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Mimivirus fibrils are intriguing structures that have drawn attention since their discovery. Although still under investigation, the function of fibrils may be related to host cell adhesion.

Background and Aim: Molecular approaches to diagnose respiratory viruses have provided an opportunity for early and subclinical pathogen detection, particularly in samples from the upper respiratory tract. This study aimed to investigate the presence of herpesviruses, particularly equid herpesvirus (EHV)-2 and EHV-5, in samples from the lower respiratory tract of healthy racehorses from Southern Brazil. Materials and Methods: Samples from the lower respiratory tract (i.e., bronchoalveolar lavage fluid [BALF]) were assessed by video endoscopy, cytological evaluation of BALF, and tracheal aspirates (TA), along with quantitative polymerase chain reaction (qPCR), to detect equine herpesvirus infection in the lower respiratory tract samples and compare corresponding cytological and endoscopic findings. Results: At least one abnormality per horse during endoscopy examination was observed, including, but not limited to, mucous secretion in the airways and pharyngeal lymphoid hyperplasia. The presence of EHV-2 and/or EHV-5 was detected by qPCR in 3/10 animals. One horse was positive for EHV-2 alone, one for EHV-5 alone, and one for both. Conclusion: To the authors' knowledge, this is the first molecular detection of EHV-2 and EHV-5 in Brazilian thoroughbred horses. These findings may provide new insights into the epidemiology of EHV-2 and EHV-5 in Brazilian horses, evidencing the importance of the molecular investigation, early detection, and prevention of respiratory diseases. State Jockey Club, Paran Published version The authors are thankful to State Jockey Club, Paran?, Brazil, or providing the necessary facilities for the study. The authors did not receive any funds for this study.

Studies related to the prevalence of leptospirosis in the semiarid region showed that even during long periods of drought, the disease has a remarkable frequency in herds in the region. It is a neglected disease and the extent of its effects in the Brazilian semiarid region is not known. The dynamics of this agent is well studied in the urinary tract, however, there are not many studies regarding the genital tract in female goats. Observing this scenario, the present work aimed to diagnose Leptospira spp. in female goats kept in the Brazilian semiarid region by means of serological, molecular and isolation techniques. Blood samples, vaginal fluid, urine and fragments of organs from the genitourinary tract were collected from 40 goats destined for slaughter. Microscopic agglutination test (MAT) was used as a serological technique, with a battery of 24 serovars. The Polymerase Chain Reaction (PCR) of the vaginal fluid, urine and organ fragments was performed, as well as the bacterial growth of these same products in a selective medium. Isolation positive samples were subjected to PCR. It was observed that two (5%) animals were serologically positive for the Pyrogenes serogroup. A total of 29 (72.5%) animals were PCR positive, with DNA present in 51/160 (31.8%) samples from the genital tract and 34/120 (28.3%) from the urinary tract, with no statistical difference. For bacterial growth, 22/40 (55%) animals were positive for growth, with morphology being observed in 19/160 (11.8%) for the genital tract and 16/120 (13.3%) for the urinary tract, with no statistical difference. Two uterus samples showed 99% similarity with L. interrogans after sequencing. Thus, female goats kept under semiarid conditions were positive for Leptospira spp, with positive samples from both the urinary and genital tracts, which possible is an alternative way of adapting and maintaining the agent for severe and adverse conditions.

Made available in DSpace on 2021-06-25T10:24:47Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-01-01 Haemotropic mycoplasmas (haemoplasmas) are small pleomorphic bacteria infecting erythrocytes of several mammalian species, including human beings. No study to date has focused on the risk of bacteria exposure in hunting activities, particularly in natural environments of highly tick-infested areas. Accordingly, the present study aimed to assess haemoplasma occurrence in the complex encompassing wild boars, hunting dogs and hunters of Brazil. A total of 38/65 (58.5%) wild boars and 94/159 (59.1%) dogs were positive by qPCR for at least one haemoplasma. All 25 hunters were negative. Dogs with high hunting frequency were 2.4 more likely to be infected. Sequencing revealed a probable novel haemoplasma species in wild boars. Although exposure to haemoplasma species was present, the study herein found no evidence of cross-species transmission. Department of Comparative Pathobiology Purdue University Department of Cell and Molecular Biology Universidade Federal do Paraná Biotechnology Institute Universidade Estadual Paulista (UNESP) Department of Preventive Medicine Universidade de São Paulo Department of Medicine Universidade Estadual de Ponta Grossa Department of Veterinary Medicine Universidade Federal do Paraná Department of Parasitology Institute of Biological Sciences Universidade Federal de Minas Gerais Biotechnology Institute Universidade Estadual Paulista (UNESP)

Bovine leptospirosis causes economic losses and raises public health concerns. It is possible that there are peculiarities in the epidemiology of leptospirosis in regions with a semiarid climate, such as the Caatinga biome in Brazil, where the climate is hot and dry, and the etiological agent require alternative routes of transmission. This study aimed to close knowledge gaps to the diagnosis and epidemiology of Leptospira spp. infection in cows from the Caatinga biome, Brazil. Samples of the blood, urinary tract (urine, bladder and kidney) and reproductive tract (vaginal fluid, uterus, uterine tube, ovary and placenta) were collected from 42 slaughtered cows. Diagnostic tests included were the microscopic agglutination test (MAT), polymerase chain reaction (PCR) and bacterial isolation. Anti-Leptospira spp. antibodies were found in 27 (64.3%) of the animals analyzed using MAT at a 1:50 dilution (cut-off 50), while 31 (73.8%) animals had at least one organ/fluid where the presence of Leptospira spp. DNA was identified, and 29 animals (69%) were positive at bacteriological culture. The highest sensitivity values for MAT were obtained at the cut-off point of 50. In conclusion, even under hot and dry climate conditions, it is possible that Leptospira spp. can spread through alternative routes such as venereal transmission; moreover, a cut-off of 50 is recommended for the serological diagnosis of cattle from the Caatinga biome.

Leptospirosis is poorly studied in small ruminants raised in field semiarid conditions. In this study we compared serological, bacteriological and molecular diagnostic methods in ewes maintained in field Brazilian semiarid conditions. Blood, vaginal fluid and urine samples were collected from 60 Morada Nova ewes raised in a semi-intensive system in the Brazilian semiarid. Diagnostic tests performed were microscopic agglutination test (MAT), polymerase chain reaction (PCR) and bacterial isolation. Anti-Leptospira sp. antibodies were found in eight (13.33%) animals analyzed by MAT at reciprocal titer 25 (cut-off 25), while Leptospira sp. DNA was detected in urine or vaginal fluid of 56 animals (93.33%). There was growth of leptospires in 10 urine cultures and in 11 vaginal fluid cultures, however, two of urine (2/60-3.33%) and eight cultures of vaginal fluid (8/60-13.33%) were confirmed by PCR. Two samples of vaginal fluid (one of each animal) were submitted to sequencing demonstrating 99% similarity with L. santarosai and L. interrogans. The highest MAT sensitivities were obtained with reciprocal titer 25 (cut-off 25) compared to 50 and 100. The performance of different diagnostic techniques for leptospirosis in ewes raised in field semiarid conditions allowed a better evaluation of the herd, as well as made it possible to identify carrier animals. Genital route may be important for efficient transmission and without dependence on environmental factors in ewes from semiarid, as well as it's highlighted that titer 1:25 in serology was more efficient, indicating its use in ewes in field semiarid conditions.

