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Central nervous system (CNS) infections comprise life-threatening clinical conditions in domestic species, and are commonly related to severe sequelae, disability, or high fatality rates. A set of bacterial pathogens have been identified in central nervous infections in livestock and companion animals, although the most of descriptions are restricted to case reports and a lack of comprehensive studies involving CNS-related bacterial infections have been focused on a great number of domestic species. In this scenario, we retrospectively investigated selected epidemiological data, clinical findings, bacteriological culture, and in vitro susceptibility patterns of 136 nonrepetitive neurologic cases in domestic species (2005-2021). Bacterial isolates were recovered from 25% (34/136) of the cerebrospinal fluid (CSF) sampled. The isolates were obtained from cattle (9/136 = 6.6%), dogs (7/136 = 5.1%), horses (6/136 = 4.4%), goats (3/136 = 2.2%), pigs (3/136 = 2.2%), sheep (3/136 = 2.2%), cats (2/136 = 1.5%), and asinine (1/136 = 0.7%). Among animals with bacterial isolation, Staphylococcus aureus (6/34 = 17.6%), Escherichia coli (5/34 = 14.7%), Staphylococcus beta-hemolytic (5/34 = 14.7%), and Trueperella pyogenes (3/34 = 8.8%) were predominant, in addition to a miscellaneous of other bacteria isolated in minor frequency, e.g., Corynebacterium pseudotuberculosis, Enterobacter cloacae, Mannheimia haemolytica, Pseudomonas aeruginosa, and Streptococcus equi subsp. equi. In vitro susceptibility tests of isolates revealed that amoxicillin/clavulanic acid (11/13 = 84.6%), cephalexin (9/11 = 81.8%), and florfenicol (9/12 = 75%) were the most effective antimicrobials. Conversely, isolates exhibited resistance mainly to tetracycline (6/10 = 60%), penicillin (6/11 = 54.5%), and trimethoprim/sulfamethoxazole (5/11 = 45.5%). Also, multidrug resistance to ≥ 3 classes of antimicrobials was found in 23.5% (8/34) strains. Data relative to the outcome was available in 79.4% (27/34) of animals that had bacterial isolation, and from these, the lethality rate was 92.6% (25/27). Incoordination (14/34 = 41.2%), recumbency (11/34 = 32.4%), apathy (10/34 = 29.4%), anorexia (9/34 = 26.5%), blindness (7/34 = 20.6%), seizure (6/34 = 17.6%), limb paresis (5/34 = 14.7%), head-pressing (4/34 = 11.8%), and nystagmus (3/34 = 8.8%) were the most frequent clinical signs. A variety of bacterial pathogens were identified in the CSF of domestic species showing neurologic signs, with a predominance of staphylococci, streptococci, and enterobacteria. High lethality of cases, poor in vitro efficacy of conventional antimicrobials, and a high in vitro multidrug resistance pattern of isolates were seen. Our results contribute to etiological characterization, antimicrobial resistance patterns, and clinical-epidemiological findings of bacterial infections in domestic species with neurological signs.

