Spread of Phlebotominae in temperate climates: province of Córdoba, Argentina.

We report the presence of the competent vector for Leishmania spp, Migonemyia migonei, and the Evandromyia cortelezzii-sallesi complex south of its known distribution in the central temperate region of Argentina, in the province of Córdoba. The persistence of this phlebotomine in the northern border of the province, its association with a case of cutaneous leishmaniasis, and the new record in the outskirts of the city of Córdoba, the second most populated in the country, strengthens the need for regular vector surveillance and a case detection-sensitive health system in vulnerable regions, even in temperate climates.

Systematic Review of Impacts of Educational Interventions to Control Breeding Sites of Aedes aegypti and Aedes albopictus Mosquitoes.

Community participation is a critical element in the management of Aedes aegypti and Aedes albopictus breeding sites. Many educational interventions have been conducted to encourage prevention and elimination of breeding sites among different community actors, such as government-run programs for vector surveillance aimed at preventing and eliminating breeding sites at the household level within a community. Getting people involved in prevention and elimination of vector breeding sites in their communities requires communication and social mobilization strategies to promote and reinforce those prevention actions that, in turn, should be effective from the entomological standpoint. Articles published in English, Spanish, and Portuguese, were reviewed to assess whether educational interventions targeting Ae. aegypti and Ae. albopictus were effective in reducing entomological indicators or in improving practices to prevent the presence of or eliminate breeding sites. The most widely used indicators were larval indices and the practices associated with reducing/eliminating breeding sites. We found that using a community-based approach adapted to eco-epidemiological and sociocultural scenarios explains the reduction of entomological indicators by educational interventions. Those who design or implement educational interventions should strengthen the evaluation of those interventions using qualitative approaches that provide a more complete picture of the social context and the barriers and facilitators to implementing vector control. Engaging school children in cross-sectorial collaboration involving the health and education spheres promotes the participation of the community in vector surveillance and reduces the risk of arboviral disease transmission.

Culicidae (Diptera) selection of humans, chickens and rabbits in three different environments in the province of Chaco, Argentina.

Studies were conducted to determine the selection of humans, chickens and rabbits by Culicidae in three different environments in the province of Chaco, Argentina. Mosquitoes were collected fortnightly using cylindrical metal traps containing animal bait (chickens and rabbits). The mosquitoes were collected between June 2001-May 2002. During the same period and with the same frequency, mosquitoes biting the human operators of the traps were collected during the first 15 min of exposure within different time intervals: from 09:00 am-11:00 am, 01:00 pm-03:00 pm, 05:00 pm-07:00 pm and 09:00 pm-10:00 pm. A total of 19,430 mosquitoes of 49 species belonging to 10 genera were collected. Culex species mainly selected chicken bait and Wyeomyia species selected rabbit bait. Ochlerotatus and Psorophora species were more abundant in rabbit-baited traps. Anopheles triannulatus, Coquillettidia nigricans, Ochlerotatus scapularis, Mansonia titillans and Psorophora albigenu showed a strong attraction for human bait. The Anopheles, Coquillettidia, Culex and Mansonia species were more active between 05:00 pm-09:00 pm, while Ochlerotatus, Psorophora, Haemagogus and Wyeomyia were most active from 09:00 am-07:00 pm. This study provides additional information about the biology and ecology of arbovirus vectors in Chaco.

COI barcode versus morphological identification of Culex (Culex) (Diptera: Culicidae) species: a case study using samples from Argentina and Brazil.

Sequences of the cytochrome c oxidase subunit I (COI) mitochondrial gene from adults of 22 Culex (Culex) species from Argentina and Brazil were employed to assess species identification and to test the usefulness of COI for barcoding using the best close match (BCM) algorithm. A pairwise Kimura two-parameter distance matrix including the mean intra and interspecific distances for 71 COI barcode sequences was constructed. Of the 12 COI lineages recovered in the Neighbour-joining topology, five confirmed recognised morphological species (Cx. acharistus, Cx. chidesteri, Cx. dolosus, Cx. lygrus and Cx. saltanensis) with intraspecific divergences lower than 1.75%. Cx. bilineatus is formally resurrected from the synonymy of Cx. dolosus. Cx. maxi , Cx. surinamensis and the Coronator group species included were clustered into an unresolved lineage. The intraspecific distance of Cx. pipiens (3%) was almost twice the interspecific between it and Cx. quinquefasciatus (1.6%). Regarding the BCM criteria, the COI barcode successfully identified 69% of all species. The rest of the sequences, approximately 10%, 18% and 3%, remained as ambiguously, mis and unidentified, respectively. The COI barcode does not contain enough information to distinguish Culex (Cux.) species.

