Genetic diversity and geographic distribution of parasitic flies of the Philornis torquans complex in Argentina.

Philornis Meinert 1890 (Diptera: Muscidae) is a genus of flies that parasitize birds in the Neotropical region. The characteristics of the host-parasite interactions and its consequences may depend on the Philornis species involved, and thus precise identification of these parasites is crucial for the interpretation of ecological and epidemiological studies. However, morphological identification of Argentine Philornis species is elusive while molecular evidence points towards the existence of a complex of cryptic species or lineages undergoing a speciation process, which were named the 'Philornis torquans complex'. Herein the authors extended the current knowledge on the systematics and biogeography of parasitic Philornis flies from Argentina, analysing samples collected in several ecoregions, including the Atlantic Forest, Iberá Wetlands, Open Fields and Grasslands, Espinal, Pampa, Dry Chaco, Humid Chaco, Delta and Paraná River Islands, Monte of Plains and Plateaus. The results of the present study strengthen the evidence on previously described Philornis genotypes using four genetic markers (ITS2, COI, ND6, 12S rRNA). The authors report new patterns of occurrence and describe the presence of a novel genotype of subcutaneous Philornis. In addition, the present study unveils ecological niche differences among genotypes of the Philornis torquans complex in southern South America.

Environmental variables determining the distribution of an avian parasite: the case of the Philornis torquans complex in South America.

Philornis flies are the major cause of myiasis in nestlings of Neotropical birds, being of major concern in geographically-restricted and endangered bird species. Despite its relevance for the conservation of birds, there is little information about the environmental dimensions determining Philornis spp. geographical range. By using maximum entropy, we identified for the first time the macro-environmental variables constraining the abiotic niche of the P. torquans complex in South America, and provided a model map of its potential distribution based on environmental suitability. We identified the minimum temperature of the coldest month as the most relevant variable, associated with the largest decrease in habitat suitability in Brazil and northern South America. Furthermore, the mean temperature of the warmest quarter limited suitability mostly along with the Andean range. In addition, humidity and moisture are influential factors in most of Argentina, northern Chile, and coastal Peru. The geographical projection suggests that environments in most of central-eastern Argentina, and in a broad area in central Chile, are suitable for the presence of the P. torquans complex. Besides providing information about the ecology of Philornis spp., this study represents a tool for bird conservation and a reference for future work on the distribution of this genus.

Dynamics of Exposure to Rickettsia parkeri in Cattle in the Paraná River Delta, Argentina.

Several cases of human rickettsiosis caused by Rickettsia parkeri were recently documented in the Paraná River delta of Argentina, where the tick vector is Amblyomma triste Koch. As cattle suffer recurrent A. triste infestations, they are at risk of becoming infected with R. parkeri Herein we investigated the dynamics of R. parkeri and its A. triste vector in a herd of beef cattle. Cattle were followed for 18 mo and samples were analyzed for the presence of antibodies against four Rickettsia species (R. parkeri, Rickettsia bellii, Rickettsia amblyommii, and Rickettsia felis) and also for the presence of rickettsial DNA. Additionally, cattle were examined for attached ticks and questing adult ticks were collected. All ticks were analyzed for the presence of rickettsial DNA. No evidence of rickettsemia was found in any cow, but the high R. parkeri infection rate documented in A. triste both questing in the study area (13.9%) and feeding on cattle (19.8%) and the identification of antibodies against R. parkeri antigen in 90% of cattle are evidence that infection is taking place. Altogether, our data suggest that A. triste ticks are capable of naturally exposing cattle to R. parkeri However, the progress of R. parkeri infection and its impact on bovine health and production remain to be established.

First report of Trypanosoma evansi in a canine in Argentina.

An Argentinian Dogo which suffered from anorexia, lymphadenopathy, cachexia and paresis of the hind limbs was diagnosed with trypanosomiasis in Argentina in 2013. In this study, we describe the clinical profile and its evolution as well as the molecular method employed to identify and quantify Trypanosoma evansi.

Detection of Trypanosoma evansi infection in wild capybaras from Argentina using smear microscopy and real-time PCR assays.

Trypanosoma evansi is a flagellated protozoan that parasitizes a wide variety of mammals, occasionally including humans. In South America, it infects horses, cattle, buffaloes, dogs and wild mammals, causing a disease known as "Mal de Caderas", which results in important economic losses due to a wide range of pathological expressions. Argentina represents the southern limit of its distribution. The capybara (Hydrochoerus hydrochaeris) is a large rodent found in tropical to temperate freshwater wetlands of South America. As capybaras infected with T. evansi present no clinical signs of disease, withstanding high parasitaemia, this species was proposed as a reservoir host. In this study we investigated the prevalence and parasitaemic intensity of T. evansi in samples obtained from 60 free-ranging capybaras of Esteros del Iberá (Corrientes province, northeastern Argentina) using smear microscopy and real-time PCR assays. All the cases of capybaras infected with T. evansi were found during one of the years studied, with no evidence of seasonality. The overall infection prevalence was 10%, but between years it ranged from 0% to 17% (in 2011). This is the first confirmation of T. evansi infection in Argentina by molecular biology techniques. Our results showed no differences between the methods used to detect the presence of T. evansi in capybaras, which indicates that simple methods like microscopy can generate important data on the ecoepidemiology of this parasite. Both techniques used in this study represent a viable tool for ecoepidemiological studies, and can be used to produce good estimates of prevalence and parasitaemic level of the infection, which inform for the implementation of strategies for the control of the disease.

First record of the avian ectoparasite Philornis downsi Dodge & Aitken, 1968 (Diptera: Muscidae) in Argentina.

