval source management plans aimed at controlling this invasive malaria vector in Ethiopia.

Building the vector in: construction practices and the invasion and persistence of Anopheles stephensi in Jigjiga, Ethiopia.

Anopheles stephensi is a major vector of malaria in Asia and the Arabian Peninsula, and its recent invasion into Africa poses a major threat to malaria control and elimination efforts on the continent. The mosquito is well adapted to urban environments, and its presence in Africa could potentially lead to an increase in malaria transmission in cities. Most of the knowledge about An stephensi ecology in Africa has been generated from studies conducted during the rainy season, when vectors are most abundant. Here, we provide evidence from the peak of the dry season in the city of Jigjiga in Ethiopia, and report An stephensi immature stages infesting predominantly in water reservoirs made to support construction operations (ie, in construction sites or associated with brick-manufacturing businesses). Political and economic changes in Ethiopia (particularly the Somali Region) have fuelled an unprecedented construction boom since 2018 that, in our opinion, has been instrumental in the establishment, persistence, and propagation of An stephensi via the year-round availability of perennial larval habitats associated with construction. We argue that larval source management during the dry season might provide a unique opportunity for focused control of An stephensi in Jigjiga and similar areas.

Analysis of the Knockdown Resistance Locus (kdr) in Anopheles stephensi, An. arabiensis, and Culex pipiens s.l. for Insight Into the Evolution of Target-site Pyrethroid Resistance in Eastern Ethiopia.

The malaria vector, Anopheles stephensi, which is typically restricted to South Asia and the Middle East, was recently detected in the Horn of Africa. Addressing the spread of this vector could involve integrated vector control that considers the status of insecticide resistance of multiple vector species in the region. Previous reports indicate that the knockdown resistance mutations (kdr) in the voltage-gated sodium channel (vgsc) are absent in both pyrethroid-resistant and pyrethroid-sensitive An. stephensi in eastern Ethiopia; however, similar information about other vector species in the same areas is limited. In this study, kdr and the neighboring intron were analyzed in An. stephensi, An. arabiensis, and Culex pipiens s.l. collected between 2016 and 2017 to determine the evolutionary history of kdr in eastern Ethiopia. A sequence analysis revealed that all of Cx. pipiens s.l. (N = 42) and 71.6% of the An. arabiensis (N = 67) carried kdr L1014F, which is known to confer target-site pyrethroid resistance. Intronic variation was only observed in An. stephensi (six segregating sites, three haplotypes), which was previously shown to have no kdr mutations. In addition, no evidence of non-neutral evolutionary processes was detected at the An. stephensi kdr intron, thereby further supporting the target-site mechanism not being a major resistance mechanism in this An. stephensi population. Overall, these results show key differences in the evolution of target-site pyrethroid/dichlorodiphenyltrichloroethane resistance mutations in populations of vector species from the same region. Variations in insecticide resistance mechanism profiles between eastern Ethiopian mosquito vectors may lead to different responses to insecticides used in integrated vector control.

Epidemiological, Entomological, and Climatological Investigation of the 2019 Dengue Fever Outbreak in Gewane District, Afar Region, North-East Ethiopia.

Dengue Fever (DF) is an important arthropod-borne viral infection that has repeatedly occurred as outbreaks in eastern and northeastern Ethiopia since 2013. A cross-sectional epidemiological outbreak investigation was carried out from September to November 2019 on febrile patients (confirmed malaria negative) who presented with suspected and confirmed DF at both public and private health facilities in Gewane District, Afar Region, northeastern Ethiopia. Entomological investigation of containers found in randomly selected houses belonging to DF-positive patients was undertaken to survey for the presence of Aedes larvae/pupae. A total of 1185 DF cases were recorded from six health facilities during the 3-month study period. The mean age of DF cases was 27.2 years, and 42.7% of cases were female. The most affected age group was 15−49 years old (78.98%). The total case proportions differed significantly across age groups when compared to the population distribution; there were approximately 15% and 5% higher case proportions among those aged 15−49 years and 49+ years, respectively. A total of 162 artificial containers were inspected from 62 houses, with 49.4% found positive for Aedes aegypti larva/pupae. Aedes mosquitoes were most commonly observed breeding in plastic tanks, tires, and plastic or metal buckets/bowls. World Health Organization entomological indices classified the study site as high risk for dengue virus outbreaks (House Index = 45.2%, Container Index = 49.4%, and Breteau Index = 129). Time series climate data, specifically rainfall, were found to be significantly predictive of AR (p = 0.035). Study findings highlight the importance of vector control to prevent future DF outbreaks in the region. The scarcity of drinking water and microclimatic conditions may have also contributed to the occurrence of this outbreak.

