RESUMEN Introducción: la necesidad de contar con técnicas de laboratorio rápidas, sensibles y específicas para la detección de patógenos de difícil diagnóstico llevó a la creación del Laboratorio de Biología molecular en el Centro Provincial de Higiene, Epidemiología y Microbiología de Villa Clara. Objetivo: caracterizar la evolución y la importancia de la Microbiología molecular en el Laboratorio del Centro Provincial de Higiene, Epidemiología y Microbiología de Villa Clara de junio de 2018 a diciembre de 2023. Métodos: estudio descriptivo transversal realizado en el Laboratorio de Biología Molecular de Villa Clara en el período comprendido de junio de 2018 a diciembre de 2023 a partir del total de muestras procesadas por reacción en cadena de la polimerasa, se emplearon varias plataformas tecnológicas y diversos estuches comerciales. Resultados: en 2018 se implementó el diagnóstico molecular con dos plataformas, Rotor Gene-Q® y Gene Xpert® MTB/RIF, y en 2021, se dispuso de cuatro, al introducir el SLAN y la plataforma COBAS 4800®. El laboratorio adquirió relevancia por la llegada de la pandemia por el nuevo coronavirus de 2019: se procesaron 511 225 muestras de exudado nasofaríngeo y 14 568 determinaciones para el diagnóstico de los virus de inmunodeficiencia humana, de hepatitis C y B y del papiloma humano que se realizaron en la plataforma COBAS 4800®. En el año 2023 se fortaleció la capacidad de detección de patógenos y se identificó la cocirculación comunitaria de influenza A y B, del virus parainfluenza tipo 4, el coronavirus OC 43, el SARS-CoV-2 y el dengue. Conclusiones: el aumento progresivo de la capacidad resolutiva del Laboratorio de Biología molecular, junto a la implementación de nuevas plataformas diagnósticas, ha permitido fortalecer la detección de patógenos de interés clínico y perfeccionar la vigilancia centinela virológica.

ABSTRACT Introduction: the need to have rapid, sensitive and specific laboratory techniques for the detection of pathogens that are difficult to diagnose led to the creation of the Molecular Biology Laboratory at the Provincial Center of Hygiene, Epidemiology and Microbiology of Villa Clara. Objective: to characterize the evolution and importance of Molecular Microbiology in the Laboratory of the Provincial Center of Hygiene, Epidemiology and Microbiology of Villa Clara from June 2018 to December 2023. Methods: cross-sectional descriptive study carried out at the Villa Clara Molecular Biology Laboratory in the period from June 2018 to December 2023 based on the total samples processed by polymerase chain reaction, several technological platforms and various kits were used. commercial. Results: In 2018, molecular diagnosis was implemented with two platforms, Rotor Gene-Q® and Gene Xpert® MTB/RIF, and in 2021, four were available, when the SLAN and the COBAS 4800® platform were introduced. The laboratory became relevant due to the arrival of the 2019 new coronavirus pandemic: 511,225 nasopharyngeal exudate samples and 14,568 determinations were processed for the diagnosis of human immunodeficiency viruses, hepatitis C and B, and human papilloma carried out on the COBAS 4800® platform. In 2023, the pathogen detection capacity was strengthened and the community cocirculation of influenza A and B, parainfluenza virus type 4, the OC 43 coronavirus, SARS-CoV-2 and dengue was identified. Conclusions: the progressive increase in the resolution capacity of the Molecular Biology Laboratory, together with the implementation of new diagnostic platforms, has allowed us to strengthen the detection of pathogens of clinical interest and improve virological sentinel surveillance.

Abstract Diarrheagenic Escherichia coli comprises a heterogeneous group of pathotypes or pathogenic variants that share phenotypic characteristics with marked differences in virulence genes, colonization sites, pathogenesis, clinical presentation, and epidemiology of infection. The most studied pathotypes are Shiga toxin-producing E.coli (STEC), enterotoxigenic E.coli (ETEC), enteropathogenic E.coli (EPEC), enteroaggregative E.coli (EAEC), and enteroinvasive E.coli (EIEC). The objective of the study was to characterize the isolates of diarrheagenic E.coli from an outpatient pediatric population with diarrhea attended in two public hospitals from Buenos Aires, Argentina. Diarrheagenic E.coli pathotypes were investigated by amplifying characteristic virulence gene fragments: intimin (eae), heat-labile toxin (lt), heat-stable toxins (stp, sth), invasion plasmid antigen H (ipaH), transcriptional activator R (aggR) and Shiga toxins (stx1, stx2). Molecular subtyping of isolates was performed using PFGE (XbaI). Diarrheagenic E.coli was detected in 14% (84/601) of cases. The EAEC pathotype was prevalent, while ETEC, STEC, EPEC and EIEC were found in a lower proportion. EAEC isolates exhibited a high degree of genetic diversity. All pathotypes were found in children under 5years of age, while only EAEC, EIEC and ETEC were detected in the older population. Future studies that include the characterization of isolates from a greater number of genes and populations from other geographical areas will be necessary to determine the relevance of diarrheagenic E.coli in Argentina.

Resumen Escherichia coli diarreogénica abarca un grupo heterogéneo de patotipos o variantes patogénicas que comparten características fenotípicas con marcadas diferencias en genes de virulencia, sitios de colonización, patogenia, presentación clínica y epidemiología de la infección. Los patotipos más estudiados son E.coli productora de toxina Shiga (STEC), E.coli enterotoxigénica (ETEC), E.coli enteropatogénica (EPEC), E.coli enteroagregativa (EAEC) y E.coli enteroinvasiva (EIEC). El objetivo del estudio fue caracterizar los aislamientos de E.coli diarreogénica provenientes de población pediátrica ambulatoria con diarrea, atendida en dos hospitales públicos de Buenos Aires, Argentina. Los patotipos de E.coli diarreogénica se investigaron mediante la amplificación de fragmentos de genes de virulencia característicos: intimina (eae), toxina termolábil (lt), toxinas termoestables (stp, sth), antígeno plasmídico de invasiónH (ipaH), activador transcripcional R (aggR) y toxinas Shiga (stx1, stx2). La subtipificación molecular de aislamientos se realizó mediante PFGE (XbaI). E.coli diarreogénica fue detectada en el 14% (84/601) de los casos. El patotipo EAEC fue prevalente, mientras que ETEC, STEC, EPEC y EIEC fueron hallados en menor proporción. Los aislamientos de EAEC presentaron un alto grado de diversidad genética. Todos los patotipos fueron hallados en niños menores de 5años, mientras que solamente EAEC, EIEC y ETEC fueron detectados en población de mayor edad. Futuros estudios que incluyan la caracterización de aislamientos a partir de un mayor número de genes y población de otras áreas geográficas serán necesarios para determinar la relevancia de E.coli diarreogénica en Argentina.

Resumo O estudo de pesquisa de campo foi realizado na temporada (2017). A soja e as ervas daninhas foram amostradas semanalmente de forma aleatória. Tripes adultos foram identificados e contados. A detecção do vírus isolado e a incidência natural foram determinadas usando transmissão mecânica, gama de hospedeiros, DAS-ELISA, RT-PCR. A incidência natural de tripes em indivíduos foi detectada dependendo da % de SVNV em tripes e hospedeiros infestantes. Dez espécies de tripes foram associadas a plantas de soja no campo. A espécie mais abundante foi T. tabaci, com média de 256,5 número médio de indivíduos, seguida por F. occidentalis (142,5) e N. variabilis (86,6 / número médio de indivíduos). Catorze espécies de tripes ocorreram em 5 culturas de leguminosas e 41 espécies de plantas daninhas dentro de campos de soja. O maior número médio de 40,6 indivíduos foi registrado em Ammi majus. Enquanto o mais baixo, 3,3 número médio de indivíduos, foi no Urtica urens. Apenas 21 espécies de plantas diagnosticadas foram suscetíveis à infecção com SVNV. G. max e Vigna radiate foram os maiores percentuais de infecção, 80%, seguidos por V. unguilata e N. benthamiana, 75%. O isolado egípcio neste estudo mostrou um alto grau de similaridade e está intimamente relacionado ao TSWV do Egito (DQ479968) e ao TCSV dos EUA (KY820965), com identidade de sequência de nucleotídeos de 78%. Quatro espécies de tripes transmitiram SVNV (F. fusca 4,0%, F. schultzei 4,3%, F. tritici 3,3% e N. variabilis 68,0% de transmissão). Tanto C. phaseoli quanto M. sjostedti podem adquirir o vírus, mas não podem transmiti-lo. As seguintes espécies, T. tabaci, F. occidentalis, S. dorsallis e T. palmi não podem adquirir ou transmitir SVNV. A incidência de SVNV no campo, iniciada no final de julho, aumentou gradativamente de 12,7 para 71,3% no final da temporada. Em conclusão, poucos indivíduos de tripes invadiram a cultura da soja e são suficientes para transmitir alta taxa de SVNV dentro da cultura. Além disso, várias espécies de vetores também abundam em ervas daninhas, que são as principais fontes dos vírus da soja transmitidos às lavouras. Essas informações podem ser importantes para controlar e reduzir a incidência de infecção por SVNV.

