Black soldier fly larvae oil (Hermetia illucens L.) calcium salt enhances intestinal morphology and barrier function in laying hens.

This study aimed to determine the influence of black soldier fly larvae oil calcium salt (BSFLO-SCa) supplementation on performance, jejunal histomorphology and gene expression of tight junctions and inflammatory cytokines in laying hens. A total of 60 ISA Brown laying hens (40 wk of age) were divided into 3 treatment groups, including a control group fed a basal diet (T0) and basal diets supplemented with 1% (T1) and 2% (T2) of BSFLO-SCa. Each treatment group consisted of 5 replicates with 4 laying hens each. Results showed that 1% and 2% BSFLO-SCa supplementation significantly reduced (P < 0.05) feed conversion ratio (FCR), while egg weight (EW) increased (P < 0.05). The inclusion with 2% increased (P < 0.05) both egg production (HDA) and mass (EM). The addition of 1% and 2% BSFLO-SCa significantly increased (P < 0.05) villus height (VH) and villus width (VW), while crypt depth (CD) significantly increased (P < 0.05) with 2% BSFLO-SCa. The tight junction and gene expression of claudin-1 (CLDN-1), junctional adhesion molecules-2 (JAM-2), and occludin (OCLN) were significantly upregulated (P < 0.05) with 2% BSFLO-SCa. The pro-inflammatory cytokines and gene expression of interleukin-6 (IL-6) was significantly downregulated (P < 0.05) with the addition of BSFLO-SCa, while gene expression of interleukin-18 (IL-18), toll-like receptor 4 (TLR-4), and tumor necrosis factor-α (TNF-α) were downregulated with 2% BSFLO-SCa. On the other hand, the anti-inflammatory cytokines and gene expression of interleukin-13 (IL-13) and interleukin-10 (IL-10) were significantly upregulated (P < 0.05) at 2% BSFLO-SCa. In conclusion, dietary supplementation with 2% BSFLO-SCa improved productivity, intestinal morphology and integrity by upregulating tight junction-related protein of gene expression of laying hens. In addition, supplementation with BSFLO-SCa enhanced intestinal immune responses by upregulating anti-inflammatory and downregulating pro-inflammatory cytokine gene expression.

The cluster digging behavior of larvae confers trophic benefits to fitness in insects.

Collective behaviors efficiently impart benefits to a diversity of species ranging from bacteria to humans. Fly larvae tend to cluster and form coordinated digging groups under crowded conditions, yet understanding the rules governing this behavior is in its infancy. We primarily took advantage of the Drosophila model to investigate cooperative foraging behavior. Here, we report that Drosophila-related species and the black soldier fly have evolved a conserved strategy of cluster digging in food foraging. Subsequently, we investigated relative factors, including larval stage, population density, and food stiffness and quality, that affect the cluster digging behavior. Remarkably, oxygen supply through the posterior breathing spiracles is necessary for the organization of digging clusters. More importantly, we theoretically devise a mathematical model to accurately calculate how the cluster digging behavior expands food resources by diving depth, cross-section area, and food volume. We found that cluster digging behavior approximately increases 2.2 fold depth, 1.7-fold cross-section area, and 1.9 fold volume than control groups, respectively. Amplification of food sources significantly facilitates survival, larval development, and reproductive success of Drosophila challenged with competition for limited food resources, thereby conferring trophic benefits to fitness in insects. Overall, our findings highlight that the cluster digging behavior is a pivotal behavior for their adaptation to food scarcity, advancing a better understanding of how this cooperative behavior confers fitness benefits in the animal kingdom.

Potential global distribution of Aleurocanthus woglumi considering climate change and irrigation.