Malaria prevalence, knowledge, attitude, and practice among febrile patients attending Chagni health center, Northwest Ethiopia: a cross-sectional study.

BACKGROUND: Ethiopia has achieved considerable progresses in the prevention and control of malaria in the past decades; hitherto it is a formidable health concern and socio-economic impediment. This study aimed at assessing the magnitude, knowledge, attitudes and practices towards malaria among febrile patients attending Chagni health center, northwest Ethiopia. METHODS: Health facility-based cross-sectional study was conducted to estimate the prevalence of malaria and KAP towards malaria among febrile patients at Chagni health center in Chagni Town during September 2017 to February 2018. In order to determine the magnitude of malaria, finger prick blood samples were collected and thick and thin smears were prepared and microscopically examined for the presence of malaria parasites. A pre-tested structured questionnaire was also applied to assess KAP of suspected malaria patients, attending the health center. Data were analyzed using SPSS version 20.0. RESULTS: Prevalence of malaria among febrile patients, who visited the sampled health facility, was 7.3%. Of these, Plasmodium falciparum, P. vivax, and mixed infections accounted for 55, 44.3 and 0.7% of the cases, respectively. This study also revealed that 97% of the respondents had ever heard about malaria and recognized it as a serious health problem. Mosquito bite was identified as the main malaria transmission. Taking drug (86.3%), use of mosquito nets (73.3%), drain stagnated water (68%), and house spay with insecticides (66%) were mentioned as the main malaria prevention methods. Mosquito net coverage and utilizations in the prior night were 98 and 75%, respectively. Indoor residual spraying (IRS) coverage was 99%, of which 77.5% of study participants' houses have been sprayed in the last 6 months. CONCLUSIONS: The current study revealed that prevalence of malaria among febrile illnesses in the study area was relatively low (7.3%) with a high proportion of P. falciparum. Besides, participants had adequate knowledge, encouraging attitudes, and good practices about prevention and control of malaria. However, some misconceptions on malaria disease, its transmission, and prevention have been noted that actually require due attention by the concerned stakeholders. The findings of this study could be used as important inputs for the implementation of effective malaria prevention and control methods, including community health education programs, and scaling up coverage of evidence-based interventions.

Physico-chemical characteristics of honey produced by stingless bees (Meliponula beccarii) from West Showa zone of Oromia Region, Ethiopia.

The study was investigated to characterize the physico-chemical properties of honey produced from stingless bees (Meliponula beccarii) in Oromia Region of Ethiopia. Honey samples were collected from underground soils through an excavation of natural nests from four representative districts (Wolmera, Jeldu, Toke Kutaye and Chalyia). About 120 honey samples (0.35-1.5 lit volumes) were collected and pooled for each respective area (district) for analysis. Principal component analysis (PCA) of pollen data and GPS were used to categorize the botanical origin and geographical location of honey sample sources, respectively. Consecutively, the physico-chemical characters were evaluated for the moisture content (MC), electrical conductivity (EC), pH, ash, total acidity (TA), hydroxyl methyl furfuraldhyde (HMF), insoluble materials (IM), and proline. The study results showed that the mean values of parameters for honey samples were: MC (29 ± 1.45), EC (0.22 ± 0.03), pH (3.73 ± 0.13), Ash (0.41 ± 1.1), TA (7.3 ± 0.36), HMF (18 ± 3.45), IM (0.70 ± 0.3) and proline (214.5 ± 15). There was no mean significance differences (P > 0.05) observed among the sampled districts for the analyzed parameters, except for proline. The values were compared against the standard physico-chemical parameters of Apis mellifera honey of Ethiopia and these values significantly varied for moisture content, total acidity and HMF. This is the first study investigated on composition property of stingless bee honey in Ethiopia and recall for further study on its nutritional and medicinal values so as to create a specific standardization.

