Infection by human polyomaviruses JCPyV and BKPyV in blood donors of Argentina.

BACKGROUND AND OBJECTIVES: A spectrum of blood-borne infectious agents may be transmitted through transfusion of blood components from asymptomatic donors. Despite the persistence of polyomaviruses in blood cells, no studies have been conducted in Argentina to assess the risk of transfusion infection. MATERIALS AND METHODS: We investigated BKPyV and JCPyV in 720 blood donors, using polymerase chain reaction (PCR) for a region of T antigen common to both viruses. Positive T-antigen samples were subjected to two additional PCR assays targeting the VP1 region. Viral genotypes were characterized by phylogenetic analysis. RESULTS: Polyomaviruses were detected in 1.25% (9/720) of the blood samples selected; JCPyV was identified in 0.97% (7/720) and BKPyV in 0.28% (2/720) of them. Phylogenetic analysis showed that the JCPyV sequences clustered with 2A genotype and Ia of BKPyV. CONCLUSION: This study describes for the first time the prevalence of polyomavirus DNA in blood donors of Córdoba, Argentina. The polyomavirus DNAemia in healthy populations suggests that those viruses are present in blood components eligible for transfusion. Therefore, the epidemiological surveillance of polyomavirus in blood banks might be incorporated into haemovigilance programmes, to determine the infectious risk and implement newer interventions to ensure the safety of blood supplies, if required.

Seronegative human T-cell lymphotropic virus 1 carriers in blood banks: A potential viral source for silent transmission?

BACKGROUND AND OBJECTIVES: Transfusion-transmitted viruses count among the greatest threats to blood safety. In Argentina, current laws oblige testing all donated blood for the presence of antibodies against human T-cell lymphotropic viruses 1 and 2 (HTLV-1/2). In endemic zones of the country, a high rate of seronegative HTLV-1 individuals with clear evidence of infection because of symptoms and/or presence of tax sequences of HTLV-1 and/or IgG anti-Tax antibodies has been recently described. Migration from endemic to nonendemic zones of Argentina is very frequent. MATERIALS AND METHODS: During a 1-year period, in the blood bank of Córdoba city, we performed molecular screening of all donors who were born in or arose from endemic zones for HTLV-1/2 in Argentina and neighbouring countries. RESULTS: By screening 219 bp of HTLV-1/2 tax gene, 0.6% (2/317) of the blood donors proved to be positive for HTLV-1 tax sequence. One of the donors presented anti-Tax antibodies, demonstrating the transcriptional activity of the tax gene, and the other donor was also positive for LTR and pol gene sequences. The HTLV-1 genetic analysis of the LTR sequence determined that it belonged to the Cosmopolitan subtype HTLV-1aA. CONCLUSION: These findings suggest potential limitations of some currently approved screening assays for HTLV-1 detection applied in some donor populations and the possibility of an HTLV-1 seronegative carrier state with the potential for silent transmission by blood.

Establishment of the first platelet-donor registry in Argentina.

BACKGROUND: Platelet transfusions are necessary to prevent and treat haemorrhages in thrombocytopenic patients or those with severe platelet dysfunction. In Latin American countries, including Argentina, blood supplies from voluntary non-remunerated blood donors remain dependent on family replacement donors, since altruistic repeat donors are exceptional and platelet donors are very scarce. The aim of this study was to recruit a group of frequent, voluntary, altruistic blood donors and determine their human platelet antigen (HPA)-genotype in order to establish the first registry of HPA-typed voluntary platelet donors in Argentina. MATERIAL AND METHODS: In this study, we invited and recruited voluntary blood donors who attended the Fundación Banco Central de Sangre between July 2016 and July 2017. DNA was extracted from K2EDTA anticoagulated whole blood and genotyping was performed by polymerase chain reaction, using sequence-specific primers to type the HPA-1 to -6, -9 and -15 systems. A subset of samples was also tested using a commercial HPA-TYPE kit. Donors were invited to join the National Register of Haematopoietic Stem Cell Donors of Argentina. RESULTS: A cohort of 500 platelet donors was recruited and characterised and a database with their personal information, including their genotype for the most relevant HPA alloantigens, was created. Eight of the 500 donors (1.6%) were HPA-1a negative. HPA allelic variants -4b, -6b and -9b were detected for the first time in our population. There was 100% concordance between our in-house assay and the commercial kits in the subset of 150 donor samples assayed in parallel. DISCUSSION: The efforts made to recruit, characterise and register voluntary platelet donors will provide the first sustainable source of HPA and human leukocyte antigen-typed platelets for compatible transfusions in the country. Remarkably, we identified a higher percentage of HPA-1a-negative donors than previously detected in the Argentinean population.