Abstract Field survey study was conducted season (2017). Soybeans and weeds were weekly sampled randomly. Thrips adults were identified and counted. Detection of the virus isolate and the natural incidence was determined using; Mechanical transmission, host range, DAS-ELISA, RT-PCR. The natural incidence thrips individuals was detected depending on the SVNV% in thrips individuals and weeds hosts. Ten thrips species were associated with soybean plants in the field. The most abundant species was T. tabaci, average 256.5 average no.of individuals, followed by F. occidentalis (142.5 average no. of individuals), then N. variabilis (86.6/ average no. of individuals). Fourteen thrips species occurred on 5 legumes field crops and 41 weed plant species within soybean field. The highest average number 40.6.of individuals were recorded on Ammi majus. While the lowest one 3.3 average no. of individuals were on Urtica urens. Only 21diagnostic plant species were susceptible to infection with SVNV. G. max and Vigna radiate, were the highest percentage of infection 80% followed by V. unguilata & N. benthamiana, 75%. Egyptian isolate of Soybean vein necrosis virus (SVNV) in this study showed a high degree of similarity and it is closely related to TSWV from Egypt (DQ479968) and TCSV from USA (KY820965) with nucleotide sequence identity of 78%. Four thrips species transmitted SVNV (F. fusca 4.0%, F. schultzei 4.3%, F. tritici 3.3% and N. variabilis 68.0% transmission). Both C. phaseoli and M. sjostedti can acquire the virus but unable to transmit it. The following species; T. tabaci, F. occidentalis, S. dorsallis and T. palmi cannot acquire or transmit SVNV. The incidence of SVNV in the field started by the end of July then increased gradualy from 12.7 to 71.3% by the end of the season. In conclusion, few thrips individuals invaded soybean crops are enough to transmit high rate of SVNV within the crop. Furthermore, several vector species are also abundant on weeds, which are the major sources of soybean viruses transmitted to the crops. This information might be important for control and reduce the incidence of SVNV infection.

Abstract Toxoplasma gondii and Neospora caninum are two closely related protozoans that infect a wide range of animals, including birds. However, the occurrence of N. caninum and T. gondii in seabirds is unknown. Therefore, this study aimed to determine the presence of T. gondii and N. caninum DNA in tissue samples of seabirds. Tissue samples of the pectoral muscles, heart, and brain were collected from 47 birds along the coastline of Santa Catarina State, SC, Brazil. The DNA was extracted from the tissues and screened using nested-PCR (nPCR) targeting internal transcribed spacer 1 (ITS1). T. gondii DNA was detected in tissues from seven seabirds (7/47, 14.8%), kelp gull (Larus dominicanus) (5/21), and Manx shearwater (Puffinus puffinus) (2/8). N. caninum DNA was detected in tissues of nine seabirds (9/47, 19.1%), the kelp gull (L. dominicanus) (4/21), Manx shearwater (P. puffinus) (2/8), neotropic cormorant (Phalacrocorax brasilianus) (1/4), brown booby (Sula leucogaster) (1/5), and white-chinned petrel (Procellaria aequinoctialis) (1/1); however, no co-infection was observed. In conclusion, this study showed the circulation of N. caninum and T. gondii in seabirds along the coastline of Santa Catarina State. Further studies are required to clarify the role of these birds in the epidemiology of neosporosis and toxoplasmosis. animals However N T unknown Therefore muscles heart 4 State SC Brazil nestedPCR nested PCR nPCR (nPCR ITS1. ITS1 ITS . (ITS1) 7/47, 747 7 (7/47 14.8%, 148 14.8% , 14 8 14.8%) Larus dominicanus 5/21, 521 5/21 5 21 (5/21) Puffinus puffinus 2/8. 28 2/8 2 (2/8) 9/47, 947 9 (9/47 19.1%, 191 19.1% 19 19.1%) L. L (L 4/21, 421 4/21 (4/21) P. P (P 2/8, Phalacrocorax brasilianus 1/4, 1/4 (1/4) Sula leucogaster 1/5, 15 1/5 (1/5) whitechinned white chinned Procellaria aequinoctialis 1/1 11 (1/1) however coinfection co infection observed conclusion toxoplasmosis (ITS1 7/47 74 (7/4 14.8 52 5/2 (5/21 2/ (2/8 9/47 94 (9/4 19.1 42 4/2 (4/21 1/ (1/4 (1/5 (1/1 (ITS 7/4 (7/ 14. 5/ (5/2 (2/ 9/4 (9/ 19. 4/ (4/2 (1/ 7/ (7 (5/ (2 9/ (9 (4/ (1 ( (5 (4

Resumo Toxoplasma gondii e Neospora caninum são dois protozoários intimamente relacionados que infectam uma ampla gama de animais, incluindo-se aves. Entretanto, a ocorrência de N. caninum e T. gondii em aves marinhas é desconhecida. Portanto, este estudo objetivou determinar a presença de DNA de T. gondii e N. caninum em amostras de tecidos, coletadas de 47 aves marinhas. Amostras de tecidos dos músculos peitorais, coração e cérebro foram coletadas de 47 aves marinhas mortas ao longo da costa do estado de Santa Catarina, SC, Brasil. O DNA foi extraído dos tecidos e testado, usando-se a reação em cadeia da polimerase nested (nPCR), tendo em vista o espaçador interno transcrito 1 (ITS1). O DNA de T. gondii foi detectado em tecidos de sete aves marinhas (7/47, 14,8%), como gaivota (Larus dominicanus) (5/21) e bobo-pequeno (Puffinus puffinus) (2/8). O DNA de N. caninum foi detectado em tecidos de nove aves marinhas (9/47, 19,1%); gaivota (L. dominicanus) (4/21), bobo-pequeno (P. puffinus) (2/8), corvo-marinho (Phalacrocorax brasilianus) 1/4), atobá-pardo (Sula leucogaster) (1/5), e pardela-preta (Procellaria aequinoctialis) (1/1). No entanto, não foi observada coinfecção. Em conclusão, este estudo demonstrou a circulação de N. caninum e T. gondii em aves marinhas ao longo da costa de Santa Catarina, Brasil. Mais estudos são necessários para esclarecer o papel dessas aves na epidemiologia da neosporose e da toxoplasmose. animais incluindose incluindo se Entretanto N T desconhecida Portanto 4 peitorais Catarina SC Brasil testado usandose usando nPCR, nPCR , (nPCR) ITS1. ITS1 ITS . (ITS1) 7/47, 747 7 (7/47 14,8%, 148 14,8% 14 8 14,8%) Larus dominicanus 5/21 521 5 21 (5/21 bobopequeno bobo pequeno Puffinus puffinus 2/8. 28 2/8 2 (2/8) 9/47, 947 9 (9/47 19,1% 191 19 19,1%) L. L (L 4/21, 421 4/21 (4/21) P. P (P 2/8, corvomarinho corvo marinho Phalacrocorax brasilianus 1/4, 1/4 1/4) atobápardo atobá pardo Sula leucogaster 1/5, 15 1/5 (1/5) pardelapreta pardela preta Procellaria aequinoctialis 1/1. 11 1/1 (1/1) entanto coinfecção conclusão toxoplasmose (nPCR (ITS1 7/47 74 (7/4 14,8 5/2 52 (5/2 2/ (2/8 9/47 94 (9/4 19,1 42 4/2 (4/21 1/ (1/5 (1/1 (ITS 7/4 (7/ 14, 5/ (5/ (2/ 9/4 (9/ 19, 4/ (4/2 (1/ 7/ (7 (5 (2 9/ (9 (4/ (1 ( (4

