Culicidae assemblages of artificial containers and possible biotic interactions affecting Aedes albopictus (Diptera: Culicidae) in Argentina.

In Argentina, the distribution of Aedes albopictus (Skuse) is limited to two provinces with a subtropical climate and few records. This study aims to describe and compare assemblies of Culicidae that breed in artificial containers in two areas with different degrees of urbanisation where Ae. albopictus is present and to evaluate possible biotic interactions. We sampled container larval habitats of an urban (Eldorado city) and a rural environment (Colonia Aurora village). We performed generalized linear mixed models to evaluate which variables (containers characteristics or environment) are associated with the presence and abundance of Ae. albopictus, Aedes aegypti Linneaus and Culex quinquefasciatus Say, and the presence of mosquito predators (Lutzia bigoti (Bellardi) and Toxorhynchites spp.). Also, the relationship between the most abundant species was quantified in each environment using Hurlbert's C8 association coefficient. Ae. aegypti was the most abundant species in the urban environment, while Cx. quinquefasciatus and Ae. albopictus were the most abundant in the rural area. Predators were more present in the rural environment and affected the abundance of Aedes mosquitoes. Regarding the C8 index, Ae. aegypti was negatively associated with Ae. albopictus in the urban area, whereas in the rural area these species presented a significantly positive relationship. These results show that in urban environments the high abundance of Ae. aegypti could be affecting the Asian tiger mosquito as evidenced by local studies of food larval competition. Also a greater presence of predators could be affecting Ae. albopictus in rural environments.

Controlling Rotavirus-associated diarrhea: Could single-domain antibody fragments make the difference?

Group A Rotavirus (RVA) remains a leading cause of severe diarrhea and child mortality. The variable domain of camelid heavy chain antibodies (VHH) display potent antigen-binding capacity, have low production costs and are suitable for oral therapies. Two sets of anti-RVA VHHs have been developed: ARP1-ARP3; 2KD1-3B2. Here, we explore the potential of both sets as a prevention strategy complementary to vaccination and a treatment option against RVA-associated diarrhea in endangered populations. Both sets have been expressed in multiple production systems, showing extensive neutralizing capacity against strains of RVA in vitro. They were also tested in the neonatal mouse model with various degrees of success in preventing or treating RVA-induced diarrhea. Interestingly, mitigation of the symptoms was also achieved with freeze-dried ARP1, so that it could be applied in areas where cold chains are difficult to maintain. 3B2 was tested in a pre-clinical trial involving gnotobiotic piglets where it conferred complete protection against RVA-induced diarrhea. ARP1 was used in the first clinical trial for anti-RVA VHHs, successfully reducing stool output in infants with RVA diarrhea, with no detected side effects.

City puzzles: Does urban land scape affect genetic population structure in Aedes aegypti?

Cities usually offer a suitable environment for the dengue vector Aedes aegypti, providing oviposition sites, accessibility to human hosts and nectar meals. However, large urban centres are highly heterogeneous environments, forming a patched landscape that could affect Ae. aegypti population dynamics and dispersal. Here, we performed a genome-wide analysis using Rad-seq data from 99 Ae. aegypti specimens collected in three areas within Buenos Aires city with varying levels of urbanization/land use: highly urbanized Area 1, intermediate Area 2 and poorly urbanized Area 3. We found an inverse association between urbanization levels and spatial genetic structure. Populations from highly urbanized Area 1 did not present genetic structure whereas two and three clusters were detected in Areas 2 and 3, respectively. In the case of Area 3, initial analyses showed separation in clusters was mostly due to elevated consanguinity within sites although three clusters were still detected after closely related individuals were discarded. Mosquitoes around each site displayed a high degree of isolation, evidencing a close dependence between the vector and human dwellings. Interestingly, specimens from distant boroughs (within the limits of the city) and the city's outskirts formed a single cluster with inner city sites (Area 1), highlighting the role of passive transport in shaping population structure. Genetic distances were poorly correlated with geographic distances in Buenos Aires, suggesting a stronger influence of passive than active dispersal on population structure. Only Area 2 displayed a significant isolation-by-distance pattern (p = 0.046), with males dispersing more than females (p = 0.004 and p = 0.016, respectively). Kinship analyses allowed us to detect full-siblings located 1.5 km apart in Area 1, which could be due to an extreme event of active female dispersal. Effective population size was higher in Area 2 confirming that cemeteries represent highly favourable environments for Ae. aegypti and need to be specifically targeted. Our results suggest that control programs should take into account urban landscape heterogeneity in order to improve vector control.