First Description of Seronegative HTLV-1 Carriers in Argentina.

In some areas of Argentina endemic for human T-lymphotropic virus type 1 (HTLV-1), tropical spastic paraparesis is frequent in subjects who lack antibodies against the virus; however, the relevance of this seronegative status in the country has not been investigated. In neighboring countries, HTLV-1 seronegative status has been described in patients with different diseases; however, data regarding features of seronegative HTLV-1 carriers are scarce. We investigated the seronegative status in 124 relatives of 28 HTLV-1 infected subjects from an endemic area in Northwest Argentina. Blood samples and clinical/epidemiological data were collected. Human T-lymphotropic virus type 1 infection was diagnosed by serology and long terminal repeat (LTR) sequence, env and tax gene detection. IgG anti-Tax HTLV-1 antibody, tax gene sequence, and DNA proviral load were also evaluated. Seventy-five percent of the 124 relatives were negative for HTLV-1/2 antibodies; 35.5% were also negative by molecular assays and 64.5% were negative for HTLV-1 LTR and env sequences, but positive for two sequences of HTLV-1 tax gene. Also, 35.7% of these subjects had IgG anti-Tax antibodies. The seronegative HTLV-1 status was significantly associated with male gender, youth, and sensory symptoms/autonomic nervous system dysfunction. High rates of seronegative symptomatic and asymptomatic HTLV-1 carriers in Argentina are described. The evidence highlights that HTLV-1 prevalence may be underestimated worldwide. Larger cohort studies are required to assess disease outcome in these seronegative subjects. Also, the findings emphasize the limitations of ongoing screening assays for diagnosis and blood safety. Therefore, algorithms for HTLV-1 diagnosis should include not only serological but also molecular assays.

Detección y caracterización molecular de Chlamydia psittaci y Chlamydia abortus en psitácidos mascotas en la provincia de Buenos Aires, Argentina / Detection and molecular characterization of Chlamydia psittaci and Chlamydia abortus in psittacine pet birds in Buenos Aires province, Argentina

In order to determine the presence and genetic diversity of Chlamydia spp. in the north-eastern area of Buenos Aires province, Argentina, conjunctival, oropharyngeal, cloacal swab and tissues were collected from a total of 90 psittacine pet birds of different age and clinical manifestations. Through molecular methods, Chlamydiaceae was detected in 30% (27/90) of the samples, out of which 70.3% (19/27) were positive for Chlamydia psittaci and 14.9% (4/27) for Chlamydia abortus. Nine C. psittaci positive samples were genotyped by ompA gene sequences, 8 clustered within genotype A and 1 within genotype B. A significant association was observed between the presence of Chlamydia spp. and the manifestation of clinical signs compatible with chlamydiosis, as well as with the age of the birds (younger than one year old). This report contributes to the improvement of our understanding of chlamydial agents in our country.

Con el objetivo de determinar la presencia de Chlamydia spp. en psitácidos del área noreste de la provincia de Buenos Aires y conocer su diversidad genética, se recolectaron y analizaron mediante métodos moleculares hisopados conjuntivales, orofaríngeos, cloacales y tejidos de un total de 90 psitácidos de diferentes edades y con diversas manifestaciones clínicas. El 30% (27/90) de las muestras procesadas fueron positivas para Chlamydiaceae; el 70,3% (19/27) de estas resultaron positivas para Chlamydia psittaci y el 14,9% (4/27) para Chlamydia abortus. Nueve muestras positivas para C. psittaci fueron genotipificadas por secuenciación del gen ompA: 8 correspondieron al genotipo Ay una al genotipo B. Se observó una asociación significativa entre la presencia de Chlamydia spp. y la manifestación de signos clínicos compatibles con clamidiosis, como así también con la edad de las aves (menores de un ano). Este informe contribuye a mejorar nuestro conocimiento de los agentes clamidiales en nuestro país.

Detection and molecular characterization of Chlamydia psittaci and Chlamydia abortus in psittacine pet birds in Buenos Aires province, Argentina.

In order to determine the presence and genetic diversity of Chlamydia spp. in the north-eastern area of Buenos Aires province, Argentina, conjunctival, oropharyngeal, cloacal swab and tissues were collected from a total of 90 psittacine pet birds of different age and clinical manifestations. Through molecular methods, Chlamydiaceae was detected in 30% (27/90) of the samples, out of which 70.3% (19/27) were positive for Chlamydia psittaci and 14.9% (4/27) for Chlamydia abortus. Nine C. psittaci positive samples were genotyped by ompA gene sequences, 8 clustered within genotype A and 1 within genotype B. A significant association was observed between the presence of Chlamydia spp. and the manifestation of clinical signs compatible with chlamydiosis, as well as with the age of the birds (younger than one year old). This report contributes to the improvement of our understanding of chlamydial agents in our country.

