Anti-Wolbachia therapy for onchocerciasis & lymphatic filariasis: Current perspectives

Onchocerciasis and lymphatic filariasis (LF) are human filarial diseases belonging to the group of neglected tropical diseases, leading to permanent and long-term disability in infected individuals in the endemic countries such as Africa and India. Microfilaricidal drugs such as ivermectin and albendazole have been used as the standard therapy in filariasis, although their efficacy in eliminating the diseases is not fully established. Anti-Wolbachia therapy employs antibiotics and is a promising approach showing potent macrofilaricidal activity and also prevents embryogenesis. This has translated to clinical benefits resulting in successful eradication of microfilarial burden, thus averting the risk of adverse events from target species as well as those due to co-infection with loiasis. Doxycycline shows potential as an anti-Wolbachia treatment, leading to the death of adult parasitic worms. It is readily available, cheap and safe to use in adult non-pregnant patients. Besides doxycycline, several other potential antibiotics are also being investigated for the treatment of LF and onchocerciasis. This review aims to discuss and summarise recent developments in the use of anti-Wolbachia drugs to treat onchocerciasis and LF.

Abundance and diversity of black flies (Diptera: Simuliidae) in rivers of the Andean Eastern Hills of Bogotá (Colombia), and its relationship with water stream physicochemical variables / Abundancia y diversidad de moscas negras (Díptera: Simuliidae) en ríos de los Cerros Orientales Andinos de Bogotá (Colombia), y su relación con variables fisicoquímicas de las corrientes de agua / Abundancia e diversidade de moscas negras (Diptera: Simuliidae) em rios dos Cerros Orientais Andinos de Bogotá (Colômbia), e sua relação com variáveis físico-químicas das correntes de água

Abstract Black flies are abundant benthic organisms in well-oxygenated running water and are considered effective bioindicators of water quality. Information on the ecology of these organisms at the species level is important, since up to now information has mainly been available on a family level. The aim of this study was to evaluate the composition of black flies and their relationships to a group of physical and chemical factors in four small rivers of the Eastern Hills around Bogotá, Colombia. These headwaters are protected by the Empresa de Acueducto y Alcantarillado de Bogotá. Black fly larvae and pupae were collected during four sampling periods during the dry season to the early rainy season of 2012. Multivariate methods were used to determine the presence of each species in relation to dissolved oxygen, nitrates, pH, temperature, and water velocity. PCA ordination revealed a physicochemical environment with a tendency towards a certain homogeneity in the four rivers studied. The DCA ordination confirmed that in La Vieja River the G. ortizi complex dominated while S. muiscorum was dominant in the other three rivers. Similarly, according to the NMDS, the composition of black flies in Arzobispo and Chorro de Padilla rivers was similar, while that of El Delirio and La Vieja rivers were different, especially in this last river. The rivers had low species richness (four species and one species complex). Simulium muiscorum was negatively correlated with dissolved oxygen, temperature and current velocity while S. ignescens was associated with slightly more oxygenated waters and S. furcilatum with faster currents. The low species diversity and richness for the four rivers is consistent with previous reports of low Neotropical diversity of black flies. Results showed that simuliid species could possibly be good indicators of the environmental conditions of Eastern Hills rivers around Bogotá.

