Oviposition dynamics of Aedes aegypti in Central Argentina.

Aedes (Stegomyia) aegypti (L.) (Diptera: Culicidae) is the vector of multiple arboviruses. To evaluate the association between environmental factors and the oviposition activity of Ae. aegypti in Argentina, data on the presence and abundance of eggs were collected using ovitraps, between September of 2018 and May of 2019, in the cities of Villa María, Río Cuarto and Salsipuedes (Córdoba province, Argentina). We analysed the relationships between oviposition and five environmental factors: Temperature, precipitation, vegetation cover, human population density and distance to sites with a potential high density of larval habitats, like cemeteries and trash dumps. Environmental factors' data were collected using satellite image products. The oviposition activity was randomly distributed in three cities. Using generalized linear mixed models, we show that the house where each ovitrap was placed was a source of variability in oviposition, suggesting the relevance of microsite factors and the importance of domestic control actions. Ae. aegypti oviposition was positively correlated with night-time temperature of the previous 3 weeks, and in a context-dependent manner, it was positively correlated with human population density, vegetation cover and precipitation. The consistency and magnitude of these relationships varied between cities, indicating that oviposition is related to a complex system of environmental variables.

Arbovirus serosurvey (Orthobunyavirus, Flavivirus, and Alphavirus) in a draft horse population from Santa Fe, Argentina (2013-2016).

In the present study, we serosurveyed the exposure of 222 draft horses to different arboviruses in the city of Santa Fe, Argentina. Plaque reduction neutralization tests confirmed exposure to Fort Sherman virus (FSV), Saint Louis encephalitis virus (SLEV), West Nile virus (WNV), and Río Negro virus (RNV). Apparently, Western and Eastern equine encephalitis viruses did not circulate in the population tested. The confirmation of five seroconversions for WNV, FSV, and SLEV and the association between prevalence and age are indicative of recent circulation. These results highlight the importance of considering draft horses in arboviral surveillance in urban and rural areas of developing countries.

Effects of rainfall on Culex mosquito population dynamics.

The dynamics of a mosquito population depends heavily on climatic variables such as temperature and precipitation. Since climate change models predict that global warming will impact on the frequency and intensity of rainfall, it is important to understand how these variables affect the mosquito populations. We present a model of the dynamics of a Culex quinquefasciatus mosquito population that incorporates the effect of rainfall and use it to study the influence of the number of rainy days and the mean monthly precipitation on the maximum yearly abundance of mosquitoes Mmax. Additionally, using a fracturing process, we investigate the influence of the variability in daily rainfall on Mmax. We find that, given a constant value of monthly precipitation, there is an optimum number of rainy days for which Mmax is a maximum. On the other hand, we show that increasing daily rainfall variability reduces the dependence of Mmax on the number of rainy days, leading also to a higher abundance of mosquitoes for the case of low mean monthly precipitation. Finally, we explore the effect of the rainfall in the months preceding the wettest season, and we obtain that a regimen with high precipitations throughout the year and a higher variability tends to advance slightly the time at which the peak mosquito abundance occurs, but could significantly change the total mosquito abundance in a year.

Detection of Orthobunyavirus in mosquitoes collected in Argentina.

Bunyamwera virus (BUNV) (Bunyaviridae, genus Orthobunyavirus, serogroup Bunyamwera) is considered an emerging pathogen for humans and animals in American countries. The CbaAr-426 strain of BUNV was recovered from mosquitoes Ochlerotatus albifasciatus (Diptera: Culicidae) collected in Córdoba province (Argentina), where serological studies detected high seroprevalences in humans and animals. Molecular detection of Orthobunyavirus was performed in mosquitoes collected in Córdoba province. Seventeen mosquito pools of Oc. albifasciatus, Ochlerotatus scapularis and Culex quinquefasciatus (Diptera: Culicidae) showed positive results; four of these positive pools, all of Oc. scapularis, were sequenced. All amplicons grouped with BUNV in the Bunyamwera serogroup. The findings highlight the circulation of BUNV in Córdoba province and represent the first report of BUNV-infected Oc. scapularis mosquitoes in Argentina.

Evaluation of the cytotoxicity, genotoxicity and apoptotic induction of an aqueous extract of Achyrocline satureioides (Lam.) DC.