Made available in DSpace on 2022-04-29T08:38:43Z (GMT). No. of bitstreams: 0 Previous issue date: 2022-02-01 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Mimiviruses are giant viruses of amoeba that can be found in association with virophages. These satellite-like viruses are dependent on the mimivirus viral factory to replicate. Mimiviruses can also be associated with linear DNA molecules called transpovirons. Transpovirons and virophages are important drivers of giant virus evolution although they are still poorly studied elements. Here, we describe the isolation and genomic characterization of a mimivirus/virophage/transpoviron tripartite system from Brazil. We analyzed transmission electron microscopy images and performed genome sequencing and assembly, gene annotation, and phylogenetic analysis. Our data confirm the isolation of a lineage A mimivirus (1.2 Mb/1012 ORFs), called mimivirus argentum, and a sputnik virophage (18,880 bp/20 ORFs). We also detected a third sequence corresponding to a transpoviron from clade A (6365 bp/6 ORFs) that presents small terminal inverted repeats (77 nt). The main genomic features of mimivirus argentum and of its virophage/transpoviron elements corroborates with what is described for other known elements. This highlights that this triple genomic and biological interaction may be ancient and well-conserved. The results expand the basic knowledge about unique and little-known elements and pave the way to future studies that might contribute to a better understanding of this tripartite relationship. Laboratório de Vírus Departamento de Microbiologia Universidade Federal de Minas Gerais (UFMG), MG Laboratório de Virologia Departamento de Microbiologia e Imunologia Instituto de Biotecnologia Universidade Estadual Paulista (Unesp), Alameda das Tecomarias s/n, Chácara Capão Bonito, SP Laboratório de Virologia Departamento de Microbiologia e Imunologia Instituto de Biotecnologia Universidade Estadual Paulista (Unesp), Alameda das Tecomarias s/n, Chácara Capão Bonito, SP CNPq: 302081/2018-6

Zika virus (ZIKV) was discovered in 1947 with the virus isolation from Rhesus monkey (Macaca mulatta) in Uganda forest, Africa. Old World Primates are involved in a sylvatic cycle of maintenance of this arbovirus, however a limited knowledge about the role of New World primates in ZIKV transmission cycles has been established.This work aimed to investigate the presence of enzootic circulation of ZIKV in New World Primates from three Brazilian states: São Paulo, Paraíba, and Paraná.We analyzed 100 non-human primate samples collected in 2018 and 2020 from free-ranging and captive environments from São Paulo (six municipalities belonging to Sorocaba region), Paraíba (João Pessoa municipality), and Paraná (Foz do Iguaçu municipality) using reverse transcriptase quantitative polymerase reaction (RT-qPCR) assays, indirect enzyme-linked immunosorbent assay (ELISA), and plaque reduction neutralization test (PRNT).All samples (n = 141) tested negative for the presence of ZIKV genome from tissue and blood samples. In addition, all sera (n = 58) from Foz do Iguaçu' non-human primates (NHPs) were negative in serological assays.No evidence of ZIKV circulation (molecular and serological) was found in neotropical primates. In addition, the absence of antibodies against ZIKV suggests the absence of previous viral exposure of NHPs from Foz do Iguaçu-PR.

Made available in DSpace on 2022-04-28T19:46:25Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-10-01 This study focused on the epidemiological characterization and spatial distribution of bat shelters concerning livestock animal rabies in Paraná State, southern Brazil. A spatiotemporal cluster analysis was performed based on rabies-positive cases and the Desmodus rotundus shelters. A total of 1742 suspect rabies cases submitted for diagnosis from 2011 to 2017 were analyzed; 481 (27.61%) were positive, and 1261 (72.39%) were negative by direct immunofluorescence and biological testing in mice. Out of the positive samples, 413/481 (85.8%) was bovine, 44/481 (9.1%) equine, 6/481 (1.2%) sheep, 5/481 (1.0%) bubaline, and 14/481 (2.9%) were bats. From 22 Regional Units of Agricultural Health, the northeast 129 (26.82%) and central 86 (17.88%) units had the highest recurrence rates of positive cases. Paraná State was continuously endemic for livestock rabies, with the highest caseload seen in the southern-central regions, which was associated with the highest number of vampire bat shelters and natural geographical characteristics favoring bat housing. There was a decrease in the number of rabies cases in livestock in 2013 and 2014. Spatiotemporal analyses of point process mapping and control of D. rotundus shelters and suspected livestock rabies cases in the study area were steady and statistically correlated. However, as bats may travel up to 35-40 km to prey on cattle clusters, bat shelter locations may not be the most sensitive measure of exposure. Furthermore, future studies should consider landscape features such as altitude as potential associated risk factors. Rabies vaccination of livestock and bat hematophagous shelters identification combined with bat control is recommended to increase the efficacy of preventive measures, particularly in natural geographic characteristics favoring local bat housing. Graduate Program in Cellular and Molecular Biology Federal University of Paraná Agency of Agricultural Defense of Paraná-ADAPAR Program Surveillance and Prevention of Transmissible Nervous Syndromes in Production Animals Department of Preventive Veterinary Medicine and Animal Health University of São Paulo Institute of Biotechnology UNESP-Univ. Estadual Paulista Campus de Botucatu Department of Comparative Pathobiology College of Veterinary Medicine Purdue University Department of Veterinary Medicine Federal University of Paraná Department of Comparative Pathobiology Purdue University Institute of Biotechnology UNESP-Univ. Estadual Paulista Campus de Botucatu

Made available in DSpace on 2022-04-29T08:35:28Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-10-01 Financiadora de Estudos e Projetos Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Center for Information Technology Center for Scientific Review National Institutes of Health Office of Extramural Research, National Institutes of Health Office of Research Infrastructure Programs, National Institutes of Health Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ) The emergence of several SARS-CoV-2 lineages presenting adaptive mutations is a matter of concern worldwide due to their potential ability to increase transmission and/or evade the immune response. While performing epidemiological and genomic surveillance of SARS-CoV-2 in samples from Porto Ferreira—São Paulo—Brazil, we identified sequences classified by pangolin as B.1.1.28 harboring Spike L452R mutation, in the RBD region. Phylogenetic analysis revealed that these sequences grouped into a monophyletic branch, with others from Brazil, mainly from the state of São Paulo. The sequences had a set of 15 clade defining amino acid mutations, of which six were in the Spike protein. A new lineage was proposed to Pango and it was accepted and designated P.4. In samples from the city of Porto Ferreira, P.4 lineage has been increasing in frequency since it was first detected in March 2021, corresponding to 34.7% of the samples sequenced in June, the second in prevalence after P.1. Also, it is circulating in 30 cities from the state of São Paulo, and it was also detected in one sample from the state of Sergipe and two from the state of Rio de Janeiro. Further studies are needed to understand whether P.4 should be considered a new threat. Laboratório de Estudos Genômicos Departamento de Biologia Instituto de Biociências Letras e Ciências Exatas (IBILCE) Universidade Estadual Paulista (Unesp) São José do Rio Preto Instituto de Biotecnologia Universidade Estadual Paulista (Unesp) Laboratório de Bioinformática Laboratório Nacional de Computação Científica (LNCC) Laboratório de Síntese e Estudos de Biomoléculas (LaSEBio) Departamento de Bioquímica e Química Orgânica Instituto de Química Universidade Estadual Paulista (Unesp) Laboratório de Química Medicinal Síntese Orgânica e Modelagem Molecular (LaQMedSOMM) Departamento de Bioquímica e Química Orgânica Instituto de Química Universidade Estadual Paulista (Unesp) Laboratório de Pesquisas em Virologia (LPV) Departamento de Doenças Dermatológicas Infecciosas e Parasitárias Faculdade de Medicina de São José do Rio Preto (FAMERP) Laboratório de Medicina Veterinária Preventiva Aplicada Departamento de Medicina Veterinária Faculdade de Zootecnia e Engenharia de Alimentos (FZEA) Universidade de São Paulo (USP) Departamento de Análises Clínicas Faculdade de Ciências Farmacêuticas (FCFAR) Universidade Estadual Paulista (Unesp) Laboratório de Microbiologia Molecular Instituto de Ciências da Saúde Universidade Feevale Novo Hamburgo Laboratório de Estudos Genômicos Departamento de Biologia Instituto de Biociências Letras e Ciências Exatas (IBILCE) Universidade Estadual Paulista (Unesp) São José do Rio Preto Instituto de Biotecnologia Universidade Estadual Paulista (Unesp) Laboratório de Síntese e Estudos de Biomoléculas (LaSEBio) Departamento de Bioquímica e Química Orgânica Instituto de Química Universidade Estadual Paulista (Unesp) Laboratório de Química Medicinal Síntese Orgânica e Modelagem Molecular (LaQMedSOMM) Departamento de Bioquímica e Química Orgânica Instituto de Química Universidade Estadual Paulista (Unesp) Departamento de Análises Clínicas Faculdade de Ciências Farmacêuticas (FCFAR) Universidade Estadual Paulista (Unesp) Financiadora de Estudos e Projetos: 01.20.0029.000462/20 FAPESP: 20/04836-0 CNPq: 303170/2017-4 CNPq: 404096/2020-4 Center for Information Technology: CREATE-NEO 1 U01 AI151807-01 Center for Scientific Review: CREATE-NEO 1 U01 AI151807-01 National Institutes of Health: CREATE-NEO 1 U01 AI151807-01 Office of Extramural Research, National Institutes of Health: CREATE-NEO 1 U01 AI151807-01 Office of Research Infrastructure Programs, National Institutes of Health: CREATE-NEO 1 U01 AI151807-01 FAPERJ: E-26/202.903/20