First detection of Rio Negro virus (Venezuelan equine encephalitis complex subtype VI) in Córdoba, Argentina.

Rio Negro virus (RNV) (Venezuelan equine encephalitis subtype VI) circulates only in Argentina; in northern provinces, isolates have been obtained from mosquitoes and rodents since 1980 and have been associated with acute febrile illness in humans. However, no studies of RNV have been performed in the central area of the country. We carried out molecular and serological detection of RNV in Córdoba, a province of the central part of the country, in mosquitoes and humans, respectively. One mosquito pool tested positive for alphavirus RNA by reverse transcriptase-nested polymerase chain reaction (RT-nested PCR). Subsequent sequencing determined that this alphavirus grouped with RNV. Serological studies detected antibodies to RNV in one human serum sample, which was obtained during the same period that RNV was detected using the aforementioned molecular methods. This is the first report of RNV circulation in the central area of Argentina, indicating an expansion of its original distribution. These results highlight the importance of strengthening surveillance procedures in endemic areas, as well as in new regions where RNV may emerge.

Classification of immature mosquito species according to characteristics of the larval habitat in the subtropical province of Chaco, Argentina.

To classify mosquito species based on common features of their habitats, samples were obtained fortnightly between June 2001-October 2003 in the subtropical province of Chaco, Argentina. Data on the type of larval habitat, nature of the habitat (artificial or natural), size, depth, location related to sunlight, distance to the neighbouring houses, type of substrate, organic material, vegetation and algae type and their presence were collected. Data on the permanence, temperature, pH, turbidity, colour, odour and movement of the larval habitat's water were also collected. From the cluster analysis, three groups of species associated by their degree of habitat similarity were obtained and are listed below. Group 1 consisted of Aedes aegypti. Group 2 consisted of Culex imitator, Culex davisi, Wyeomyia muehlensi and Toxorhynchites haemorrhoidalis separatus. Within group 3, two subgroups are distinguished: A (Psorophora ferox, Psorophora cyanescens, Psorophora varinervis, Psorophora confinnis, Psorophora cingulata, Ochlerotatus hastatus-oligopistus, Ochlerotatus serratus, Ochlerotatus scapularis, Culex intrincatus, Culex quinquefasciatus, Culex pilosus, Ochlerotatus albifasciatus, Culex bidens) and B (Culex maxi, Culex eduardoi, Culex chidesteri, Uranotaenia lowii, Uranotaenia pulcherrima, Anopheles neomaculipalpus, Anopheles triannulatus, Anopheles albitarsis, Uranotaenia apicalis, Mansonia humeralis and Aedeomyia squamipennis). Principal component analysis indicates that the size of the larval habitats and the presence of aquatic vegetation are the main characteristics that explain the variation among different species. In contrast, water permanence is second in importance. Water temperature, pH and the type of larval habitat are less important in explaining the clustering of species.

Gonoactivity of Culex (Culex) (Diptera: Culicidae) Mosquitoes During Winter in Temperate Argentina.

The subgenus Culex L. includes species involved in summer-autumn arbovirus transmission but studies during winter are scarce in temperate Argentina. Female specimens were collected host-seeking at dry-ice-baited traps during autumn-winter-spring at two sites in Córdoba City during 2016 and 2017. The specimens were morphologically identified and dissected to determine the follicular developmental stage (gonotrophic activity). Females with advanced follicular stages (≥III) were subjected to molecular procedures to confirm or re-identify previous morphological identification. Five species (Culex apicinus Philippi (Diptera: Culicidae), Culex dolosus (Lynch-Arribálzaga) (Diptera: Culicidae), Culex maxi Dyar (Diptera: Culicidae), Culex pipiens pipiens L. (Diptera: Culicidae), and Culex quinquefasciatus Say (Diptera: Culicidae)) were collected and found gonoactive during winter; showing that a high proportion of Culex (Culex) females remain reproductively active during the unfavorable season for mosquito populations. Among them, it is worth noting the collection of Cx. quinquefasciatus, vector of the St. Louis encephalitis virus (endemic in the city), a specimen of Cx. p. pipiens, and a hybrid of Cx. p. pipiens/Cx. quinquefasciatus (during autumn). The study of this community during winter should continue because a high gonoactive female proportion with advanced follicular stages was found: 29.12 and 13.07% in 2016 and 2017, respectively. Local studies such as this one provide evidence about ornithophilic Culex species with active year-round life cycles, species that could favor arbovirus overwintering.