Resumen Las moscas negras son organismos bénticos abundantes en corrientes de agua bien oxigenadas y se consideran bioindicadores efectivos de la calidad de agua. La información sobre la ecología de estos organismos a nivel de especie es importante, ya que hasta el momento, ha estado disponible principalmente solo a nivel de familia. El objetivo de este estudio fue evaluar la composición de moscas negras y sus relaciones con un grupo de factores físicos y químicos en cuatro ríos pequeños de los Cerros Orientales alrededor de Bogotá, Colombia. Estos nacimientos están protegidos por la Empresa de Acueducto y Alcantarillado de Bogotá, Colombia. Se colectaron larvas y pupas de moscas negras a lo largo de cuatro períodos de muestreo, desde la estación seca hasta la estación lluviosa de 2012. Se utilizaron métodos multivariados para determinar la presencia de cada especie en relación con: oxígeno disuelto, nitratos, pH, temperatura y velocidad del agua. La ordenación por PCA reveló un ambiente fisicoquímico con tendencia hacia la homogeneidad en los cuatro ríos estudiados. La ordenación por DCA confirmó que en el río La Vieja dominó el complejo G. orti%i, mientras que S. muiscorum fue dominante en los otros tres. De igual forma de acuerdo con el análisis NMDS, la composición de moscas negras en los ríos Arzobispo y Chorro de Padilla fue similar, mientras que en los ríos El Delirio y La Vieja fue diferente, especialmente en este último. Los ríos presentaron baja riqueza de especies (cuatro especies y un complejo de especies). Simulium muiscorum se correlacionó negativamente con el oxígeno disuelto, temperatura y velocidad de la corriente, mientras que S. ignescens estuvo asociado con aguas ligeramente más oxigendas y S. furcilatum, con corrientes más rápidas. La baja diversidad y riqueza para los cuatro ríos es consistente con reportes previos de baja diversidad neotropical de moscas negras. Los resultados muestran que las especies de simúlidos podrían ser buenos indicadores de las condiciones ambientales de los ríos de los Cerros Orientales de Bogotá.

Resumo As moscas negras são organismos bentônicos abundantes em correntes de água bem oxigenadas e se consideram bioindicadores efetivos da calidade de água. A informação sobre a ecologia de estes organismos quanto a espécie é importante, pois até o momento, há informação somente em nível de família. O objetivo de este estudo foi avaliar a composição de moscas negras e suas relações com um grupo de fatores físicos e químicos em quatro rios pequenos dos Cerros Orientais nas redondezas de Bogotá, Colômbia. As nascentes estão protegidas pela Empresa de Acueducto y Alcantarillado de Bogotá, Colômbia. Foram coletadas larvas e poupas de moscas negras ao longo de quatro períodos de amostragem, desde a estação seca até a estação chuvosa de 2012. Foram utilizados métodos multivariados para determinar a presença de cada espécie em relação com: oxigênio dissolvido, nitratos, pH, temperatura e velocidade da água. A ordenação por PCA revelou um ambiente físico-químico com tendência à homogeneidade nos quatro rios estudados. A ordenação por DCS confirmou que no rio La Vieja dominou o complexo G. orti%i, enquanto que S. muiscorum foi dominante nos outros três rios. Igualmente, de acordo com a análise NMDS, a composição de moscas negras nos rios Arzobispo e Chorro de Padilla foi similar, enquanto que nos rios El delírio e La Vieja foi diferente, em especial neste último. Os rios apresentaram baixa riqueza de espécies (quatro espécies e um complexo de espécies). S. muiscorum se correlacionou negativamente com o oxigênio dissolvido, temperatura e velocidades da corrente, enquanto que S. ignescens foi associado com águas ligeiramente mais oxigenadas e S. furcilatum, com correntes mais rápidas. A baixa diversidade e riqueza para os quatro rios é consistente com reportes prévios de baixa diversidade neotropical de moscas negras. Os resultados mostram que espécies de Simulium poderiam ser boas indicadoras das condições ambientais dos rios dos Cerros Orientais de Bogotá.

Water pollution and distribution of the black fly (Diptera: Simuliidae) in the Atlantic Forest, Brazil / Contaminación del agua y distribución de la mosca negra (Díptera: Simuliidae) en el bosque Atlán&tico, Brasil