Epidemiological and molecular characteristics of Chlamydia psittaci from 8 human cases of psittacosis and 4 related birds in Argentina / Características epidemiológicas y moleculares de Chlamydia psittaci provenientes de 8 casos humanos de psitacosis y de 4 aves relacionadas en la Argentina

In Argentina, the epidemiological and molecular characteristics of Chlamydia psittaci infections are still not sufficiently known. A total of 846 respiratory and 10 ocular samples from patients with suspected human psittacosis were tested for C. psittaci from January 2010 to March 2015. Four samples of birds related to these patients were also studied. Forty-eight samples were positive for C. psittaci by a nested PCR. The molecular characterization of twelve C. psittaci PCR-positive samples received in the National Reference Laboratory INEI-ANLIS "Dr. Carlos G. Malbrán", Buenos Aires, Argentina was performed. Eight positive samples from humans and four from birds were genotyped by ompA gene sequencing. C. psittaci genotype A was found in all human samples and in the related birds. This report contributes to our increasing knowledge of the epidemiological and molecular characteristics of C. psittaci to conduct effective surveillance of its zoonotic infections.

En la Argentina, aún no se conocen suficientemente las características epidemiológicas y moleculares de las infecciones por Chlamydia psittaci. Entre enero del 2010 y marzo del 2015 se estudiaron 846 muestras respiratorias y 10 oculares de pacientes con sospecha de psitacosis para la búsqueda de C. psittaci. También se estudiaron 4 muestras de aves relacionadas con estos pacientes. De ese total, 48 muestras fueron positivas para C. psittaci mediante una reacción en cadena de la polimerasa (PCR) anidada. Posteriormente, se realizó en el INEI-ANLIS «Dr. Carlos G. Malbrán¼ la caracterización molecular de 12 muestras positivas para C. psittaci, 8 de humanos y 4 de aves, que fueron genotipificadas por secuenciación del gen ompA. C. psittaci genotipo A se encontró en todas esas muestras. Este informe contribuye a mejorar nuestro conocimiento de las características epidemiológicas y moleculares de C. psittaci para lograr una vigilancia efectiva de la zoonosis que produce.

Epidemiological and molecular characteristics of Chlamydia psittaci from 8 human cases of psittacosis and 4 related birds in Argentina.

In Argentina, the epidemiological and molecular characteristics of Chlamydia psittaci infections are still not sufficiently known. A total of 846 respiratory and 10 ocular samples from patients with suspected human psittacosis were tested for C. psittaci from January 2010 to March 2015. Four samples of birds related to these patients were also studied. Forty-eight samples were positive for C. psittaci by a nested PCR. The molecular characterization of twelve C. psittaci PCR-positive samples received in the National Reference Laboratory INEI-ANLIS "Dr. Carlos G. Malbrán", Buenos Aires, Argentina was performed. Eight positive samples from humans and four from birds were genotyped by ompA gene sequencing. C. psittaci genotype A was found in all human samples and in the related birds. This report contributes to our increasing knowledge of the epidemiological and molecular characteristics of C. psittaci to conduct effective surveillance of its zoonotic infections.

Silent dissemination of HTLV-1 in an endemic area of Argentina. Epidemiological and molecular evidence of intrafamilial transmission.

BACKGROUND: Molecular and epidemiological studies of transmission routes and risk factors for infection by HTLV-1 are extremely important in order to implement control measures, especially because of the high prevalence of HTLV-1 in several regions of the world. San Salvador de Jujuy, Northwest Argentina, is a highly endemic area for HTLV-1 and foci of tropical spastic paraparesis/HTLV-1-associated myelopathy. OBJECTIVE: To gain further insight into the role of intrafamilial transmission of HTLV-1 in a highly endemic region in Argentina. METHOD: Cross-sectional study in Northwest Argentina. Epidemiological data and blood samples were collected from 28 HTLV-1 infected subjects (index cases) and 92 close relatives/cohabitants. HTLV-1 infection was diagnosed by detection of antibodies and proviral DNA. The LTR region was sequenced and analyzed for genetic distances (VESPA software), in addition to determination and identification of polymorphisms to define HTLV-1 family signatures. RESULTS: Fifty seven of the 120 subjects enrolled had antibodies against HTLV-1 and were typified as HTLV-1 by PCR. The prevalence rate of HTLV-1 infection in family members of infected index cases was 31.52% (29/92). The infection was significantly associated with gender, age and prolonged lactation. Identity of LTR sequences and presence of polymorphisms revealed high prevalence of mother-to-child and interspousal transmission of HTLV-1 among these families. CONCLUSION: There is an ongoing and silent transmission of HTLV-1 through vertical and sexual routes within family clusters in Northwest Argentina. This evidence highlights that HTLV-1 infection should be considered as a matter of public health in Argentina, in order to introduce preventive measures as prenatal screening and breastfeeding control.