Urban Populations of Aedes aegypti (Diptera: Culicidae) From Central Argentina: Dispersal Patterns Assessed by Bayesian and Multivariate Methods.

Aedes aegypti (L.), the main vector of dengue and other arboviruses, was declared eradicated from Argentina in 1964; however, in 1987, it was detected again and nowadays it occurs in most of the country territory. To understand the transmission of vector-borne diseases, knowledge of the dispersal of vector populations is essential to evaluate the risk of pathogen transmission. We conducted a population genetic analysis of Ae. aegypti in 20 neighborhoods from Córdoba, the second largest city in Argentina, using 10 microsatellite loci. High genetic differentiation and the absence of an isolation by distance pattern was found using Weir and Cockerham's θ. Bayesian and multivariate clustering analyses showed that the studied sites included individuals with high membership coefficients (Q) in their populations, individuals with membership in another cluster, and admixed individuals. Individuals with high Q in clusters different from the population in which they were collected strongly suggests that passive transport is important in shaping the Ae. aegypti dispersal pattern in Córdoba city. Knowing the genetic structure of Ae. aegypti populations and their dispersal patterns would contribute to the implementation of vector control programs.

Culex saltanensis morphological redescription of the immature and adult stages.

Culex (Culex) saltanensis is redescribed in the adult, pupal, and larval stages. The male genitalia, pupae, and 4th-stage larvae are illustrated. Information about distribution, bionomics, and taxonomy is included. Culex saltanensis adults are separated from the closely related species Cx. (Cux.) coronator on the basis of morphometric characters.

Infection by UNA virus (Alphavirus; Togaviridae) and risk factor analysis in black howler monkeys (Alouatta caraya) from Paraguay and Argentina.

A neutralizing antibody (NTAb) survey on UNA and Mayaro viruses in black howler monkeys (Aloutta caraya) from subtropical regions of Argentina and Paraguay was carried out. Risk factors for infection in monkeys were analyzed. No positive sera for Mayaro virus were detected. A prevalence of 73% (61/84) of NTAb against UNAV was detected with titers ranging from 20 to 1280. According to the statistical analysis performed, the monkey's age was a significant risk factor, but not the origin or sex. This is the first report of Alouatta caraya infection by UNAV and the first record of its activity in Paraguay.

[Laboratory susceptibility tests of Aedes aegypti and Culex quinquefasciatus larvae to the entomopathogenic nematode Heterorhabditis bacteriophora].

INTRODUCTION: Aedes aegypti is the vector of dengue, yellow fever, Zika and Chikungunya viruses, and Culex quinquefasciatus is the vector of St. Louis and West Nile encephalitis viruses. OBJECTIVE: To evaluate infectivity of Heterorhabditis bacteriophora N4 in C. quinquefasciatus and A. aegypti larvae under laboratory conditions. MATERIALS AND METHODS: Thirty second-instar larvae of the two mosquito species were exposed each to different doses (0:1, 1:1, 5:1, 15:1, 100:1, 500:1, 750:1 and 1,500:1) of nematode infective juveniles. Four replications per dose were performed. RESULTS: Parasitism varied between 2.5 and 80 % in C. quinquefasciatus, and between 4.2 and 92.5 % in A. aegypti, with significant differences between doses (p<0.0001). DL50 were: 160.8 infective juveniles per larva for C. quinquefasciatus and 113.6 infective juveniles per larva for A. aegypti. In C. quinquefasciatus, 4 to 6 % of the infective juveniles developed to adults and in A. aegypti, 12- 61 %. In A. aegypti the emergence of new infective juveniles occurred with 100:1, 500:1, 750:1 and 1,500:1 infective juveniles per larva, and in C. quinquefasciatus, with 1,500:1 infective juveniles per larva. Melanization of infective juveniles was observed in both mosquito species. CONCLUSION: The susceptibility of these mosquito species to parasitism of an indigenous isolate of H. bacteriophora in the laboratory was demonstrated. Heterorhabditis bacteriophora N4 could be an efficient biological control agent.