Funda??o de Amparo ? Pesquisa do Estado de S?o Paulo (2014/13532-3). S?o Paulo State University. Institute for Biotechnology. Botucatu, SP, Brazil. Vales do Jequitinhonha e Mucuri Federal University. Agrarian Sciences Institute. Una?, MG, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil S?o Paulo State University. Institute for Biotechnology. Botucatu, SP, Brazil. Universidade Federal de Minas Gerais. Virology Laboratory. Belo Horizonte, MG, Brasil. Mato Grosso Federal University. Virology and Rickettsiosis Laboratory. Cuiab?, GO, Brazil. Mato Grosso Federal University. Virology and Rickettsiosis Laboratory. Cuiab?, GO, Brazil. EMBRAPA Pantanal. Corumb?, MT, Brazil. S?o Paulo State University. Institute for Biotechnology. Botucatu, SP, Brazil. S?o Paulo State University. Institute for Biotechnology. Botucatu, SP, Brazil. Abstract: Equine infectious anemia virus (EIAV) is a persistent lentivirus that causes equine infectious anemia (EIA). In Brazil, EIAV is endemic in the Pantanal region, and euthanasia is not mandatory in this area. All of the complete genomic sequences from field viruses are from North America, Asia, and Europe, and only proviral genomic sequences are available. Sequences from Brazilian EIAV are currently available only for gag and LTR regions. Thus, the present study aimed for the first time to sequence the entire EIAV genomic RNA in naturally infected horses from an endemic area in Brazil. RNA in plasma from naturally infected horses was used for next-generation sequencing (NGS), and gaps were filled using Sanger sequencing methodology. Complete viral genomes of EIAV from two horses were obtained and annotated (Access Number: MN560970 and MN560971). Putative genes were analyzed and compared with previously described genes, showing conservation in gag and pol genes and high variations in LTR and env sequences. Amino acid changes were identified in the p26 protein, one of the most common targets used for diagnosis, and p26 molecular modelling showed surface amino acid alterations in some epitopes. Brazilian genome sequences presented 88.6% nucleotide identity with one another and 75.8 to 77.3% with main field strains, such as EIAV Liaoning, Wyoming, Ireland, and Italy isolates. Furthermore, phylogenetic analysis suggested that this Brazilian strain comprises a separate monophyletic group. These results may help to better characterize EIAV and to overcome the challenges of diagnosing and controlling EIA in endemic regions

Background: The emergence of the Brazilian variant of concern, Gamma lineage (P.1), impacted the epidemiological profile of COVID-19 cases due to its higher transmissibility rate and immune evasion ability. Methods: We sequenced 305 SARS-CoV-2 whole-genomes and performed phylogenetic analyses to identify introduction events and the circulating lineages. Additionally, we use epidemiological data of COVID-19 cases, severe cases, and deaths to measure the impact of vaccination coverage and mortality risk. Results: Here we show that Gamma introduction in São José do Rio Preto, São Paulo, Brazil, was followed by the displacement of seven circulating SARS-CoV-2 variants and a rapid increase in prevalence two months after its first detection in January 2021. Moreover, Gamma variant is associated with increased mortality risk and severity of COVID-19 cases in younger age groups, which corresponds to the unvaccinated population at the time. Conclusions: Our findings highlight the beneficial effects of vaccination indicated by a pronounced reduction of severe cases and deaths in immunized individuals, reinforcing the need for rapid and massive vaccination.

The emergence of the new Brazilian variant of concern, P.1 lineage (Gamma), raised concern about its impact on the epidemiological profile of COVID-19 cases due to its higher transmissibility rate and immune evasion ability. Using 272 whole-genome sequences combined with epidemiological data, we showed that P.1 introduction in São José do Rio Preto, São Paulo, Brazil, was followed by the displacement of eight circulating SARS-CoV-2 variants and a rapid increase in prevalence two months after its first detection. Our findings support that the P.1 variant is associated with an increase in mortality risk and severity of COVID-19 cases in younger aged groups, which corresponds to the unvaccinated population at the time. Moreover, our data highlight the beneficial effects of vaccination indicated by a pronounced reduction of severe cases and deaths in immunized individuals, reinforcing the need for rapid and massive vaccination.

Made available in DSpace on 2022-05-01T08:15:09Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-08-01 Soil is the principal habitat and reservoir of fungi that act on ecological processes vital for life on Earth. Understanding soil fungal community structures and the patterns of species distribution is crucial, considering climatic change and the increasing anthropic impacts affecting nature. We evaluated the soil fungal diversity in southeastern Brazil, in a transitional region that harbors patches of distinct biomes and ecoregions. The samples originated from eight habitats, namely: Semi-deciduous forest, Brazilian savanna, pasture, coffee and sugarcane plantation, abandoned buildings, owls’ and armadillos’ burrows. Forty-four soil samples collected in two periods were evaluated by metagenomic approaches, focusing on the high-throughput DNA sequencing of the ITS2 rDNA region in the Illumina platform. Normalized difference vegetation index (NDVI) was used for vegetation cover analysis. NDVI values showed a linear relationship with both diversity and richness, reinforcing the importance of a healthy vegetation for the establishment of a diverse and complex fungal community. The owls’ burrows presented a peculiar fungal composition, including high rates of Onygenales, commonly associated with keratinous animal wastes, and Trichosporonales, a group of basidiomycetous yeasts. Levels of organic matter and copper influenced all guild communities analyzed, supporting them as important drivers in shaping the fungal communities’ structures. Department of Chemical and Biological Sciences Institute of Biosciences São Paulo State University ‘Júlio de Mesquita Filho’ Center for Tropical Medicine Faculty of Medicine University of Brasília (UnB), Brasília, Distrito Federal Department of Biostatistics Plant Biology Parasitology and Zoology Institute of Biosciences São Paulo State University ‘Júlio de Mesquita Filho’ Institute for Biotechnology São Paulo State University ‘Júlio de Mesquita Filho’ Laboratory of Molecular Biology Medical School São Paulo State University ‘Júlio de Mesquita Filho’ Department of Chemical and Biological Sciences Institute of Biosciences São Paulo State University ‘Júlio de Mesquita Filho’ Department of Biostatistics Plant Biology Parasitology and Zoology Institute of Biosciences São Paulo State University ‘Júlio de Mesquita Filho’ Institute for Biotechnology São Paulo State University ‘Júlio de Mesquita Filho’ Laboratory of Molecular Biology Medical School São Paulo State University ‘Júlio de Mesquita Filho’

Made available in DSpace on 2022-04-28T19:43:26Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-07-01 Universidade Federal do Paraná Institute of Biotechnology and Bioengineering, Isfahan University of Technology Purdue University Homeless persons have been considered as one of the most susceptible populations to sexually transmitted infections. In Brazil, these population experienced an increase of 140% from 2012 to 2020. Accordingly, the present study aimed to assess the seroprevalence of anti-Treponema pallidum, anti-HCV, anti-HIV antibodies, and the risk factors associated with homeless persons in a daytime attendance shelter of São Paulo city during the syphilis epidemic in Brazil. Blood samples of 116 volunteers and epidemiological data were conveniently collected in the shelter from June through August 2018. Detection of syphilis, HCV, and HIV antibodies was performed by chemiluminescent microparticle immunoassay (CMIA). CMIA-reagent samples for anti-T. pallidum antibodies were confirmed by Venereal Disease Research Laboratory (VDRL) non-treponemal test. VDRL non-reagent samples were confirmed by treponemal rapid immunochromatographic test. A rapid immunoblot assay confirmed seropositivity to HIV. Overall, anti-T. pallidum antibodies were observed in 29/116 (25.0%), anti-HCV antibodies in 4/116 (3.4%), and anti-HIV antibodies in 2/116 (1.7%) individuals, both co-infected with anti-T. pallidum antibodies. Associated risk factors for syphilis in homeless persons were being born or previously living in another city (p = 0.043) and becoming homeless due to family conflicts (p = 0.035). Besides homeless vulnerability, worldwide shortage of benzathine penicillin supply and increasing of syphilis testing access through rapid testing in primary health care services may have also impacted disease spreading at the time. The prevalence of syphilis found herein is the highest worldwide to date in this population. Federal University of Paraná Department of Veterinary Medicine College of Veterinary Science Federal University of Paraná Clinics Hospital Clinical Analysis Laboratory Unit Federal University of Paraná Department of Cellular and Molecular Biology College of Cellular and Molecular Biology State University of Ponta Grossa Department of Nursing and Public Health São Paulo State University Institute of Biotechnology, Botucatu University of São Paulo Department of Medical Microbiology Purdue University Department of Comparative Pathobiology Federal University of Paraná Department of Veterinary Medicine Brazil São Paulo State University Institute of Biotechnology, Botucatu

