Domingo Javier Liotta

Epidemiological aspects of cutaneous leishmaniasis in the Iguazú falls area of Argentina

Over the last three decades the incidence of American cutaneous leishmaniasis (ACL) has increased sharply in Argentina and throughout the world. In the Iguazú Falls area, on the border between Brazil and Paraguay, the incidence of human ACL has risen since 2004. Most of the 36 cases of human ACL reported until 2005 have involved males over 15 years old (75%) infected during deforestation to establish individual farms. Captures carried out in primary forest, periurban areas, and deforested land sites yielded 18,438 sand flies belonging to 13 species; the most prevalent species were Lutzomyia (Nyssomyia) whitmani (87.4%) and Lutzomyia (Mygonemyia.) migonei (7.6%). Cluster analysis was used to group traps according to species and abundance of sand flies. The group of traps located in recently deforested places, in pig and chicken dwellings of houses where ACL cases had been reported in the past, and at one house with an active ACL case, had the highest abundance of Lu. whitmani and Lu. whitmani+Lu. migonei as well as the highest ratio of Lu. whitmani/Lu. migonei. Leishmania sp. infections, both in Lu. whitmani, in Lu. quinquefer, and in smears from human cases were detected by DNA kinetoplast amplification using a generic PCR protocol. The risk of ACL outbreak in the Iguazú Falls area is still associated with economic and leisure activities in primary-secondary forest, including deforestation, rural settlements, fishing, hunting, and ecotourism. In addition, the risk of periurban transmission seems likely, and this is discussed within the framework of surveillance and prevention strategies.

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.

Detection of Leishmania infantum in naturally infected Lutzomyia longipalpis (Diptera: Psychodidae: Phlebotominae) and Canis familiaris in Misiones, Argentina: the first report of a PCR-RFLP and sequencing-based confirmation assay

In this study, a genotypification of Leishmania was performed using polimerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and sequencing techniques to identify species of Leishmania parasites in phlebotomine sand flies and dogs naturally infected. Between January-February of 2009, CDC light traps were used to collect insect samples from 13 capture sites in the municipality of Posadas, which is located in the province of Misiones of Argentina. Sand flies identified as Lutzomyia longipalpis were grouped into 28 separate pools for molecular biological analysis. Canine samples were taken from lymph node aspirates of two symptomatic stray animals that had been positively diagnosed with canine visceral leishmaniasis. One vector pool of 10 sand flies (1 out of the 28 pools tested) and both of the canine samples tested positively for Leishmania infantum by PCR and RFLP analysis. PCR products were confirmed by sequencing and showed a maximum identity with L. infantum. Given that infection was detected in one out of the 28 pools and that at least one infected insect was infected, it was possible to infer an infection rate at least of 0.47% for Lu. longipalpis among the analyzed samples. These results contribute to incriminate Lu. longipalpis as the vector of L. infantum in the municipality of Posadas, where cases of the disease in humans and dogs have been reported since 2005.

Human papillomavirus (HPV) detection and Papanicolaou cytology in low-resource women in Posadas city, Misiones, Argentina.

The objective of this study was to determine the prevalence of HPV infection and cervical lesions present in women who attended a health center in a low-resource area of the city of Posadas, Misiones, Argentina. Cervical cell samples (n = 163) were processed for Papanicolaou cytology and HPV-PCR tests. Socio-cultural risk factors were estimated using the odds ratio (OR, CI 95 %). Cervical lesions were detected in 14.7 % of women. The general prevalence of HPV infection was of 38 %. The most common types among the total population were HPV-16 (9.8 %) and HPV-33 (9.3 %). HPV-16 was detected in association with 29.2 % and 6.5 % of women with and without cervical lesions, respectively, the OR being 5.3 (1.8-15.8). Risk factors for HPV-16 infection were a smoking habit and a history of previous sexually-transmitted diseases. These data are important for the implementation of prevention programs, including an appropriate introduction of vaccination and the baseline for virological surveillance in the vaccine era.