Molecular characterization of Chlamydia pneumoniae in animals and humans from Argentina: Genetic characterization of Chlamydia pneumoniae.

In this study, genetic diversity of Chlamydia pneumoniae was investigated and the relationships between sequences amplified of different sources, clinical conditions and geographical regions of central Argentina were established. Samples amplified were similar to human C. pneumoniae patterns and show the high clonality of the population.

Mutation detection of E6 and LCR genes from HPV 16 associated with carcinogenesis.

Human papillomavirus (HPV) is responsible for one of the most frequent sexually transmitted infections. The first phylogenetic analysis was based on a LCR region fragment. Nowadays, 4 variants are known: African (Af-1, Af-2), Asian-American (AA) and European (E). However the existence of sub-lineages of the European variant havs been proposed, specific mutations in the E6 and LCR sequences being possibly related to persistent viral infections. The aim of this study was a phylogenetic study of HPV16 sequences of endocervical samples from Cordoba, in order to detect the circulating lineages and analyze the presence of mutations that could be correlated with malignant disease. The phylogenetic analysis determined that 86% of the samples belonged to the E variant, 7% to AF-1 and the remaining 7% to AF-2. The most frequent mutation in LCR sequences was G7521A, in 80% of the analyzed samples; it affects the binding site of a transcription factor that could contribute to carcinogenesis. In the E6 sequences, the most common mutation was T350G (L83V), detected in 67% of the samples, associated with increased risk of persistent infection. The high detection rate of the European lineage correlated with patterns of human migration. This study emphasizes the importance of recognizing circulating lineages, as well as the detection of mutations associated with high-grade neoplastic lesions that could be correlated to the development of carcinogenic lesions.

Genetic diversity of Chlamydia among captive birds from central Argentina.

To study the occurrence of Chlamydia spp. and their genetic diversity, we analysed 793 cloacal swabs from 12 avian orders, including 76 genera, obtained from 80 species of asymptomatic wild and captive birds that were examined with conventional nested polymerase chain reaction and quantitative polymerase chain reaction. Chlamydia spp. were not detected in wild birds; however, four species (Chlamydia psittaci, Chlamydia pecorum, Chlamydia pneumoniae and Chlamydia gallinacea) were identified among captive birds (Passeriformes, n = 20; Psittaciformes, n = 15; Rheiformes, n = 8; Falconiformes n = 2; Piciformes n = 2; Anseriformes n = 1; Galliformes n = 1; Strigiformes n = 1). Two pathogens (C. pneumoniae and C. pecorum) were identified simultaneously in samples obtained from captive birds. Based on nucleotide-sequence variations of the ompA gene, three C. psittaci-positive samples detected were grouped into a cluster with the genotype WC derived from mammalian hosts. A single positive sample was phylogenetically related to a new strain of C. gallinacea. This report contributes to our increasing understanding of the abundance of Chlamydia in the animal kingdom.

Molecular evidence of Chlamydophila pneumoniae infection in reptiles in Argentina.

In the central area of Argentina, the epidemiological and molecular characteristics of Chlamydophila pneumoniae infections in reptiles are still unknown. A nested polymerase chain reaction of the rpoB gene was used to detect C. pneumoniae in cloacal swab samples from 19 reptiles at a recreational area. Eleven (57.89%) reptiles were positive; the sequencing and phylogenetic analysis confirmed the presence of this bacterium. Neither C. pneumoniae DNA in the caregivers pharynges nor IgM antibodies anti-C. pneumoniae in their serum samples were detected; however, caregivers presented very high titers of IgG anti-C. pneumoniae. The detection of C. pneumoniae DNA in reptiles demonstrated the circulation of this agent in the recreational area and could be responsible for the exacerbated immune response of the personnel handling the reptiles, which suggests a potential zoonotic cycle. This is the first report of the detection of C. pneumoniae in reptiles in Argentina.