Visceral leishmaniasis is a parasitic zoonosis that mainly affects poorest and most vulnerable populations, and domestic dogs are considered to be the main source of infection to the vector and therefore humans. However, several studies have investigated the role of other vertebrate hosts in the disease cycle. In this context, the aim of the present study was to conduct a survey of Leishmania infantum infection in donkeys and mules living in a semiarid region of Brazil. Whole blood sampled from 72 equids (65 donkeys and 7 mules) was used to perform molecular diagnosis using the real-time polymerase chain reaction (qPCR) technique. A total of 25% of the samples (18/72) were positive through qPCR, but there were no significant differences between the species (donkeys or mules), sex (male or female) and abandonment situation of the animals (yes or no). Donkeys and mules living under semiarid conditions have high frequency of L. infantum infection. It is therefore worth assigning importance to these species in the epidemiological cycle of visceral leishmaniasis, either as potential reservoirs or just as an abundant food source for vectors. Resumo A leishmaniose visceral é uma zoonose parasitária que afeta principalmente populações mais pobres e vulneráveis, e os cães domésticos são considerados as principais fontes de infecção para o vetor e, portanto, para os humanos. Porém diversos estudos têm pesquisado o papel de outros hospedeiros vertebrados no ciclo da doença. Neste contexto, objetivou-se realizar um levantamento da infecção por Leishmania infantum em asininos e muares, vivendo em região semiárida do Brasil. Foi utilizado sangue total de 72 equídeos (65 asininos e 7 muares) para a realização de diagnóstico molecular por meio da técnica de Reação em Cadeia de Polimerase em Tempo Real (qPCR). Um total de 25% das amostras (18/72) resultaram positivas na qPCR, porém não houve diferença significativa entre as espécies (asininos e muares), sexo (macho e fêmea) e situação de abandono dos animais (sim ou não). Asininos e muares, vivendo em condições semiáridas, apresentam alta frequência de infecção por L. infantum, sendo válido atribuir importância a essas espécies no ciclo epidemiológico da leishmaniose visceral, seja como um reservatório em potencial, seja apenas como uma fonte alimentar abundante para os vetores.

Made available in DSpace on 2021-06-25T11:07:48Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-01-01 Even in the adverse environmental conditions of the semiarid region, leptospires can survive and spread by alternative routes of transmission, such as sexual in ewes, however, there is no data on rams. Thus, the present study aimed to evaluate the use of serological, molecular and microbial tools applied to diagnosis of Leptospira sp. Infection in rams maintained in semiarid conditions. Biological samples of urinary (urine, kidney and bladder) and genital (vas deferens, epididymis tail and vesicular gland) tracts were collected from 40 slaughtered rams for polymerase chain reaction (PCR) and bacterial isolation, as well as blood samples for antibody detection through microscopic serum agglutination test (MAT). Anti-Leptospira antibodies were found in five (12.5%) animals with antibody titer of 50 and 2 (5%) for the titer 100 for serogroups Pyrogenes, Ballum, Icterohaemorrhagiae and Australis. Leptospira sp. DNA was found in PCR of organs and urine of 30 (75%) animals. Overall, 240 fragments of organs from the urinary and genital tracts and urine were evaluated, with 93 (38.7%) positive samples, being 48/120 (40%) for the urinary tract and 45/120 (37.5%) for the genital. There was no statistically significant difference between the tracts. A bladder sample was sent for sequencing and showed 99% similarity with L. interrogans. Of the 240 cultures evaluated, 59 (24.5%) had leptospire growth, being that 23 (39%) were confirmed in PCR. Considering the PCR of organs and urine and bacterial growth as gold standards, the cut-off 50 in MAT showed greater sensitivity when compared to cut-off 100, regardless of the material used. The great proportion of leptospiral DNA in organs, urine and culture and bacterial growth from the genital tracts reinforce its importance as an extra-renal site and highlights the possible role of rams in venereal transmission, as well as the sensitivity of the cut-off 50 suggested its adoption in the serology of rams maintained in semiarid conditions. Universidade Federal de Campina Grande (UFCG) Centro de Saúde e Tecnologia Rural (CSTR), Av. Universitária, s/n, Santa Cecília Universidade de São Paulo (USP), Av. Prof. Almeida Prado, 1280 Universidade Estadual Paulista (Unesp), Av. Prof. Mário Rubens Guimarães Montenegro, s/n, campus de Botucatu Universidade Federal da Paraíba (UFPB) Centro de Ciências Agrárias (CCA), Rodovia BR 079, Km 02 Universidade Estadual Paulista (Unesp), Av. Prof. Mário Rubens Guimarães Montenegro, s/n, campus de Botucatu

Made available in DSpace on 2021-06-25T11:07:24Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-11-05 Seroprevalence of Toxoplasma gondii has been extensively studied in a variety of different human populations. However, no study has focused on homeless populations. Accordingly, the present study aimed to assess the seroprevalence of anti-T. gondii antibodies and the risk factors associated in homeless persons from homeless shelter of São Paulo city, southeastern Brazil. In addition, anti-HIV antibodies and associated risk of T. gondii and HIV coinfection have been evaluated. Anti-T. gondii antibodies were detected by indirect fluorescent antibody test. In addition, anti-HIV levels were tested by chemiluminescence enzyme immunoassay, with positive samples confirmed by rapid immunoblot assay. Overall, IgG anti-T. gondii seropositivity was found in 43/120 (35.8%) homeless persons, with endpoint titers varying from 16 to 1,024. The only two pregnant women tested were negative for IgM by chemiluminescence enzyme immunoassay, with normal parturition and clinically healthy newborns in both cases. There were no statistical differences in the risk factors for anti-T. gondii serology (p > 0.05). Anti-HIV seropositivity was found in 2/120 (1.7%) homeless persons, confirmed as HIV-1. One HIV seropositive individual was also sero-reactive to IgG anti-T. gondii, and both were negative to IgM anti-T. gondii. This is the first study that reports the serosurvey of T. gondii in homeless persons worldwide. Despite the limited sample size available in the present study, our findings have shown that the prevalence of anti-T. gondii antibodies in homeless persons herein was lower than the general population, probably due to homeless diet habit of eating mainly processed food intake. No statistical differences were found regarding risk factors for anti-T. gondii exposure in homeless persons. Future studies should be conducted to fully establish risk factors for anti-T. gondii exposure in homeless persons. Department of Veterinary Medicine Graduate College of Veterinary Science Federal University of Paraná Clinical Analysis Laboratory Unit Clinics Hospital Federal University of Paraná Department of Veterinary Hygiene and Public Health São Paulo State University Graduate College of Cellular and Molecular Biology Federal University of Paraná Department of Nursing and Public Health State University of Ponta Grossa AACT Research Consulting LTDA Institute of Biotechnology São Paulo State University (UNESP) Department of Medical Microbiology University of São Paulo Department of Comparative Pathobiology Purdue University Department of Veterinary Medicine Federal University of Paraná Department of Veterinary Hygiene and Public Health São Paulo State University Institute of Biotechnology São Paulo State University (UNESP)

Background: Respiratory conditions are the leading cause of training disruption in racing horses. Molecular approaches to diagnose respiratory viruses have provided an opportunity for early and subclinical pathogen detection, particularly in samples from the upper respiratory tract. Gammaherpesvius (EHV-2 and EHV-5) have variable presentations in horses. However, the infection can be asymptomatic and act as a co-factor for the development of other diseases. In this descriptive observational study, 10 healthy, young horses at regular training in Southern Brazil underwent clinical examination, videoendoscopy of the respiratory system, cytological evaluation of TA (tracheal aspirate) and BALF (bronchoalveolar lavage fluid), along with qPCR, in order to evaluate the presence of EHV-2 and EHV-5 in lower respiratory tract samples and compare with correspondent cytological and endoscopical findings. Results: At least one abnormality per horse during endoscopy examination was observed, including, but not limited to, mucous secretion in the airways and pharyngeal lymphoid hyperplasia. The presence of EHV-2 and EHV-5 was detected by qPCR in three out of ten animals. One horse was positive for EHV-2 alone, one for EHV-5 alone, and one was positive for both viruses. No videoendoscopic finding correlated with each other neither predicts gammaherpesvirus status (positive or negative test). Additionally, there was no relationship between the percentage of cells in both TA and BALF and the probability to test positive for herpesvirus.Conclusions: To the authors’ knowledge, this is the first molecular detection of EHV-2 and 5 in Brazilian Thoroughbred horses. These findings may provide new insights into the epidemiological situation of EHV-2 and 5 in Brazilian athletic young horses, evidencing the importance of the molecular investigation, early detection, and prevention of respiratory diseases.