Genetic characterization and clinical implications of human papillomavirus type 16 (HPV16) variants from northeastern Argentina

BACKGROUND Human papillomavirus type 16 (HPV16) plays a central role in the development of cervical cancer. Worldwide studies indicate the existence of HPV16 variants that show different geographic distributions and oncogenic potential. OBJECTIVE Our goal was to describe the genetic variation of HPV16 isolates identified in urban women with different grades of cervical lesions living in northeastern Argentina. STUDY DESIGN We analyzed 116 HPV16-positive cervical samples (16 NLIM, 62 L-SIL, 16 H-SIL and 22 cervical cancer) from patients attending health centers in Misiones (Argentina) during 2006-13. HPV16 isolates were genetically characterized through PCR amplification and direct sequencing of 364 bp within the long control region, and the resulting sequences classified into variants based on phylogenetic analysis (lineages A, B, C and D). A potential association between HPV16 variants and lesion grade was evaluated through an odds ratio (OR) test. A temporal framework for the origin of HPV16 variants was assessed through coalescence analysis (BEAST v 1.7.5). RESULTS Phylogenetic analysis of HPV16 sequences showed that 92.1% of the samples clustered with lineage A, and 6.9% to lineage D. HPV16 variants from lineage D were more frequently associated with high-grade lesions and cancer (HSIL+) than lineage A variants at an OR of 13.8 (1.6-117.0). The time to most common recent ancestor (tMCRA) of all variants was 119,103 years before present (HPD 95%=48,486-197,239), a date consistent with the time frame for modern human evolution. CONCLUSION Our results suggest that HPV16 variants from lineage D may represent an additional risk factor for the development of cervical cancer in women living in northeastern Argentina. This study provides new information about viral isolates present in Argentina that will contribute to the monitoring of HPV16 infection in the vaccine era.

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.

Assessment of the role of small mammals in the transmission cycle of tegumentary leishmaniasis and first report of natural infection with Leishmania braziliensis in two sigmodontines in northeastern Argentina

To contribute to the knowledge of the role of small mammals in the transmission cycle of tegumentary leishmaniasis caused by Leishmania braziliensis, we studied the small mammal community and its temporal and spatial association with phlebotominae, as well as small mammal infection by Leishmania spp. by PCR-RFLP analyses in an endemic area of northeastern Argentina. Ten small mammal samplings were conducted (2007–2009, 7506 Sherman trap nights and 422 cage trap nights). In two of these samplings, 16 capture stations each one consisting of a CDC light trap to capture phlebotominae, two to four Sherman traps and two cage traps were placed. We found co-occurrence of phlebotominae and small mammal captures in four stations, which were all the stations with small mammal captures and yielded 97% (2295 specimens, including 21 gravid females) of the total phlebotominae captures, suggesting that small mammals may provide a potential source of blood for phlebotominae females. One Didelphis albiventris and two Rattus rattus were associated with high captures of Nyssomyia whitmani, vector of L. braziliensis in the study area. The PCR-RFLP analyses confirm the presence of L. braziliensis in two sigmodontine small mammals (Akodon sp. and Euryoryzomys russatus) for the first time in Argentina, to our knowledge.

Mitochondrial DNA ancestry, HPV infection and the risk of cervical cancer in a multiethnic population of northeastern Argentina

Background Misiones Province in northeastern Argentina is considered to be a region with a high prevalence of HPV infection and a high mortality rate due to cervical cancer. The reasons for this epidemiological trend are not completely understood. To gain insight into this problem, we explored the relationship between mitochondrial DNA (mtDNA) ancestry, HPV infection, and development of cervical lesions/cancer in women from the city of Posadas in Misiones Province. Methods Two hundred and sixty-one women, including 92 cases of patients diagnosed with cervical lesions and 169 controls, were analyzed. mtDNA ancestry was assessed through HVS1 sequencing, while the detection and typing of HPV infection was conducted through nested multiplex PCR analysis. Multivariate logistic regression was conducted with the resulting data to estimate the odds ratios (ORs) adjusted by socio-demographic variables. Results The study participants showed 68.6% Amerindian, 26.1% European and 5.3% African mtDNA ancestry, respectively. Multiple regression analysis showed that women with African mtDNAs were three times more likely to develop a cervical lesion than those with Native American or European mtDNAs [OR of 3.8 (1.2–11.5) for ancestry and OR of 3.5 (1.0–12.0) for L haplogroups], although the associated p values were not significant when tested under more complex multivariate models. HPV infection and the development of cervical lesions/cancer were significant for all tested models, with the highest OR values for HPV16 [OR of 24.2 (9.3–62.7)] and HPV-58 [OR of 19.0 (2.4–147.7)]. Conclusion HPV infection remains a central risk factor for cervical cancer in the Posadas population. The potential role of African mtDNA ancestry opens a new avenue for future medical association studies in multiethnic populations, and will require further confirmation in large-scale studies.