Achyrocline satureioides is widely consumed as infusion or aperitif and shows important therapeutic properties. Previously, we reported absence of genotoxicity of cold aqueous extract (CAE) of A. satureioides by Allium test. However, one test cannot predict the genotoxic effects of a substance. Thus, the aim of this work was to investigate cytotoxicity, genotoxicity and apoptotic ability of CAE of A. satureioides. In addition, CAE was chemically characterized. The cytotoxicity was evaluated by Trypan blue and MTT assays. The apoptotic capacity was evaluated by Hoechst staining and DNA fragmentation-analysis. The genotoxicity was studied by comet assay (CA) and micronucleus test. The identification and quantification of flavonoids were performed by HPLC-ESI-MS/MS. The cytotoxicity studies indicated low toxicity of CAE. In addition, CAE did not induce apoptotic effects on human PBMCs. CAE did not show genotoxicity in vitro against Vero cells, at 10-50 µg/mL. CAE did not induce in vivo genotoxic effects, but it showed at high concentrations cytotoxicity by micronucleus assay. CAE presented flavonoids such as quercetin, 3-O-methylquercetin and luteolin. In conclusion, A. satureioides at popularly concentrations used, in aperitif or infusion, can be consumed safely because did not show any cytotoxic or genotoxic effects.

Cocirculation of Rio Negro Virus (RNV) and Pixuna Virus (PIXV) in Tucumán province, Argentina.

Venezuelan equine encephalitis complex includes viruses considered emerging pathogens for humans and animals in the Americas. Two members of this complex have been detected previously in Argentina: Rio Negro Virus (RNV), detected in mosquitoes from Chaco province and rodents from Formosa province, and Pixuna Virus (PIXV), detected in mosquitoes from Chaco province. To carry out surveillance studies in other parts of the country, detection of a 195-bp fragment of alphaviruses by RT-nested PCR was performed in mosquito samples from San Miguel de Tucumán city. Four pools resulted positive and three were sequenced. Two amplicons grouped with RNV and one with PIXV. This is the first report of viral activity of members of the Venezuelan equine encephalitis complex in north-eastern Argentina.

[Prevalence of Flavivirus antibodies in Alouatta caraya primate autochthonous of Argentina]. / Prevalencia de anticuerpos para Flavivirus en el primate Alouatta caraya autoctono de la Argentina.

Flavivirus constitute a human health problem in our country. Primates are known to participate in the maintenance of Dengue and Yellow Fever viruses. However, these animals play a role which still remains to be determined in the maintenance of other viruses with potential pathogenicity for human beings and/or animals. Detección of antibodies was performed for different flavivirus in 105 sera samples of Alouatta caraya primates by the hemagglutination inhibition (HI) test. The neutralization (NT) test confirmed only infections caused by St. Louis Encephalitis (SLE) virus with a high prevalence in HI (35.23%) and NT (32.38%) antibodies. No antibody titres indicative of infections by Yellow Fever, Dengue and Bussuquara viruses were registered. Infection by the liheus virus could not be confirmed in these primates. There is a need for studies to detect new or reemergent viral infections in Argentina and the role that these primates could play in the maintenance of such infections.

Prevalencia de anticuerpos para Flavivirus en el primate Alouatta caraya autoctono de la Argentina. / [Prevalence of Flavivirus antibodies in Alouatta caraya primate autochthonous of Argentina]

Flavivirus constitute a human health problem in our country. Primates are known to participate in the maintenance of Dengue and Yellow Fever viruses. However, these animals play a role which still remains to be determined in the maintenance of other viruses with potential pathogenicity for human beings and/or animals. Detección of antibodies was performed for different flavivirus in 105 sera samples of Alouatta caraya primates by the hemagglutination inhibition (HI) test. The neutralization (NT) test confirmed only infections caused by St. Louis Encephalitis (SLE) virus with a high prevalence in HI (35.23

) and NT (32.38

) antibodies. No antibody titres indicative of infections by Yellow Fever, Dengue and Bussuquara viruses were registered. Infection by the liheus virus could not be confirmed in these primates. There is a need for studies to detect new or reemergent viral infections in Argentina and the role that these primates could play in the maintenance of such infections.

The role of mononuclear blood cells in experimental Junin virus spread to the central nervous system.

The neuroinvasiveness of Junin virus depends on the viral strain, animal species, and age. The role of infected blood cells in hematogenous Junin virus spread to the central nervous system (CNS) was studied by determining the growth in pheripheral mononuclear cells and brain tissue of Candid 1 and XJCL3 laboratory strains, in Calomys musculinus and guinea pigs. The present study demonstrated that Junin virus replicates in circulating peripheral lymphocytes and macrophages of 11-day-old guinea pigs and 6 +/- 1-day-old Calomys musculinus. Moreover, the observation that mononuclear phagocyte depletion did block Junin virus neuroinvasion firmly indicates that the cellular viremia (circulating monocytes) is one of the mechanisms of Junin virus attenuated strains spread to the CNS in animal hosts.