Made available in DSpace on 2022-05-01T08:44:43Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-01-01 The present study investigated the serum microscopic agglutination test (MAT) among 203 bovine bulls with reproduction by natural means, without apparent signs of orchitis or inflammation of accessory reproductive glands. Simultaneously, the semen of all bulls was subjected to sperm viability analysis and PCR based on the 16S rRNA gene. PCR-positive results of semen samples were confirmed by sequencing. A modified seminal plasma agglutination (MSPA) test, replacing the blood serum of all bulls in the MAT with seminal plasma was performed as well. Eight (8/203 = 3.9%) semen samples from bulls were considered nonviable (necrospermia and azoospermia) without relation to the PCR diagnosis. No agglutinin titers were identified in MSPA test. A high frequency (132/203 = 65%) of leptospiral agglutinin titers was identified in the MAT, particularly for the Sejroe serogroup (Hardjo CTG, 100/203 = 49.3%; Wolffi 74/203 = 36.4%; Guaricura 72/203 = 35.5%; and Hardjoprajitno 56/203 = 27.6%). Three (3/203 = 1.5%) semen samples of bulls were positive in the PCR, but these results were not confirmed by sequencing. The high frequency of serovars from the Sejroe serogroup typically adapted to bovines indicates the need for measures for the prophylaxis/control of the pathogen on the sampled farms. Discrepancies among the MAT, sperm viability, and molecular detection of leptospires in semen highlight the need for a combination of methods to diagnose leptospirosis in bovine bulls. To our knowledge, modified seminal plasma agglutination is described for the first time here to investigate anti-Leptospira antibodies produced locally in the genital tract in the diagnosis of bovine leptospirosis among bulls that reproduce by natural means. Department of Animal Production and Preventive Veterinary Medicine School of Veterinary Medicine an Animal Science São Paulo State University - UNESP Santo Amaro University - UNISA Faculty of Veterinary Medicine University of Oeste Paulista Technology Faculty - FATEC APTA Beef Cattle Center Institute of Animal Science University of São Paulo - USP Department of Animal Production and Preventive Veterinary Medicine School of Veterinary Medicine an Animal Science São Paulo State University - UNESP

Made available in DSpace on 2022-04-29T08:31:35Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-01-01 We evaluated the use of MALDI-TOF MS for the identification of 3 major, dairy-associated Prototheca species, namely, Prototheca bovis (formerly P. zopfii genotype 2), P. blaschkeae, and P. ciferrii (formerly P. zopfii genotype 1). The MALDI-TOF MS spectra established for those species were introduced into the reference spectra library of the Bruker Biotyper MALDI-TOF MS analysis software. Next, 31 Prototheca isolates from Holstein cows with mastitis, from herds located in the midwestern area of São Paulo State, Brazil, were subjected to MALDI-TOF MS profiling. MALDI-TOF MS allowed identification of 22 of 27 P. bovis and 3 of 4 P. blaschkeae isolates with scores >2.0, with 5 of 27 P. bovis and 1 of 4 P. blaschkeae isolates identified only to the genus level. With our extended algae database, MALDI-TOF MS can contribute to quick and effective speciation of Prototheca from mastitis cases. Department of Animal Nutrition and Production School of Veterinary Medicine and Animal Science University of São Paulo-USP Department of Medical Microbiology Institute of Microbiology Faculty of Biology University of Warsaw Department of Animal Production and Preventive Veterinary Medicine School of Veterinary Medicine and Animal Sciences São Paulo State University-UNESP Department of Animal Production and Preventive Veterinary Medicine School of Veterinary Medicine and Animal Sciences São Paulo State University-UNESP

Caseous lymphadenitis is a well-known disease caused by Corynebacterium pseudotuberculosis affecting small ruminants with small significance to human health because of its minor zoonotic potential. In both cases, few treatment options are available and conventional antimicrobial therapy is commonly refractory due to development of pyogranulomatous reactions, bringing great interest in discovering novel therapeutics for more suitable approaches. Dideoxynucleotides presented antibacterial action against various bacteria but were never described for C. pseudotuberculosis. Hypothesizing the antimicrobial action of 2',3'-dideoxiadenosine (ddATP) against C. pseudotuberculosis, we performed for the first time an investigation of its minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) in the ATCC® 19,410 strain and a well-characterized clinical isolate of C. pseudotuberculosis. We also assessed potential synergism with penicillin. ddATP showed a growth delay effect for C. pseudotuberculosis at 2 µmol/mL and a MIC and MBC of 4 µmol/mL against the ATCC® 19,410 strain, but not for the clinical strain. An antimicrobial effect was observed when using concentrations lower than the MIC of ddATP associated with penicillin for both strains tested. Our data suggest the potential of nucleotide analogs, especially adenosine, and its combination with penicillin, as a possible novel treatment for C. pseudotuberculosis-induced infections, and contributes with knowledge regarding alternative drugs to treat C. pseudotuberculosis infections.