Citrus blackfly, Aleurocanthus woglumi Ashby (Hemiptera: Aleyrodidae), is an important agricultural quarantine pest, causing substantial economic losses to citrus and many other cultivated crops. Aleurocanthus woglumi is found in tropical and subtropical regions but is presently unknown in Europe and the Mediterranean Basin. We used CLIMEX to model the potential distribution of A. woglumi under an historical climate scenario (centred on 1995), including a spatially explicit irrigation scenario. We found that A. woglumi could potentially invade the Mediterranean Basin, and south-east Asia, including Australia. There is potential for it to invade most of sub-Saharan Africa. Irrigation is revealed as an important habitat factor affecting the potential distribution of A. woglumi, increasing its potential range by 53% in Asia. Under a future climate scenario for 2050, its potential distribution increased across all continents except Africa, where potential range expansion due to relaxation of cold stresses was limited, and was offset by range decrease due to lethal heat or dry stress. As global climates warm, Europe is likely to face a substantial increase in the area at risk of establishment by A. woglumi (almost doubling under the 2050 irrigation scenario). The biosecurity threat from A. woglumi is significant in current citrus production areas and poses a challenge to biosecurity managers and risk analysts.

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.

Evaluating the diagnostic test accuracy of molecular xenomonitoring methods for characterising the community burden of Onchocerciasis.

BACKGROUND: Molecular xenomonitoring (MX), the detection of parasite nucleic acid in the vector population, is recommended for onchocerciasis surveillance in elimination settings. However, the sensitivity of MX for detecting onchocerciasis-positive communities has not previously been evaluated. MX may have additional applications for control programmes but its utility is restricted by a limited understanding of the relationship between MX results and human prevalence. METHODS: We conducted a systematic review of studies reporting the prevalence of Onchocerca volvulus DNA in wild-caught Simulium spp. flies (MX rate) and corresponding prevalence of microfilaria (mf) in humans. We evaluated the sensitivity of MX for detecting onchocerciasis-positive communities and describe the characteristics of studies with reduced sensitivity. We conducted a linear regression to evaluate the relationship between mf prevalence and MX rate. RESULTS: We identified 15 relevant studies, with 13 studies comprising 34 study communities included in the quantitative analyses. Most communities were at advanced stages towards elimination and had no or extremely low human prevalence. MX detected positive flies in every study area with >1% mf prevalence, with the exception of one study conducted in the Venezuelan Amazonian focus. We identified a significant relationship between the two measurements, with mf prevalence accounting for half of the variation in MX rate (R2 0.50, p<0.001). CONCLUSION: MX is sensitive to communities with ongoing onchocerciasis transmission. It has potential to predict human mf prevalence, but further data is required to understand this relationship, particularly from MX surveys conducted earlier in control programmes before transmission has been interrupted.

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.

Atypical behavior of a black fly species connects cavity-nesting birds with generalist blood parasites in an arid area of Spain.

BACKGROUND: The feeding behavior of bloodsucking insects determines the transmission, distribution, host spectrum and evolution of blood parasites in the wild. Conventional wisdom suggests that some vector groups (e.g. black flies, family Simuliidae) are consistently exophagous daytime biters. We aimed to understand more about the exceptions to this pattern by combining targeted trapping and molecular identification of parasites in vectors. METHODS: In this study, we collected black flies in nest boxes used by European rollers Coracias garrulus in southeastern Spain. We molecularly analyzed 434 individual insects, identifying the black fly species caught in the nest boxes, their potential vertebrate blood meals, and the haemosporidian parasite lineages that they carried. RESULTS: Only one black fly species, Simulium rubzovianum, appeared to enter the nest boxes of rollers. Among the trapped specimens, 15% contained vertebrate DNA, which always belonged to rollers, even though only half of those specimens were visibly engorged. Furthermore, 15% of all black flies contained Leucocytozoon lineages, indicating previous feeding on avian hosts but probably not on infected adult rollers. The known vertebrate hosts of the recorded Leucocytozoon lineages suggested that large and/or abundant birds are their hosts. Particularly represented were cavity-nesting species breeding in the vicinity, such as pigeons, corvids and owls. Open-nesting species such as thrushes and birds of prey were also represented. CONCLUSIONS: Our data strongly suggest that S. rubzovianum bites uninfected roller nestlings and infected individuals of other species, potentially incubating adults, inside nest boxes and natural cavities. This simuliid does not appear to have a strong preference for specific host clades. Contrary to the general pattern for the group, and possibly enhanced by the harsh environmental conditions in the study area, this black fly appeared to intensively use and may even have a preference for confined spaces such as cavities for feeding and resting. Preferences of vectors for atypical microhabitat niches where hosts are less mobile may enable social and within-family transmission and parasite speciation in the long term. At the same time, a lack of host preference in concentrated multispecies communities can lead to host switches. Both processes may be underappreciated driving forces in the evolution of avian blood parasites.