[Presence of antibodies against Venezuelan equine encephalitis virus subtype VI in patients with acute febrile illness]. / Presencia de anticuerpos contra el virus de la encefalitis equina venezolana subtipo VI en pacientes con enfermedad aguda febril.

In Argentina, there is no record of human cases produced by Dengue virus (Flavivirus), but Paraguay and Brasil (neighbouring countries) have notified human outbreaks of Dengue Haemorrhagic Fever. In this report, we inform the serological results of a limited human outbreak of a Dengue-like acute illness that occurred in General Belgrano Island, Formosa, Argentina in April 1989. This island is 35 km far from Clorinda city of Paraguay river, with a human population of 150 inhabitants. The weather of this area is humid with abundant rainfall, favouring mosquitoes proliferation. Two samples of serum from 28 human notified cases were studied using hemagglutination inhibition test (HI), complement fixation (CF), and plaque reduction neutralization (NT) test in Vero cell cultures. All tested sera were negative to Dengue, St. Louis encephalitis, Yellow Fever, Bussuquara, Rocio, Eastern and Western Equine Encephalitis arboviruses as well as Influenza and Rubella viruses. By contrast, infection with Venezuelan equine encephalitis virus (VEE), subtype VI-AG80-663 strain was demonstrated (34.5% positive by HI, 39.1% by CF and 51.6% by NT). Seroconversion was detected by NT in six cases and only five were positive by CF. The 26.8% of the sera reacted also with VEE subtype I AB by NT. Considering that no cross reaction were detected in NT with these two subtypes, our results suggest that both viruses are concomitantly circulating in the studied area. Furthermore, the seroconversions detected with AG80-663 strain firmly indicate that during the outbreak this virus subtype was circulating in the island, although we could not assure that it was the causal agent of the acute disease.

Presencia de anticuerpos contra el virus de la encefalitis equina venezolana subtipo VI en pacientes con enfermedad aguda febril / [Presence of antibodies against Venezuelan equine encephalitis virus subtype VI in patients with acute febrile illness]

In Argentina, there is no record of human cases produced by Dengue virus (Flavivirus), but Paraguay and Brasil (neighbouring countries) have notified human outbreaks of Dengue Haemorrhagic Fever. In this report, we inform the serological results of a limited human outbreak of a Dengue-like acute illness that occurred in General Belgrano Island, Formosa, Argentina in April 1989. This island is 35 km far from Clorinda city of Paraguay river, with a human population of 150 inhabitants. The weather of this area is humid with abundant rainfall, favouring mosquitoes proliferation. Two samples of serum from 28 human notified cases were studied using hemagglutination inhibition test (HI), complement fixation (CF), and plaque reduction neutralization (NT) test in Vero cell cultures. All tested sera were negative to Dengue, St. Louis encephalitis, Yellow Fever, Bussuquara, Rocio, Eastern and Western Equine Encephalitis arboviruses as well as Influenza and Rubella viruses. By contrast, infection with Venezuelan equine encephalitis virus (VEE), subtype VI-AG80-663 strain was demonstrated (34.5

positive by HI, 39.1

by CF and 51.6

by NT). Seroconversion was detected by NT in six cases and only five were positive by CF. The 26.8

of the sera reacted also with VEE subtype I AB by NT. Considering that no cross reaction were detected in NT with these two subtypes, our results suggest that both viruses are concomitantly circulating in the studied area. Furthermore, the seroconversions detected with AG80-663 strain firmly indicate that during the outbreak this virus subtype was circulating in the island, although we could not assure that it was the causal agent of the acute disease.