ABSTRACT The occurrence of Toxoplasma gondii, Neospora caninum, Ehrlichia canis and Babesia vogeli in dogs from the municipality of Cândido Sales, Bahia, was investigated. Total and peripheral blood samples were obtained from 131 dogs. Blood smears were performed to check for morulae. Toxoplasma gondii and N. caninum infections were determined by an Indirect Immunofluorescence Reaction. Nested-PCR and PCR were used for the diagnoses of E. canis and B. vogeli infections, respectively. Additionally, the risk factors associated with infection by these agents were analyzed. The frequency of infection was 70.2% for N. caninum, 67.9% for T. gondii and 37.4% for E. canis. None of the dogs tested positive for B. vogeli. Morulae of Ehrlichia spp. were identified in two animals and the piroplasm in one animal. Age (> 3 years) was considered a risk factor for infections by E. canis and N. caninum and the rural habitat for infections by N. caninum. Co-infections were frequent, mainly with N. caninum and T. gondii (45.03% of dogs). Ehrlichia canis infection was significantly associated with N. caninum and T. gondii infection. These findings indicated a high occurrence of T. gondii, N. caninum, and E. canis in the studied region. Sales Bahia investigated 13 morulae N Reaction NestedPCR Nested E B respectively Additionally analyzed 702 70 2 70.2 679 67 9 67.9 T 374 37 4 37.4 spp animal > ( years Coinfections Co frequent 45.03% 4503 45 03 (45.03 . dogs) region 1 7 70. 6 67. 37. 45.03 450 0 (45.0 45.0 (45. 45. (45 (4

RESUMO Avaliou-se a ocorrência de infecções por Toxoplasma gondii, Neospora caninum, Ehrlichia canis e Babesia vogeli em cães do município de Cândido Sales - BA. Para isso, foram obtidas amostras de sangue total e periférico de 131 cães. Esfregaços sanguíneos foram confeccionados para verificação de mórulas. Infecções por T. gondii e N. caninum foram determinadas pela reação de imunofluorescência indireta. O diagnóstico das infecções por E. canis e B. vogeli foi feito por meio de Nested-PCR e PCR, respectivamente. Adicionalmente, foram analisados os fatores de risco associados à infecção por esses agentes. Verificou-se frequência de infecção de 70,2% para N. caninum, 67,9% para T. gondii e de 37,4% para E. canis. Nenhum cão foi positivo para B. vogeli. Mórulas de Ehrlichia spp. foram identificadas em dois animais, e piroplasma em um. A idade (acima de 3 anos) foi fator de risco para infecções por E. canis e N. caninum, e hábitat rural para N. caninum. Coinfecções foram frequentes, principalmente por N. caninum e T. gondii (45,03%). Infecção por E. canis foi associada significativamente às infecções por N. caninum e T. gondii. Os achados indicam elevada ocorrência de T. gondii, N. caninum, e E. canis na região. Avaliouse Avaliou se BA isso 13 mórulas T N indireta E B NestedPCR Nested PCR respectivamente Adicionalmente agentes Verificouse Verificou 702 70 2 70,2 679 67 9 67,9 374 37 4 37,4 spp animais um acima anos frequentes 45,03%. 4503 45,03% . 45 03 (45,03%) região 1 7 70, 6 67, 37, 450 45,03 0 (45,03% 45,0 (45,03 45, (45,0 (45, (45 (4 (

Abstract Despite an economy based mostly on agriculture, literature on viral diseases of plants is scarce in Paraguay. Only recently, researches on plant viruses took an impulse resulting in a precise identification of many of them affecting plants either cultivated or not. To provide reliable information regarding plant viruses present in Paraguay, an annotated list of them was prepared, covering descriptions from 1920 to present day. There have been some important outbreaks with severe yield losses in crops as cucurbits, citrus, sesame, bean, maize, peanuts and tomato. Many of older descriptions are included for their historical significance, but most identifications made require confirmation. On the other hand, recent descriptions have been completed, based on several assays, especially molecular characterization. This list is organized alphabetically following scientific names of the plant species found naturally infected by viruses, with comments about symptoms, geographical distribution, incidence, identification procedures, and other information, with due literature references. It is based on a compilation of publications made on plant virus diseases in Paraguay. Described virus species, in a total of 38 recognized by ICTV, belonging to 17 different genera (Alphaendornavirus, Ampelovirus, Begomovirus, Benyvirus, Carlavirus, Cilevirus, Closterovirus, Comovirus, Cucumovirus, Dichorhavirus, Fabavirus, Luteovirus, Ophiovirus, Orthotospovirus, Potexvirus, Potyvirus and Tobamovirus), besides two unclassified, and four unidentified. There is also a case of viroid described in Citrus spp. Infections caused by potyviruses are the most numerous. These viruses were described in more than 40 plant species, belonging to 18 botanical families. Because of crop diversity and richness in native flora, many more viruses must be present in Paraguay, which future works will certainly reveal, especially with the increase in manpower involving researches, especially cooperative with foreign centers, on plant viruses, which has been very limited until now. Also, knowledge on existing viruses may have relevance in understanding their epidemiology and provide the basis for their control strategies and quarantine measures, to avoid new variants of existing viruses or new viruses being introduced. agriculture Paraguay recently not prepared 192 day cucurbits citrus sesame bean maize tomato significance confirmation hand completed assays characterization symptoms distribution incidence procedures references 3 ICTV 1 Alphaendornavirus, Alphaendornavirus (Alphaendornavirus Ampelovirus Begomovirus Benyvirus Carlavirus Cilevirus Closterovirus Comovirus Cucumovirus Dichorhavirus Fabavirus Luteovirus Ophiovirus Orthotospovirus Potexvirus Tobamovirus, Tobamovirus , Tobamovirus) unclassified unidentified spp numerous 4 families flora reveal centers now Also measures introduced 19