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.

The Mbam drainage system and onchocerciasis transmission post ivermectin mass drug administration (MDA) campaign, Cameroon.

BACKGROUND: The impact of large scale Mass Drug Adminstration (MDA) of ivermectin on active onchocerciasis transmission by Simulium damnosum, which transmits the parasite O. volvulus is of great importance for onchocerciasis control programmes. We investigated in the Mbam river system area, the impact of MDA of ivermectin on entomological indices and also verify if there are river system factors that could have favoured the transmission of onchocerciasis in this area and contribute to the persistence of disease. We compared three independent techniques to detect Onchocerca larvae in blackflies and also analyzed the river system within 9 months post-MDA of ivermectin. METHOD: Simulium flies were captured before and after 1, 3, 6 and 9months of ivermectin-MDA. The biting rate was determined and 41% of the flies dissected while the rest were grouped into pools of 100 flies for DNA extraction. The extracted DNA was then subjected to O-150 LAMP and real-time PCR for the detection of infection by Onchocerca species using pool screening. The river system was analysed and the water discharge compared between rainy and dry seasons. PRINCIPAL FINDINGS: We used human landing collection method (previously called human bait) to collect 22,274 adult female Simulium flies from Mbam River System. Of this number, 9,134 were dissected while 129 pools constituted for molecular screening. Overall biting and parous rates of 1113 flies/man/day and 24.7%, respectively, were observed. All diagnostic techniques detected similar rates of O. volvulus infection (P = 0.9252) and infectivity (P = 0.4825) at all monitoring time points. Onchocerca ochengi larvae were only detected in 2 of the 129 pools. Analysis of the river drainage revealed two hydroelectric dams constructed on the tributaries of the Mbam river were the key contributing factor to the high-water discharge during both rainy and dry seasons. CONCLUSION: Results from fly dissection (Microscopy), real-time PCR and LAMP revealed the same trends pre- and post-MDA. The infection rate with animal Onchocerca sp was exceptionally low. The dense river system generate important breeding sites that govern the abundance of Simulium during both dry and rainy seasons.

Optimization of Slash and Clear Community-Directed Control of Simulium damnosum Sensu Stricto in Northern Uganda.

Onchocerciasis, caused by infection with Onchocerca volvulus, has been targeted for elimination by 2030. Currently, onchocerciasis elimination programs rely primarily on mass distribution of ivermectin. However, ivermectin alone may not be sufficient to achieve elimination in some circumstances, and additional tools may be needed. Vector control has been used as a tool to control onchocerciasis, but vector control using insecticides is expensive and ecologically detrimental. Community-directed removal of the trailing vegetation black fly larval attachment sites (slash and clear) has been shown to dramatically reduce vector biting densities. Here, we report studies to optimize the slash and clear process. Conducting slash and clear interventions at Simulium damnosum sensu stricto breeding sites located within 2 km of afflicted communities resulted in a 95% reduction in vector biting. Extending slash and clear further than 2 km resulted in no further decrease. A single intervention conducted at the first half of the rainy season resulted in a 97% reduction in biting rate, whereas an intervention conducted at the end of the rainy season resulted in a 94% reduction. Vector numbers in any of the intervention villages did not fully recover by the start of the following rainy season. These results suggest that slash and clear may offer an inexpensive and effective way to augment ivermectin distribution in the effort to eliminate onchocerciasis in Africa.

Blood parasites in vectors reveal a united blackfly community in the upper canopy.