Larval Competition Between Aedes albopictus and Aedes aegypti (Diptera: Culicidae) in Argentina: Coexistence and Implications in the Distribution of the Asian Tiger Mosquito.

Aedes aegypti (Linnaeus) and Aedes albopictus (Skuse) are worldwide vectors of dengue and yellow fever viruses. These species coexist in many countries and the biotic interactions between them can influence their abundances and distributions. In Argentina, Ae. aegypti is widely distributed in the north and center regions of the country, with temperate and subtropical climate, while both are sympatric only in the northeastern area of the subtropical region. Interspecific and intraspecific larval competition for food was evaluated to assess if their interaction influences on patterns of abundance and distribution. Finite rates of increase and survivorship for each species were estimated and the effects of mosquito density ratio and detritus availability were determined. The Lambda (λ´) index of population performance of both showed there is no competitive exclusion pattern. However, survival of Ae. albopictus was negatively affected by the presence of Ae. aegypti. These results suggest one possible explanation for the codominance pattern of both species display in rural regions of the southernmost distribution of Ae. albopictus in South America. They also show Ae. aegypti as a potential biotic barrier for the expansion of Ae. albopictus as was reported in regions of the United States.

Genome-Wide Screening of Aedes aegypti (Culicidae: Diptera) Populations From Northwestern Argentina: Active and Passive Dispersal Shape Genetic Structure.

Aedes aegypti is the primary vector of arboviruses of great impact on human health. Our goal was to assess the spatial genetic structure of Ae. aegypti at the regional and local levels in Northwestern Argentina, an area with high prevalence of dengue fever. We analyzed 59 Ae. aegypti individuals collected from six locations in Northwestern Argentina using nuclear genome-wide Single Nucleotide Polymorphisms (SNPs) generated with double digest Restriction-site Associated DNA Sequencing. We also performed an entomological survey in 70 households in the cities of Orán and Tartagal. An analysis at the regional level indicated that the populations of Ae. aegypti in Northwestern Argentina are spatially structured and present a significant IBD pattern. Our results suggest that passive transport of eggs/immature stages, in both northward and southward directions, plays an important role in structuring Ae. aegypti populations at a regional scale and also as a source for the introduction of novel genetic variants through migration events into established populations. At a local level, we found neither spatial genetic structure nor significant isolation by distance (IBD) in Tartagal, indicating high gene flow within the city and active dispersal. In contrast, samples from Orán formed two clusters with a significant IBD pattern, although weaker than that at a regional level. Both populations showed signs of recent bottleneck events, probably coincident with past eradication campaigns. The entomological survey revealed a high prevalence of Ae. aegypti in both cities, although significantly higher in Tartagal.

Anti-VP6 VHH: An Experimental Treatment for Rotavirus A-Associated Disease.