Resumo A pesar de una economía basada principalmente en la agricultura, la literatura sobre enfermedades virales de las plantas es escasa en Paraguay. Sólo recientemente se han impulsado las investigaciones sobre los virus de plantas, lo que ha permitido identificar con precisión muchos de ellos que afectan a plantas cultivadas o no. Para brindar información confiable sobre los virus de plantas presentes en el Paraguay, se elaboró una lista comentada de los mismos, abarcando descripciones desde 1920 hasta la actualidad. Se han producido algunos focos importantes con severas pérdidas de rendimiento en cultivos de cucurbitáceas, cítricos, sésamo, frijol, maíz, maní y tomate. Muchas de las descripciones más antiguas se incluyen por su importancia histórica, pero la mayoría de las identificaciones realizadas requieren confirmación. Por otro lado, las descripciones recientes han sido completadas, basadas en varios ensayos, especialmente de caracterización molecular. Esta lista está organizada alfabéticamente siguiendo los nombres científicos de las especies de plantas que se encontraron naturalmente infectadas por virus, con comentarios sobre síntomas, distribución geográfica, incidencia, procedimientos de identificación y otras informaciones, con las debidas referencias bibliográficas. Se basa en una recopilación de publicaciones realizadas sobre enfermedades virales de plantas en Paraguay. Especies de virus descritas, en un total de 38 reconocidas por el ICTV, pertenecientes a 17 géneros diferentes (Alphaendornavirus, Ampelovirus, Begomovirus, Benyvirus, Carlavirus, Cilevirus, Closterovirus, Comovirus, Cucumovirus, Dichorhavirus, Fabavirus, Luteovirus, Ophiovirus, Orthotospovirus, Potexvirus, Potyvirus y Tobamovirus), además de dos sin clasificar y cuatro sin identificar. También existe un caso de un viroide descrito en Citrus spp. Las infecciones causadas por potyvirus son las más numerosas. Estos virus fueron descritos en más de 40 especies de plantas, pertenecientes a 18 familias botánicas. Debido a la diversidad de cultivos y la riqueza de la flora nativa, muchos más virus deben estar presentes en Paraguay, lo que seguramente revelarán trabajos futuros, especialmente con el aumento de la mano de obra involucrada en investigaciones, en cooperación con centros extranjeros, sobre virus de plantas, que ha sido muy limitada hasta el momento. Además, el conocimiento sobre los virus existentes puede ser relevante para comprender su epidemiología y proporcionar una base para sus estrategias de control y medidas de cuarentena, para evitar la introducción de nuevas variantes de virus existentes o nuevos virus. agricultura Paraguay no mismos 192 actualidad cucurbitáceas cítricos sésamo frijol maíz tomate histórica confirmación lado completadas ensayos molecular síntomas geográfica incidencia informaciones bibliográficas descritas 3 ICTV 1 Alphaendornavirus, Alphaendornavirus (Alphaendornavirus Ampelovirus Begomovirus Benyvirus Carlavirus Cilevirus Closterovirus Comovirus Cucumovirus Dichorhavirus Fabavirus Luteovirus Ophiovirus Orthotospovirus Potexvirus Tobamovirus, Tobamovirus , Tobamovirus) spp numerosas 4 botánicas nativa futuros extranjeros momento Además cuarentena 19

ABSTRACT Wild birds have great prominence in the transmission of diseases to humans, mainly due to their ease of access to human populations, raising concerns about the potential impact of that proximity on public health. The present study reports ARV and PBV occurrence in wild birds from an environmental protection area in the Amazon biome, in Pará state, Brazil. We obtained 155 fecal specimens from 47 species of wild birds for RNA extraction, ARV and PBV detection utilizing molecular methods, nucleotide sequencing, and phylogenetic analysis. ARV prevalence was 0.6% (1/155), being positive in an individual of Myrmotherula longipennis, and PBV prevalence was 1.29% (2/155), affecting two individuals of Guira guira. The PBV strains were assigned to genogroup I based on phylogenetic analysis, and they shared a strong phylogenetic link with strains isolated from different geographic locations. The ARV strain was more closely related to strains that had previously circulated in the same region. The presence of ARV and PBV in this habitat suggests that infection cycles of these agents occur naturally in the wild ecosystem, potentially triggering transmission events between bird species and humans. This is the first study on ARV detection in wild birds in Brazil and the first report on the occurrence of PBV in wild Guira guira. Additional studies are required to determine the epidemiology, origin, evolution, and emergence of new potentially pathogenic viruses in the Amazon. humans populations health biome state 15 4 extraction methods sequencing analysis 06 0 6 0.6 1/155, 1155 1/155 , 1 (1/155) longipennis 129 29 1.29 2/155, 2155 2/155 2 (2/155) guira locations region ecosystem epidemiology origin evolution 0. 115 1/15 (1/155 12 1.2 215 2/15 (2/155 11 1/1 (1/15 1. 21 2/1 (2/15 1/ (1/1 2/ (2/1 (1/ (2/ (1 (2 (

RESUMO As aves silvestres têm grande destaque na transmissão de doenças ao homem, principalmente pela facilidade de acesso à população humana, levantando preocupações sobre o potencial impacto dessa proximidade no contexto da saúde pública. O presente estudo relata a ocorrência de ARV e PBV em aves silvestres de uma área de proteção ambiental no bioma Amazônico. Obtivemos 155 espécimes fecais de 47 espécies de aves silvestres, que foram submetidos a extração de RNA, detecção de ARV e PBV por técnicas moleculares, sequenciamento nucleotídico e análise filogenética. A prevalência de infecção por ARV foi 0,6% (1/155), sendo positivo em um indivíduo de Myrmotherula longipennis, e a prevalência de PBV foi 1,29% (2/155), afetando dois insdivíduos de Guira guira. A análise filogenética agrupou as cepas de PBV no genogrupo I, mostrando intensa relação filogenética com cepas isoladas de outras regiões geográficas. A cepa de ARV esteve mais relacionada filogeneticamente a cepas previamente circulantes na mesma região. Este é o primeiro estudo sobre detecção de ARV em aves silvestres no Brasil e o primeiro relato sobre a ocorrência de PBV em Guira guira. A circulação de ARV e PBV nesse ambiente demonstra que os ciclos desses agentes estão ocorrendo de forma natural no ecossistema silvestre, o que pode desencadear possíveis eventos de transmissão entre as espécies aviárias e humanos. Estudos adicionais são necessários para determinar a epidemiologia, origem, evolução e surgimento de novos vírus potencialmente patogênicos na Amazônia. homem humana pública Amazônico 15 4 RNA moleculares 06 0 6 0,6 1/155, 1155 1/155 , 1 (1/155) longipennis 129 29 1,29 2/155, 2155 2/155 2 (2/155) guira I geográficas região silvestre humanos epidemiologia origem Amazônia 0, 115 1/15 (1/155 12 1,2 215 2/15 (2/155 11 1/1 (1/15 1, 21 2/1 (2/15 1/ (1/1 2/ (2/1 (1/ (2/ (1 (2 (

ABSTRACT: Clostridioides difficile is a strictly anaerobic, spore-forming Gram-positive bacterium associated with diarrhea, known as C. difficile infection (CDI). In domestic animals, C. difficile is considered an important pathogen mostly in pigs and horses, but there are also reports in other domestic species. In wild animals, the epidemiology of C. difficile is largely unknown, and the role of the bacterium as a cause of diarrhea is unclear. The aim of this study was to determine the prevalence of C. difficile in the feces of wild animals referred to the Center of Medicine and Research in Wild Animals (CEMPAS). Fecal samples obtained from 100 animals of 34 different species were subjected to qPCR for the detection of the C. difficile 16S rRNA gene and two major toxin genes (tcdA and tcdB) and to anaerobic bacterial isolation. A total of 63 animals (63%) were positive for C. difficile by qPCR, and 16 isolates were recovered. The opossum (Didelphis spp.) had the highest number of positive animals in both tests (from 21 samples, 19 were qPCR positive, and four isolates were recovered). Three toxigenic strains (RT 002, 004, and 014), all previously described as infecting humans and animals, were isolated in the following species: bearded dragon (Pogona vitticeps), pampas fox (Lycalopex vetulus), and marmoset (Callithrix sp.). The presence of C. difficile in the feces of wild animals highlights the importance of wildlife as potential carriers of infection for production animals or humans. ABSTRACT sporeforming spore forming Grampositive Gram C CDI. CDI . (CDI) horses unknown unclear CEMPAS. CEMPAS (CEMPAS) 10 3 S tcdA tcdB isolation 6 63% (63% 1 recovered Didelphis spp. spp 2 recovered) RT 002 004 014, 014 , 014) Pogona vitticeps, vitticeps vitticeps) Lycalopex vetulus, vetulus vetulus) Callithrix sp.. sp sp. sp.) (CDI (CEMPAS (63 00 01 (6 0 (

