Demographic patterns of human antibody levels to Simulium damnosum s.l. saliva in onchocerciasis-endemic areas: An indicator of exposure to vector bites

Background In onchocerciasis endemic areas in Africa, heterogenous biting rates by blackfly vectors on humans are assumed to partially explain age- and sex-dependent infection patterns with Onchocerca volvulus. To underpin these assumptions and further improve predictions made by onchocerciasis transmission models, demographic patterns in antibody responses to salivary antigens of Simulium damnosum s.l. are evaluated as a measure of blackfly exposure. Methodology/Principal findings Recently developed IgG and IgM anti-saliva immunoassays for S. damnosum s.l. were applied to blood samples collected from residents in four onchocerciasis endemic villages in Ghana. Demographic patterns in antibody levels according to village, sex and age were explored by fitting generalized linear models. Antibody levels varied between villages but showed consistent patterns with age and sex. Both IgG and IgM responses declined with increasing age. IgG responses were generally lower in males than in females and exhibited a steeper decline in adult males than in adult females. No sex-specific difference was observed in IgM responses. Conclusions/Significance The decline in age-specific antibody patterns suggested development of immunotolerance or desensitization to blackfly saliva antigen in response to persistent exposure. The variation between sexes, and between adults and youngsters may reflect differences in behaviour influencing cumulative exposure. These measures of antibody acquisition and decay could be incorporated into onchocerciasis transmission models towards informing onchocerciasis control, elimination, and surveillance.

Onchocerca volvulus transmission in the Mbam valley of Cameroon following 16 years of annual community-directed treatment with ivermectin, and the description of a new cytotype of Simulium squamosum

Background The onchocerciasis focus surrounding the lower Mbam and Sanaga rivers, where Onchocerca volvulus is transmitted by Simulium damnosum s.l. (Diptera: Simuliidae), was historically the largest in the southern regions of Cameroon. Annual community-directed treatment with ivermectin (CDTI) has been taking place since 2000, but recent studies have shown that new infections are occurring in children. We aimed to investigate blackfly biting and O. volvulus transmission rates along the lower Mbam river 16 years after the formal onset of annual CDTI. Methods Black flies were collected for three consecutive days each month between July 2016 and June 2017 at two riverside villages and two inland sites situated 4.9 km and 7.9 km from the riverside. Specimens collected at each site were dissected on one of the three collection days each month to estimate parity rates and O. volvulus infection rates, while the remaining samples were preserved for pool screening. Results In total, 93,573 S. damnosum s.l. black flies were recorded biting humans and 9281 were dissected. Annual biting rates of up to 606,370 were estimated at the riverside, decreasing to 20,540 at 7.9 km, while, based on dissections, annual transmission potentials of up to 4488 were estimated at the riverside, decreasing to 102 and 0 at 4.9 km and 7.9 km, respectively. However, pool screening showed evidence of infection in black flies at the furthest distance from the river. Results of both methods demonstrated the percentage of infective flies to be relatively low (0.10–0.36%), but above the WHO threshold for interruption of transmission. In addition, a small number of larvae collected during the dry season revealed the presence of Simulium squamosum E. This is the first time S. squamosum E has been found east of Lake Volta in Ghana, but our material was chromosomally distinctive, and we call it S. squamosum E2. Conclusions Relatively low O. volvulus infection rates appear to be offset by extremely high densities of biting black flies which are sustaining transmission along the banks of the lower Mbam river. Graphical Abstract

“Slash and Clear”, a Community-Based Vector Control Method to Reduce Onchocerciasis Transmission by Simulium sirbanum in Maridi, South Sudan: A Prospective Study

Background: High ongoing Onchocerca volvulus transmission was recently documented in Maridi County, South Sudan. To complement community-directed treatment with ivermectin (CDTI) as the main onchocerciasis control strategy, we initiated a community-based vector control method “slash and clear” at the Maridi dam, a Simulium damnosum s.l. breeding site, to reduce O. volvulus transmission. Methods: Simulium damnosum s.l. biting rates were collected before and during the twenty months following the “slash and clear” intervention using the human landing catches. Black flies were dissected to measure parity rates before and twelve months after the intervention. Larvae and pupae of S. damnosum s.l. were collected from the dam for morphological and chromosomal characterization to identify the cytospecies involved. Results: Biting rates of S. damnosum s.l. close to the Maridi dam spillway decreased by >90% post-“slash and clear” for more than six months. Twelve months after the “slash and clear” intervention, the reduction in biting rates was still at <50% (p = 0.0007). Parity rates reduced from 13% pre-“slash and clear” (November 2019) to 5.6% post-“slash and clear” (November 2020). Larvae collected from the dam were identified as Simulium sirbanum. Conclusion: The “slash and clear” method was found to be an effective and cheap community-based method to reduce black fly biting rates caused by S. sirbanum. When repeated at least annually together with a high CDTI coverage, this intervention has the potential to considerably accelerate onchocerciasis elimination.