A five year trend analysis of malaria prevalence in Guba district, Benishangul-Gumuz regional state, western Ethiopia: a retrospective study.

BACKGROUND: In Ethiopia, malaria is a serious public health concern and has great impact on socio-economy. The trend analysis of malaria data from health facilities is useful for understanding its transmission dynamics and implementing evidence-based malaria control strategies. The aim of this study was to determine the trends of malaria infection in Guba district, western Ethiopia. METHODS: A retrospective study was undertaken at Mankush Health Centre, western Ethiopia. All malaria cases reported from 2014 to 2018 were carefully reviewed from the laboratory record books to determine the trends of malaria morbidity. Data were analyzed using SPSS version 20.0. RESULTS: In total, 16,964 malaria suspects were diagnosed using microscopy over the last 5 years, of which 8658 (51.04%) were confirmed positive cases. Plasmodium falciparum, P. vivax, and mixed infection (both species) accounted for 75.2, 24.5 and 0.28% of the cases, respectively. Males patients were more affected (n = 5028, 58.1%) than female ones (n = 3630, 41.9%). Of the total confirmed cases, 60.4% were age group of subjects (≥ 15 years) followed by 22.6% of 5-14 years and 15.9% of under 5 years. High malaria prevalence was observed in spring (September to November) season, while the least was observed in autumn (March to May) with the prevalence of 45.6 and 11.5%, respectively. CONCLUSIONS: The study demonstrated that malaria is a public health concern, in which P. falciparum is the predominant species followed by P. vivax. Therefore, the district health bureau and other concerned stakeholders should strength evidence-based malaria control and prevention interventions to interrupt disease transmission and eventual reduction malaria of malaria cases in Guba district.

Environmental Factors Associated with Larval Habitats of Anopheline Mosquitoes (Diptera: Culicidae) in Metema District, Northwestern Ethiopia.

BACKGROUND: Malaria is one of the major public health concerns in Ethiopia. Control options available for the management of malaria, include case detection, personal protection and larval source management. Effective control of Anopheles larvae largely depends on understanding of the habitats of the vectors. The aims of this study were to identify the breeding habitats of mosquitoes and characterize the larval habitats in Gende Wuha Town in northwestern Ethiopia. METHODS: Different aquatic habitats were sampled and characterized for anopheline larvae from November 2012 to June 2013. RESULTS: In total, 2784 larvae of Anopheles mosquitoes were collected from various breeding habitats. Microscopic identification of the III and IV instars revealed the presence of seven Anopheles species. Of the Anopheles spp, Anopheles gambiae s.l. (80%) was the most predominant species in the study area. Spearman correlation coefficient results also determined that the density of An. gambiae s.l. increased significantly with habitat temperature (r= 0.346, p< 0.01). Significantly higher An. gambiae s.l. larvae were obtained in non-shaded habitats (z= -3.120, p< 0.05) when compared with shaded habitats. CONCLUSION: The current study demonstrated An. gambiae s.l., the principal malaria vector in the country, is the predominant species in the larval sampling habitats. It was also noted the importance of edge of stream as larva breeding habitats for this species during the dry season of the year. Therefore, attention should be given for this breeding habitat for control of the vector during dry season.