BACKGROUND: The behaviour of blood-sucking arthropods is a crucial determinant of blood protozoan distribution and hence of host-parasite coevolution, but it is very challenging to study in the wild. The molecular identification of parasite lineages in vectors can be a useful key to understand the behaviour and transmission patterns realised by these vectors. METHODS: In this study, we collected blackflies around nests of three raptor species in the upper forest canopy in central Europe and examined the presence of vertebrate DNA and haemosporidian parasites in them. We molecularly analysed 156 blackfly individuals, their vertebrate blood meals, and the haemosporidian parasite lineages they carried. RESULTS: We identified nine species of Simulium blackflies, largely belonging to the subgenera Nevermannia and Eusimulium. Only 1% of the collected specimens was visibly engorged, and only 4% contained remains of host DNA. However, in 29% of the blackflies Leucocytozoon lineages were identified, which is evidence of a previous blood meal on an avian host. Based on the known vertebrate hosts of the recorded Leucocytozoon lineages, we can infer that large and/or abundant birds, such as thrushes, crows, pigeons, birds of prey, owls and tits are the main targets of ornithophilic blackflies in the canopy. Blackfly species contained similar proportions of host group-specific parasite lineages and thus do not appear to be associated with particular host groups. CONCLUSIONS: The Leucocytozoon clade infecting thrushes, crows, and pigeons present in most represented blackfly species suggests a lack of association between hosts and blackflies, which can increase the probability of host switches of blood parasites. However, the composition of the simuliid species differed between nests of common buzzards, goshawks and red kites. This segregation can be explained by coinciding habitat preferences between host and vector, and may lead to the fast speciation of Leucocytozoon parasites. Thus, subtle ecological preferences and lack of host preference of vectors in the canopy may enable both parasite diversification and host switches, and enforce a habitat-dependent evolution of avian malaria parasites and related haemosporidia.

Blackfly fever and dermatitis caused by Simulium kiritshenkoi: a human case report in Iran.

BACKGROUND: Besides the considerable role of blackflies to the transmission of many disease agents, these flies considered as annoying biting pests of wildlife, livestock, poultry, and humans. There are few literature reports of blackfly fever following Simulium spp. This study describes a case of blackfly fever and dermatitis following blackflies numerous bites in Iran. CASE PRESENTATION: The present report describes a 25-year-old man that was attacked by numerous flies while fishing and camping near Namrood river in Firuzkuh County, Tehran Province, Iran. Pruritic dermatitis with marked edema appeared mainly on the hands and legs and subsequently, the patient's condition worsened with swollen lymph nodes, joints aching, and 40 °C fever. The patient's clinical signs and symptoms were alleviated by injection of intramuscular Dexamethasone Phosphate (DEXADIC®) 8 mg/2 ml after 24 h. CONCLUSIONS: This study reported a human case with blackfly fever and dermatitis following numerous bites of Simulium kiritshenkoi, for the first time in Iran.

Optimization of Hermetia illucens (L.) egg laying under different nutrition and light conditions.

The black soldier fly Hermetia illucens is gaining growing interest as a tool for the valorisation of bio-waste in a circular economy perspective. Although a wide variety of studies are available for larvae rearing, the indoor breeding of adults still presents a great challenge for industrial purposes. This study was designed to assess the simultaneous influence of 3 different light sources (the Mix of LED UV:blue:green 1:1:3, White LED, Neon light) and 3 types of nutrition (sugar and water, only water, no sugar no water) on adult performances, obtaining different egg production parameters that included the number and weight of the egg masses and single eggs laid by the females, the duration of pre-oviposition and oviposition period, the adult life span, the hatchability of the eggs. Our results showed that production parameters are influenced mainly by nutrition rather than light factor, although light plays an important secondary role. Moreover, the presence of sugar positively affects the egg production (12,93-27,10 mg eggs/female) and increases oviposition period (18,2-31,8 days) and adult lifespan (20,79-27,11 days). Light sources also affect egg production parameters, with the exposure to the Mix of LEDs resulting in the best performance of flies. Results obtained from this study are very useful for the design and management of an efficient industrial black soldier fly mass rearing process.