RESUMO: O Clostridioides difficile é uma bactéria Gram-positiva estritamente anaeróbica e formadora de esporos, associada à diarreia e conhecida como infecção por C. difficile (CID). Em animais domésticos, o C. difficile é considerado um patógeno importante principalmente em porcos e cavalos, mas também há relatos em outras espécies domésticas. Em animais selvagens, a epidemiologia do C. difficile é amplamente desconhecida, e o papel da bactéria como causa de diarreia não está claro. O objetivo deste estudo foi determinar a prevalência do C. difficile nas fezes de animais selvagens encaminhados ao Centro de Medicina e Pesquisa em Animais Selvagens (CEMPAS). Amostras de fezes obtidas de 100 animais de 34 espécies diferentes foram submetidas à qPCR para a detecção do gene 16S rRNA do C. difficile e dois principais genes de toxina (tcdA e tcdB), além de isolamento bacteriano anaeróbico. Um total de 63 animais (63%) foram positivos para C. difficile por qPCR, e 16 isolados foram recuperados. O gambá (Didelphis spp.) apresentou o maior número de animais positivos em ambos os testes (de 21 amostras, 19 foram positivas na qPCR, e quatro isolados foram recuperados). Três cepas toxigênicas (RT 002, 004 e 014), todas previamente descritas como infectando humanos e animais, foram isoladas nas seguintes espécies: dragão barbado (Pogona vitticeps), raposa-pampas (Lycalopex vetulus) e sagui (Callithrix sp.). A presença de C. difficile nas fezes de animais selvagens destaca a importância da vida selvagem como potencial portadora de infecção para animais de produção ou seres humanos. RESUMO Grampositiva Gram positiva esporos C CID. CID . (CID) domésticos cavalos domésticas desconhecida claro CEMPAS. CEMPAS (CEMPAS) 10 3 S tcdA tcdB, tcdB , tcdB) anaeróbico 6 63% (63% 1 recuperados Didelphis spp. spp 2 amostras recuperados) RT 002 00 014, 014 014) Pogona vitticeps, vitticeps vitticeps) raposapampas raposa pampas Lycalopex vetulus Callithrix sp.. sp sp. sp.) (CID (CEMPAS (63 0 01 (6 (

ABSTRACT: The Caatinga biome is unique to Brazil, with unfavorable environmental characteristics for the survival of Leptospira spp. However, recent studies have shown high positivity at PCR (polymerase chain reaction) in small ruminants. There are no Leptospira spp. studies based on sample calculation in pigs in the Caatinga. The aim of this study was to assess the importance of pigs in the spread of leptospirosis in the Caatinga biome. Overall, 200 biological samples (urine, blood, vaginal fluid, and tissues of reproductive and urinary tracts) were collected from 40 slaughtered sows, and MAT (microscopic agglutination test) and PCR tests were carried out to detect anti-Leptospira spp. antibodies and the agent’s DNA, respectively. The serological analysis showed a positivity rate of 5% (2/40), and the PCR identified Leptospira spp. DNA in 62.5% (25/40) of the animals. Only 2.5% (1/40) of the animals were positive for both techniques. The detected serogroups were Australis (50%) and Bataviae (50%), with antibody titers of 25 and 50. Leptospira spp. DNA was detected in 40% (16/40) of the reproductive tract samples, 32.5% (13/40) of the urinary tract, 32.5% (13/40) of the vaginal fluid and 30% (12/40) of the urine. There was no agreement (Kappa <0) between PCR samples from the genital tract vs. urinary tract or serological results. Genetic sequencing of one urine and one urinary tract tissue sample revealed 99% identity with L. borgpetersenii. The results indicate that leptospirosis is a concern in pigs in the context of Caatinga, with a high prevalence of infection detected by different diagnostic methods. The molecular analysis revealed a considerable proportion of infected animals. The findings emphasize the importance of a multifaceted approach in the diagnosis of leptospirosis in pigs, with a focus on the use of genital tract samples for the diagnosis of leptospirosis in this animal species, providing valuable insights for the control and prevention of this disease in both animals and the zoonotic context. Finally, the detection of leptospires in the genital tract indicates a possibility of male-female transmission in the venereal context. ABSTRACT Brazil spp However polymerase reaction ruminants Overall 20 urine, (urine blood tracts 4 sows microscopic test antiLeptospira anti agents agent s respectively 5 2/40, 240 2/40 , 2 (2/40) 625 62 62.5 25/40 2540 (25/40 2.5 1/40 140 1 (1/40 techniques 50% 50 (50% 50%, 16/40 1640 16 (16/40 325 32 32.5 13/40 1340 13 (13/40 30 12/40 1240 12 (12/40 Kappa <0 0 vs 99 L borgpetersenii methods species Finally malefemale male female 24 2/4 (2/40 6 62. 25/4 254 (25/4 2. 1/4 14 (1/4 (50 16/4 164 (16/4 3 32. 13/4 134 (13/4 12/4 124 (12/4 < 9 2/ (2/4 25/ (25/ 1/ (1/ (5 16/ (16/ 13/ (13/ 12/ (12/ (2/ (25 (1 ( (16 (13 (12 (2

RESUMO: O bioma Caatinga é único no Brasil, com características ambientais desfavoráveis à sobrevivência de Leptospira spp. Porém, estudos recentes demonstraram alta positividade na PCR (reação em cadeia da polimerase) em pequenos ruminantes. Não existem estudos para a infecção por Leptospira spp. baseados em cálculo amostral em suínos na Caatinga. O objetivo deste estudo foi avaliar a importância dos suínos na disseminação da leptospirose no bioma Caatinga. Foram coletadas 200 amostras biológicas (urina, sangue, fluido vaginal e tecidos do trato reprodutivo e urinário) de 40 porcas abatidas e realizados testes SAM (teste de soroaglutinação microscópica) e PCR para detecção de anticorpos anti-Leptospira spp. e DNA do agente, respectivamente. A análise sorológica mostrou taxa de positividade de 5% (2/40) e a PCR identificou o DNA de Leptospira spp. em 62,5% (25/40) dos animais. Apenas 2,5% (1/40) dos animais foram positivos para ambas as técnicas. Os sorogrupos detectados foram Australis (50%) e Bataviae (50%), com títulos de anticorpos de 25 e 50. O DNA de Leptospira spp. foi detectado em 40% (16/40) das amostras do trato reprodutivo, 32,5% (13/40) do trato urinário, 32,5% (13/40) do fluido vaginal e 30% (12/40) de urina. Não houve concordância (Kappa <0) entre amostras de PCR do trato genital vs. trato urinário ou resultados sorológicos. O sequenciamento genético de uma amostra de urina e de uma amostra de tecido do trato urinário revelou 99% de identidade com L. borgpetersenii. Os resultados obtidos indicam que a leptospirose representa uma preocupação em suínos no contexto da Caatinga, com alta prevalência de infecção detectada por diferentes métodos diagnósticos, bem como análises moleculares revelaram proporção considerável de animais infectados. Os resultados enfatizam a importância de uma abordagem multifacetada no diagnóstico da leptospirose em suínos, com foco no uso de amostras do trato genital para o diagnóstico da leptospirose nesta espécie animal, fornecendo informações valiosas para o controle e prevenção desta doença em animais e no contexto zoonótico. Por fim, a detecção de leptospiras no trato genital indica possibilidade de transmissão macho-fêmea no contexto venéreo. RESUMO Brasil spp Porém reação polimerase ruminantes 20 urina, (urina sangue 4 teste microscópica antiLeptospira anti agente respectivamente 5 2/40 240 2 (2/40 625 62 62,5 25/40 2540 (25/40 2,5 1/40 140 1 (1/40 técnicas 50% 50 (50% 50%, , 16/40 1640 16 (16/40 325 32 32,5 13/40 1340 13 (13/40 30 12/40 1240 12 (12/40 Kappa <0 0 vs sorológicos 99 L borgpetersenii diagnósticos infectados animal zoonótico fim machofêmea macho fêmea venéreo 2/4 24 (2/4 6 62, 25/4 254 (25/4 2, 1/4 14 (1/4 (50 16/4 164 (16/4 3 32, 13/4 134 (13/4 12/4 124 (12/4 < 9 2/ (2/ 25/ (25/ 1/ (1/ (5 16/ (16/ 13/ (13/ 12/ (12/ (2 (25 (1 ( (16 (13 (12

ABSTRACT Introduction: In Cuba, a country with a low incidence of tuberculosis (TB), there is no information about of the dynamics of transmission of the disease for two decades. Objective: Determine the genetic patterns and sublineages of Mycobacterium tuberculosis complex (MTBC) isolates circulating in 2009 and their relationship with the epidemiological data of the patients. Methods: A retrospective descriptive study was carried out in Cuba with 178 MTBC isolates. Spoligotyping and mycobacterial interspersed repetitive-unit-variable-number tandem-repeat (MIRU-VNTR) typing with 24loci were performed. For statistical analysis, non-parametric methods, analysis of variance and homogeneity test, were used. Results: Spoligotyping produced 39 spoligotypes. The S, Beijing, LAM and Haarlem sublineages prevailed. The clustering rate was 75.84% and the Hunter-Gaston discriminatory index (HGDI): 0.8734. MIRU-VNTR typing with 24-loci defined 154 genetic patterns: six grouped 30 isolates and 148 showed unique patterns. The clustering rate was 14.60% and the HGDI: 0.9926. There was a predominance of sublineages by region: S, Beijing and LAM in the West, Center and East, respectively. Conclusions: The dynamics of TB transmission in Cuba in 2009 is reported, inferring that this occurs in a limited manner and belonging to risk groups does not favor transmission. It serves to evaluate a megaproject of the Global Fund to reduce the transmission of TB in this country. The population genetic structure of MTBC resembles that of an Ibero-American country, with the exception of the high frequency of the Beijing and S sublineages.