Rabies is a viral zoonotic disease present in two thirds of all countries, and causes the death of one person every 10 minutes (~70,000 deaths/year). The reservoirs of the Rabies Virus (RABV) are bats and canids, and it has also been found in other animals, including Cerdocyon thous (crab-eating fox) and Pseudalopex vetulus (hoary fox). Here we used Next Generation Sequencing (NGS), phylogenetic, and in vitro/in vivo analyses, to characterized the genome of a new subtype of RABV circulating in foxes of the Northeastern region of Brazil. We verified that although these variants were similar to existing strains from wild canids and domestic canines from Brazil, the samples contained escape mutants, suggesting that it was a heterogeneous virus population. In all, we used several molecular techniques to characterize a new RABV strain circulating in wild-foxes in Northeastern Brazil, and verified still manifested its notorious pathogenic characteristics.

Made available in DSpace on 2020-12-12T01:14:28Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-07-01 Carrier animals are considered key in the transmission cycle of leptospirosis. Although investigations have been carried out on several species, the role of pigs in the epidemiology of the disease is still poorly studied in the semi-arid region. Thus, the objective of this study was to determine the presence of Leptospira spp. in the genitourinary tract of pigs intended for slaughter. Fifty pigs were used: adults and unvaccinated. Samples of the kidney, urine, and vaginal fluid were collected for the molecular detection of Leptospira spp. and blood samples for the serological test. The molecular test was performed using the polymerase chain reaction (PCR), and the serological test was performed with the microscopic agglutination test (MAT). Samples with DNA amplification were submitted to genetic sequencing. Twenty (40%) animals were found with anti-Leptospira spp. antibodies, and the majority of the reactions (50%) occurred for the serogroup Tarassovi. Leptospiral DNA was found in the tissue of 11 (22%) pigs. The gene from a urine sample was sequenced and showed similarity to L. borgpetersenii. The results evidenced a high rate of porcine carriers; therefore, they appear to be important sources of agent infection, being potential transmitters of the disease to other animal species and man. Federal University of Campina Grande (UFCG), Av. Universitária, no number, Santa Cecília Paulista State University (Unesp) Paulista State University (Unesp)

Made available in DSpace on 2020-12-12T02:39:42Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-07-01 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) It is possible that there are peculiarities in the epidemiology of leptospirosis in regions with a semiarid climate, where the environment is often adverse, allowing the occurrence of alternative transmission routes. The objective of the work was to generate contributions to the diagnosis and epidemiology of Leptospira sp. infection in sheep reared in semiarid conditions, using serological, molecular and microbiological techniques for diagnosis in dry and rainy seasons. Samples of blood, vaginal fluid, urine, bladder, kidney, uterus, uterine tube, ovary and placenta were collected from 104 sheep (52 animals per season – dry and rainy) slaughtered in the Brazilian semiarid. Diagnostic tests performed were microscopic agglutination test (MAT), polymerase chain reaction (PCR) and bacterial isolation. Anti-Leptospira sp. antibodies were found in 26 (25%) of the animals analyzed by MAT at 1:50 dilution (cut-off 50), while 69 (66.3%) animals had at least one organ/fluid with the presence of Leptospira sp. DNA. Overall, PCR was performed on 758 fragments of organs/fluids from the genital and urinary tracts, and 519 (68.5%) samples tested positive. PCR-positivity was statistically different in dry (46.2%) and rainy (11.5%) seasons for vaginal fluid. It was possible to perform the DNA sequencing in nine samples with 99% similarity to L. interrogans and recovery of viable strains in three samples of vaginal fluid. Regardless of the biological material used in PCR to detect carrier animals and the season, the highest MAT sensitivity values were obtained with cut-off 50 compared to 100. The results obtained indicate that, even in the adverse environmental conditions of the semiarid region, leptospires may survive and propagate by alternative routes of transmission, such as sexual, and the presence of PCR-positive genital tracts in ewes suggests that sexual transmission may play an important role in the epidemiology of the disease in sheep in Brazilian semiarid. In addition, it is suggested the use of titer 50 as cut-off point at serology in semiarid conditions. Centro de Saúde e Tecnologia Rural (CSTR) Universidade Federal de Campina Grande (UFCG), Av. Universitária, s/n, Santa Cecília Departamento de Ciências Veterinárias (DCV) Universidade Federal da Paraíba (UFPB), PB-079, s/n, Cidade Universitária Universidade Estadual Paulista (Unesp), Av. Prof. Mário Rubens Guimarães Montenegro, s/n, campus de Botucatu Universidade Estadual Paulista (Unesp), Av. Prof. Mário Rubens Guimarães Montenegro, s/n, campus de Botucatu CNPq: 302222/2016-2 CNPq: 423836/2018-8

Made available in DSpace on 2020-12-10T17:40:40Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-03-06 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Coordination for the Improvement of Higher Education Personnel Leptospirosis is a zoonotic disease of global importance caused by an obligate aerobic spirochaete that infects a wide variety of domestic and wild animals. Natural hosts are asymptomatic or show moderate signs of the disease. Accidental hosts develop a severe, often lethal, form of the disease. All young southern tamanduas died suddenly at the zoo in the city of Joao Pessoa, Brazil. The animals were found dead without any noticeable clinical signs. Necropsy revealed extensive haemorrhage in the subcutaneous tissues, kidneys, lungs in addition to the presence of red fluid in the thoracic, abdominal and pericardial cavities. Histopathology of kidneys exhibited acute interstitial nephritis and tubular necrosis. Immunohistochemical staining revealed typical leptospiral wavy forms and aggregates in the lumen of several kidney tubules and lungs. Pathological and molecular investigations confirmedLeptospira interrogansinfection. The adult tamanduas did not present with clinical alterations. To our knowledge, this investigation is the first study to report that leptospirosis should be considered as a possible cause of death in tamanduas. This article warns of the risks of anthropization with respect to Leptospira transmission to tamanduas, other animals and humans. Univ Fed Paraiba, Areia, Paraiba, Brazil Univ Fed Campina Grande, Patos de Minas, Paraiba, Brazil Univ Estadual Paulista, Botucatu, SP, Brazil Univ Fed Santa Maria, Santa Maria, RS, Brazil Parque Zoobot Arruda Camara, Joao Pessoa, Paraiba, Brazil Univ Estadual Paulista, Botucatu, SP, Brazil CNPq: 429862/2016-4

Made available in DSpace on 2021-06-25T10:18:29Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-01-01. Added 1 bitstream(s) on 2021-07-15T15:04:36Z : No. of bitstreams: 1 S0103-84782020001200454.pdf: 850016 bytes, checksum: 77e5fcaa5c1275c720ca5b83a4e453a6 (MD5) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) We used 12 tegu lizards (Tupinambis merianae) from northeastern Brazil, and we reported nine (75%) Leptospira sp. PCR-positive animals and six (50%) seropositive. Leptospira sp. DNA sequencing revealed 99% similarity with L. interrogans. Our findings indicated that this species may play a role in the transmission of human leptospirosis. Universidade Federal de Campina Grande (UFCG) Universidade Federal da Paraíba (UFPB) Universidade Estadual Paulista (Unesp) Universidade Estadual Paulista (Unesp) CNPq: 404039/2016-2

Made available in DSpace on 2018-12-11T16:54:21Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-07-09 Although some studies in sheep have indicated leptospire colonization of the genital tract, further studies are needed to clarify the role of genital carriers in this species. Thus, this study aimed to evaluate the colonization of pathogenic leptospires in the genital and urinary tract of slaughtered sheep. Fifty-seven adult, female woolless sheep destined for slaughter were used. Renal (n = 57), bladder (n = 57), ovary (n = 34), uterine tube (n = 44), and uterus (n = 33) samples were collected for molecular detection of Leptospira sp. DNA, and blood samples (n = 57) for serological testing. The molecular testing was performed using polymerase chain reaction (PCR), and the serological testing was performed using microscopic serum agglutination test (MAT). Samples with amplifying DNA were subjected to genetic sequencing. In total, leptospiral DNA was found in the tissues of 44 (77.2%) sheep, whereas only nine animals were positive on both PCR and MAT; there was slight agreement between PCR and MAT techniques (k = 0.0268; p = 0.684). In 61 (54.9%) genital tract and in five (4.4%) urinary tract samples, the leptospiral DNA was detected, with significant difference (p < 0.001). The genes of one sample from the uterine tube and another from the bladder were sequenced and demonstrated 99% similarity to Leptospira interrogans. Anti-Leptospira antibodies were detected in 11 (19.3%) of the tested animals. The results reinforce the importance of the genital tract as an extra-renal site of colonization, suggesting the possibility of venereal transmission in sheep. Universidade Federal de Campina Grande (UFCG), Av. Universitária, s/n, Santa Cecília Universidade Estadual Paulista (Unesp) Instituto de Biotecnologia Universidade Estadual Paulista (Unesp) Instituto de Biotecnologia