The Galabat-Metema cross-border onchocerciasis focus: The first coordinated interruption of onchocerciasis transmission in Africa.

BACKGROUND: Onchocerciasis transmission across international borders is not uncommon, yet a coordinated cross border stops mass drug administration (MDA) decision has not been documented. METHODS/PRINCIPLE FINDINGS: The Galabat-Metema focus involves neighboring districts on the border between Sudan and Ethiopia. Mass drug administration (MDA) was provided once and subsequently twice per year in this focus, with twice-per-year beginning in Ethiopia's Metema subfocus in 2016 and in the Sudan's Galabat subfocus in 2008. Ov16 ELISA-based serosurveys were conducted in 6072 children under 10 years of age in the Metema subfocus in 2014, and 3931 in the Galabat in 2015. Between 2014 and 2016, a total of 27,583 vector Simulium damnosum flies from Metema and 9,148 flies from Galabat were tested by pool screen PCR for Onchocerca volvulus O-150 DNA. Only 8 children were Ov16 seropositive (all in the Metema subfocus); all were negative by skin snip PCR. The upper limit of the 95% confidence interval (UCL) for Ov16 seropositive was <0.1% for the overall focus and 0.14 positive fly heads per 2000 (UCL = 0.39/2000). However, an entomological 'hotspot' was detected on the Wudi Gemzu river in Metema district. The hotspot was confirmed when 4 more positive fly pools were found on repeat testing in 2017 (1.04 L3/2000 flies (UCL = 2.26/2000). Information exchange between the two countries led to stopping MDA in a coordinated fashion in 2018, with the exception of the hotspot at Wudi Gemzu, where MDA with ivermectin was increased to every three months to hasten interruption of transmission. CONCLUSION: Coordinated stop MDA decisions were made by Sudan and Ethiopia based on data satisfying the World Health Organization's criteria for interruption of onchocerciasis transmission. Definitions of entomological 'hotspots' and buffer zones around the focus are proposed.

Ecology of black flies (Diptera: Simuliidae) in streams of northern and southern Thailand: Factors associated with larval and pupal distributions.

The ecology of black flies in Thailand was investigated, based on 19,451 larvae and pupae collected from 65 stream sites in 10 northern provinces during the rainy, cool, and hot seasons, and 1,906 larvae and pupae collected from 18 sites in 9 southern provinces during the cool season. Twenty-seven black fly species were identified from northern Thailand, of which 26 were found in the cool season, when richness was greatest. Significant regressions between species richness and elevation fit a unimodal model in the rainy season but a linear model in the cool and hot seasons. Twenty-two species occurred in all seasons. Species in the subgenera Gomphostilbia and Nevermannia were most common in the hot season, whereas species in the subgenus Simulium were predominant in the cool season. Some species (e.g., S. nakhonense) were geographically widespread, whereas others (e.g., S. chaliowae and S. weji) were restricted to particular localities. Eighteen species and species complexes were found in southern Thailand. The S. tani complex was the most widely distributed taxon, occurring at 66.7% of the sites in the South. Ecological analyses revealed that water temperature, elevation, conductivity, dissolved oxygen, and stream size were among the significant factors associated with the distributions of black flies in both regions of Thailand-the same factors associated with simuliid distributions in other areas of the world.

Biting Rates and Onchocerca Infectivity Status of Black Flies from the Simulium damnosum Complex (Diptera: Simuliidae) in Osun State, Nigeria.