Species A Rotaviruses (RVA) remain a leading cause of mortality in children under 5 years of age. Current treatment options are limited. We assessed the efficacy of two VP6-specific llama-derived heavy chain antibody fragments (VHH) -2KD1 and 3B2- as an oral prophylactic and therapeutic treatment against RVA-induced diarrhea in a neonatal mouse model inoculated with virulent murine RVA (ECw, G16P[16]I7). Joint therapeutic administration of 2KD1+3B2 (200 µg/dose) successfully reduced diarrhea duration, RVA infection severity and virus shedding in feces. While the same dose of 2KD1 or 3B2 (200 µg) significantly reduced duration of RVA-induced diarrhea, 2KD1 was more effective in diminishing the severity of intestinal infection and RVA shedding in feces, perhaps because 2KD1 presented higher binding affinity for RVA particles than 3B2. Neither prophylactic nor therapeutic administration of the VHH interfered with the host's humoral immune response against RVA. When 2KD1 (200 µg) was administered after diarrhea development, it also significantly reduced RVA intestinal infection and fecal shedding. Host antibody responses against the oral VHH treatment were not detected, nor did viral escape mutants. Our findings show that oral administration of anti-VP6 VHH constitute, not only an effective prophylactic treatment against RVA-associated diarrhea, but also a safe therapeutic tool against RVA infection, even once diarrhea is present. Anti-VP6 VHH could be used complementary to ongoing vaccination, especially in populations that have shown lower immunization efficacy. These VHH could also be scaled-up to develop pediatric medication or functional food like infant milk formulas that might help treat RVA diarrhea.

Hacia el control de la diarrea por rotavirus A: ¿podrían los nanoanticuerpos VHH marcar la diferencia? / Controlling rotavirus-associated diarrhea: could single-domain antibody fragments make the difference?

Los rotavirus del grupo A (RVA) constituyen la principal causa de diarrea grave y mortalidad infantil. La porción variable de los anticuerpos de cadena pesada derivados de camélidos presentan una amplia capacidad de unión antigénica (reconocen sitios antigénicos no accesibles a los anticuerpos tradicionales, con elevada afinidad) tienen bajos costos de producción y resultan ideales para las terapias orales. A la fecha, se desarrollaron 2 pares de nanoanticuerpos VHH contra RVA: ARP1-ARP3 y 2KD1-3B2. En este trabajo, exploramos el potencial de ambos grupos de nanoanticuerpos como estrategias de prevención complementarias a la vacunación y como una opción de tratamiento frente a la diarrea asociada a RVA en poblaciones de riesgo. Ambos pares de nanoanticuerpos fueron expresados en diferentes sistemas de producción y mostraron amplia capacidad neutralizante contra diversas cepas de RVA in vitro. También fueron usados en el modelo de ratón lactante, en el que evidenciaron distintos grados de éxito en la prevención o el tratamiento de la diarrea inducida por RVA. Es interesante destacar que la mitigación de los síntomas también se logró con ARP1 liofilizado y conservado, por lo que podría ser utilizado en áreas donde es difícil mantener la cadena de frío. Asimismo, 3B2 fue testeado en una prueba preclínica utilizando como modelo al cerdo gnotobiótico, al cual confirió completa protección contra la diarrea inducida por RVA. ARP1 fue usado en la primera prueba clínica de nanoanticuerpos VHH contra RVA, donde redujo significativamente las deposiciones en pacientes pediátricos con diarrea positivos para RVA, sin evidenciar ninguna reacción adversa

Group A Rotavirus (RVA) remains a leading cause of severe diarrhea and child mortality. The variable domain of camelid heavy chain antibodies (VHH) display potent antigen-binding capacity, have low production costs and are suitable for oral therapies. Two sets of anti-RVA VHHs have been developed: ARP1-ARP3; 2KD1-3B2. Here, we explore the potential of both sets as a prevention strategy complementary to vaccination and a treatment option against RVA-associated diarrhea in endangered populations. Both sets have been expressed in multiple production systems, showing extensive neutralizing capacity against strains of RVA in vitro. They were also tested in the neonatal mouse model with various degrees of success in preventing or treating RVA-induced diarrhea. Interestingly, mitigation of the symptoms was also achieved with freeze-dried ARP1, so that it could be applied in areas where cold chains are difficult to maintain. 3B2 was tested in a pre-clinical trial involving gnotobiotic piglets where it conferred complete protection against RVA-induced diarrhea. ARP1 was used in the first clinical trial for anti-RVA VHHs, successfully reducing stool output in infants with RVA diarrhea, with no detected side effects