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.

May captive populations of Greater Rhea (Rhea americana) act as genetic reservoirs in Argentina?

The Greater Rhea (Rhea americana) is a characteristic bird of the Argentine Pampas. Despite the increasing farming interest of this ratite, their natural populations are progressively decreasing in size and range. The object of this study was to evaluate the status of captive populations as potential genetic reservoirs. Using Inter-Simple Sequence Repeats as molecular markers, levels of genetic variability of F1 individuals from two captive populations were estimated and compared with those of wild populations in the same region. The captive populations were polymorphic for 12.22 and 13.33% of the loci, with a genetic diversity of 0.050. Differences with wild populations were not significant (z=1.79; P>0.05). Therefore, captive populations of rheas in Argentina should not be overlooked as genetic reservoir and source of individuals for reinforcement of natural populations, through reintroduction and translocation.

Genetic relationships among populations of Aedes aegypti from Uruguay and northeastern Argentina inferred from ISSR-PCR data.

Aedes aegypti (L.) (Diptera: Culicidae), the main vector of yellow fever and dengue viruses, was eradicated from Argentina between 1955 and 1963, but reinvaded the country in 1986. In Uruguay, the species was reintroduced in 1997. In this study we used highly polymorphic inter-simple sequence repeats (ISSR) markers to analyse the genetic structure of Ae. aegypti populations from Uruguay and northeastern Argentina to identify possible colonization patterns of the vector. Overall genetic differentiation among populations was high (F(ST) = 0.106) and showed no correlation with geographic distance, which is consistent with the short time since the reintroduction of the species in the area. Differentiation between pairs of Argentine populations (F(ST) 0.072 to 0.221) was on average higher than between Uruguayan populations (F(ST)-0.044 to 0.116). Bayesian estimation of population structure defined four genetic clusters and most populations were admixtures of two of them: Mercedes and Treinta y Tres (Uruguay) were mixtures of clusters 1 and 3; Salto (Uruguay) and Paraná (Argentina) of clusters 1 and 4; Fray Bentos (Uruguay) of clusters 2 and 3, and Gualeguaychú (Argentina) of clusters 2 and 3. Posadas and Buenos Aires in Argentina were fairly genetically homogeneous. Our results suggest that Ae. aegypti recolonized Uruguay from bordering cities in Argentina via bridges over the Uruguay River and also from Brazil.

Current resistance status to temephos in Aedes aegypti from different regions of Argentina.

In Argentina, more than 25,000 cases of dengue were reported in the summer of 2009, even in provinces where the disease was formerly absent. We analysed the susceptibility levels to the larvicide temephos in seven populations of Aedes aegypti, the primary vector of dengue, collected during summer 2007/2008, using the susceptible Rockefeller strain as a control. Although no control failures were observed during the experiment, a majority of the lethal concentration and resistance ratio values indicate an incipient resistance. An integrative program to monitor the resistance of Ae. aegypti to insecticides is needed in the country.

Two different routes of colonization of Aedes aegypti in Argentina from neighboring countries.

Aedes aegypti L. (Diptera, Culicidae) is the main vector of dengue and yellow fever. In Argentina, the species was apparently eradicated approximately in 1964; by 1986, it was reintroduced. To identify different gene pools in geographical populations of the species and to ascertain the possible routes of colonization, we analyzed the diversity of mitochondrial DNA haplotypes in 572 specimens from Argentina and neighboring countries. We found that the restriction fragment length polymorphism-polymerase chain reaction screening of a large DNA fragment including the A+T-rich region was the best strategy to reconstruct the colonization pattern ofAe. aegypti in Argentina. Twenty haplotypes were recognized; levels of genetic similarity varied among populations from different geographical locations. The haplotype network constructed on the basis of genetic distances showed three well differentiated groups. Two of them exhibited a well defined spatial distribution and populations in these groups presented an isolation-by-distance pattern. The persistence of relictual populations after the last eradication campaigns would explain the high levels of haplotype diversity and the presence of exclusive haplotypes in urban centers from northwestern Argentina. Eastern Argentine populations showed one prevalent haplotype, also predominant in Brazil and Paraguay. Our results highlight the need for efficient surveys and control campaigns, given the strong effect of land trade on genetic exchange among mosquito populations from Argentina and neighboring countries where dengue is endemic.

Recent range expansion and low levels of contemporary gene flow in Calomys musculinus: its relationship with the emergence and spread of Argentine haemorrhagic fever.