Evidencia molecular de la infección por Chlamydophila pneumoniae en reptiles de Argentina / Molecular evidence of Chlamydophila pneumoniae infection in reptiles in Argentina

En la región central de Argentina, las características epidemiológicas y moleculares de las infecciones por Chlamydophila pneumoniae en reptiles son desconocidas. Para detectar C. pneumoniae, se usó la reacción en cadena de la polimerasa anidada que amplifica el gen rpoB en muestras de hisopado cloacal de 19 reptiles. Once (57,89 %) reptiles resultaron positivos. La secuenciación y el análisis filogenético corroboraron la presencia de esta bacteria. No se detectó ADN de C. pneumoniae en la faringe ni IgM anti-C. pneumoniae en el suero de los cuidadores; sin embargo, ellos presentaron títulos muy elevados de IgG anti-C. pneumoniae. La detección de ADN de C. pneumoniae en los reptiles demostró la circulación de este agente en el centro recreativo donde se realizó este estudio, lo que podría explicar la exacerbada respuesta inmunitaria en los cuidadores; este hallazgo sugiere la presencia de un potencial ciclo zoonótico. Se reporta aquí por primera vez la detección de C. pneumoniae en reptiles en Argentina

In the central area of Argentina, the epidemiological and molecular characteristics of Chlamydophila pneumoniae infections in reptiles are still unknown. A nested polymerase chain reaction of the rpoB gene was used to detect C. pneumoniae in cloacal swab samples from 19 reptiles at a recreational area. Eleven (57.89%) reptiles were positive; the sequencing and phylogenetic analysis confirmed the presence of this bacterium. Neither C. pneumoniae DNA in the caregivers'pharynges nor IgM antibodies anti-C. pneumoniae in their serum samples were detected; however, caregivers presented very high titers of IgG anti-C. pneumoniae. The detection of C. pneumoniae DNA in reptiles demonstrated the circulation of this agent in the recreational area and could be responsible for the exacerbated immune response of the personnel handling the reptiles, which suggests a potential zoonotic cycle. This is the first report of the detection of C. pneumoniae in reptiles in Argentina

Detection of the ompA gene of Chlamydophila pecorum in captive birds in Argentina.

Bacteria belonging to the family Chlamydiaceae cause a broad spectrum of diseases in a wide range of hosts, including humans, other mammals and birds. However, very little is known about chlamydial infections in birds in our region. In the present study, we examined 28 clinically normal birds in illegal captivity that were confiscated in the province of C6rdoba, Argentina. The objective was to detect Chlamydophila spp. in cloacal swabs by genetic analysis of the ompA gene. Nested-PCR of the ompA gene identified five samples as Chlamydophila pecorum and the sequence analysis demonstrated the presence of the ompA gene of C. pecorum in these birds. On the other hand, Chlamydophila psittaci was not detected. These birds could be either asymptomatic reservoirs or subclinical carriers of C. pecorum. This is the first report of the detection of C. pecorum in Argentina.

Detection of the ompA gene of Chlamydophila pecorum in captive birds in Argentina / Detección del gen ompA de Chlamydophila pecorum en aves en cautiverio, en la Argentina

Bacteria belonging to the family Chlamydiaceae cause a broad spectrum of diseases in a wide range of hosts, Including humans, other mammals and birds. However, very little is known about chlamydial infections in birds in our region. In the present study, we examined 28 clinically normal birds In illegal captivity that were confiscated in the province of Córdoba, Argentina. The objective was to detect Chlamydophila spp. in cloacal swabs by genetic analysis of the ompA gene. Nested-PCR of the ompA gene identified five samples as Chlamydophila pecorum and the sequence analysis demonstrated the presence of the ompA gene of C. pecorum In these birds. On the other hand, Chlamydophila psittaci was not detected. These birds could be either asymptomatic reservoirs or subclinical carriers of C. pecorum. This is the first report of the detection of C. pecorum in Argentina.

Las bacterias que pertenecen a la familia Chlamydiaceae causan un extenso espectro de enfermedades en una amplia gama de huéspedes, incluidos los seres humanos, otros mamíferos y aves. Sin embargo, se sabe muy poco acerca de las infecciones por clamidias en aves de nuestra reglón. Esta Investigación examinó 28 aves clínicamente normales mantenidas en cautiverio ¡legal, que fueron confiscadas en Córdoba, Argentina. El objetivo fue detectar Chlamydophila spp. en hisopados cloacales por análisis del gen ompA. La PCR anidada del gen ompA reveló la presencia de Chlamydophila pecorum en cinco muestras. El análisis de secuencias demostró la presencia del gen ompA de C. pecorum en estas aves. Por el contrario, Chlamydophila psittaci no se detectó. Estas aves pueden ser reservónos asintomáticos o portadores subclínlcos de C. pecorum. Este es el primer informe de la detección de C. pecorum en la Argentina.