Distribution of Fly Pests in a Farm in Plateau State, Nigeria

An evaluation of fly pests of cattle was conducted in Federal College of Animal Health and Production Technology (FCAH&PT) Farm, Chaha Village, Jos South Local Government Area, Plateau State, Nigeria between August and October, 2020. Samples were collected from the farm weekly for three months. A total of nine hundred and nineteen (919) adult dipterous flies comprising fifteen species in eight genera across the three sub-orders were caught and identified using aerial sweeping hand net, light trapping and aerial insecticide spray. Phlebotomus papaltasi (44.18%) was the most prevalent while Simulium damnosum (0.22%) was the least. Others were Musca domestica (17.73%), Stomoxys calcitrans (11.43%), Culicoides species (10.99%), Musca autumnalis (10.23%), mosquitoes – Anopheles gambiae/Culex quequinfasciatus (4.89%) and Tabanus species (0.33%). The haematophagous flies (72.04%) out-numbered the non-haematophagous flies (27.96%) and the number of females collected/caught 780 (84.87%) were higher than the males 139 (15.13%). The highest collections were made in October (353 representing 38.41%) while September had the lowest (263 /28.62%). The study established the presence of fly pests in the farm and recommends that appropriate control measures should be put in place to forestall any disease outbreak.

Seasonal Larval Abundance of the Blackfly, Simulium damnosum Complex and the Physico-chemical Properties of their Breeding Sites in Oji River System, Enugu State, Nigeria

Seasonal larval abundance of the blackfly, Simulium damnosum complex, and the physico-chemical properties of their breeding sites in Oji river system was carried out, with the aim of finding out the season that promotes the breeding of S. damnosum complex and the effect of the physico-chemical properties of the breeding sites of S. damnosum complex on larval abundance. Larvae of S. damnosum complex were collected monthly from forty-eight (48) breeding sites in four sampling units for three years, in dry and wet seasons. The physico-chemical properties of the breeding sites of S. damnosum complex studied were copper, dissolved oxygen, magnesium, nitrate, pH, phosphate, potassium, temperature, and water velocity. A total of six hundred and ninety one (691) larvae of S. damnosum complex were collected in the Oji river system in both seasons. Wet season catch was more and comprised 82% of the total catch, while dry season catch comprised 18% of the total catch. Dry and wet seasons catches in the sampling units were found to differ statistically. All the physico-chemical properties of the breeding sites of S. damnosum complex determined showed positive correlations with total larval abundance in both seasons, except copper which showed no correlation with total larval abundance in the dry season. Wet season promotes larval abundance of S. damnosum complex, and the physico-chemical properties of the breeding sites of S. damnosum complex play some roles (in varying degrees) in larval development.

Identification of the Sibling Species of the Blackfly, Simulium damnosum Complex Using their Polytene Chromosomal Banding Patterns in Oji River System, Enugu State, Nigeria

Identification of the sibling species of the blackfly, S. damnosum complex using their polytene chromosomal banding patterns in Oji river system was carried out with the aim of accurately identifying the particular sibling species of S. damnosum complex involved in the transmission of onchocerciasis in Oji-river LGA and environ. Simulium damnosum complex larvae were collected monthly in forty eight (48) breeding sites in four sampling units for three years in the Oji river system in wet and dry seasons. Collected larvae were preserved in cold Carnoy’s solution prior to processing. Salivary gland chromosomes were extracted, processed and analyzed microscopically for polytene chromosomal banding patterns. Three sibling species of Simulium damnosum complex that breed in the Oji river system were identified in varying degrees. They included S. squamosum, S. yahense and S. damnosum sensu stricto (s.s.). Variation in the occurrence of the sibling species was statistically significant. Dry and wet season catches for all the sibling species in the sampling units were found to differ statistically. The identified sibling species of S. damnosum complex are known to be vectors of Onchocerca volvulus, the causative agent of onchocerciasis, a disease highly prevalent in Oji-river LGA and environ.

