Urban environmental clustering to assess the spatial dynamics of Aedes aegypti breeding sites.

The identification of Aedes aegypti breeding hotspots in urban areas is crucial for the rational design of control strategies against this disease vector. Remote sensing and geographic information systems offer valuable tools for mapping habitat suitability of a given area. However, predicting species occurrences by means of probability distribution maps based on transversal entomological surveys has limited utility for local authorities. The aim of the present study was to carefully examine the temporal evolution of the number of houses infested with immature stages of Ae. aegypti in each individual neighbourhood and to explore the value of producing environmental clusters generated with information provided by remotely sensed variables to explain the observed differential temporal behaviour. Entomological surveys were conducted between 2011 and 2013 throughout a small town in Argentina registering the number of houses with containers harbouring immature stages of Ae. aegypti. A SPOT 5 satellite image was used to obtain land cover variables, which were subsequently submitted to k-means partitioning for grouping neighbourhoods into four environmental clusters. Finally, a generalized linear model was fitted showing that the number of houses found to be positive for Ae. aegypti was jointly affected by the interaction between environmental clusters and the year of sampling. Moreover, the number of positive houses in one of the clusters was 9.5 times higher (P<0.005, SE=0.37) in 2013 than in 2012, but we did not observe any other statistically significant increases.

Unprecedented plant species loss after a decade in fragmented subtropical Chaco Serrano forests.

Current biodiversity loss is mostly caused by anthropogenic habitat loss and fragmentation, climate change, and resource exploitation. Measuring the balance of species loss and gain in remaining fragmented landscapes throughout time entails a central research challenge. We resurveyed in 2013 plant species richness in the same plots of a previous sampling conducted in 2003 across 18 forest fragments of different sizes of the Chaco Serrano forest in Argentina. While the area of these forest remnants was kept constant, their surrounding forest cover changed over this time period. We compared plant species richness of both sampling years and calculated the proportion of species loss and gain at forest edges and interiors. As in 2003, we found a positive relationship between fragment area and plant richness in 2013 and both years showed a similar slope. However, we detected a net decrease of 24% of species' richness across all forest fragments, implying an unprecedentedly high rate and magnitude of species loss driven mainly by non-woody, short-lived species. There was a higher proportion of lost and gained species at forest edges than in forest interiors. Importantly, fragment area interacted with percent change in surrounding forest cover to explain the proportion of species lost. Small forest fragments showed a relatively constant proportion of species loss regardless of any changes in surrounding forest cover, whereas in larger fragments the proportion of species lost increased when surrounding forest cover decreased. We show that despite preserving fragment area, habitat quality and availability in the surroundings is of fundamental importance in shaping extinction and immigration dynamics of plant species at any given forest remnant. Because the Chaco Serrano forest has already lost 94% of its original cover, we argue that plant extinctions will continue through the coming decades unless active management actions are taken to increase native forest areas.

Neuronal Degeneration in Mice Induced by an Epidemic Strain of Saint Louis Encephalitis Virus Isolated in Argentina.

Saint Louis encephalitis virus (SLEV) is a neglected flavivirus that causes severe neurological disorders. The epidemic strain of SLEV, CbaAr-4005, isolated during an outbreak in Córdoba city (Argentina), causes meningitis and encephalitis associated with neurological symptoms in a murine experimental model. Here, we identified the affected brain areas and the damage triggered by this neurotropic arbovirus. We performed a detailed analysis of brain neurodegeneration associated with CbaAr-4005 SLEV infection in mice. The motor cortex, corpus striatum and cerebellum were the most affected structures. Neurodegeneration was also found in the olfactory bulb, thalamus, hypothalamus, hippocampus, and hindbrain. SLEV infection triggered brain cell apoptosis as well as somatodendritic and terminal degeneration. In addition, we observed massive excitotoxic-like degeneration in many cortical structures. Apoptosis was also detected in the neuroblastoma cell line N2a cultured with SLEV. The results evidenced that SLEV CbaAr-4005 infection induced severe degenerative alterations within the central nervous system of infected mice, providing new information about the targets of this flavivirus infection.

Alphaviruses: Serological Evidence of Human Infection in Paraguay (2012-2013).