A molecular analysis of sand fly blood meals in a visceral leishmaniasis endemic region of northwestern Ethiopia reveals a complex host-vector system.

BACKGROUND: Visceral leishmaniasis (VL, or "kala-azar") is a major cause of disability and death, especially in East Africa. Its vectors, sand flies (Diptera: Psychodidae: Phlebotominae), are poorly controlled and guarded against in these regions, owing in part to a lack of understanding about their feeding behavior. METHODS: A total of 746 freshly fed female sand flies were collected in five population centers in Kafta Humera (northwestern Ethiopia), where VL is endemic. Flies were collected from habitats that ranged from inside houses to open fields, using light traps and sticky traps. Sources of sand fly blood meals were identified using enzyme-linked immunosorbent assays (ELISA) and DNA amplification with reverse-line blot analysis (PCR-RLB); 632 specimens were screened using ELISA, 408 of which had identifiable blood meals, and 114 were screened using PCR-RLB, 53 of which yielded identifications. Fly species determinations were based on morphology, and those specimens subjected to PCR-RLB were also screened for Leishmania parasites using conventional PCR to amplify the nuclear marker ITS1 (internal transcribed spacer 1) with Leishmania-specific primers. RESULTS: More than three-fourths of all sand flies collected were Phlebotomus orientalis, and the remaining portion was comprised of nine other species. Nearly two-thirds of P. orientalis specimens were collected at village peripheries. The most common blood source for all flies was donkey (33.9% of all identifications), followed by cow (24.2%), human (17.6%), dog (11.8%), and goat or sheep (8.6%); mixtures of blood meals from different sources were found in 28.2% of all flies screened. Unidentified blood meals, presumably from wildlife, not domestic animals, were significantly higher in farm fields. Leishmania parasites were not detected in any of the 114 flies screened, not surprising given an expected infection rate of 1-5 out of 1,000. Meals that included a mixture of human and cow blood were significantly more frequent relative to all cow meals than human blood meals were to non-cow meals, suggesting a zoopotentiative interaction between cows and humans in this system. CONCLUSIONS: Habitat and host preferences of sand fly vectors in Kafta Humera confirmed the finding of previous reports that the main vector in the region, Phlebotomus orientalis, is a highly opportunistic feeder that prefers large animals and is most commonly found at village peripheries. These results were similar to those of a previous study conducted in a nearby region (Tahtay Adiabo), except for the role of cattle on the prevalence of human blood meals. Preliminary examinations of blood meal data from different settings point to the need for additional surveys and field experiments to understand the role of livestock on biting risks.

First detection of Anopheles stephensi Liston, 1901 (Diptera: culicidae) in Ethiopia using molecular and morphological approaches.

Malaria is a major public health concern in Ethiopia. With the increase in malaria cases in the Somali Region of Ethiopia, understanding the distribution and identifying the species of malaria vectors is vital to public health. Here we report the first detection of Anopheles stephensi in Ethiopia, a malaria vector typically found in the Middle East, the Indian subcontinent, and China, but recently found in Djibouti. An entomological investigation was conducted during November to December 2016 in Kebri Dehar town of the Ethiopian Somali Regional State as ancillary work for Anopheles spp. surveillance. Mosquito larvae were collected from water reservoirs. Larvae were reared in the laboratory to the adult stage and identified morphologically. PCR and sequencing of cytochrome oxidase 1 (COI) and internal transcribed spacer 2 (ITS2) loci were performed. Basic Local Alignment Search Tool (BLAST) was used to compare sample sequences to sequences in the NCBI nucleotide database for species identification. To further analyze the relationship between the specimen we collected in Kebri Dehar and other Anopheles samples available in Genbank, phylogenetic analysis was performed using a maximum likelihood approach. Molecular and morphological results confirm specimens were An. stephensi. The closest high scoring hit was for all specimens was for the An. stephensi sequence. Independent phylogenetic analyses of COI and ITS2 sequences revealed in both cases strong bootstrap (100) support for our sequence forming a clade with other An. stephensi sequences to the exclusion of any other species of Anopheles. In conclusion, Anopheles stephensi is present in Kebri Dehar town in Ethiopia. These findings highlight the need for additional research to examine the role of An. stephensi in malaria transmission in Ethiopia.