Human immune response against salivary antigens of Simulium damnosum s.l.: A new epidemiological marker for exposure to blackfly bites in onchocerciasis endemic areas

Background Simulium damnosum sensu lato (s.l.) blackflies transmit Onchocerca volvulus, a filarial nematode that causes human onchocerciasis. Human landing catches (HLCs) is currently the sole method used to estimate blackfly biting rates but is labour-intensive and questionable on ethical grounds. A potential alternative is to measure host antibodies to vector saliva deposited during bloodfeeding. In this study, immunoassays to quantify human antibody responses to S. damnosum s.l. saliva were developed, and the salivary proteome of S. damnosum s.l. was investigated. Methodology/Principal findings Blood samples from people living in onchocerciasis-endemic areas in Ghana were collected during the wet season; samples from people living in Accra, a blackfly-free area, were considered negative controls and compared to samples from blackfly-free locations in Sudan. Blackflies were collected by HLCs and dissected to extract their salivary glands. An ELISA measuring anti-S. damnosum s.l. salivary IgG and IgM was optimized and used to quantify the humoral immune response of 958 individuals. Both immunoassays differentiated negative controls from endemic participants. Salivary proteins were separated by gel-electrophoresis, and antigenic proteins visualized by immunoblot. Liquid chromatography mass spectrometry (LC–MS/MS) was performed to characterize the proteome of S. damnosum s.l. salivary glands. Several antigenic proteins were recognized, with the major ones located around 15 and 40 kDa. LC–MS/MS identified the presence of antigen 5-related protein, apyrase/nucleotidase, and hyaluronidase. Conclusions/Significance This study validated for the first time human immunoassays that quantify humoral immune responses as potential markers of exposure to blackfly bites. These assays have the potential to facilitate understanding patterns of exposure as well as evaluating the impact of vector control on biting rates. Future studies need to investigate seasonal fluctuations of these antibody responses, potential cross-reactions with other bloodsucking arthropods, and thoroughly identify the most immunogenic proteins.

Biting behaviour and infectivity of Simulium damnosum complex with Onchocerca parasite in Alabameta, Osun State, Southwestern, Nigeria

A longitudinal study was carried out to investigate species composition, seasonal abundance, parity and transmission potential of Simulium damnosum complex in Alabameta community in Osun State, Southwestern, Nigeria. Adult Simulium damnosum complex were collected along Owena River, Alabameta, by two dark complexioned vector collectors from 07:00hr to 18:00hr weekly using collecting tubes from November 2014 to April 2015. The flies were morphologically identified and dissected for the purpose of detecting Onchocerca parasite using dissecting microscope. The Monthly Biting Rate (MBR) of flies was determined using World Health Organization standard formula. A total of four hundred and forty flies were collected during the study period with all of them identified as forest species of Simulium damnosum complex. There was significant variation in monthly collection of the flies with the month of November having the highest number of flies (194) (44%) while the month of April recorded the lowest number of flies (31) (7%) (p<0.05). The morning biting peak (09hr - 11hr) (137) was higher than the evening biting peak (15hr -17hr) (64) (p<0.05) while nulliparous flies (294) (67%) were more abundant than the parous flies (146) (33%) (p<0.05). There was absence of infection (zero infectivity) of the flies (p<0.05). The zero infectivity in the flies may plausibly indicate the possibility of zero transmission of Onchocerca parasite in the community which if sustained over a period of time may signify the possibility of onchocerciasis elimination. Also, the presence of forest species of the flies reduces the risk of resident’s intense exposure to blinding savannah strain of onchocerciasis.

An entomological study of onchocerciasis vectors, Simulium damnosum s.l., in Kinshasa, Democratic Republic of Congo

Background Onchocerciasis, a parasitic disease caused by the filarial worm Onchocerca volvulus transmitted through the bite of Simulium (black flies), is a cause of global concern, with the African population being majorly affected. This study focused on the bite rates, bite cycle, and transmission potential of Simulium damnosum s.l. in two sites with river blindness outbreaks in Kinshasa, DRC: Gombe (S1) and Mont-Ngafula at Kimwenza (S2). Methods From August 1, 2019, to July 31, 2020, we captured adult female black flies near breeding sites along the Congo River at S1 and Lukaya Valley at S2. Collections using human baits at the two sites were conducted for five days/month. Results A total of 6082 black flies of species Simulium squamosum (classified based on other entomological surveys) were captured during the study period. The daily cycle of aggression revealed two peaks: one between 8 and 9 a.m. and the other between 4 and 5 p.m. Low bite rates were observed between 11 a.m. and 1 p.m. The annual biting rate/person reached 13,463 in S1 and 23,638 in S2, with a total of 37,101 bites/person. The average daily biting rate, 37 ± 10 and 69 ± 23 bites/day/person in S1 and S2, respectively, did not differ significantly (P = 0.8901). The high density of the host population can disrupt the transmission of vector-borne diseases by diluting the transmission indices. There was no evidence of onchocerciasis transmission at the study sites because of inadequate laboratory facilities in the DRC. Various larval supports have been identified: at Gombe: aquatic plants, plastic bags, dead leaves, and rocks; at Kimwenza: Ledermaniella ledermanii (the most abundant species at the site), plastic bags, artificial waste, and aquatic plants. Conclusions The study provides further evidence for the need for alternative strategies to eliminate the parasite in the formerly hyper-endemic foci.