Made available in DSpace on 2018-12-11T17:37:41Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-07-03 The aim of the present study was to describe the strategies of the control of an outbreak of leptospiral infection in dairy cattle in Maranhão State, Northeastern Brazil. In the period from January to July 2015, 18 (17%) out of 106 cows presented abortion, six (5.7%) stillbirth, and 12 (11.3%) repeated estrus, totaling 24 animals with reproductive problems. The diagnosis of leptospirosis was based on serology (microscopic agglutination test—MAT), bacteriological culture, and polymerase chain reaction (PCR). Antibiotic therapy, vaccination protocols, and changes in management practices were suggested as control measures. Of all animals on the farm (n = 280), 136 (48.6%) were seropositive for at least one serovar of Leptospira sp. No pure leptospiral culture was obtained. Eight of the animals with reproductive problems yielded positive PCR results (vaginal fluid of seven animals and urine and vaginal fluid of one animal). Genetic sequencing of a vaginal fluid/urine PCR-positive sample revealed Leptospira borgpetersenii. One year after the adoption of control measures, no reproductive problems were observed. Thus, leptospirosis probably caused the reproductive failures in the herd, and the control and prevention measures implemented were efficient in controlling the disease. Centro de Saúde e Tecnologia Rural (CSTR) Universidade Federal de Campina Grande (UFCG), Av. Universitária, s/n, Santa Cecília Centro de Ciências Agrárias (CCA) Universidade Federal da Paraíba (UFPB), Rodovia BR 079 - Km 12 Universidade Estadual Paulista (Unesp), Av. Prof. Mário Rubens Guimarães Montenegro, s/n, Campus de Botucatu Universidade Estadual Paulista (Unesp), Av. Prof. Mário Rubens Guimarães Montenegro, s/n, Campus de Botucatu

Made available in DSpace on 2022-04-29T08:37:21Z (GMT). No. of bitstreams: 0 Previous issue date: 2022-02-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) COVID-19 has posed a worldwide public health challenge affecting millions of people in different countries. Rapid and efficient detection of SARS-CoV-2 is essential for pandemic control. Reverse Transcription quantitative PCR (RT-qPCR) of nasopharyngeal swabs is the gold standard method for the virus detection, but the high demand for tests has substantially increased the costs and reduced the availability of reagents, including genetic material purification kits. Thus, the present study aimed to compare two bead-based RNA extraction methods (an in-house and a commercial kit) from nasopharyngeal swabs and RT-qPCR detection of SARS-CoV-2. Twenty-five positive and five negative nasopharyngeal swab samples were subjected to extraction of nucleic acids using both methods in an automated platform. Both protocols revealed a high correlation between Cycle Quantifications (Cqs) (r = 0.99, p < 0.0001). In addition, the in-house kit was 89.5 % cheaper when compared to the mean cost of commercial RNA extraction kits. The results show that the in-house protocol is an affordable and reliable option for RNA extraction for SARS-CoV-2 detection from nasopharyngeal swabs. São Paulo State University (UNESP) Institute of Biotechnology (IBTEC) São Paulo State University (UNESP) Department of Animal Production and Preventive Veterinary Medicine School of Veterinary Medicine and Animal Science São Paulo State University (UNESP) Department of Veterinary Clinics School of Veterinary Medicine and Animal Science São Paulo State University (UNESP) Institute of Biotechnology (IBTEC) São Paulo State University (UNESP) Department of Animal Production and Preventive Veterinary Medicine School of Veterinary Medicine and Animal Science São Paulo State University (UNESP) Department of Veterinary Clinics School of Veterinary Medicine and Animal Science FAPESP: 19/18581-6

Made available in DSpace on 2021-06-25T10:29:21Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-10-01 Food safety is essential to the development of a society, and foodborne diseases affect thousands of people worldwide every year. Among the main foodborne disease agents, Salmonella stands out as the most often associated with hospitalizations and deaths. Pork products are an important source of nutrients for the population, considered the most consumed meat in the world. Several reports revealed the presence of Multidrug-Resistant (MDR) Salmonella strains along the pork production chain, and the occurrence of elements that allow for the transference of resistance genes between bacterial populations is a public health concern. The present work aimed to evaluate the resistance genes and integrons in MDR Salmonella strains isolated from swine mesenteric lymph nodes. Nine strains had the genome sequenced with the Illumina NextSeq system. The results were submitted to the Resfinder server for evaluation of acquired genes and point mutations related to AMR and the presence and sequences of integrons were determined with a BLAST search. Different resistance genes for the same antimicrobial class were present in each strain. In two samples, resistance genes for olaquindox, a growth promoter outlawed in Brazil since 2004, were found. All observed phenotypic resistance had a genetic basis. Five isolates harbored class 1 integrons, which were classified in three distinct Integron Profiles (IPs). The IPs were also reported in animal source food and human clinical isolates. The diversity of resistance genes for the same antimicrobial class in each strain and the presence of Class 1 Integrons, reflect the high selection pressure that these bacterial lineages face along the pork production chain, and reinforces the role of animal production as a reservoir and disseminator of AMR genes. São Paulo State University (Unesp) Institute for Biotechnology, Tecomarias Av. São Paulo State University (Unesp) Institute for Biotechnology, Tecomarias Av.

Made available in DSpace on 2022-05-01T06:02:16Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-07-01 The rapid development of efficacious and safe vaccines against coronavirus disease 2019 (COVID-19) has been instrumental in mitigating the transmission of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Moreover, the emergence of SARS-CoV-2 variants raised concerns on the efficacy of these vaccines. Herein, we report two cases of breakthrough infections with the P1 variant in patients vaccinated with CoronaVac, which is one of the two vaccines authorized for emergency use in the Brazilian immunization program. Our observations suggest that the vaccine reduced the severity of the disease and highlight the potential risk of illness following vaccination and subsequent infection with the P1 variant as well as for continued efforts to prevent and diagnose infection in vaccinated persons. Laboratório de Pesquisas em Virologia (LPV) Medicine School of São José do Rio Preto (FAMERP) Hospital de Base Biotecnology Institute São Paulo State University (UNESP) Intitute of Biosciences Languages and Exact Sciences São Paulo State University (UNESP) Department of Veterinary Medicine FZEA-USP University of São Paulo (USP) Department of Pathology University of Texas Medical Branch, 301 University Blvd Sealy Center for Vector-Borne and Zoonotic Diseases University of Texas Medical Branch, 301 University Blvd Center for Biodefense and Emerging Infectious Diseases University of Texas Medical Branch, 301 University Blvd Center for Tropical Diseases University of Texas Medical Branch, 301 University Blvd Institute for Human Infection and Immunity University of Texas Medical Branch, 301 University Blvd Biotecnology Institute São Paulo State University (UNESP) Intitute of Biosciences Languages and Exact Sciences São Paulo State University (UNESP)

Leptospirosis is a bacterial zoonoses distributed worldwide. It can undertake any animalspecies, maintaining the agent in the nature via renal chronical infection of carriers animals,which can secrete the organism through the urine into the environment, being able to infectother animals and the human. Regarding Leptospira spp., diagnostic techniques have helpedunderstand important eco-epidemiological aspects such as environmental serovar distributionand new hosts, in addition to improve the clinical diagnosis of the disease. This review focuseson present diverse techniques used in leptospirosis diagnosis to biological samples, includingsome studies on practical applications for the molecular detection of Leptospira spp.,reinforcing the importance of the knowledge and the choice of the correct technique in thediagnosis of the disease.

Porcine virus type 2 (PCV2) is an emergent virus found in commercial pig farms and may cause clinical or subclinical infection. Wild pigs such as collared peccary (Tayassu tajacu) and white-lipped peccary (Tayassu pecari) may also be infected by PCV2. Accordingly, the aim of this study was to molecularly identify PCV2 in whole blood samples of captive peccaries (16 collared and 6 white-lipped) by conventional and quantitative PCR (qPCR). Although the housekeeping gene (?-actin) DNA was successfully amplified, all 22 peccaries tested negative for PCV2 by both molecular methods. Previous studies have shown no antibodies against PCV2 in 49 free-range collared peccaries of Southern Brazil but PCV2 DNA was detected by PCR in tissue samples of 9/10 (90.0%) collared and 3/3 (100%) white-lipped free-ranging peccaries from Central and Northern Brazil. In conclusion, although PCV2 may be endemic in free ranging wild pigs of Central and Northern Brazil, lack of serological and molecular PCV2 evidence (in whole blood) of both captive and free-range wild pigs may indicate low risk of disease in Southern Brazil.