Abstract:Black flies have medical importance because some species are vectors of the Onchocerciasis and Mansonelosis, nevertheless, their ecology and potential use as bioindicators is still poorly studied in the Neotropical Region. In Brazil, bioindicators use is strongly focused in a multimetrical ecological index approach; this way, we investigated the black fly spatial distribution, in relation to abiotic factors correlated to water quality, to provide baseline information for their utilization as standalone indicators of lotic systems integrity. We have tested the hypothesis that environmental changes related to urbanization, lead to decreased abundance and loss in the number of species of the black fly fauna. The sampling was conducted in 10 urban and 10 preserved streams during the dry season (August-September) of 2012, in the mountainous region of Teresópolis, State of Rio de Janeiro, Brazil. The streams were characterized for their environmental integrity conditions and physico-chemical properties of water. In each stream, five different rapid points were sampled in a section of 50 meters, 10 meters apart from each other. The black flies were sampled with a kick-net sampler on rocky substrates. The material was separated and the larvae were sorted in morphotypes, and later, the final instar specimens were dissected and identified with the help of taxonomical literature at species level. A total abundance of 488 larvae from nine species were collected, 5 (1.02 %) in extremely impacted streams, 470 (96.31 %) in intermediate streams and 13 (2.66 %) in preserved streams. The visual evaluation (HII) differed in rela&tion to the water physico-chemical evaluation, in which more variation in the characterization of the sampling sites was observed. In Canonical Correspondence Analysis Simulium subpallidum, S. inscrustatumand S. pertinaxwere significantly associated with intermediate values of most of the variables, and then to intermediate impacted sites. On the other hand, Lutzsimulium hirticosta, S. subnigrumand Simuliumsp. A were associated to low values of chemical variables, and then to more preserved sites. Most studies on the bioindicator potential of Simuliidae have suggested an approach to agricultural impacts, while our results, on the other hand, showed that Simuliidae species were present in streams with intermediate urban pollution impacts, but absent in heavily impacted sites. Thus, our data suggested that some species are associated to more pristine breeding sites, such as L. hirticostaand Simuliumsp. A, while others may be good bioindicators of moderately impacted streams, such as S. pertinax, S. subnigrumand S. subpallidum.

Resumen:Aunque las moscas negras tienen importancia médica debido a que algunas de sus especies son vectores de la Oncocercosis y Mansonelosis, su ecología y su potencial como bioindicadores es aún poco estudiado en la Región Neotropical. En Brasil el uso de bioindicadores se centra principalmente en un índice ecológico multimétrico, en este estudio investigamos la distribución espacial de las moscas negras y su relación con los factores abióticos, para su utilización como indicadores de la integridad de los sis&temas lóticos. En este sentido, hemos probado la hipótesis de que los cambios ambientales relacionados con la urba&nización conducen a la disminución de la abundancia y la pérdida de especies de la mosca negra. El muestreo se llevó a cabo en la región montañosa del Estado de Río de Janei&ro, Brasil, en 10 zonas urbanas y 10 arroyos conservados durante la estación seca (agosto-septiembre) de 2012. Los arroyos se caracterizaron por sus condiciones de integridad del ambiente y las propiedades físico-químicas de agua. Las moscas negras se muestrearon en sustratos rocosos con un kick-net sampler.El material se separó y las larvas se ordenaron por morfotipos y más tarde los especímenes en último estadio se disectaron e identificaron con la ayuda de la literatura taxonómica a nivel de especie. Una abundancia total de 488 larvas de nueve especies fueron recolectadas, 5 (1.02 %) en arroyos muy afectados, 470 (96.31 %) en intermedios y 13 (2.66 %) en conservados. La evaluación visual (HII) difiere de la físico-química del agua, en la que se observó una mayor variación. En el Análisis Correspon&dencia Canónica, Simulium subpallidum, S. inscrustatumy S. pertinaxestaban asociadas significativamente con sitios de impacto intermedio, para la mayoría de las variables. Por otro lado, Lutzsimulium hirticosta, S. subnigrumy Simuliumsp. A se asocian con valores bajos de las varia&bles químicas, relacionado a sitios más conservados. La mayoría de los estudios sobre el potencial bioindicador de Simuliidae tienden a enfocarse en los impactos agrícolas, mientras que nuestros resultados, por otro lado, sugieren que las especies de Simuliidae están presentes en arroyos con impactos intermedios de contaminación urbana, pero ausentes en los sitios fuertemente afectados. Por lo tanto, nuestros datos sugieren que algunas especies tales como L. hirticostay Simuliumsp. A están asociadas a lugares de reproducción más prístinos, mientras que otras pueden ser bioindicadores de arroyos moderadamente afectados, como S. pertinax, S. subnigrumy S. subpallidum.

