Infection of Aedes mosquitoes by native Wolbachia in urban cemeteries of Southern Mexico / Infección de mosquitos Aedes con Wolbachia nativa en cementerios urbanos del sur de México

Abstract: Objective: To evaluate the prevalence of Wolbachia infections in Aedes spp. field populations from cemeteries of Southern Mexico. Materials and methods: Six cemeteries were selected to be sampled in the central part of the Soconusco region, Chiapas. Aedes albopictus and Ae. aegypti mosquitoes were collected during the rainy season of 2015. Females were analyzed individually by PCR to determine the presence of Wolbachia. Results: A field overall prevalence of 38% was found; only Ae. albopictus mosquitoes were positive. Conclusion: Local strains of Wolbachia were detected and have the potential to be applied as a biological method for vector control.

Resumen: Objetivo: Evaluar la presencia de Wolbachia en poblaciones de campo de Aedes spp. en cementerios del Sur de México. Material y métodos: Se seleccionaron seis cementerios como sitios de colecta para las poblaciones silvestres de Aedes albopictus y Ae. aegypti, en la región del Soconusco, Chiapas, durante la época de lluvias 2015. Se determinó la infección por Wolbachia en hembras individuales por PCR. Resultados: Se obtuvo una infección de 38% por Wolbachia en Ae. albopictus. Conclusión: Existen cepas locales de Wolbachia en los mosquitos y poseen el potencial de aplicarse como medida de control biológico de vectores.

Detection of filarial parasites and Wolbachia endosymbiotic bacteria in mosquitoes by PCR

Wuchereria Bancroft; the principal etiologic agent of lymphatic filariasis is mosquito dependant in the biological transmission. Dirofilariasis is essentially a disease of canines which can also be trans-mitted to humans by culicine mosquitoes. Wolbachia are Rickettsia-like, obligatory intracellular bacteria that infect the reproductive and somatic tissues of some arthropods and nematodes. Our study aimed to identify the possible association between filarial parasites and Wolbachia by single and multiplex PCR. 1600 female mosquitoes collected from: four localities in Assiut Governorate, Egypt were microscopically identified and divided into 64 mosquito pools according to their genera and collection site. Single PCR was firstly conducted on mosquitoes followed by multiplex PCR for simultaneous detection of the three filarial parasites [Wucheraria bancrofti, Dirofilaria immitis, and Dirofilaria repens] and Wolbachia in mosquitoes. The results indicated that: Out of 64 mosquito pools, 8 pools were positive for Wuchererio bancrofti with estimated rate of infection [ERI 0.53%], 3 for each of Dirofilaria immitis and Dirofilaria repens [ERI 0.19%] and 6 pools were positive for Wolbachia [ERI 0.39%], five of them with filarial parasites [W. Bancrofti, D. immitis and D repens] [83.3%], with a significant P. value [< 0.05]. We concluded that there was a strong association between the presence of Wolbachia and filarial parasites that should be considered during the treatment of patients with filarial diseases by the use of specific anti-Wolbachia antibiotic in addition to the usual anti-filarial drugs

Detection of Wolbachia pipientis, including a new strain containing the wsp gene, in two sister species of Paraphlebotomus sandflies, potential vectors of zoonotic cutaneous leishmaniasis

Individual, naturally occurring Phlebotomus mongolensis and Phlebotomus caucasicus from Iran were screened for infections with the maternally inherited intracellular Rickettsia-like bacterium Wolbachia pipientis via targeting a major surface protein gene (wsp). The main objective of this study was to determine if W. pipientis could be detected in these species. The sandflies were screened using polymerase chain reaction to amplify a fragment of the Wolbachia surface protein gene. The obtained sequences were edited and aligned with database sequences to identify W. pipientis haplotypes. Two strains of Wolbachia were found. Strain Turk 54 (accession EU780683) is widespread and has previously been reported in Phlebotomus papatasi and other insects. Strain Turk 07 (accession KC576916) is a novel strain, found for first time in the two sister species. A-group strains of W. pipientis occur throughout much of the habitat of these sandflies. It is possible that Wolbachia is transferred via horizontal transmission. Horizontal transfer could shed light on sandfly control because Wolbachia is believed to drive a deleterious gene into sandflies that reduces their natural population density. With regard to our findings in this study, we can conclude that one species of sandfly can be infected with different Wolbachia strains and that different species of sandflies can be infected with a common strain.