Multiple pathogens may cause and sustain respiratory infection in horses, either alone or as co-infection, leading to disruption and interruption of training, racing and other competitions. Molecular approaches have successfully provided opportunity for pathogen involvement and characterization, particularly from samples of horse upper respiratory tract. Accordingly, the aim of this study was to molecularly assess bronchoalveolar lavage fluid (BALF) samples from 10 healthy training horses at the Jockey Club of Parana State, southern Brazil. Virus-specific real-time polymerase chain reaction (qPCR) assays were applied as panel for concomitant detection of Equine Adenovirus, Equine Influenza Virus, Equine Herpesvirus Type 1 (EHV-1), Type 2 (EHV-2), Type 4 (EHV-4), and Type 5 (EHV-5), Equine Rhinitis A Virus, Equine Rhinitis B Virus, and Streptococcus equi subsp. equi. Amplicons from positive samples were amplified and sequenced to confirm the virus identification. Results have confirmed the first molecular detection of EHV-2 and EHV-5 in healthy training horses of Brazil, with an overall 3/10 (30%) EHV positivity. Two horses were positive for at least one EHV, and one was positive for both EHV-2 and EHV-5. No horse presented clinical signs of respiratory disease at the time or following samplings. These findings may provide new insights on the epidemiological situation of EHV-2 and EHV-5 in Brazil, highlighting the importance of molecular detection and active surveillance in clinically healthy horses, which may act as spreading infection source for younger equines.

Made available in DSpace on 2020-12-12T01:11:17Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-04-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Hemotropic mycoplasmas (hemoplasmas) are bacteria distributed worldwide and affect domestic and wildlife animals and human beings. Hemoplasmas have been described infecting hematophagous and non-hematophagous bats; however, transmission risk and zoonotic potential in vampire bats remain to be fully established. This study aimed to evaluate the presence of hemotropic mycoplasma species in free-ranging bats from this area using a universal PCR protocol for hemoplasmas. Accordingly, ten blood samples were collected from six male common vampire bats (Desmodus rotundus), two male hairy-legged vampire bats (Diphylla ecaudata), and two female non-hematophagous Pallas's mastiff bats (Molossus sp.) from the Curitiba's region, Paraná State, Southern Brazil. A total of eight (8/10) blood samples were positive byconventional PCR; five (5/6) Desmodus rotundus, two (2/2) Diphylla ecaudata, and one (1/2) Molossus sp. bats. The analyses of the partial sequence of the 16S rDNA gene suggest that the hemoplasma detected in Desmodus rotundus in South Brazil has a high identity compared to the hemoplasma circulating in vampire bats from Central and South America. Engineering and Exact Sciences Center Universidade Estadual do Oeste do Paraná Department of Veterinary Medicine Universidade Federal do Paraná, R. dos Funcionários, 1540 Department of Preventive Veterinary Medicine Universidade Estadual de Londrina Department of Cell and Molecular Biology Universidade Federal do Paraná, Av. Cel. Francisco H. dos Santos, s/n Department of Comparative Pathobiology College of Veterinary Medicine Purdue University Biotechnology Institute Universidade Estadual Paulista (UNESP), Al. das Tecomarias, s/n, Chácara Capão Bonito Biotechnology Institute Universidade Estadual Paulista (UNESP), Al. das Tecomarias, s/n, Chácara Capão Bonito FAPESP: 2014/23159-8

Leptospirosis has been investigated in several species of wild animals. The white-eared opossum (Didelphis albiventris) is a mammal common in the brazilian semi-arid, so, this study aimed to investigate its role in the occurrence of the leptospirosis in the region Northeast of Brazil. 12 animals were used, from which samples were collected for the attempt of isolation, molecular detection and serological examination. There was no microbial growth, nor were any anti-Leptospira sp. antibodies found in the serological samples. The PCR detected leptospiric DNA in the central nervous system (CNS) of five animals (41.7 %). The gene in one of the samples was sequenced and showed identity with Leptospira interrogans. The presence of Leptospira sp. in the CNS of Didelphis albiventris does not allow the characterization of the studied animals as reservoirs with potential for transmission of the pathogen in the region, however it represents a site that needs to be further investigated.

Made available in DSpace on 2018-12-11T16:53:47Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-06-01 Background: Bat rabies surveillance data and risk factors for rabies spillover without human cases have been evaluated in Curitiba, the ninth biggest city in Brazil, during a 6-year period (2010-2015). A retrospective analysis of bat complaints, bat species identification and rabies testing of bats, dogs and cats has been performed using methodologies of seasonal decomposition, spatial distribution and kernel density analysis. Results: Overall, a total of 1003 requests for bat removal have been attended to, and 806 bats were collected in 606 city locations. Bat species were identified among 13 genera of three families, with a higher frequency of Nyctinomops in the central-northern region and Molossidae scattered throughout city limits. Out of the bats captured alive, 419/806 (52.0%) healthy bats were released due to absence of human or animal contacts. The remaining 387/806 (48.0%) bats were sent for euthanasia and rabies testing, which resulted in 9/387 (2.32%) positives. Linear regression has shown an increase on sample numbers tested over time (regression: y = 2.02 + 0.17×; p < 0.001 and r2 = 0.29), as well as significant seasonal variation, which increases in January and decreases in May, June and July. The Kernel density analysis showed the center-northern city area to be statistically important, and the southern region had no tested samples within the period. In addition, a total of 4769 random and suspicious samples were sent for rabies diagnosis including those from dogs, cats, bats and others from 2007 to 2015. While all 2676 dog brains tested negative, only 1/1136 (0.088%) cat brains tested positive for rabies. Conclusion: Only non-hematophagous bats were collected during the study, and the highest frequency of collections occurred in the center-northern region of the city. Rabies spillover from bats to cats may be more likely due to the registered exposure associated with cats' innate hunting habits, predisposing them to even closer contact with potentially infected bats. Although associated with a very low frequency of rabies, cats should always be included in rabies surveillance and vaccination programs. Federal University of Parana Graduate Program in Cellular and Molecular Biology City Secretary of Health Zoonoses Control Center University of São Paulo Department of Preventive Veterinary Medicine and Animal Health Institute of Biotechnology Botucatu São Paulo UNESP - Univ. Estadual Paulista Campus de Botucatu Federal University of Paraná Department of Veterinary Medicine, Rua dos Funcionários, 1540 Institute of Biotechnology Botucatu São Paulo UNESP - Univ. Estadual Paulista Campus de Botucatu

Made available in DSpace on 2019-10-06T16:02:52Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-12-01 National Institutes of Health In Africa, Old World Primates are involved in the maintenance of sylvatic circulation of ZIKV. However, in Brazil, the hosts for the sylvatic cycle remain unknown. We hypothesized that free-living NHPs might play a role in urban/periurban ZIKV dynamics, thus we undertook an NHP ZIKV investigation in two cities in Brazil. We identified ZIKV-positive NHPs and sequences obtained were phylogenetically related to the American lineage of ZIKV. Additionally, we inoculated four C. penicillata with ZIKV and our results demonstrated that marmosets had a sustained viremia. The natural and experimental infection of NHPs with ZIKV, support the hypothesis that NHPs may be a vertebrate host in the maintainance of ZIKV transmission/circulation in urban tropical settings. Further studies are needed to understand the role they may play in maintaining the urban cycle of the ZIKV and how they may be a conduit in establishing an enzootic transmission cycle in tropical Latin America. São José do Rio Preto School of Medicine (FAMERP) Avenida Brigadeiro Faria Lima 5416 CEP: 15090-000, Vila São Pedro Laboratório de Vírus - Universidade Federal de Minas Gerais (UFMG) Avenida Antônio Carlos 6627 CEP: 31270-901 Pampulha Center for Drug Research and Development Instituto de Ciências Biológicas Universidade Federal de Minas Gerais (UFMG) Avenida Antônio Carlos 6627 CEP: 31270-901 Pampulha Instituto Adolfo Lutz (IAL) Avenida Dr. Arnaldo 351 − 7 Andar Sala 706 CEP: 01246-000 Pacaembú Laboratory of Wildlife Comparative Pathology Department of Pathology School of Veterinary Medicine and Animal Sciences University of São Paulo (LAPOCM-FMVZ-USP) Avenida Orlando Marques de Paiva 87 CEP: 05508-270 Department of Biology Institute of Biosciences Letters and Exact Sciences – São Paulo State University São José do Rio Preto – (IBILCE/UNESP), Rua Cristóvão Colombo, 2265, CEP: 15054-000 Epidemiological Surveillance Departament of São José do Rio Preto Avenida Romeu Strazzi 199 CEP: 15084-010 Vila Sinibaldi São Paulo State University (Unesp) Institute for Biotechnology Alameda das Tecomarias s/n CEP: 18607-440 Chácara Capão Bonito Department of Biochemistry and Molecular Biology University of Texas Medical Branch 301 University Blvd Centro de Controle de Zoonoses Belo Horizonte Council Rua Édna Quintel 173 CEP: 31270-705 São Bernardo Faceres Medical School Avenida Anísio Haddad 6751 CEP: 15090-305 Jardim Francisco Fernandes Department of Epidemiology School of Public Health of the University of São Paulo Avenida Dr. Arnaldo 715 CEP: 01246-904 Department of Pathology and Center of Biodefense and Emerging Infectious Diseases Center for Tropical Diseases Institute for Human Infections and Immunity University of Texas Medical Branch 301 University Blvd Department of Biology Institute of Biosciences Letters and Exact Sciences – São Paulo State University São José do Rio Preto – (IBILCE/UNESP), Rua Cristóvão Colombo, 2265, CEP: 15054-000 São Paulo State University (Unesp) Institute for Biotechnology Alameda das Tecomarias s/n CEP: 18607-440 Chácara Capão Bonito National Institutes of Health: R24AI120942