Presencia de anticuerpos contra el virus de la encefalitis equina venezolana subtipo VI en pacientes con enfermedad aguda febril. / [Presence of antibodies against Venezuelan equine encephalitis virus subtype VI in patients with acute febrile illness]

In Argentina, there is no record of human cases produced by Dengue virus (Flavivirus), but Paraguay and Brasil (neighbouring countries) have notified human outbreaks of Dengue Haemorrhagic Fever. In this report, we inform the serological results of a limited human outbreak of a Dengue-like acute illness that occurred in General Belgrano Island, Formosa, Argentina in April 1989. This island is 35 km far from Clorinda city of Paraguay river, with a human population of 150 inhabitants. The weather of this area is humid with abundant rainfall, favouring mosquitoes proliferation. Two samples of serum from 28 human notified cases were studied using hemagglutination inhibition test (HI), complement fixation (CF), and plaque reduction neutralization (NT) test in Vero cell cultures. All tested sera were negative to Dengue, St. Louis encephalitis, Yellow Fever, Bussuquara, Rocio, Eastern and Western Equine Encephalitis arboviruses as well as Influenza and Rubella viruses. By contrast, infection with Venezuelan equine encephalitis virus (VEE), subtype VI-AG80-663 strain was demonstrated (34.5

positive by HI, 39.1

by CF and 51.6

by NT). Seroconversion was detected by NT in six cases and only five were positive by CF. The 26.8

of the sera reacted also with VEE subtype I AB by NT. Considering that no cross reaction were detected in NT with these two subtypes, our results suggest that both viruses are concomitantly circulating in the studied area. Furthermore, the seroconversions detected with AG80-663 strain firmly indicate that during the outbreak this virus subtype was circulating in the island, although we could not assure that it was the causal agent of the acute disease.

Experimental neuroinvasiveness of wild and laboratory Junin virus strains.

The neuroinvasiveness of Candid 1 and XJCL3 laboratory strains and CbalV4454 and CbaFHA5069 wild strains of Junin virus was studied in albino mice, guinea pigs, and a South American wild rodent, Calomys musculinus (Cm), of different ages inoculated by a non-neural route. Infectivity in brain, blood and organs, as well as lethality, were determined. The results with the 3 hosts indicate that Junin virus neuroinvasiveness is virus-strain-dependent, host species- and age-dependent, with the Candid 1 strain proving to be the least neuroinvasive of the strains studied. The lethal efficiency index (log PFU/LD50) in 2-day old albino mice and the neuroinvasiveness index (Log PFU/ND50) in 6 +/- 1 day-old Cm of the various strains using the intraperitoneal (ip) route could therefore be useful markers of Junin virus neuroinvasiveness. Moreover, different patterns of infection were established using the results of the presence of infectious virus in brain and viraemia in the 3 hosts. In nearly all cases, virus neuroinvasion was present without detectable viraemia (virus in plasma). Current evidence leads to the assumption that virus might reach the brain associated with the white cells in blood (undetectable by conventional isolation methods) or by another possible mechanism of neuroinvasion which is not haematogenous.

Rapid vascular clearance of two strains of Junin virus in Calomys musculinus: selective macrophage clearance.

Clearance of Junin (JUN) virus strains with different virulence for Calomys musculinus (Cm) was followed using the Candid #1 virulent and CbaFHA 5069 attenuated strains. In addition, virulent virus albino mice (AM) were included as control host and Venezuelan equine encephalitis (VEE-VI) virus as control virus. The virus inoculum (Vo) and the blood samples (Vt) obtained at different times post-inoculation (p.i.) were titrated on Vero cells and the cleared plaque forming-units (PFU) were calculated as the log Vt/Vo. In Cm both JUN virus strains were cleared rapidly (within 5 min the Candid #1 strain and within 10 min the CbaFHA 5069 strain); meanwhile, VEE-VI virus could be recovered from blood until 30 min p. i. Furthermore, JUN and VEE-VI viruses showed the same behaviour in Am as in Cm. We conclude that the JUN virus strains of different virulence for Cm did not show differences in their clearance from the blood of these animals. Moreover, the rapid clearance observed was independent of the animal host and viral dose.

Neurovirulence of wild and laboratory Junin virus strains in animal hosts.