INTRODUCTION: Alphaviruses can produce febrile illness and encephalitis in dead-end hosts such as horses and humans. Within this genus, the Venezuelan Equine Encephalitis virus (VEEV) complex includes pathogenic epizootic subtypes and enzootic subtypes that are not pathogenic in horses (except subtype IE, Mexican strains), although they can cause febrile symptoms in humans. The Rio Negro virus (RNV-VEEV subtype VI) circulates in Argentina, where it was associated with undifferentiated febrile illness. Mayaro (MAYV) and Una (UNAV) viruses belong to a different group, the Semliki Forest virus complex, with confirmed circulation. OBJECTIVE: The present study aimed to determine RNV, MAYV, and UNAV seroprevalences by plaque reduction neutralization test in 652 samples of Paraguayan individuals mainly from the Central Department, between years 2012 and 2013. METHODS: Samples with antibodies titer >1:20 against RNV were also tested for Mosso das Pedras-subtype IF, subtype IAB, and Pixuna (PIXV)-subtype IV viruses that belongs to VEEV antigenic complex. RESULTS: The overall seroprevalence of RNV was 3.83%, and for UNAV it was 0.46%, and no neutralizing antibodies were detected against MAYV in the studied population. Two of the twenty-seven heterotypic samples were positive for PIXV. The 50.1% of neutralizing antibody titers against RNV were high (equal to or greater than 1/640), suggesting recent infections. The effect of age on the prevalence of RNV was negligible. CONCLUSIONS: These results bring new information about neglected alphaviruses in South America, and these data will serve as the basis for future studies of seroprevalence of other VEEV, and studies to search potential hosts and vectors of these viruses in the region.

Pixuna virus modifies host cell cytoskeleton to secure infection.

Pixuna virus (PIXV) is an enzootic member of the Venezuelan Equine Encephalitis Virus complex and belongs to the New World cluster of alphaviruses. Herein we explore the role of the cellular cytoskeleton during PIXV replication. We first identified that PIXV undergoes an eclipse phase consisting of 4 h followed by 20 h of an exponential phase in Vero cells. The infected cells showed morphological changes due to structural modifications in actin microfilaments (MFs) and microtubules (MTs). Cytoskeleton-binding agents, that alter the architecture and dynamics of MFs and MTs, were used to study the role of cytoskeleton on PIXV replication. The virus production was significantly affected (p < 0.05) after treatment with paclitaxel or nocodazole due to changes in the MTs network. Interestingly, disassembly of MFs with cytochalasin D, at early stage of PIXV replication cycle, significantly increased the virus yields in the extracellular medium (p < 0.005). Furthermore, the stabilization of actin network with jasplakinolide had no effect on virus yields. Our results demonstrate that PIXV relies not only on intact MTs for the efficient production of virus, but also on a dynamic actin network during the early steps of viral replication.

Tissue tropism of Saint Louis encephalitis virus: Histopathology triggered by epidemic and non-epidemic strains isolated in Argentina.

Saint Louis encephalitis virus (SLEV) reemerged in South America, and caused encephalitis outbreaks at the beginning of the 21st century. To enhance our knowledge about SLEV virulence, we performed comparative pathogenesis studies in Swiss albino mice inoculated with two different variants, the epidemic strain CbaAr-4005 and the non-epidemic strain CorAn-9275. Only the infection of mice with SLEV strain CbaAr-4005 resulted in high viremia, invasion of peripheral tissues including the lungs, kidney, and spleen, and viral neuroinvasion. This was associated with inflammatory pathology in the lungs, spleen, and brain as well as morbidity and mortality. In contrast, neither signs of desease nor viral replication were observed in mice infected with strain CorAn-9275. Interestingly, important loss of B cells and development of altered germinal centers (GC) were detected in the spleen of mice infected with strain CbaAr-4005, whereas mice infected with SLEV CorAn-9275 developed prominent GC with conserved follicular architecture, and neutralizing antibodies.

Silent circulation of St. Louis encephalitis virus prior to an encephalitis outbreak in Cordoba, Argentina (2005).

St. Louis encephalitis virus is a complex zoonoses. In 2005, 47 laboratory-confirmed and probable clinical cases of SLEV infection were reported in Córdoba, Argentina. Although the causes of 2005 outbreak remain unknown, they might be related not only to virological factors, but also to ecological and environmental conditions. We hypothesized that one of the factors for SLE reemergence in Córdoba, Argentina, was the introduction of a new SLEV genotype (SLEV genotype III), with no previous activity in the area. In order to evaluate this hypothesis we carried out a molecular characterization of SLEV detections from mosquitoes collected between 2001 and 2004 in Córdoba city. A total of 315 mosquito pools (11,002 individuals) including 12 mosquitoes species were analyzed. Overall, 20 pools (8 mosquitoes species) were positive for SLEV. During this study, genotypes II, V and VII were detected. No mosquito pool infected with genotype III was detected before the 2005 outbreak. Genotype V was found every year and in the 8 sampled sites. Genotypes II and VII showed limited temporal and spatial activities. We cannot dismiss the association of genotype II and V as etiological agents during the outbreak. However, the silent circulation of other SLEV strains in Córdoba city before the 2005 outbreak suggests that the introduction of genotype III was an important factor associated to this event. Not mutually exclusive, other factors such as changes in avian hosts and mosquitoes vectors communities, driven by climatic and environmental modifications, should also be taken into consideration in further studies.