High prevalence and lack of diversity of Wolbachia pipientis in Aedes albopictus populations from Northeast Brazil

The use of Wolbachia as a tool to control insect vectors has recently been suggested. In this context, studies on the prevalence and diversity of this bacterium in wild populations are relevant. Here, we evaluated the diversity of two Wolbachiagenes (ftsZ and wsp) and the prevalence of this endosymbiont in wild Aedes albopictus. Using semi-nested polymerase chain reaction, our results showed that 99.3 percent of the individuals were superinfected with Wolbachia. In regards to genetic diversity, the two genes showed no variation within or among mosquito populations. An analysis of other Wolbachia markers may help to clarify the relationship between insect and endosymbiont.

Wolbachia screening in spiders and assessment of horizontal transmission between predator and prey

Recent studies have revealed that the prevalence of Wolbachia in arthropods is attributable not only to its vertical transmission, but also to its horizontal transfer. In order to assess the horizontal transmission of Wolbachia between predator and prey, arthropods belonging to 11 spider families and six insect families were collected in the same field of rice. The distribution of Wolbachia in these arthropods was detected by diagnostic PCR amplification of the wsp (Wolbachia outer surface protein gene) and 16S rDNA genes. Nurscia albofasciata Strand (Araneae: Titanoecidae), Propylea japonica Thunberg (Coleoptera: Coccinellidae), Paederus fuscipes Curtis (Coleoptera: Staphylinidae), and Nilaparvata lugens Stal (Homoptera: Delphacidae) were infected with Wolbachia. This is the first report of infection of N. albofasciata and P. fuscipes by Wolbachia. No direct evidence indicated the existence of horizontal transmission of Wolbachia between predator and prey.

Wolbachia in two populations of Melittobia digitata Dahms (Hymenoptera: Eulophidae) / Wolbachia en dos poblaciones de Melittobia digitata Dahms (Hymenoptera: Eulophidae)

We investigated two populations of Melittobia digitata Dahms, a gregarious parasitoid (primarily upon a wide range of solitary bees, wasps, and flies), in search of Wolbachia infection. The first population, from Xalapa, Mexico, was originally collected from and reared on Mexican fruit fly pupae, Anastrepha ludens Loew (Diptera: Tephritidae); the other, from Athens, Georgia, was collected from and reared on prepupae of mud dauber wasps, Trypoxylon politum Say (Hymenoptera: Crabronidae). PCR studies of the ITS2 region corroborated that both parasitoid populations were the same species; this potentially provides a useful molecular taxonomic profile since females of Melittobia species are superficially similar. Amplification of the Wolbachia surface protein gene (wsp) confirmed the presence of this endosymbiont in both populations. Sequencing revealed that the Wolbachia harbored in both populations exhibited a wsp belonging to a unique subgroup (denoted here as Dig) within the B-supergroup of known wsp genes. This new subgroup of wsp may either belong to a different strain of Wolbachia from those previously found to infect Melittobia or may be the result of a recombination event. In either case, known hosts of Wolbachia with a wsp of this subgroup are only distantly related taxonomically. Reasons are advanced as to why Melittobia - an easily reared and managed parasitoid - holds promise as an instructive model organism of Wolbachia infection amenable to the investigation of Wolbachia strains among its diverse hosts.