COMPORTAMIENTO DE OVIPOSICIÓN Y CARACTERIZACIÓN DE LA MICROESTRUCTURA CORIONICA DE LOS HUEVOS DE Simulium dinelliii (Joan) (Diptera: Simuliidae) / Oviposition Behavior and Microstructure of the Eggs of Simulium dinellii (Joan) (Diptera: Simuliidae)

Little is known about the oviposition habits and egg structure of Neotropical members of the Simuliidae family. The oviposition behavior of Simulium dinellii (Joan) was observed near at Otún Quimbaya, Colombia. Small groups of females hovered about 2 cm above the water surface to oviposit. Some of these females had been capture and the eggs were obtained. These eggs were counted, measured and processed by scanning electron microscopy. Subtriangular eggs had a latticework of polygons on the endochorionic surface. Chorionic differences between the eggs of S. dinellii and the few previously studied species suggest potential taxonomic value of egg microstructure.

El conocimiento sobre los hábitos de oviposición y estructura de los huevos de la familia Simuliidae es muy escaso. En este estudio se observó el comportamiento de oviposición de Simulium dinellii (Joan) en quebradas cercanas al Santuario de fauna y flora Otún Quimbaya, Colombia. El cual consistió en la formación de pequeños grupos de hembras que sobrevolaban a 2 cm de la superficie del agua para ovipositar, algunas de estas hembras fueron colectadas, obteniendo así los huevos, los cuales fueron contados, medidos y procesados por Microscopia Electrónica de Barrido con el fin de caracterizar la estructura endocoriónica. Los huevos presentaron una forma subtriangular, con una entramado poligonal sobre la superficie. Las características morfológicas observadas en los huevos de S. dinellii fueron diferentes a las previamente reportadas en otras especies de la familia Simuliidae, sugiriendo un posible valor taxonómico de la microestructura del huevo.

Fauna of Simuliidae (Diptera) from the state of Espírito Santo, Brazil: distribution, new records and list of species

Fauna of Simuliidae (Diptera) from the state of Espírito Santo, Brazil: Distribution, new records and list of species. The fauna of Simuliidae in Brazil is composed of 87 species, 17 of which are recorded from the state of Espírito Santo. Entomological collections were carried out in 2010-2011 with the objective of increasing the knowledge of the species richness of this family in the state. Ninety three rivers and streams were sampled, each collection being carried out in a 50m transect. During the study period 30 species were collected, 13 of which represent new records for the state, 12 of the genus Simulium and one of the genus Lutzsimulium. Among these new state records one, Simulium lobatoi, also represents a new record from southeastern Brazil. The other newly recorded species are: Lutzsimulium hirticosta, Simulium distinctum, Simulium exiguum, Simulium oyapockense, Simulium botulibranchium, Simulium petropoliense, Simulium clavibranchium, Simulium rappae, Simulium minusculum, Simulium dinellii, Simulium ochraceum and Simulium scutistriatum.

Mansonella ozzardi in Amazonas, Brazil: prevalence and distribution in the municipality of Coari, in the middle Solimões River

This study investigated some epidemiological aspects of the Mansonella ozzardi in municipality of Coari, Amazonas. Clinical symptoms were correlated with the filarial infection and the parasitic infection rates (PIR) were estimated in simuliid vectors. The general M. ozzardi human prevalence rate was 13.3 percent (231/1733), of which 10.2 percent (109/1069) were from the urban area and 18.4 percent (122/664) from the rural area. The prevalence rates were higher in men (14.5 percent urban and 19.7 percent rural) than in women (6.7 percent urban and 17.2 percent rural) and occurred in most age groups. The indices of microfilaremics were higher in people > 51 years old (26.9 percent urban and 61.5 percent rural). High prevalence rates were observed in retired people (27.1 percent urban area), housewives and farmer (41.6 percent and 25 percent, respectively, in rural area). The main clinical symptoms were joint pains and sensation of leg coldness. Only Cerqueirellum argentiscutum (Simuliidae) transmits M. ozzardi in this municipality (PIR = 5.6 percent urban and 7.1 percent rural). M. ozzardi is a widely distributed parasitic disease in Coari. Thus, temporary residency in the region of people from other localities involved with the local gas exploitation might be a contributing factor in spreading the disease.