Some aspects of entomological determinants of Phlebotomus orientalis in highland and lowland visceral leishmaniasis foci in northwestern Ethiopia.

Visceral leishmaniasis (VL) is one of the major public health problems in northwest Ethiopia, mainly in Libo-Kemkem and Metema districts, where Phlebotomus orientalis is the most probable vector of the disease. The aim of this study was to determine the physiological age, host preference and vectorial potential of P. orientalis in the highland and lowland foci of the region. Sand flies were collected using CDC light traps between May 2011 and April 2012 in Libo-Kemkem and October 2012 and September 2013 in Metema from household compounds, farm field and mixed forest. Females belonging to Phlebotomus were dissected for physiological age determination and Leishmania detection and isolation. Leishmania infections in sand flies were investigated using molecular methods. Freshly fed Phlebotomus females were tested to identify blood meal sources using PCR-RLB and ELISA. A total of 1149 (936 from Libo-Kemkem and 213 from Metema) blood unfed female P. orientalis were dissected for age determination. The parity rate was 45.6% and 66.2% in Libo-Kemkem and Metema, respectively. None of 798 female P. orientalis dissected (578 from Libo-Kemkem and 220 from Metema) was infected with Leishmania parasites. A total of 347 P. orientalis specimens collected from Libo-Kemkem were processed using PCR, of which 10 (2.8%) specimens were found with DNA of Leishmania spp. Of a total 491 freshly fed female P. orientalis analyzed for blood meal origins by RLB-PCR and ELISA, 57.6% (67.8% from Libo-Kemkem and 49.8% from Metema) were found to contain bovine blood while 4.9% (3.7% from Libo-Kemkem and 5.7% from Metema) were of human blood. In conclusion, the present study showed parity difference between the two populations of P. orientalis and that both populations have strong zoophilic behavior. Based on the presented evidences, the species is strongly implicated as a vector of kala-azar in both areas. Therefore, vector control should be a component of a strategy to manage visceral leishmaniasis in both study areas.

A fine scale eco-epidemiological study on endemic visceral leishmaniasis in north ethiopian villages.

Visceral Leishmaniasis (VL) is a disseminated protozoan infection caused by Leishmania donovani that affects almost half a million people annually. In Northern Ethiopia, VL is common in migrant agricultural laborers returning from the lowland sesame fields of Metema and Humera. Recent VL foci have emerged in resident rural populations near the town. In the current study, we evaluate multilevel entomological, epidemiological and ecological factors associated with infection and disease through fine-scale eco-epidemiological analyses in three villages. Satellite images showed that villages constructed in or close to vertisols, were likely to become endemic for VL. Vertisols or black-cotton soil, are characterized by high contents of smectitic clay minerals, which swell when hydrated and shrink upon desiccation, causing extensive deep cracking during the dry season. The population densities of Phlebotomus orientalis, the vector, were negatively correlated with distance from vertisols and persons living close to vertisols were more likely to be bitten by sand flies, as evidenced by sero-positivity to Ph. orientalis saliva. Apparent (albeit non-significant) clustering of VL cases and abundant asymptomatic infections close to vertisols, suggest anthroponotic transmission around houses located close to vertisols. Comparable rates of male and female volunteers, mostly under 15 years of age, were infected with L. donovani but a significantly higher proportion of males succumbed to VL indicating a physiological gender-linked male susceptibility. Our data suggest that the abundant infected persons with high parasitemias who remain asymptomatic, may serve as reservoir hosts for anthroponotic transmission inside villages. Only limited insights on the transmission dynamics of L. donovani were gained by the study of environmental factors such as presence of animals, house structure and vegetation cover.