Made available in DSpace on 2022-04-28T19:44:39Z (GMT). No. of bitstreams: 0 Previous issue date: 2022-03-01 Brucellosis is a zoonotic disease with a global impact. Brucellasuis is one of the most pathogenic species to humans, requiring different measures for the control and/or eradication of the disease. The serological investigation for brucellosis was performed in pigs, horses, dogs, and cattle on a farm with a history of abortion in sows and necropsy of a boar with severe necrosuppurative orchitis. One sow, two cows, and two dogs reveled positive to Rose Bengal Test (RBT), although only the sow had a confirmatory outcome in 2-mercaptoethanol (2-ME). The 2-ME-positive sow was euthanized and microbiological culture of lymph nodes and liver followed by biochemical characterization allowed phenotypic characterization of Brucellasuis biotype 1. PCR multiplex Bruce-ladder and Suis-ladder enabled molecular confirmation, respectively, of Brucellasuis and biotype 1. The transmission aspects of B.suis to pigs and other domestic species, the combination of diagnostic procedures to diagnosis, as well as human health concerns of brucellosis are discussed. Department of Animal Production and Preventive Veterinary Medicine Faculty of Veterinary Medicine and Zootechnics State University of Sao Paulo, SP Department of Veterinary Clinical Science Federal University of Jataí, GO

Made available in DSpace on 2022-05-01T12:09:41Z (GMT). No. of bitstreams: 0 Previous issue date: 2022-03-01 Klebsiella species, particularly K. pneumoniae, are well-known opportunistic enterobacteria related to complexity of clinical infections in humans and animals, commonly refractory to conventional therapy. The domestic animals may represent a source of the pathogenic and multidrug-resistant Klebsiella species to humans. Nevertheless, most studies involving Klebsiella-induced infections in domestic animals are restricted to case reports or outbreaks. We retrospectively investigated selected epidemiological data, clinical aspects, and in vitro susceptibility pattern of 697 non-repetitive Klebsiella infections in livestock and companion species (1997–2019). The isolates were obtained from different clinical disorders from dogs (n = 393), cattle (n = 149), horses (n = 98), cats (n = 27), pigs (n = 22), sheep (n = 5), goats (n = 2), and buffalo (n = 1), except four isolates from subclinical bovine mastitis. Urinary (223/697 = 32%), enteric (117/697 = 16.8%), mammary (85/697 = 12.2%), reproductive (85/697 = 12.2%), and respiratory disorders (67/697 = 9.6%) were the most common clinical manifestations. Other miscellaneous clinical pictures (116/697 = 16.6%) included abscesses, otitis, hepatitis, conjunctivitis, pyodermitis, sepsis, and encephalitis. Norfloxacin (183/245 = 74.7%) and gentamicin (226/330 = 68.5%) were the most effective antimicrobials. High in vitro resistance of the isolates was seen to ampicillin (326/355 = 91.8%), amoxicillin/clavulanic acid (25/62 = 40.3%), and trimethoprim/sulfamethoxazole (100/252 = 39.7), and multidrug resistance to ≥ 3 classes of antimicrobials was found in 20.4% (142/697) isolates. Wide variety of clinical manifestations of Klebsiella-induced infections was observed, with a predominance of urinary, enteric, mammary, reproductive, and respiratory tract disorders, reinforcing opportunistic behavior of agent. Poor in vitro efficacy was observed to some conventional antimicrobials and ~ 20% of isolates exhibited resistance pattern, reinforcing the need for proper use of drugs on therapy approaches in domestic animals to avoid multidrug-resistant bacteria, an emergent global concern. Department of Animal Production and Preventive Veterinary Medicine Sao Paulo State University-UNESP, SP School of Veterinary Medicine and Animal Science São Paulo State University-UNESP, SP Department of Veterinary Medicine Faculty of Veterinary Medicine and Animal Sciences Antonio Nariño University Technology Faculty FATEC, SP Department of Animal Production and Preventive Veterinary Medicine Sao Paulo State University-UNESP, SP School of Veterinary Medicine and Animal Science São Paulo State University-UNESP, SP