Culicidae (Diptera) community structure, spatial and temporal distribution in three environments of the province of Chaco, Argentina.

The aim of this work was to study the composition of the Culicidae community in three environments with different degrees of anthropic disturbance in the province of Chaco. The nonparametric richness estimator ACE was calculated to measure the completeness of the sampling in each environment, and the diversity of each environment (α-diversity) were estimated. The composition, abundance and uniformity of species from the different environments were compared by means of range-species curves. ß-diversity was measured as species complementarity, which allowed us to know the degree of dissimilarity among the environments. The synanthropic index was estimated, identifying urban environment (synanthropic) species, eusynanthropic species, and wild species. The influence of climatic factors (mean temperature, relative humidity and rainfall) on the monthly and annual variations of the identified mosquito species was analysed using multilevel Poisson models with over-dispersion. The wild environment showed higher diversity and the semi-urban environment higher species richness. Despite having lower S and abundance, the urban environment showed greater diversity than semi-urban environment, although it also showed lower completeness, which means S could increase. Anthropogenic disturbance can lead to the elimination and/or modification or substitution of habitats, with the subsequent loss of richness in the more sensitive species of Culicidae fauna, although the conditions are also favourable for the settling of opportunistic and exotic species well-adjusted to disturbed environments. Most of the species abundances were positively influenced by the analysed climatic variables. The study area showed a rich Culicidae fauna of public health significance, with a risk of pathogen transmission, suggesting the need of further studies and the febrile and entomological surveillance.

UNA virus: first report of human infection in Argentina.

Una virus (UNAV), Togaviridae family, is widely distributed in South America, where infections have been detected in mosquitoes and vertebrate hosts (humans, birds and horses). We analyzed human sera from Córdoba inhabitants aged 44 to 89 years and using a neutralization test, we found a prevalence of UNAV antibodies of 3.8% (3/79). The low titers detected suggest past infections probably acquired in rural areas of the Province of Córdoba (central Argentina). None sera were found positive for MAYV neutralizing antibodies. This is the first report of human infections by UNAV in Argentina.

[Main breeding-containers for Aedes aegypti and associated culicids, Argentina]. / Principales criadero para Aedes aegypti y culícidos asociados, Argentina.

Breeding containers for Aedes (Stegomyia) aegypti were identified in two cities of Chaco Province (northeast Argentina): Presidencia Roque Saenz Peña and Machagai. All water-retaining recipients found in house backyards capable to retain water were classified according to their type and size, counted and checked. Aedes aegypti and Culex quinquefasciatus were the most frequently collected species, being also found Cx. maxi, Cx. saltanensis and Ochlerotatus scapularis. Tires and car batteries represented the most important type of container where immature forms of culicids could be found. Rain was an important factor for Ae. aegypti proliferation, as well as the widespread habit of the population of keeping useless containers at home, which allows the development of culicids.

Landscape determinants of Saint Louis encephalitis human infections in Córdoba city, Argentina during 2010.

Saint Louis encephalitis virus (SLEV) is endemic in Argentina. During 2005 an outbreak occurred in Córdoba. From January to April of 2010 a new outbreak occurred in Córdoba city with a lower magnitude than the one reported in 2005. Understanding the association of different landscape elements related to SLEV hosts and vectors in urban environments is important for identifying high risk areas for human infections, which was here evaluated. The current study uses a case-control approach at a household geographical location, considering symptomatic and asymptomatic human infections produced by SLEV during 2010 in Córdoba city. Geographical information systems and logistic regression analysis were used to study the distribution of infected human cases and their proximity to water bodies, vegetation abundance, agricultural fields and housing density classified as high/low density urban constructions. Population density at a neighborhood level was also analyzed as a demographic variable. Logistic regression analysis revealed vegetation abundance was significantly (p<0.01) associated with the presence of human infections by SLEV. A map of probability of human infections in Córdoba city was derived from the logistic model. The model highlights areas that are more likely to experience SLEV infections. Landscape variables contributing to the outbreak were the proximity to places with vegetation abundance (parks, squares, riversides) and the presence of low density urban constructions, like residential areas. The population density analysis shows that SLEV infections are more likely to occur when population density by neighborhood is lower. These findings and the predictive map developed could be useful for public health surveillance and to improve prevention of vector-borne diseases.