The geographic distribution of haplotype diversity in the rodent Calomys musculinus, sampled from 16 wild populations of Argentina, was analysed on two geographical scales. The species is the natural reservoir of the Junin virus, the etiological agent of the Argentine haemorrhagic fever (AHF). In all, 24 composite haplotypes were recognised in the mtDNA D-loop region. Haplotypes 1 and 2, internal in the network, were the most frequent and were present in almost all populations. The absence of large genetic gaps between widely distributed haplotypes, the existence of exclusive haplotypes in more than 50% of the sampled populations and the absence of isolation by distance at a macrogeographical scale are in support of the hypothesis of a recent range expansion of the populations of the Humid Pampa, with low to moderate current gene flow. The dispersal of this opportunistic species would have been favoured by the explosive increments in density after agriculture was introduced. When only nearby populations within the endemic area of AHF were considered, a pattern of isolation by distance was detected. At present, genetic drift appears to be the main force acting to randomly differentiate C. musculinus populations, which would also lead to random differentiation of Junin virus strains and a reduction in the virulence of the pathogen in 'historic' AHF areas. The knowledge of migration patterns of the reservoir populations facilitates reliable prediction of the potential spread of the human disease.

Distribution of genetic variability in populations of two chromosomal races of Dichroplus pratensis (Melanoplinae, Acrididae) and their hybrid zone.

We examined, through allozyme electrophoresis, the genetic structure of populations of the acridid grasshopper Dichroplus pratensis from two chromosomal races (Northern and Southern) and their hybrid zone in Argentina. No fixed alleles for any particular race were found, although genetic differentiation among parental races was significant (0 = 0.044, 95% CI: 0.004-0.068). Hybrid populations are genetically more similar to the Southern race (0 = 0.008, 95% CI: -0.005-0.018) than to Northern ones (0 = 0.018, 95% CI: 0.002-0.030). Differential viability or fertility of hybrids, or asymmetry in mating preferences in favour of one particular cross would cause a higher proportion of matings between hybrid individuals and those from the Southern race. This would explain the high genetic similarity between those groups, in spite of their geographical vicinity with northern race populations.

Restricted gene flow in Calomys musculinus (Rodentia, Muridae), the natural reservoir of Junin virus.

Random amplified polymorphic DNA polymerase chain reaction (RAPD-PCR) markers were used to evaluate the relative contribution of gene flow as a determinant of the population genetic structure of the wild rodent Calomys musculinus (the reservoir of Argentine hemorrhagic fever [AHF]) in central Argentina. One hundred eighty-seven individuals from 13 populations (9 of them from the endemic zone of AHF and 5 from areas outside it) were analyzed using 78 polymorphic RAPD loci. Genetic variation within each population was high; each individual was characterized by a unique RAPD phenotype. C. musculinus populations showed a moderate to high genetic subdivision and a random pattern of differentiation. Populations separated by the same geographic distance showed very different degrees of genetic divergence. The results indicate that populations of C. musculinus have colonized their present ranges relatively recently and differentiation by genetic drift has proceeded faster than homogenization by gene flow at the macrogeographic scale analyzed (10-700 km).

Genetic relationships among Aedes aegypti (Diptera: Culicidae) populations from Argentina using random amplified polymorphic DNA polymerase chain reaction markers.

Random amplified polymorphic DNA polymerase chain reaction (RAPD-PCR) polymorphism was analyzed in five Aedes aegypti (L.) populations from Argentina and one from Puerto Rico to estimate levels of intraspecific polymorphism and genetic relatedness. Allele frequencies were estimated assuming that RAPD products segregate as dominants and that genotype frequencies at those loci are in Hardy-Weinberg equilibrium. Mean expected heterozygosity (He) was 0.350; F(ST) values were significant at all loci except one, supporting the usefulness of the fragments used here to discriminate among populations. Rogers' genetic similarity between samples ranged from 0.806 to 0.621. The population from Puerto Rico was the most different from the Argentina populations. Considering that Ae. aegypti eggs, larvae, and pupae can be transported easily, relationships among the Argentinian populations may reflect the routes and intensity of commercial transit.

Inheritance of random amplified polymorphic DNA (RAPD-PCR) markers and their use in population genetic studies of Calomys musculinus (Rodentia, Muridae), the reservoir of Argentine hemorrhagic fever.