RESUMEN Introducción: En Cuba, país con baja incidencia de tuberculosis (TB), no existe información sobre la dinámica de transmisión de la enfermedad desde hace dos décadas. Objetivo: Determinar los patrones genéticos y sublinajes de los aislados del complejo Mycobacterium tuberculosis (MTBC) circulantes en el año 2009 y su relación con los datos clínico-epidemiológicos de los pacientes. Métodos: Se realizó un estudio descriptivo retrospectivo en Cuba con 178 aislamientos de MTBC. Se realizó el espoligotipaje y el tipaje con unidades repetitivas intercaladas de micobacterias con repetición en tándem de número variable (MIRU-VNTR) con 24loci. Para el análisis estadístico, se utilizaron métodos no paramétricos, análisis de varianza y prueba de homogeneidad. Resultados: El espoligotipaje produjo 39 espoligotipos, prevaleciendo los sublinajes S, Beijing, LAM y Haarlem. El rango de agrupamiento fue del 75,84 % y el índice discriminatorio de Hunter-Gaston (HGDI): 0,8734. El tipaje por MIRU-VNTR con 24loci definió 154 patrones genéticos: seis agruparon 30 aislados y 148 mostraron patrones únicos. El rango de agrupamiento fue del 14,60 % y el HGDI: 0,9926. Hubo predominio de los sublinajes por región: S, Beijing y LAM en Occidente, Centro y Este, respectivamente. Conclusiones: Se reporta la dinámica de transmisión de la TB en Cuba en el año 2009, infiriéndose que esta ocurre de manera limitada y la pertenencia a grupos de riesgo no favorece la transmisión. Sirve para evaluar un megaproyecto del Fondo Mundial para reducir la transmisión de la TB en este país. La estructura genética poblacional de MTBC se asemeja a la de un país iberoamericano, con la excepción de la alta frecuencia de los sublinajes Beijing y S.

Abstract Sarcoptic mange or scabies is a contagious parasitic skin disease that affects a wide range of domestic and wildlife species. A cross-sectional study was carried out to determine the prevalence, molecular identification, and characteristics of sarcoptic mange in vicuñas. A total of 3,274 vicuñas were examined. Following ritual harvesting events (“chaccus”) in 13 Andean communities. The presence of mange mites was determined by the skin scraping technique and confirmed by PCR analysis using specific primers for the ITS2 gene of Sarcoptes scabiei. The presence of mange mites was also confirmed by microscopy using samples taken from wallows. A data collection form was used to register the characteristics of the vicuñas sampled. The prevalence of sarcoptic mange was 4.9% (95% CI: 4.1 - 5.6%). All samples from wallows tested positive (9/9). Importantly, the presence of the species S. scabiei was molecularly confirmed. Adult females with regular body condition were more susceptible to sarcoptic mange, although the lesions were mild. This study confirms the presence of S. scabiei in semi-captive vicuñas and points to the possible role of wallows in the dissemination of Sarcoptes mites. crosssectional cross sectional identification 3274 3 274 3,27 examined chaccus (“chaccus” 1 communities ITS sampled 49 4 9 4.9 95% 95 (95 CI 41 4. 5.6%. 56 5.6% . 5 6 5.6%) 9/9. 99 9/9 (9/9) Importantly S mild semicaptive semi captive 327 27 3,2 (“chaccus (9 5.6 9/ (9/9 32 2 3, ( 5. (9/

Resumo A sarna sarcóptica é uma doença parasitária contagiosa da pele que afeta uma ampla gama de espécies domésticas e selvagens. Um estudo transversal foi realizado para determinar a prevalência, identificação molecular e características da sarna sarcóptica em vicunhas. Foram avaliadas 3.274 vicunhas, capturadas durante o "Chaccus", em 13 comunidades rurais alto andinas do Peru. A presença do ácaro da sarna foi determinada pelo raspado de pele e confirmada pela técnica de PCR, com iniciadores específicos para o gene ITS2 de Sarcoptes scabiei e por microscopia nos chafurdeiros. Foi preenchida uma ficha de coleta de dados com as características das vicunhas analisadas. A prevalência de sarna sarcóptica em vicunhas foi de 4,9% (95% IC: 4,1 - 5,6%). Todas as amostras de chafurda testadas (9/9) foram positivas, e a presença de S. scabiei foi confirmada molecularmente. Fêmeas adultas com condições corporais regulares foram mais suscetíveis à sarna sarcóptica, embora as lesões fossem leves. O estudo confirma a presença de S. scabiei em vicunhas em semicativeiro de comunidades camponesas alto andinas do Peru que sofrem de sarna. Destaca-se, ainda, o papel das chafurdas das vicunhas na disseminação da sarna. selvagens 3274 3 274 3.27 Chaccus, Chaccus , "Chaccus" 1 PCR ITS chafurdeiros analisadas 49 4 9 4,9 95% 95 (95 IC 41 4, 5,6%. 56 5,6% . 5 6 5,6%) 9/9 99 (9/9 positivas S molecularmente leves Destacase, Destacase Destaca se, se Destaca-se ainda 327 27 3.2 "Chaccus (9 5,6 9/ (9/ 32 2 3. ( 5,

ABSTRACT Introduction: The CVA24 antigenic variant is the causal agent of epidemics of acute hemorrhagic conjunctivitis that has prevailed to date. To study the transmission dynamics and evolution of this variant, we molecularly characterized Cuban isolates obtained in 5 epidemic periods between 1986 and 2009. Methods: Using the BEAST v1.8.4 program, it was performed the phylogenetic and phylodynamic analysis of the partial genomic regions of 54 Cuban sequences and 83 sequences from 17 countries for the 3C genomic region and 35 Cuban sequences with 95 sequences from 18 countries for the VP1 region. They were analyzed the amino acid changes of the Cuban sequences and they were compared the sequences obtained from conjunctival exudates and feces. Results: It was identified that each epidemic period was caused by different genetic variants, and homologous to variants that circulated in the Americas, Africa, and Asia, and that could circulate up to 2 years prior to the epidemic peaks. The virus showed a stable effective population over time, increasing in pandemic periods, with a nucleotide substitution rate (substitutions/site/year) of 4.39 × 10-3 for the 3C region and 5.80 × 10-3 for VP1. They were obtained 19 viral transmission routes for the 3C genomic region and 25 routes for VP1. They were inferred the routes of introduction of the virus to Cuba in the epidemic periods of the years 1986, 1997, 2003 and 2008-2009 by the ancestral relationships of the Cuban sequences with sequences of destination countries of some routes, endorsed by viral transmission networks. Amino acid changes in both genomic regions could be involved in the evolution of CVA24v and it was obtained evidence that supports the use of feces to identify this viral variant. The conclusions were new knowledge in the molecular epidemiology of CVA24v that circulated in Cuba and the world for 23 years after its global emergence.

