Identification and evolutionary analysis of papillomavirus sequences in New World monkeys (genera Sapajus and Alouatta) from Argentina.

OBJECTIVE: In this study, we investigated the occurrence of papillomavirus (PV) infection in non-human primates (NHPs) in northeastern Argentina. We also explored their evolutionary history and evaluated the co-speciation hypothesis in the context of primate evolution. METHODS: We obtained DNA samples from 57 individuals belonging to wild and captive populations of Alouatta caraya, Sapajus nigritus, and Sapajus cay. We assessed PV infection by PCR amplification with the CUT primer system and sequencing of 337 bp (112 amino acids) of the L1 gene. The viral sequences were analyzed by phylogenetic and Bayesian coalescence methods to estimate the time to the most common recent ancestor (tMRCA) using BEAST, v1.4.8 software. We evaluated viral/host tree congruence with TreeMap v3.0. RESULTS: We identified two novel putative PV sequences of the genus Gammapapillomavirus in Sapajus spp. and Alouatta caraya (SPV1 and AcPV1, respectively). The tMRCA of SPV1 was estimated to be 11,941,682 years before present (ybp), and that of AcPV1 was 46,638,071 ybp, both before the coalescence times of their hosts (6.4 million years ago [MYA] and 6.8 MYA, respectively). Based on the comparison of primate and viral phylogenies, we found that the PV tree was no more congruent with the host tree than a random tree would be (P > 0.05), thus allowing us to reject the model of virus-host coevolution. CONCLUSION: This study presents the first evidence of PV infection in platyrrhine species from Argentina, expands the range of described hosts for these viruses, and suggests new scenarios for their origin and dispersal.

Leishmania infantum DNA detected in phlebotomine species from Puerto Iguazú City, Misiones province, Argentina.

In Puerto Iguazú City, Argentina, human and canine Visceral Leishmaniasis cases have been recorded since the year 2010, with Leishmania infantum as the etiological agent and Lutzomyia longipalpis as its main vector. In the present study, polymerase chain reaction and sequencing were used to detect L. infantum DNA in 3.9% of the female sandflies captured in Puerto Iguazú City. This is the first report of L. infantum DNA detection in Micropygomyia quinquefer, and the second one in Lu. longipalpis and Nyssomyia whitmani for Argentina. Although the detection of Leishmania DNA itself is not enough to determine a Phlebotomine species as a vector, these results are significant in setting the direction of further investigations of vectorial competence and capacity, necessary to define the roles of different sandflies species as specific or permissive vectors in the transmission VL cycle.

Leishmania (Viannia) DNA detection by PCR-RFLP and sequencing in free-ranging owl monkeys (Aotus azarai azarai) from Formosa, Argentina.

American Cutaneous Leishmaniasis (ACL) is caused by protozoan parasites of the Leishmania genus, and transmitted by females of the Phlebotominae family. The role of wild and domestic hosts in the cycle of Leishmania is still unknown. ACL is endemic in the province of Formosa where Nyssomyia neivai was the most abundant species in several captures and 31 cumulative ACL human cases were reported between 2005 and 2011 in the province. The present report describes the detection, by PCR-RFLP and confirmed by sequencing, of subgenus Leishmania (Viannia) DNA in four free-ranging owl monkeys (Aotus azarai azarai) from Formosa Province. The sequence amplified was the mini-exon gene present in tandem repeats in all species of the Leishmania genus from buffy coat samples. The absence of inhibitors in the samples was checked by a ß-globin protocol originally designed to amplify the human ß-globin gene. However, other free-ranging primates were found with natural infections of L. (V) braziliensis complex and Leishmania (Viannia) subgenus by parasitological means in America. To the best of our knowledge, there are no published reports on detection of subgenus Leishmania (Viannia) DNA by PCR-RFLP in argentinean free-ranging primates. Additional eco-epidemiological and parasitological studies are necessary to confirm owl monkeys, or any other natural infected mammal species detected by PCR, as a reservoir, incidental host or to propose it as an animal model for research on this topic.

TNF promoter SNP variation in Amerindians and white-admixed women from Misiones, Argentina.

The aim of this study is to describe genetic variation in the TNF promoter in the ethnically diverse population of Misiones, north-eastern Argentina. We analysed 210 women including 66 Amerindians of the Mbya-Guarani ethnic group and 144 white-admixed individuals from urban and rural areas of Misiones. Their DNA samples were surveyed for TNF polymorphisms -376 A/G, -308 A/G -244 A/G and -238 A/G by PCR amplification and direct sequencing and for the Amerindian marker -857 C/T by real-time PCR. Our main findings are as follows:(i) a distinctive pattern of Single Nucleotide Polymorphism (SNP) distribution among these groups, (ii) genetic differentiation between the Mbya-Guarani and the white-admixed populations (P < 0.05), (iii) lower gene diversity (~0.05) in Mbya-Guarani compared with the white-admixed group (~0.21); and (iv) linkage disequilibrium between the -376A and -238A SNPs in white-admixed populations. These data highlight the principal role of population history in establishing present-day genetic variation at the TNF locus and provide a framework for undertaking ethnographic and disease association studies in Misiones.

Human papillomavirus cervical infection and associated risk factors in a region of Argentina with a high incidence of cervical carcinoma.

OBJECTIVE: To assess the prevalence and potential risk factors associated with human papillomavirus (HPV) cervical infection among women residing in a region of northeastern Argentina with a high incidence of cervical cancer. METHODS: A case-control study of 330 women participating in a cervical cytological screening program conducted in Posadas city, Misiones, Argentina, from February 1997 to November 1998 was carried out. Standardized questionnaires were administered, and clinical examination including colposcopy was performed. Fresh endocervical specimens for HPV DNA detection by generic polymerase chain reaction were collected and the products typed by dot-blot hybridization. RESULTS: Human papillomavirus DNA was found in 61% of samples analyzed (185/301). Samples with normal cytology had a 43% infection rate (85/199), while those classified as low-grade squamous intraepithelial lesion, high-grade squamous intraepithelial lesion, and invasive cervical carcinoma had an infection rate of 96% (53/55), 100% (29/29), and 100% (18/18), respectively. Human papillomavirus typing showed a 64% (118/185) prevalence of type 16 among all the infected population analyzed; type 16 was detected among 49% (42/85) of infected samples with normal cytology and in an average of 74% (74/100) with abnormal cytology. Sexual behavior, residence in southern Paraguay, and history of a previous sexually transmitted diseases were the main risk factors associated with high-grade cervical lesions. CONCLUSIONS: An elevated prevalence of HPV infection was detected in this population, which also has a high incidence of cervical cancer. The broad distribution of high-risk HPV type 16 in women with normal cytology and colposcopy suggests that viral infection is an important determinant of regional cancer incidence.