The neurovirulence of Candid #1 and XJCL3 laboratory strains and CbalV4454 and CbaFHA5069 wild strains of Junin virus was studied in albino mice, guinea pigs, and a South American wild rodent, Calomys musculinus, of different ages inoculated by the intracerebral route. Infectivity in brain and organs, lethality, and neuropathological lesions were determined. The laboratory and wild strains showed similar neurovirulence only in 2-day-old mice. The neurovirulence of laboratory strains decreased with the age of the animal, and the Candid #1 strain affected only 2-day-old mice. In guinea pigs, the 2 wild strains and XJCL3 laboratory strain were neurovirulent for 11-day-old and adult animals giving moderate lymphocytic infiltration in the brain and mild lesions in the spinal cord. Virus titres from the brain and the spinal cord were lower with the XJCL3 and CbalV4454 strains than with the CbaFHA5069 strain; with the latter, virus was recovered only from the lymph nodes, the lung, kidney, liver, and spleen. The Candid #1 strain was not neurovirulent even for 11-day-old animals. In contrast, the laboratory strains were neurovirulent for Calomys musculinus, depending on the age of the animal. Virus was recovered from the brains showing lymphocyte infiltration but not from other organs. The CbaFHA5069 strain was not neurovirulent, although virus was recovered from the brain, spleen, liver, lymph nodes, and salivary glands. These results with the 3 hosts indicate that Junin virus neurovirulence is virus strain-dependent, and host species and age-dependent, with the Candid #1 strain proving the least neurovirulent of the strains studied.

[Concomitant activity of 2 bunyaviruses in horses in Argentina]. / Actividad concomitante de dos Bunyavirus en caballos de la Argentina.

A serologic survey of horses for Kairi (KRI) and Cache Valley (CV), two related Bunyaviruses, was conducted simultaneously in Cordoba and Santa Fe provinces, Argentina, during late 1983 and 1984. The prevalence of neutralizing antibodies only for KRI was 13.3% and only for CV was 40.0%; but if the total positive sera for KRI and CV were taken into account, the prevalence reached 48.3 and 75.0%, respectively. The prevalence for CV was higher than for KRI in Cordoba (p less than 0.01), but both were similar in Santa Fe province. The demonstration of seroconversion in horses of the two zones for both viruses indicates that these viruses have a concomitant activity. The infection rates (number of infections per 100 horses-month) were very high in Cordoba (4.4 and 7.1 for KRI and CV) but also in Santa Fe (2.9 and 9.5 for the two viruses respectively), without significant difference in each province. Despite this high activity, no signs of illness or death imputed to these viruses were registered, in these areas during the period of observation. This apparent absence of associated equine disease may be the consequence of the low or null virus pathogenicity or the underrecognition or underreporting of the clinical cases.

Actividad concomitante de dos Bunyavirus en caballos de la Argentina / [Concomitant activity of 2 bunyaviruses in horses in Argentina]

A serologic survey of horses for Kairi (KRI) and Cache Valley (CV), two related Bunyaviruses, was conducted simultaneously in Cordoba and Santa Fe provinces, Argentina, during late 1983 and 1984. The prevalence of neutralizing antibodies only for KRI was 13.3

and only for CV was 40.0

; but if the total positive sera for KRI and CV were taken into account, the prevalence reached 48.3 and 75.0

, respectively. The prevalence for CV was higher than for KRI in Cordoba (p less than 0.01), but both were similar in Santa Fe province. The demonstration of seroconversion in horses of the two zones for both viruses indicates that these viruses have a concomitant activity. The infection rates (number of infections per 100 horses-month) were very high in Cordoba (4.4 and 7.1 for KRI and CV) but also in Santa Fe (2.9 and 9.5 for the two viruses respectively), without significant difference in each province. Despite this high activity, no signs of illness or death imputed to these viruses were registered, in these areas during the period of observation. This apparent absence of associated equine disease may be the consequence of the low or null virus pathogenicity or the underrecognition or underreporting of the clinical cases.

Actividad concomitante de dos Bunyavirus en caballos de la Argentina. / [Concomitant activity of 2 bunyaviruses in horses in Argentina]

A serologic survey of horses for Kairi (KRI) and Cache Valley (CV), two related Bunyaviruses, was conducted simultaneously in Cordoba and Santa Fe provinces, Argentina, during late 1983 and 1984. The prevalence of neutralizing antibodies only for KRI was 13.3

and only for CV was 40.0

; but if the total positive sera for KRI and CV were taken into account, the prevalence reached 48.3 and 75.0

, respectively. The prevalence for CV was higher than for KRI in Cordoba (p less than 0.01), but both were similar in Santa Fe province. The demonstration of seroconversion in horses of the two zones for both viruses indicates that these viruses have a concomitant activity. The infection rates (number of infections per 100 horses-month) were very high in Cordoba (4.4 and 7.1 for KRI and CV) but also in Santa Fe (2.9 and 9.5 for the two viruses respectively), without significant difference in each province. Despite this high activity, no signs of illness or death imputed to these viruses were registered, in these areas during the period of observation. This apparent absence of associated equine disease may be the consequence of the low or null virus pathogenicity or the underrecognition or underreporting of the clinical cases.