Madariaga virus (MADV), the new species designation for the South American isolates of eastern equine encephalitis virus (EEEV), is genetically divergent and substantially different in ecology and pathogenesis from North American EEEV strains. We isolated and characterized a MADV isolate obtained from a horse in Brazil. Our results support previous phylogenetic studies showing there are three genetically distinct MADV lineages. The MADV isolate from Paraíba State belongs to the South American lineage III and is closely related to Peruvian, Colombian and Venezuelan isolates.

Metagenomics was used to characterize viral genomes in clinical specimens of horses with various organ-specific diseases of unknown aetiology. A novel parvovirus as well as a previously described hepacivirus closely related to human hepatitis C virus and equid herpesvirus 2 were identified in the cerebrospinal fluid of horses with neurological signs. Four co-infecting picobirnaviruses, including an unusual genome with fused RNA segments, and a divergent anellovirus were found in the plasma of two febrile horses. A novel cyclovirus genome was characterized from the nasal secretion of another febrile animal. Lastly, a small circular DNA genome with a Rep gene, from a virus we called kirkovirus, was identified in the liver and spleen of a horse with fatal idiopathic hepatopathy. This study expands the number of viruses found in horses, and characterizes their genomes to assist future epidemiological studies of their transmission and potential association with various equine diseases.

St. Louis encephalitis virus (SLEV), a member of the family Flaviviridae, genus Flavivirus, is a causative agent of encephalitis in the Americas. In Brazil, sporadic cases of SLEV infection have been reported since 1953, but the first outbreak of SLEV in Brazil was identified only in 2007, concomitant with an outbreak of dengue virus (DENV) serotype 3. This finding, along with other reports, indicates that SLEV circulation in Brazil is largely unknown, and there may be epidemiological implications of the co-circulation of SLEV, DENV and other flaviviruses in Brazil. Here, we describe the first complete genome sequence of an SLEV strain isolated from a human patient in Brazil, strain BeH 355964. Phylogenetic analysis was performed to determine the genotype of BeH 355964 using the full-length genome and envelope (E) gene sequences separately. Both analyses showed that BeH 355964 could be classified as genotype V. Although the number of single gene sequences available is greater (such as for the E gene), the phylogenetic tree based on the complete genome sequence was better supported and provided further information about the virus.

Microelectrode arrays (MEAs) are state-of-the-art devices for extracellular recording and stimulation on biological tissue. Furthermore, they are a relevant tool for the development of biomedical applications like retina, cochlear and motor prostheses, cardiac pacemakers and drug screening. Hence, research on functional cell-sensor interfaces, as well as the development of new surface structures and modifications for improved electrode characteristics, is a vivid and well established field. However, combining single-cell resolution with sufficient signal coupling remains challenging due to poor cell-electrode sealing. Furthermore, electrodes with diameters below 20 µm often suffer from a high electrical impedance affecting the noise during voltage recordings. In this study, we report on a nanocavity sensor array for voltage-controlled stimulation and extracellular action potential recordings on cellular networks. Nanocavity devices combine the advantages of low-impedance electrodes with small cell-chip interfaces, preserving a high spatial resolution for recording and stimulation. A reservoir between opening aperture and electrode is provided, allowing the cell to access the structure for a tight cell-sensor sealing. We present the well-controlled fabrication process and the effect of cavity formation and electrode patterning on the sensor's impedance. Further, we demonstrate reliable voltage-controlled stimulation using nanostructured cavity devices by capturing the pacemaker of an HL-1 cell network.

RESUMO Doenças infecciosas são as maiores responsáveis por falhas reprodutivas (FR) em cadelas, causando aborto, morte fetal e natimortalidade. Este estudo teve como objetivo investigar a associação entre agentes infecciosos, FR inexplicáveis e anemia em cadelas. Todas as amostras maternas e fetais foram negativas para a presença dos principais agentes infecciosos causadores de FR: herpes vírus canino 1, Neospora caninum, Brucella spp. e B. canis, enquanto agentes como o de Leishmania spp., parvovírus canino, Ehrlichia canis e Anaplasma platys foram encontrados em sangue materno. Coinfecções de A. platys/E. canis e A. platys/Leishmania spp. foram diagnosticadas. Os resultados indicam que os animais com anemia causadas por doenças transmitidas por vetores podem ser mais suscetíveis a sofrerem FR do que animais com valores hematológicos normais.

Made available in DSpace on 2018-12-11T17:33:44Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-08-01 The Hobi-like virus presents antigenic and molecular differences in relation to bovine virus diarrhea virus 1 and 2. The description of the complete genome of the Hobi-like virus SV757/15, isolated from a Nelore cow with gastroenteric disease in Brazil, will help in understanding the evolution and diversity of pestiviruses. Universidade Santo Amaro–UNISA Curso de Medicina Veterinária Universidade Estadual Paulista “Júlio de Mesquita Filho”–UNESP Instituto de Biociências Departamentde Medicina Veterinária Preventiva Universidade Federal de Santa Maria UFSM Faculdade de Medicina Veterinária e Zootecnia Universidade Estadual Paulista “Júlio de Mesquita Filho”– UNESP Faculdade de Medicina Veterinária e Zootecnia Universidade de São Paulo– USP Universidade Estadual Paulista “Júlio de Mesquita Filho”–UNESP Instituto de Biociências Faculdade de Medicina Veterinária e Zootecnia Universidade Estadual Paulista “Júlio de Mesquita Filho”– UNESP

Made available in DSpace on 2014-09-30T18:18:25Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-02-01Bitstream added on 2014-10-01T14:03:36Z : No. of bitstreams: 1 S0102-09352014000100001.pdf: 134074 bytes, checksum: 0d75227daf3dfeeb20a54373c2b4f89d (MD5) No Brasil existem poucos estudos sobre a ocorrência da infecção pelo vírus da imunodeficiência felina (FIV), assim como a determinação dos subtipos circulantes, o que é indispensável para o desenvolvimento de vacinas e novos testes diagnósticos. O presente trabalho investigou a ocorrência da infecção pelo FIV entre os anos de 2010 e 2011 em gatos domésticos submetidos a atendimento clínico na cidade de Pelotas e região. Amostras de sangue total de 70 animais, incluindo suspeitos (28) ou não suspeitos (42) da infecção pelo FIV, foram submetidas à reação de PCR nested. Os resultados indicaram uma frequência de infecção de 15,7% (11/70) e a análise dos fatores associados (sexo, idade e condição clínica) evidenciou uma maior ocorrência em gatos com idade superior a 10 anos e acometidos por infecções crônicas e recidivantes. Oito amostras positivas na PCR nested foram submetidas a sequenciamento genômico e somente o subtipo B foi detectado na região estudada. In Brazil there are few studies on the occurrence of the feline immunodeficiency virus (FIV) infection and its subtypes, which are essential for the development of vaccines and new diagnostic tests. The present study investigated the occurrence of the FIV infection between 2010 and 2011 in domestic cats submitted to medical attendance in the city of Pelotas and nearby area. Total blood samples of seventy cats, suspected (28) or not (42) of infection by FIV were analyzed by nested PCR in order to perform a diagnosis. The results pointed to a FIV infection frequency of 15.7% (11/70) and the analysis of the risk factors related to infection (sex, age and clinical condition) evidenced a greater occurrence in cats up to 10 years of age with chronic and recurrent infections. Eight samples found positive by nested PCR were submitted to DNA sequencing indicating that only the subtype B was detected in the studied region. Universidade Federal de Pelotas Faculdade de Veterinária Universidade Estadual Paulista Instituto de Biociências Universidade Estadual Paulista Instituto de Biociências