RESUMEN Introducción: La variante antigénica del CVA24 constituye el agente causal de epidemias de conjuntivitis hemorrágica aguda que ha prevalecido hasta la actualidad. Para estudiar la dinámica de transmisión y la evolución de esta variante, caracterizamos molecularmente aislamientos cubanos obtenidos en 5 periodos epidémicos entre 1986 y 2009. Métodos: Mediante el programa BEAST v1.8.4 se realizó el análisis filogenético y filodinámico de las regiones genómicas parciales de 54 secuencias cubanas y 83 secuencias de 17 países para la región genómica 3C y 35 secuencias cubanas con 95 secuencias de 18 países para la región VP1. Se analizaron los cambios aminoacídicos de las secuencias cubanas y se compararon las secuencias obtenidas de exudados conjuntivales y heces. Resultados: Se identificó que cada periodo epidémico fue causado por variantes genéticas diferentes, y homólogas a variantes que circularon en Las Américas, África y Asia, y que pudieron circular hasta 2 años previos a los picos epidémicos. El virus mostró una población efectiva estable en el tiempo, aumentando en los periodos pandémicos, con una tasa de sustitución nucleotídica (sustituciones/sitio/año) de 4,39 × 10-3 para la región 3C y 5,80 × 10-3 para VP1. Se obtuvieron 19 rutas de transmisión viral para la región genómica 3C y 25 rutas para VP1. Las rutas de introducción del virus a Cuba en los periodos epidémicos de los años 1986, 1997, 2003 y 2008-2009 se infirieron por las relaciones de ancestralidad de las secuencias cubanas con secuencias de países destinos de algunas rutas, avaladas por redes de transmisión viral. Los cambios aminoacídicos en ambas regiones genómicas pudieran involucrarse en la evolución del CVA24v y se obtuvieron evidencias que apoyan el uso de las heces para identificar esta variante viral. Las conclusiones fueron nuevos conocimientos en la epidemiología molecular del CVA24v que circuló en Cuba y el mundo durante 23 años a partir de su emergencia global.

ABSTRACT Introduction: Resistance to third generation cephalosporins in Enterobacteriacea is mediated by extended spectrum β-lactamase enzymes. The main families of β-lactamase enzymes are CTX-M and SHV. E. coli and its genes are spread either by clonal spread or horizontal transfer through the food chain. Previous studies demonstrated in Cuba β-lactamase enzymes CTX-M types in human strains, but there are no data in animals. The objective was to characterize commensal E. coli- third generation cephalosporins in healthy poultry and pigs. Methods: Cloacal exudates (434) were collected from birds (2013-2015) in Mayabeque and 215 rectals from pigs in western farms and slaughterhouses in (2016-2015). They were analyzed by molecular epidemiology techniques. Results: The presence of E. coli- third generation cephalosporins strains is reported in birds (62) and pigs (96). The variants blaCTX-M-1, blaCTX-M-15 were reported for the first time in Cuba in E.coli from birds and blaCTX-M-32, blaCTX-M-15 and blaCTX-M-55 in strains of pigs. The strains showed multi-resistance to other antibiotics tetracyclines and fluoroquinolones, revealed high diversity, those of birds corresponded to 23 types of sequences and different groups of plasmid incompatibility. The pig strains had 16 types of sequences, in one of them the entire genome was sequenced and the location of the blaCTX-M-32 gene in an IncX1 plasmid was verified. Coincidentally, sequence types 410 (birds) and 648 (pigs) harboring blaCTX-M-15 are detected in human clinical strains. Conclusions CTX-M-type ESBL-producing E. coli strains are widespread in poultry and pig production facilities in the western region; active horizontal transfer seems to be the main route for the dissemination of blaCTX-M genes. It is suggested to review the use of antimicrobials in animals and to strengthen good hygiene practices to minimize the risk of propagation of E.coli third generation cephalosporins clones.

RESUMEN Introducción: La resistencia a cefalosporinas de tercera generación en Enterobacteriacea es mediada por las enzimas β-lactamasas de espectro extendido. Las familias principales de enzimas β-lactamasas de espectro extendido son CTX-M y SHV. E. coli y sus genes se propagan, ya sea por diseminación clonal o transferencia horizontal a través de la cadena alimentaria. Estudios previos demostraron en Cuba tipos cefalosporinas de tercera generación CTX-M en cepas de humanos, pero no existen datos en animales. El objetivo fue caracterizar E. coli- cefalosporinas de tercera generación comensal en aves y cerdos sanos. Métodos: Se colectaron 434 exudados cloacales de aves (2013-2015) en Mayabeque y 215 rectales de cerdos en granjas y mataderos del occidente en 2016-2018. Se analizaron por técnicas de epidemiología molecular. Resultados: Se informa la presencia de 62 cepas de E.coli-RC3G en aves y 96 de cerdos. Se notificaron por primera vez en Cuba las variantes blaCTX-M-1, blaCTX-M-15 en E. coli de aves y blaCTX-M-32, blaCTX-M-15 y blaCTX-M-55 en cepas de cerdos. Las cepas mostraron multirresistencia a otros antibióticos como tetraciclinas y fluoroquinolonas, revelaron alta diversidad, las de aves correspondieron a 23 tipos de secuencias y diferentes grupos de incompatibilidad plasmídica. Las cepas de cerdos a 16 tipos de secuencias, en una de ellas se secuenció el genoma completo y se comprobó la localización del gen blaCTX-M-32 en un plásmido IncX1. Coincidentemente, se detectan en cepas clínicas de humanos los tipos de secuencias 410 (aves) y 648 (cerdos) albergando blaCTX-M-15. Conclusiones Cepas de E. coli productoras de BLEE tipo CTX-M están extendidas en instalaciones de producción de aves y cerdos en la región occidental, la transferencia horizontal activa parece ser la vía principal para la diseminación de los genes blaCTX-M. Se sugiere revisar el uso de antimicrobianos en animales y fortalecer las buenas prácticas de higiene para minimizar el riesgo de propagación de clones de E. coli R-C3G.

Resumen El cáncer colorrectal (CCR) es una enfermedad compleja determinada por factores de riesgo genéticos, ambientales y de estilo de vida. Los factores genéticos (hereditarios) tienen gran influencia en su desarrollo, sin embargo, la mayoría de los casos de CCR son esporádicos y se desarrollan gradualmente a lo largo de varios años. Los principales factores ambientales de riesgo están asociados a la vía de señalización de β-catenina, entre ellos obesidad, falta de actividad física, consumo de carnes rojas y procesadas, alcoholismo y tabaquismo. La vía está relacionada con la regulación de la homeostasis celular, autorrenovación celular durante la embriogénesis y edad adulta. La principal recomendación para evitar el desarrollo del CCR es reducir los factores de riesgo, aumentar el consumo de frutas, verduras y granos, hacer ejercicio de manera rutinaria y limitar el consumo tanto de alcohol como de tabaco. Dado que los antecedentes familiares y la presencia de un síndrome hereditario aumentan el riesgo, se sugiere hacer exámenes periódicos para detectar CCR y emplear predictores del desarrollo como los marcadores bioquímicos y moleculares, los cuales se presentan en este trabajo.

Abstract Colorectal cancer (CRC) is a complex disease, determined by genetic, environmental and lifestyle-associated risk factors. Genetic (inherited) factors have great influence on its development; however, most cases of CRC are sporadic and gradually develop over several years. The main environmental risk factors are associated with β-catenin signaling pathway, including obesity, lack of physical activity, consumption of red and processed meats, alcoholism, and smoking. The pathway is related to cell homeostasis regulation and cell self-renewal during embryogenesis and adulthood. The main recommendation for preventing the development of CRC is to reduce the risk factors, increase the consumption of fruits, vegetables and grains, exercise regularly and limit the consumption of both alcohol and tobacco. However, family history and the presence of a hereditary syndrome increase the risk, which is why carrying out periodic examinations to detect CRC is suggested, using development predictors such as biochemical and molecular markers, which are discussed in this work.