Se investigaron dos poblaciones de Melittobia digitata Dahms, un parasitoide gregario (principalmente sobre un rango amplio de abejas solitarias, avispas y moscas), en busca de infección por Wolbachia. La primera población, provenía de Xalapa, México, y fue originalmente colectada y criada sobre pupas de la Mosca Mexicana de la Fruta, Anastrepha ludens Loew (Diptera: Tephritidae). La segunda población, originaria de Athens, Georgia, fue colectada y criada sobre prepupas de avispas de barro, Trypoxylon politum Say (Hymenoptera: Crabronidae). Estudios de PCR de la región ITS2 confirmaron que ambas poblaciones del parasitoide pertenecen a la misma especie; lo que nos provee de un perfil molecular taxonómico muy útil debído a que las hembras de las diversas especies de Melittobia son superficialmente similares. La amplificación del gen de superficie de proteina (wsp) de Wolbachia confirmó la presencia de este endosimbionte en ambas poblaciones. La ejecución de la secuencia reveló que Wolbachia alojada en ambas poblaciones exibe un wsp que pertenece a un subgrupo único (denominado aquí como Dig) dentro del supergrupo B de los genes wsp conocidos. Este nuevo subgrupo de wsp podría pertenecer o a un lineaje de Wolbachia de los previamente conocidos infectando a Melittobia o podría ser el resultado de algún evento recombinante. En cualquier caso, los huéspedes conocidos de Wolbachia con un wsp en este subgrupo están relacionados taxonómicamente en forma lejana. Se presentan razones posibles del por qué Melittobia - un parasitoide fácil de criar y manipular - es prometedor como un organismo modelo conveniente para el estudio de líneas de Wolbachia entre diversos huéspedes.

Frequency of infection with A and B supergroup Wolbachia in insects and pests associated with mulberry and silkworm.

Wolbachia is a ubiquitous, Gram-negative,vertically transmitted, alpha-proteobacterium that causes an array of reproductive abnormalities including cytoplasmic incompatibility, feminization of genetic males, parthenogenesis in a number of insect species, among others. Wolbachia is now being exploited as an agent for pest and vector control. Previous surveys indicated that it is commonly seen in 16-76% of arthropods. In this paper, using polymerase chain reaction assay based on specific amplification of the ftsZ -A and -B supergroup Wolbachia gene fragments, we found that 30% of insects and pests screened were positive for Wolbachia. Among them 66.7% harbour double Wolbachia infection, while 33.3 % harbour single Wolbachia infection. These results indicate widespread infection with both double and single Wolbachia, and provide a wealth of information to exploit this endobacterium for the management of pests and vectors.

Genetic diversity of Costa Rican populations of the rice planthopper Tagosodes orizicolus (Homoptera: Delphacidae)

Tagosodes orizicolus (Homoptera: Delphacidae) is one of the main constraints of the rice production in the Neotropics. This planthopper produces severe damages as a phloem feeder, causes mechanical injury during oviposition and vectors the rice hoja blanca virus (RHBV). The main objective of this study was to determine the genetic diversity of T. orizicolus populations from three rice growing regions of Costa Rica, using RAPDs. Individuals from Guanacaste, Parrita, San Carlos and Cali-Colombia, as outgroup, were analyzed using the random primers. Phenetic relationships revealed that the Costa Rican populations were clearly separated from Cali-Colombia, sharing less than 25% similarity. Costa Rican populations were divided into two main branches separated at 30% similarity. The first branch included Guanacaste and San Carlos and the second displayed Parrita. In relation to similarity indexes within groups, the Guanacaste cluster showed the highest (over 50%) and Cali-Colombia was the most diverse (28%). The correspondence analysis confirmed the clusters of the phenogram and showed close interactions between the Parrita and San Carlos populations. The genetic separation observed could be the result of the geographic isolation among populations, but it could also be explained by the infection with the rickettsia Wolbachia pipientis. This bacterium causes cytoplasmic incompatibility in its host, which results in non-viable progeny when infected males mate with non-infected females, or when insects hosting different strains of Wolbachia mate. Then, a search for Wolbachia in previously described populations of T orizicolus was initiated. The presence of the bacteria was analyzed by PCR with 16S rDNA-specific primers for Wolbachia. The PCR analyses revealed infections of 86% in the population of San Carlos, 96% in Guanacaste, 37% in Parrita and 100% in Cali-Colombia. Crosses between individuals of T. orizicolus from Parrita and Guanacaste were performed for testing...