Made available in DSpace on 2022-04-29T08:36:20Z (GMT). No. of bitstreams: 0 Previous issue date: 2022-03-01 Tuberculosis remains one of the most important infectious diseases with well-known zoonotic nature that affect humans, wildlife, and domestic animals, including goats. Nonetheless, no intradermal tuberculin test has been standardized for caprine diagnosis of tuberculosis. The present study investigated the intradermal comparative cervical tuberculin test (ICCTT) in the diagnosis of tuberculosis among 60 goats from farms with history of tuberculosis. The cutoff applied to goats was based on a study where goats had been experimentally infected with Mycobacterium bovis and Mycobacterium avium. Clinical examination, bacteriological culture, and histopathological staining were assessed to the diagnosis. Isolates compatible with mycobacteria were subjected for molecular diagnosis based on gyrB-restriction fragment length polymorphism (RFLP) analysis and PCR restriction-enzyme analysis (PRA) of hsp65 gene by BstEII and HaeIII, namely PRA-hsp65 assay. From all goats, 60% (n = 36/60), 3.3% (n = 2/60), and 36.7% (n = 22/60) showed positive, inconclusive, and negative reactions, respectively. Out of 36 goats with ICCTT positive, 75% (n = 27/36) had isolation of mycobacteria and were detected M. bovis by gyrB-RFLP. Molecular diagnosis and histopathological findings compatible with tuberculosis showed 86.1% (n = 31/36) concordance with the ICCTT. When compared ICCTT with M. bovis isolation, gyrB-RFLP, and histopathology, the better arithmetic means of sensitivity and specificity were 2.5 mm for ICCTT compared with M. bovis isolation and gyrB-RFLP, and 4.55 mm when compared with histopathology. Both receiver operating characteristic (ROC) curves presented statistical significance (P < 0.001). The identification of other mycobacteria, e.g., M. kansasii, M. flavescens, M. avium, M. florentinum, M. lentiflavum, M. simiae, and Corynebacterium pseudotuberculosis, not influenced positive results in ICCTT. The concordance between bacteriological, histopathological, and molecular identification with ICCTT findings indicate that the tuberculin test may be used as a valuable tool for diagnosis of caprine tuberculosis and reinforce the importance of association of methods to diagnostic of the disease from animal origin. Coordination of Sustainable Rural Development Secretariat of Agriculture and Supply of State of São Paulo, SP Animal Pathology Department School of Veterinary Medicine and Animal Sciences University of São Paulo-USP, SP Department of Preventive Veterinary Medicine and Animal Science USP, SP Biological Institute of São Paulo Secretariat of Agriculture and Supply of State of São Paulo, SP Tuberculosis and Mycobacteriosies Laboratory Adolfo Lutz Institute, SP Departament of Clinics USP, SP Veterinary Medicine Academic Unit Federal University of Campina Grande UFCG, PB Department of Animal Production and Preventive Veterinary Medicine School of Veterinary Medicine and Animal Sciences São Paulo State University-UNESP, SP Department of Animal Production and Preventive Veterinary Medicine School of Veterinary Medicine and Animal Sciences São Paulo State University-UNESP, SP

Made available in DSpace on 2022-05-01T07:58:46Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-11-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG) Bloody diarrhea is a common condition in dogs, but studies evaluating the enteropathogens involved specifically in adult dogs are scarce. In the present study, stool samples from 45 adult dogs with bloody diarrhea were evaluated for the four enteric organisms mainly reported in these cases: canine parvovirus type 2 (CPV-2), Clostridioides difficile, Clostridium perfringens, and Salmonella spp. In addition, the samples were also tested for coronavirus, rotavirus, Giardia spp., and Escherichia coli pathotypes to provide a better understanding of possible co-occurrence. Vaccination status, diet, and clinical outcome were also obtained when available. CPV-2b was identified in 17 dogs (37.8%), being the most frequent cause of bloody