In order to assess the reliability of RAPD markers in the estimation of the genetic structure of natural populations of the murid rodent Calomys musculinus (reservoir of Junin virus, ethiological agent of Argentine Hemorrhagic Fever), we have analyzed the heritability of RAPD bands in 10 parents and their offspring (33 individuals). Fourteen out of a total of 119 bands obtained were absent in the parental patterns, but consistently amplified in offspring from some families. These bands can be eliminated from analyses. Overall degree of band sharing between individuals, including non-parental bands, correctly grouped members of a family in the same cluster in a UPGMA tree, with a high bootstrap percentage. Results support the usefulness of RAPDs as hereditable markers. One hundred polymorphic RAPD loci were identified in three natural populations of C. musculinus. Mean expected heterozygosity in three natural populations ranged from 0.206 to 0.220. Allele frequency based and phenotype based measures of genetic differentiation among natural populations of C. musculinus gave similar results (Weir and Cockerham's theta = 0.133; Excoffier et al.'s phi = 0.127). These values were considerably higher than those found previously using allozymes as genetic markers, and are compatible with moderate to low levels of gene flow among populations.

Allozymic polymorphism in Aedes aegypti populations from Argentina.

In Argentina, reinfestation with Aedes aegypti was registered in 1986. At present, the mosquito is well established in 10 provinces, from Buenos Aires up to the country's northern frontiers. This paper presents estimates of genetic variability in Ae. aegypti populations from central Argentina and determinations of genetic distances among them. We analyzed allozymic frequencies at 11 loci in samples from 3 localities. The proportion of polymorphic loci varied between 27.3 and 63.6. Expected mean heterozygosity ranged from 0.090 to 0.161 and Rogers' similarity among samples ranged between 0.909 and 0.958. The lack of relationship between genetic and geographic distances is in agreement with a recent colonization of the studied area. The mean Wright's coefficient FST value (0.065) indicates low levels of genetic differentiation among populations from different localities. Given the recent reinfestation with this mosquito in Argentina, the high levels of polymorphism found could indicate multiple introductions of representative samples from genetically different subpopulations.

Genetic structure of Native Andean populations from Argentina inhabiting different altitudes.

On the basis of erythrocyte and serum protein polymorphisms, we investigated the level of genetic differentiation among 3 Argentinian native populations sharing ethnic and morphological characteristics but inhabiting different altitudes in the Andes. Of the 15 loci studied, 8 are monomorphic. Phenotype distribution of the polymorphic systems are in agreement with Hardy-Weinberg equilibrium, except for malic enzyme (MOD), where a silent allele was found. The 2 highland populations exhibit a close genetic similarity, probably as a result of intense gene flow. The lowland population shows reduced genetic diversity, which suggests the influence of stochastic phenomena such as the founder effect.

Genetic structure of Aedes albifasciatus (Diptera: Culicidae) populations in central Argentina determined by random amplified polymorphic DNA-polymerase chain reaction markers.

The floodwater mosquito, Aedes albifasciatus (Macquart), is the main vector of western equine encephalomyelitis virus in Argentina. Previous studies on the genetic structure of this species using allozymes showed low levels of polymorphism, absence of subpopulations at distinct habitats, and moderate differentiation among localities separated up to 500 km. To examine gene flow using other genetic methods, we analyzed random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) polymorphism in 28 presumptive loci of Ae. albifasciatus from 6 populations in central Argentina. Allele frequencies were estimated assuming that RAPD products segregate as dominants and that genotype frequencies at those loci are in Hardy-Weinberg equilibrium. Expected heterozygosity ranged between 0.19 and 0.31, approximately 3 times the value obtained on the basis of the 16 allozymic loci studied previously. Four of the populations formed a single panmictic unit. Allele frequencies in populations occupying different phytogeographic regions gave significant FST values at 5 loci. Effective migration rates among populations estimated from FST ranged from 2.3 to 9.0. The results support the existence of a north-south cline.

Protein polymorphism in three South Amerindian populations.

Electrophoretic polymorphism at loci coding for blood red cells and plasma proteins was analysed in three native populations of South America: Mataco and Macá (from the Gran Chaco), and Guayaki (from the Paraguayan forest). Mean heterozygosity per locus varied from 0.024 and 0.073, the Macá being the most polymorphic population, probably reflecting admixture with the caucasoid population of Asuncion city. Although the external phenotype, blood group frequencies and ethno-linguistic affiliation of the Guayaki clearly differ from those of the Chaco tribes, Nei's genetic distances were very low and almost equidistant among the three populations. Heterogeneity of F(ST) values among polymorphic loci would suggest the action of natural selection.

Estimation of genetic divergence and gene flow between Culex pipiens and Culex quinquefasciatus (Diptera: Culicidae) in Argentina.