Species of Philornis Meinert, 1890 (Diptera, Muscidae) are Neotropical dipterans that include species with parasitic larvae which feed on nestling birds. To date, all Philornis species that have been recorded from Argentina have parasitic subcutaneous larvae. Here, for the first time for Argentina, we report the finding of Philornis downsi Dodge & Aitken, 1968, a fly with a nest-dwelling, semi-haematophagous larva. This record, from the humid Chaco ecoregion of Argentina in the nest of a saffron finch Sicalis flaveola pelzelni Sclater, substantially extends the known distribution of this species. We also report the consensus sequences of the internal transcribed spacer 1 (ITS1) and ITS2 regions of three of the specimens for future reference and comparison. Further investigation is needed to determine whether Argentina is part of the historical range of P. downsi or, alternatively, represents a recent expansion of its range, perhaps due to climatic changes or other factors of global environmental variation.

Exposure of neonatal female rats to bisphenol A disrupts hypothalamic LHRH pre-mRNA processing and estrogen receptor alpha expression in nuclei controlling estrous cyclicity.

This study examines the effects of neonatal exposure to the endocrine disruptor bisphenol A (BPA) on the neural network that controls estrous cyclicity. From postnatal day 1 (PND1) to PND7, female pups were injected with vehicle (control) or BPA (BPA.05: 0.05mg/kg-d, BPA20: 20mg/kg-d). At PND100 BPA.05-females showed alterations in estrous cyclicity and BPA20-females were incapable of producing an estradiol-induced LH surge. By real-time PCR we determined that hypothalamic expression of mature LH-releasing hormone (LHRH) mRNA was increased in BPA.05 and decreased in BPA20-females. Furthermore, unprocessed intron A-containing LHRH RNA was decreased in the cytoplasm of hypothalamic cells of both groups. Immunohistochemistry revealed that estrogen receptor alpha protein was up-regulated in anteroventral periventricular and down-regulated in arcuate nucleus of both groups. Our results show that BPA permanently disrupts hypothalamic LHRH pre-mRNA processing and steroid receptors expression in nuclei that control estrous cyclicity in adult rats.

Neonatal exposure to bisphenol A alters estrogen-dependent mechanisms governing sexual behavior in the adult female rat.

This study examines the effects of neonatal exposure to the endocrine disruptor bisphenol A (BPA) on the hypothalamic circuitry controlling the female sexual behaviors of adult rats. From postnatal day 1 (PND1) to PND7, pups were injected with corn oil (control) or BPA (BPA20: 20mg/kg-d; BPA.05: 0.05 mg/kg-d) and at PND85 the rats were bilaterally ovariectomized (OVX). At PND100, OVX-rats received estradiol alone or estradiol and progesterone to evaluate estrogen-dependent gene expression in the hypothalamus and sexual behavior. In BPA-exposed females, estrogen receptor alpha (ER alpha) expression was down-regulated in both the medial preoptic (MPN) and ventromedial nucleus (VMHvl), while repressor of estrogen receptor activity (REA) expression was up-regulated in the VMHvl. Interestingly, BPA-exposed females displayed significantly lower levels of proceptive behavior. Our results show that BPA permanently alters the hypothalamic estrogen-dependent mechanisms that govern sexual behavior in the adult female rat.

The estrogen receptor alpha sigma3 mRNA splicing variant is differentially regulated by estrogen and progesterone in the rat uterus.

The gene for estrogen receptor alpha (ER alpha) has been shown to be under complex hormonal control and its activity can be regulated by mRNA alternative splicing. Here we examined the regulation of ER alpha transcription and translation in the rat uterus by ovarian steroid hormones. We examined whether expression of ER alpha mRNA splice isoforms is hormonally regulated in ovariectomized (OVX) and cycling rats. Adult OVX female rats were treated daily with 17-beta estradiol (E2) (0.05 microg/rat or 5 microg/rat), progesterone (P4) (1 mg/rat) or a combination of both hormones for 4 days. Animals were killed 24 h after the last injection and uterine horns were removed. In order to determine whether ER alpha mRNA isoforms are differentially expressed under various physiological conditions, animals were evaluated at proestrus, estrus and diestrus. The ER alpha protein and mRNA were detected by immunohistochemistry and comparative RT-PCR analysis respectively. The presence of ER alpha mRNA isoforms was evaluated using a nested RT-PCR assay. In OVX control rats, ER alpha mRNA and protein levels were high, demonstrating a constitutive expression of the ER alpha gene in the uterus. When animals received P4 or the high dose of E2, a significant decrease in both ER alpha mRNA and protein was observed in the uterus. However, when rats were protein was treated with the low dose of E2, only the ER alpha down-regulated; no changes were observed in ER alpha mRNA expression. In addition to the full-length ER alpha mRNA, OVX control rat uteri expressed three shorter transcripts: sigma3, sigma4 and sigma3,4 (lacking exon 3, exon 4, or both 3 and 4 respectively). Surprisingly, when OVX animals were treated with P4, the low dose of E2 or a combination of both steroids, expression of the sigma3 isoform was completely abolished. During the estrous cycle, all ER alpha mRNA splicing variants were detected at proestrus and estrus. However, in diestrus, significant low levels of the sigma3 isoform were observed. In summary, our results suggest a dose-dependent relationship between E2 concentrations and the level of control in the ER alpha transcription-translation cascade. Moreover, the alternative splicing of the ER alpha primary transcript is influenced by the hormonal milieu, suggesting that these events could affect the estrogen responsiveness of the rat uterus during the estrous cycle.