ABSTRACT Background: Serrated lesions are the precursors of up to one-third of colorectal cancer (CRC) cases and share molecular and epidemiological features with interval CRC. Previous studies have reported wide variation in serrated polyp prevalence and diverse magnitude of its relationship with synchronous advanced adenomas. Objective: Describe the prevalence of serrated polyps and evaluate their association with synchronous advanced adenomas. Methods: The study is a retrospective analysis of 1208 colonoscopies performed in patients aged 45 to 75, predominantly for CRC screening. Data on the prevalence of serrated polyps subsets and advanced adenomas were collected, and multivariate analysis were performed to identify the association between serrated polyps and synchronous advanced adenomas. Results: The prevalence of clinically significant serrated polyps (CSSP), large serrated polyps (LSP), and sessile serrated adenomas (SSA) were 11.3%, 6%, and 3.7%. CSSP were associated with synchronous advanced adenomas (OR 2.121, 95%CI 1.321-3.406), regardless of proximal (OR 2.966, 95%CI 1.701-5.170) or distal (OR 1.945, 95%CI 1.081-3.499) location, while LSP (OR 2.872, 95%CI 1.425-5.787) and SSA (OR 5.032, 95%CI 2.395-10.576) were associated with proximal advanced adenomas. Conclusion: The prevalence of CSSP and advanced adenomas were alike. CSSP is a risk factor for advanced adenomas, and the strength of this association is stronger for proximal advanced adenomas. LSP and SSA are associated with proximal advanced adenomas. Background onethird one third (CRC Objective Methods 120 4 75 screening collected Results CSSP, , (CSSP) LSP, (LSP) (SSA 113 11 3 11.3% 6 6% 37 7 3.7% OR 2121 2 121 2.121 95CI CI 95 1.3213.406, 13213406 1.321 3.406 1 321 406 1.321-3.406) 2966 966 2.966 1.7015.170 17015170 1.701 5.170 701 5 170 1.701-5.170 1945 945 1.945 1.0813.499 10813499 1.081 3.499 081 499 1.081-3.499 location 2872 872 2.872 1.4255.787 14255787 1.425 5.787 425 787 1.425-5.787 5032 032 5.032 2.39510.576 239510576 2.395 10.576 395 10 576 2.395-10.576 Conclusion alike 12 (CSSP (LSP 11.3 3.7 212 2.12 9 3213 1.3213.406 1321340 1321 1.32 3406 3.40 32 40 1.321-3.406 296 96 2.96 7015 1.7015.17 1701517 1701 1.70 5170 5.17 70 17 1.701-5.17 194 94 1.94 0813 1.0813.49 1081349 1081 1.08 3499 3.49 08 49 1.081-3.49 287 87 2.87 4255 1.4255.78 1425578 1425 1.42 5787 5.78 42 78 1.425-5.78 503 03 5.03 39510 2.39510.57 23951057 2395 2.39 10576 10.57 39 57 2.395-10.57 11. 3. 21 2.1 1.3213.40 132134 132 1.3 340 3.4 1.321-3.40 29 2.9 1.7015.1 170151 1.7 517 5.1 1.701-5.1 19 1.9 1.0813.4 108134 108 1.0 349 0 1.081-3.4 28 8 2.8 1.4255.7 142557 142 1.4 578 5.7 1.425-5.7 50 5.0 3951 2.39510.5 2395105 239 2.3 1057 10.5 2.395-10.5 2. 1.3213.4 13213 13 1. 34 1.321-3.4 1.7015. 17015 51 5. 1.701-5. 1.0813. 10813 1.081-3. 1.4255. 14255 14 1.425-5. 2.39510. 239510 23 105 10. 2.395-10. 1.3213. 1.321-3. 1.7015 1.701-5 1.0813 1.081-3 1.4255 1.425-5 2.39510 23951 2.395-10 1.3213 1.321-3 1.701- 1.081- 1.425- 2.3951 2.395-1 1.321- 2.395-

RESUMO Contexto: Lesões serrilhadas são precursoras de até um terço dos casos de câncer colorretal (CCR) e compartilham características moleculares e epidemiológicas com o CCR de intervalo. Estudos prévios relataram ampla variação na prevalência de pólipos serrilhados e na magnitude da sua relação com adenomas avançados sincrônicos. Objetivo: Descrever a prevalência de pólipos serrilhados colorretais e avaliar sua associação com adenomas avançados sincrônicos. Métodos: O estudo é uma análise retrospectiva de 1208 colonoscopias realizadas em pacientes com idades entre 45 e 75 anos, predominantemente para rastreamento de CCR. Dados sobre a prevalência de subtipos de pólipos serrilhados e de adenomas avançados foram coletados, e análises multivariadas foram realizadas para identificar a associação entre pólipos serrilhados e adenomas avançados sincrônicos. Resultados: A prevalência de pólipos serrilhados clinicamente significativos (PSCS), pólipos serrilhados grandes (PSG) e adenomas sésseis serrilhados (ASS) foi de 11,3%, 6% e 3,7%, respectivamente. PSCS foram associados a adenomas avançados sincrônicos (OR 2,121, IC95% 1,321-3,406), independentemente da localização proximal (OR 2,966, IC95% 1,701-5,170) ou distal (OR 1,945, IC95% 1,081-3,499), enquanto PSG (OR 2,872, IC 95%1,425-5,787) e ASS (OR 5,032, IC95% 2,395-10,576) foram associados a adenomas avançados proximais. Conclusão: A prevalência de PSCS e de adenomas avançados foi semelhante. PSCS é um fator de risco para adenomas avançados, e a força dessa associação é maior para adenomas avançados proximais. PSG e ASS estão associados a adenomas avançados proximais. Contexto (CCR intervalo Objetivo Métodos 120 4 7 anos coletados Resultados PSCS, , (PSCS) (PSG (ASS 113 11 3 11,3% 6 37 3,7% respectivamente OR 2121 2 121 2,121 IC95 1,3213,406, 13213406 1,321 3,406 1 321 406 1,321-3,406) 2966 966 2,966 1,7015,170 17015170 1,701 5,170 701 5 170 1,701-5,170 1945 945 1,945 1,0813,499, 10813499 1,081 3,499 081 499 1,081-3,499) 2872 872 2,872 95%1,4255,787 9514255787 95%1,425 5,787 95 425 787 95%1,425-5,787 5032 032 5,032 2,39510,576 239510576 2,395 10,576 395 10 576 2,395-10,576 proximais Conclusão semelhante 12 (PSCS 11,3 3,7 212 2,12 IC9 3213 1,3213,406 1321340 1321 1,32 3406 3,40 32 40 1,321-3,406 296 96 2,96 7015 1,7015,17 1701517 1701 1,70 5170 5,17 70 17 1,701-5,17 194 94 1,94 0813 1,0813,499 1081349 1081 1,08 3499 3,49 08 49 1,081-3,499 287 87 2,87 4255 95%1,4255,78 951425578 951425 95%1,42 5787 5,78 9 42 78 95%1,425-5,78 503 03 5,03 39510 2,39510,57 23951057 2395 2,39 10576 10,57 39 57 2,395-10,57 11, 3, 21 2,1 1,3213,40 132134 132 1,3 340 3,4 1,321-3,40 29 2,9 1,7015,1 170151 1,7 517 5,1 1,701-5,1 19 1,9 1,0813,49 108134 108 1,0 349 0 1,081-3,49 28 8 2,8 95%1,4255,7 95142557 95142 95%1,4 578 5,7 95%1,425-5,7 50 5,0 3951 2,39510,5 2395105 239 2,3 1057 10,5 2,395-10,5 2, 1,3213,4 13213 13 1, 34 1,321-3,4 1,7015, 17015 51 5, 1,701-5, 1,0813,4 10813 1,081-3,4 95%1,4255, 9514255 9514 95%1, 95%1,425-5, 2,39510, 239510 23 105 10, 2,395-10, 1,3213, 1,321-3, 1,7015 1,701-5 1,0813, 1,081-3, 95%1,4255 951 95%1 95%1,425-5 2,39510 23951 2,395-10 1,3213 1,321-3 1,701- 1,0813 1,081-3 95% 95%1,425- 2,3951 2,395-1 1,321- 1,081- 2,395-