Allele frequencies at seven polymorphic loci controlling the synthesis of enzymes were analyzed in six populations of Culex pipiens L. and Cx. quinquefasciatus Say. Sampling sites were situated along a north-south line of about 2,000 km in Argentina. The predominant alleles at Mdh, Idh, Gpdh and Gpi loci presented similar frequencies in all the samples. Frequencies at the Pgm locus were similar for populations pairs sharing the same geographic area. The loci Cat and Hk-1 presented significant geographic variation. The latter showed a marked latitudinal cline, with a frequency for allele b ranging from 0.99 in the northernmost point to 0.04 in the southernmost one, a pattern that may be explained by natural selection (FST = 0.46; p < 0.0001) on heat sensitive alleles. The average value of FST (0.088) and Nm (61.12) indicated a high gene flow between adjacent populations. A high correlation was found between genetic and geographic distance (r = 0.83; p < 0.001). The highest genetic identity (I(N) = 0.988) corresponded to the geographically closest samples from the central area. In one of these localities Cx. quinquefasciatus was predominant and hybrid individuals were detected, while in the other, almost all the specimens were identified as Cx. pipiens. To verify the fertility between Cx. pipiens and Cx. quinquefasciatus from the northern- and southernmost populations, experimental crosses were performed. Viable egg rafts were obtained from both reciprocal crosses. Hatching ranged from 76.5 to 100%. The hybrid progenies were fertile through two subsequent generations.

Oligoryzomys flavescens (Rodentia, Muridae): gene flow among populations from central-eastern Argentina.

In species acting as hosts of infectious agents, the extent of gene flow between populations is of particular interest because the expansion of different infectious diseases is usually related to the dispersal of the host. We have estimated levels of gene flow among populations of the sigmodontine rodent Oligoryzomys flavescens, in which high titers of antibodies have been detected for a Hantavirus in Argentina that produces a severe pulmonary syndrome. Enzyme polymorphism was studied by means of starch gel electrophoresis in 10 populations from the area where human cases of Hantavirus have occurred. Genetic differentiation between populations was calculated from FST values with the equation Nm = [(1/FST) - 1]/4. To assess the relative importance of current gene flow and historical associations between populations, the relationship of population pairwise log Nm and log geographic distance was examined. Low FST (mean = 0.038) and high Nm (15.27) values suggest high levels of gene flow among populations. The lack of an isolation by distance pattern would indicate that this species has recently colonized the area. The northernmost population, located on the margin of a great river, shows very high levels of gene flow with the downstream populations despite the large geographic distances. Passive transport of animals down the river by floating plants would promote unidirectional gene flow. This fact and the highest mean heterozygosity of that northernmost population suggest it is a center of dispersal within the species' range.

Temporal variation of allozyme frequencies in Aedes albifasciatus (Diptera: Culicidae) from Argentina.

Aedes albifasciatus is a floodwater mosquito widely distributed in Argentina. It is important from economic and medical points of view. A 4-year survey of seasonal variation in allele frequencies in a population of this species was undertaken to determine possible changes in the genetic structure and their correlation with environmental conditions. Significant temporal variation was detected at most of the loci, but it did not follow a cyclic or seasonal pattern. Multivariate analysis of principal components showed a remarkable homogeneity of samples collected from December 1993 to April 1995 and a clear differentiation of the November 1991, March 1992, and November 1993 samples. This variation could be correlated with the magnitude of rainfall occurring in the area. Passive transport of larvae by water streams and river freshets produced by floods would have mixed larvae from breeding sites with different allele frequencies, causing the genetic differentiation observed.

Gene flow in Aedes albifasciatus (Diptera:Culicidae) from central Argentina.

To estimate gene flow among populations of the floodwater mosquito Aedes albifasciatus, allozymic frequencies were analyzed at 16 loci in samples from 6 localities in central Argentina. Expected mean heterozygosity ranged from 0.057 to 0.137. FST was significant for 5 of 11 polymorphic loci; mean value was 0.024, which gave an Nm of 50.96. Levels of genetic identity among samples were high (IN between 0.9815 and 0.9988) even between populations 500 km apart. However, there was a significant correlation between genetic and geographic distance, which indicated that although the 6 populations were at approximate equilibrium because of gene flow, this was restricted by distance. The distribution of allele frequencies possibly reflected the present status of a single large population occupying this region in the past. When geological change separated breeding areas, a moderate level of gene flow may have resulted in the current genetic structure of the populations.