Transmission of endemic St Louis encephalitis virus strains by local Culex quinquefasciatus populations in Cordoba, Argentina.

BACKGROUND: St Louis encephalitis virus (SLEV) is a re-emerging human pathogen widely distributed in the American continent. Although it is not fully understood, the SLEV transmission network may involve Culex quinquefasciatus mosquitoes as vectors and Columbidae species as hosts. METHODS: To calculate infection rates, we inoculated Cx. quinquefasciatus mosquitoes from Córdoba, Argentina by feeding them on viremic chicks. RESULTS: We observed differences in infection rate among the viral strains, the highest rate (78/87 mosquitoes, 90.8%) being seen in strain 78V-6507. After re-feeding on susceptible chicks, mosquitoes were able to transmit the virus. CONCLUSION: Our findings suggest that Cx. quinquefasciatus populations are susceptible to and able to transmit different SLEV strains.

Spatial patterns of high Aedes aegypti oviposition activity in northwestern Argentina.

BACKGROUND: In Argentina, dengue has affected mainly the Northern provinces, including Salta. The objective of this study was to analyze the spatial patterns of high Aedes aegypti oviposition activity in San Ramón de la Nueva Orán, northwestern Argentina. The location of clusters as hot spot areas should help control programs to identify priority areas and allocate their resources more effectively. METHODOLOGY: Oviposition activity was detected in Orán City (Salta province) using ovitraps, weekly replaced (October 2005-2007). Spatial autocorrelation was measured with Moran's Index and depicted through cluster maps to identify hot spots. Total egg numbers were spatially interpolated and a classified map with Ae. aegypti high oviposition activity areas was performed. Potential breeding and resting (PBR) sites were geo-referenced. A logistic regression analysis of interpolated egg numbers and PBR location was performed to generate a predictive mapping of mosquito oviposition activity. PRINCIPAL FINDINGS: Both cluster maps and predictive map were consistent, identifying in central and southern areas of the city high Ae. aegypti oviposition activity. A logistic regression model was successfully developed to predict Ae. aegypti oviposition activity based on distance to PBR sites, with tire dumps having the strongest association with mosquito oviposition activity. A predictive map reflecting probability of oviposition activity was produced. The predictive map delimitated an area of maximum probability of Ae. aegypti oviposition activity in the south of Orán city where tire dumps predominate. The overall fit of the model was acceptable (ROC=0.77), obtaining 99% of sensitivity and 75.29% of specificity. CONCLUSIONS: Distance to tire dumps is inversely associated with high mosquito activity, allowing us to identify hot spots. These methodologies are useful for prevention, surveillance, and control of tropical vector borne diseases and might assist National Health Ministry to focus resources more effectively.

Ensayos de sensibilidad de larvas de Aedes aegypti y Culex quinquefasciatus frente al nematodo Heterorhabditis bacteriophora en condiciones de laboratorio / Laboratory susceptibility tests of Aedes aegypti and Culex quinquefasciatus larvae to the entomopathogenic nematode Heterorhabditis bacteriophora

Resumen Introducción. Aedes aegypti es el vector de los virus del dengue, la fiebre amarilla, el Zika y el chikungunya, y Culex quinquefasciatus, de los virus de la encefalitis de Saint Louis y de la del Oeste del Nilo. Objetivo. Evaluar la capacidad infecciosa de Heterorhabditis bacteriophora N4 en larvas de C. quinquefasciatus y A. aegypti en el laboratorio. Materiales y métodos. Treinta larvas de segundo estadio de cada especie de mosquito se expusieron a diferentes dosis (0:1, 1:1, 5:1, 15:1, 100:1, 500:1, 750:1 y 1.500:1) de ejemplares juveniles infectivos del nematodo. Se hicieron cuatro réplicas por dosis. Resultados. El parasitismo varió entre 2,5 y 80 % en C. quinquefasciatus y entre 4,2 y 92,5 % en A. aegypti, con diferencias significativas entre las dosis (p<0,0001). Las DL50 fueron de 160,8 ejemplares juveniles infectivos por larva de C. quinquefasciatus y 113,6 por larva de A. aegypti; 4 a 6 % de los ejemplares juveniles infectivos llegaron a la fase adulta en C. quinquefasciatus, y 12 a 61 % en A. aegypti. Nuevos ejemplares juveniles infectivos aparecieron a partir de la dosis de 100:1. En A. aegypti aparecieron solo con dosis superiores a esta y, en C. quinquefasciatus, con una dosis de 1500:1. Se observó melanización de los ejemplares juveniles infectivos en las dos especies de mosquitos. Conclusión. Se estableció la sensibilidad de estas especies de mosquitos al parasitismo producido por un aislamiento autóctono de H. bacteriophora en condiciones de laboratorio y su potencial efecto como agente de control biológico.