diarrhea, including completely vaccinated adult dogs. Toxigenic C. difficile and C. perfringens netF+ were detected in 6 (13.3%) and 5 (11.1%) dogs, in some cases in a co-occurrence with other enteric organisms. Three fatal cases of salmonellosis were identified in dogs fed a raw meat-based diet, raising the risks associated with this increasing practice. Veterinary School. Universidade Federal de Minas Gerais (UFMG) UNESP-São Paulo State University - Department of Animal Production and Preventive Veterinary Medicine Universidade Federal do Rio Grande do Sul (UFRGS) Clínica Veterinária MedVet Clínica Veterinária VetMaster UNESP-São Paulo State University - Department of Animal Production and Preventive Veterinary Medicine CAPES: CAPES – Prêmio CAPES 2015 - 0774/2017 CNPq: CNPq - 406402/2018-3 FAPEMIG: FAPEMIG - APQ-00524-17

Etiology, multiple antibiotic resistance index, and acute-phase protein profile in the serum and synovial fluid of 44 horses presenting septic arthritis were investigated. Young animals up to 24 months of age not yet submitted to taming were sampled. Synovial samples from the horses were submitted to culture and bacterial isolates subjected to 18 antimicrobials to investigate the in vitro multidrug resistance pattern. Also, hematological aspects, fibrinogen, and serum levels of some acute-phase proteins (total protein, albumin, globulin, amyloid A, and C-reactive protein) were assessed. To statistical analysis, microbiological isolation were divided into four groups: G1 = animals with negative isolation, G2 = isolation of Gram-negative bacteria, G3 = isolation of Gram-positive bacteria), and G4 = isolation of fungi. Microbial isolation was obtained from 77.2% (32/44) of the synovial samples. Escherichia coli (7/44 = 15.9%), Staphylococcus aureus (5/44 = 11.4%), Streptococcus equi subsp. equi (3/44 = 6.8%), Aspergillus niger (3/44 = 6.8%), Pseudomonas aeruginosa (2/44 = 4.5%), Streptococcus equi subsp. zooepidemicus (1/44 = 2.3%), Trueperella pyogenes (1/44 = 2.3%), and Rhodococcus equi (1/44 = 2.3%) were prevalent bacterial agents. Aspergillus niger is reported by the first time as a primary agent of septic arthritis in horses. Gentamicin (84.9%), marbofloxacin (79.3%) and ceftriaxone (72.4%) were the most effective antimicrobials, whereas high resistance of the isolates (>50%) was found to erythromycin (75.9%), clarithromycin (75.9%), penicillin (69%), clindamycin (58.6%), and streptomycin (55.2%). Multiple antibiotic resistance index (≥ 0.3) was identified in 50% (16/32) of the isolates, with highest resistance in Gram-negative bacterial isolates. Serum levels of amyloid A, C-reactive protein, fibrinogen, globulin, and total protein, as well as the number of nucleated cells in the synovial fluid had values above the reference for horses, reinforcing acute-phase proteins as biomarkers in diagnosis of septic arthritis. We highlighted the diversity of microorganisms that may be involved in equine septic arthritis, the high resistance of bacterial isolates to conventional antimicrobials, the high lethality of young horses with septic arthritis, and importance of in vitro antimicrobial susceptibility pattern tests to therapeutic approaches in the disease. RESUMO: Foram investigados a etiologia, o índice de resistência múltipla aos antibióticos e o perfil de proteínas de fase aguda, no soro ou líquido sinovial, de 44 equinos com sinais clínicos compatíveis com artrite séptica. Foram amostrados animais até 24 meses de idade não submetidos a doma. As amostras de líquido sinovial dos 44 animais foram submetidas à cultura bacteriológica e fúngica, e os isolados bacterianos submetidos in vitro diante de 18 antimicrobianos visando investigar a multirresistência. Também foram avaliados os aspectos hematológicos e o fibrinogênio sérico, bem como os níveis séricos de certas proteínas de fase aguda (proteína total, albumina, globulina, amilóide sérico A e proteína C reativa). O isolamento microbiano das amostras foi dividido em quatro grupos: G1 = animais com isolamento negativo; G2 = isolamento de bactérias Gram-negativas; G3 = isolamento de bactérias