The Simulium damnosum Theobald complex transmits Onchocerca volvulus Leuckart (Spirurida: Onchocercidae), the causative agent of onchocerciasis. Recent evidence suggests that control efforts have strongly suppressed parasite populations, but vector surveillance is needed in parts of Africa where the disease remains endemic. Here, studies on biting rates and infectivity status of suspected vector species were conducted in three onchocerciasis-endemic areas, namely Iwo, Ede, and Obokun, in Osun State, Nigeria. A total of 3,035 black flies were collected between October 2014 and September 2016, and examined for parity and parasites using standard methods. A separate collection of 2,000 black flies was pool-screened for infectivity using polymerase chain reaction (PCR) amplification of the O-150 marker. Results showed that parous flies were significantly less common than nulliparous flies with overall parous rates of 8.02% in Iwo and 35.38% in Ede at the end of the study period. Obokun had a parous rate of 22.22% obtained in the first year only. None of the dissected parous flies were infected with O. volvulus and PCR assays showed no amplification of O-150 O. volvulus-specific repeats in head and body pools. However, annual biting rates exceeded the World Health Organization threshold of 1,000 bites/person/yr. Thus it appears that, with such high rates of biting, even low levels of vector infection can sustain onchocerciasis in African communities.

Effect of moisture content on greenhouse gas and NH3 emissions from pig manure converted by black soldier fly.

The effects of different moisture contents on greenhouse gas (GHG) emissions from pig manure (PM) digested by black soldier fly larvae (BSFL) as well as the accompanying changes of nitrogen and carbon contents in gaseous emissions and residues were studied. A mixture of PM and corncob at the ratio of 2.2:1 was prepared with a moisture content of 45%. Then, distilled water was added to adjust the moisture contents of the mixture to 55%, 65%, 75% and 85%, respectively. The prepared mixtures were digested by BSFL for eight days. The results indicated that BSFL could reduce CH4, N2O and NH3 emissions respectively by 72.63-99.99%, 99.68%-99.91% and 82.30-89.92%, compared with conventional composting, while CO2 emissions increased potentially due to BSFL metabolism. With increasing moisture content, the cumulative CH4 emissions increased, while cumulative NH3 emissions peaked at 55% moisture content and then decreased. Interestingly, the tendency of total cumulative CO2 emissions was consistent with that of the total weight of BSFL. The total GHG emissions were about only 1% those from of traditional composting at the optimum moisture content (75%), which was the most favorable for the growth of BSFL. The nitrogen and carbon contents of BSFL content in all treatments accounted for 1.03%-12.67% and 0.25%-4.68% of the initial contents in the raw materials, respectively. Moreover, the residues retained 71.12%-90.58% carbon and 67.91%-80.39% nitrogen of the initial raw materials. Overall, our results suggest that BSFL treatment is an environment-friendly alternative for decreasing CH4, N2O and NH3 emissions as well as reducing global warming potential (GWP).

The nutritive value of black soldier fly larvae reared on common organic waste streams in Kenya.

In Africa, livestock production currently accounts for about 30% of the gross value of agricultural production. However, production is struggling to keep up with the demands of expanding human populations, the rise in urbanization and the associated shifts in diet habits. High costs of feed prevent the livestock sector from thriving and to meet the rising demand. Insects have been identified as potential alternatives to the conventionally used protein sources in livestock feed due to their rich nutrients content and the fact that they can be reared on organic side streams. Substrates derived from organic by-products are suitable for industrial large-scale production of insect meal. Thus, a holistic comparison of the nutritive value of Black Soldier Fly larvae (BSFL) reared on three different organic substrates, i.e. chicken manure (CM), brewers' spent grain (SG) and kitchen waste (KW), was conducted. BSFL samples reared on every substrate were collected for chemical analysis after the feeding process. Five-hundred (500) neonatal BSFL were placed in 23 × 15 cm metallic trays on the respective substrates for a period of 3-4 weeks at 28 ± 2 °C and 65 ± 5% relative humidity. The larvae were harvested when the prepupal stage was reached using a 5 mm mesh size sieve. A sample of 200 grams prepupae was taken from each replicate and pooled for every substrate and then frozen at -20 °C for chemical analysis. Samples of BSFL and substrates were analyzed for dry matter (DM), crude protein (CP), ether extracts (EE), ash, acid detergent fibre (ADF), neutral detergent fibre (NDF), amino acids (AA), fatty acids (FA), vitamins, flavonoids, minerals and aflatoxins. The data were then subjected to analysis of variance (ANOVA) using general linear model procedure. BSFL differed in terms of nutrient composition depending on the organic substrates they were reared on. CP, EE, minerals, amino acids, ADF and NDF but not vitamins were affected by the different rearing substrates. BSFL fed on different substrates exhibited different accumulation patterns of minerals, with CM resulting in the largest turnover of minerals. Low concentrations of heavy metals (cadmium and lead) were detected in the BSFL, but no traces of aflatoxins were found. In conclusion, it is possible to take advantage of the readily available organic waste streams in Kenya to produce nutrient-rich BSFL-derived feed.