Abstract Introduction: Aedes aegypti is the vector of dengue, yellow fever, Zika and Chikungunya viruses, and Culex quinquefasciatus is the vector of St. Louis and West Nile encephalitis viruses. Objective: To evaluate infectivity of Heterorhabditis bacteriophora N4 in C. quinquefasciatus and A. aegypti larvae under laboratory conditions. Materials and methods: Thirty second-instar larvae of the two mosquito species were exposed each to different doses (0:1, 1:1, 5:1, 15:1, 100:1, 500:1, 750:1 and 1,500:1) of nematode infective juveniles. Four replications per dose were performed. Results: Parasitism varied between 2.5 and 80 % in C. quinquefasciatus, and between 4.2 and 92.5 % in A. aegypti, with significant differences between doses (p<0.0001). DL50 were: 160.8 infective juveniles per larva for C. quinquefasciatus and 113.6 infective juveniles per larva for A. aegypti. In C. quinquefasciatus, 4 to 6 % of the infective juveniles developed to adults and in A. aegypti, 12- 61 %. In A. aegypti the emergence of new infective juveniles occurred with 100:1, 500:1, 750:1 and 1,500:1 infective juveniles per larva, and in C. quinquefasciatus, with 1,500:1 infective juveniles per larva. Melanization of infective juveniles was observed in both mosquito species. Conclusion: The susceptibility of these mosquito species to parasitism of an indigenous isolate of H. bacteriophora in the laboratory was demonstrated. Heterorhabditis bacteriophora N4 could be an efficient biological control agent.

Daily oviposition activity of Aedes aegypti in Orán, Argentina.

The study aimed to determinate the maximum daily peak of Aedes aegypti oviposition in the city of Oran, northwestern Argentina. Biweekly samplings were taken between November 2006 and February 2007 (spring-summer). The city was divided into three areas (north, center, and south) and households were randomly selected. Two ovitraps were placed outdoors in the selected houses. Ovitraps were replaced every four hours, from morning (8 a.m.) to late afternoon (8 p.m.). The largest number of eggs was recorded between 4 p.m. and 8 p.m. (81%). These findings enhance our understanding of the vector and thus its control such as spraying during the hours of peak oviposition activity.

Bionomics of Aedes aegypti subpopulations (Diptera: Culicidae) from Argentina.

Differences in biological features of immature and adult Aedes aegypti, as well as variability in vector competence, seem consistent with the existence of genetic variation among subpopulations and adaptation to local conditions. This work aims to compare the bionomics of four Ae. aegypti subpopulations derived from different geographical regions reared under temperate conditions. Life statistics of three Ae. aegypti subpopulations from the provinces of Córdoba, Salta, and Misiones were studied based on horizontal life tables. The Rockefeller strain was used as a control. The development time required to complete the larva and pupa stages varied from 6.91 to 7.95 and 1.87 to 2.41 days, respectively. Significant differences were found in mean larval development time between the Córdoba and Orán subpopulations. The larva-pupa development time was similar in all the subpopulations. However, survival values varied significantly between the Orán and San Javier subpopulations. The proportion of emergent males did not differ from females within each subpopulation nor among them. Adult longevity was similar among the subpopulations. The average number of eggs laid by each female was significantly different. The Rockefeller strain laid a significantly greater number of eggs (463.99 eggs/female) than the rest of the subpopulations. Moreover, differences in the demographic growth parameter R(o) were detected among the four subpopulations. The differences obtained in larval development time, larva-pupa survival values, and net reproductive rates among the subpopulations might reflect underlying genetic differences as a result of colonization from different regions that probably involve adaptations to local conditions.