Anticoagulant activity in salivary gland homogenates of Thyrsopelma guianense (Diptera: Simuliidae), the primary vector of onchocerciasis in the Brazilian Amazon

In this study, anticoagulant activity was detected in salivary gland homogenates (SGHs) of Thyrsopelma guianense (Diptera: Simuliidae). The SGH yielded 1.07 ìg ± 0.03 (n = 15) of total soluble protein per pair of glands. In addition, following SDS-PAGE (12.5 percent gel) and silver nitrate staining, 12 polypeptides with molecular weights ranging from 14-69 kDa were detected in all physiological ages analyzed (12 h, 24 h, 48 h and 72 h following emergence). Coagulation bioassays showed that the SGHs had activities that interacted at all levels of coagulation (the intrinsic, extrinsic and common pathways), by extending the plasma recalcification time, prothrombin time, thrombin time. This is the first report on the activity of salivary gland proteins from the main vector of onchocerciasis in Brazil. We also suggest detailed studies on the morphology and function of the salivary glands in order to understand the role of these proteins in host/vector interactions.

Velocidade da água e a distribuição de larvas e pupas de Chirostilbia pertinax (Kollar) (Diptera, Simuliidae) e macroinvertebrados associados / Water velocity and the distribution of larvae and pupae of Chirostilbia pertinax (Kollar) (Diptera, Simuliidae) and associated macroinvertebrates

The objective of the current study was to evaluate the distribution of the immature stages of Chirostilbia pertinax (Kollar, 1832) and associated bentic macroinvertebrates in relation to water velocity. This study was carried out on the upper course of Sinos river, in Rio Grande do Sul. Seasonal collections of larvae and pupae of black flies and associated benthic fauna were carried out at five places. In all sampling sites artificial substrates were installed, remaining submersed for 14 days in places of great water flow. Before removing the substrates the flow speeds on the collection sites were measured. To assess the distinct distribution patterns of the benthic fauna, the flow speed was divided in three classes: slow (0.20 - 0.63 m/s), medium (0.64 - 1.07 m/s) and fast (1.08 - 1.50 m/s). A total of 39,598 specimens were collected, from which 33,418 were black flies and 6,180 associated macroinvertebrates. From the total black flies, 5,704 were in the last larval instar and pupae from which 828 were C. pertinax. In the distribution of this species 30.8 percent occurred in slow flow, 24.9 percent in medium flow and 44.3 percent in fast flow. The abundance of C. pertinax is inversely related to the predatory macroinvertebrates (GLM, R² = 0.207; F 2.50 = 6.536; P = 0.003). These results demonstrate that the immature stages of C. pertinax have a tendency to occupy microhabitats with less predator occurrence.

O presente trabalho objetivou investigar a relação entre a velocidade da água e a distribuição dos estágios imaturos de Chirostilbia pertinax (Kollar, 1832) e os macroinvertebrados bentônicos associados. As coletas foram realizadas em cinco pontos, no trecho superior do Rio dos Sinos, no Estado do Rio Grande do Sul. A fauna bentônica foi amostrada com substratos artificiais, que permaneceram instalados nos locais de maior corenteza d'água por períodos de 14 dias. Antes do recolhimento dos substratos foram realizadas medidas de velocidade d'água no local da sua instalação. Essas medidas foram agrupadas em três classes: baixa (0,20 a 0,63 m/s), média (0,64 a 1,07 m/s) e alta (1,08 a 1,50 m/s) para posterior relação com a distribuição da fauna coletada. Para avaliar a influência da velocidade de água sobre a distribuição dos macroinvertebrados e a abundância de C. pertinax foi realizada uma regressão múltipla. As coletas resultaram em 39.598 indivíduos, sendo 33.418 simulídeos e 6.180 macroinvertebrados associados. Entre os simulídeos 5.704 espécimes correspondem a larvas do último ínstar e pupas, dos quais 828 são de C. pertinax. Quanto à distribuição dessa espécie, 30,8 por cento ocorreram na classe de baixa velocidade d'água, 24,9 por cento na velocidade média e 44,3 por cento na alta velocidade. Foi observado que a abundância de C. petinax aumenta à medida que os macroinvertebrados predadores diminuem e os organismos não predadores aumentam (GLM, R² = 0,207; F 2,50 = 6,536; P = 0,003). Esses resultados revelam que os imaturos de C. pertinax têm uma tendência em ocupar um microhábitat com menor ocorrência de predadores.