Characterization of IgE-binding proteins in the salivary glands of Simulium nigrogilvum (Diptera: Simuliidae).

Simulium dermatitis is an IgE-mediated skin reaction in animals and humans caused by the bites of black flies. Although Simulium nigrogilvum has been incriminated as the main human-biting black fly species in Thailand, information on its salivary allergens is lacking. Salivary gland extract of S. nigrogilvum females was subjected to sodium dodecylsulfate-polyacrylamide gel electrophoresis, and the separated components were applied onto nitrocellulose membranes for immunoblotting, which was performed by probing the protein blots with sera from 17 individuals who were allergic to the bites of S. nigrogilvum. IgE-reactive protein bands were characterized further by liquid chromatography-mass spectrometry (LC-MS/MS) analysis. Nine protein bands (79, 42, 32, 25, 24, 22, 15, 13, and 11 kDa) were recognized in the serum of the subjects. Four of the nine protein bands (32, 24, 15, and 11 kDa) showed IgE reactivity in all (100%) of the tested sera, and they were identified as salivary secreted antigen 5-related protein, salivary serine protease, erythema protein, and hypothetical secreted protein, respectively. Three other proteins, salivary serine protease (25 kDa), salivary D7 secreted protein (22 kDa), and hypothetical protein (13 kDa), reacted with > 50% of the sera. The relevance of the identified protein bands as allergens needs to be confirmed by using pure recombinant proteins, either in the in vivo skin prick test or in vitro detection of the specific IgE in the serum samples of allergic subjects. This will be useful for the rational design of component-resolved diagnosis and allergen immunotherapy for the allergy mediated by the bites of black flies.

Optimization and evaluation of the Esperanza Window Trap to reduce biting rates of Simulium damnosum sensu lato in Northern Uganda.

BACKGROUND: Onchocerciasis, or river blindness, has historically been an important cause of blindness, skin disease and economic disruption in Africa and the Americas. It is caused by the filarial parasite Onchocerca volvulus, which is transmitted by black flies in the genus Simulium. Over the past decade, several international programs have been formed to control, or more recently eliminate onchocerciasis, using mass drug administration (MDA) of ivermectin. However, in many areas of Africa (particularly those which are endemic for the eyeworm, Loa loa, or where vector densities are very high) ivermectin MDA alone will not be sufficient to achieve elimination. In these situations, additional interventions may be necessary. METHODOLOGY/PRINCIPAL FINDINGS: The Esperanza Window trap (EWT), a simple trap originally developed to replace human landing collections for entomological surveillance of O. volvulus transmission was optimized, resulting in a 17-fold improvement in trap performance. The optimized trap was tested in trials in schools and in agricultural fields to determine if it could reduce vector biting locally. The traps resulted in a 90% reduction in biting in the school setting. In the field setting, results varied. In one location, the traps reduced biting by roughly 50%, while in a separate trial, the traps did not significantly reduce the biting rate. Examination of the two settings suggested that trap placement may be critical to their success. CONCLUSIONS/SIGNIFICANCE: These results suggest that the optimized EWT might be capable of reducing local vector black fly biting in areas commonly frequented by residents. Together with other recently developed methods of community directed vector control, the traps may augment ivermectin MDA, bringing the goal of onchocerciasis elimination within reach in much of Africa.