Gram-positivas e G4 = isolamento fúngico. O isolamento microbiano foi obtido em 77,2% (32/44) das amostras de líquido sinovial. Escherichia coli (7/44 = 15,9%), Staphylococcus aureus (5/44 = 11,4%), Streptococcus equi subsp. equi (3/44 = 6,8%), Aspergillus niger (3/44 = 6,8%), Pseudomonas aeruginosa (2/44 = 4,5%), Streptococcus equi subsp. zooepidemicus (1/44 = 2,3%), Trueperella pyogenes (1/44=2,3%) e Rhodococcus equi (1/44 = 2,3%) foram os patógenos mais frequentes. Aspergillus niger é relatado pela primeira vez como agente primário de artrite séptica em cavalos. Gentamicina (84,9%), marbofloxacino (79,3%) e ceftriaxona (72,4%) foram os antimicrobianos mais eficazes, enquanto elevada resistência dos isolados (>50%) foi encontrada para eritromicina (75,9%), claritromicina (75,9%), penicilina (69%), clindamicina (58,6%) e estreptomicina (55,2%). O índice de resistência múltipla aos antibióticos (≥ 0,3) foi identificado em 50% (16/32) dos isolados, com maior resistência obervada em bactérias Gram-negativas. Os níveis séricos de amilóide A, proteína C reativa, fibrinogênio, globulina e concentração total de proteínas, bem como o número de células nucleadas no líquido sinovial apresentaram valores acima da referência para equinos saudáveis, reforçando o uso destas proteínas de fase aguda como biomarcadores no diagnóstico da doença. O presente estudo enfatiza a diversidade de microrganismos que podem estar envolvidos na artrite séptica equina, a alta resistência dos isolados aos antimicrobianos convencionais, a alta mortalidade de equinos jovens com artrite séptica e a importância de realizar o tratamento da afecção com respaldo em testes de sensibilidade microbiana in vitro.

Made available in DSpace on 2020-12-10T19:53:14Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-04-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Escherichia coli is a major pathogen involved in the etiology of environmentally derived bovine mastitis and is characterized by a variety of virulence factors (VF). Mammary infections with E. coli have shown a wide range of clinical signs, causing changes in milk (score 1, or mild), abnormal appearance of milk and udder inflammation (score 2, or moderate), and abnormalities in milk, udder inflammation, and systemic signs of illness (score 3, or severe). Nevertheless, to date, the profile of the genes related to the virulence of the pathogen in mammary infections arid the severity scores of cases have not been thoroughly elucidated. Therefore, a panel of 18 virulence-encoding genes associated with extra-enteric pathogenicity of E. coli (ExPEC) were investigated in addition to in vitro swimming and swarming motility profiles and antimicrobial susceptibility/resistance patterns among 114 E. coli strains isolated from cows with clinical mastitis and different severity scores. Of 114 clinical cases, 39.5, 54.4, and 6.1% were mild, moderate, and severe, respectively. The main genes related to VF harbored by isolates were adhesins (fimH 100%; ecpA 64.0%, firnA 31.6%), serum resistance (traT 81.6%; ompT 35.1%), siderophores (irp2 9.6%), and hemolysin (hlyA 7%). Among the isolates studied, 99.1% showed in vitro resistance to bacitracin and cloxacillin, and 98.2% to lincosamin. Of the total isolates, 98.2% were considered multidrug resistant based on the multiple antimicrobial resistance index. No significant difference was observed between mean swimming (13.8 mm) and swarming (13.5 mm) motility, as well as severity scores of clinical mastitis and the ExPEC genes studied. The isolation of strains resistant to various antimicrobials, even though tested only in vitro, highlights the importance of rational use of antimicrobials for mastitis treatment. The high prevalence of the genes related to serum resistance (traT and ompT) and adhesion (ecpA) of the pathogen, in addition to main associations between the genes fimH, ecpA, and traT among cows with severity scores of 1 (15%) arid 2 (22.6%), indicates that the genes traT, ecpA, and ompT could be further studied as biomarkers of ExPEC for clinical intramammary infections. In addition, the ExPEC genes ompT (protectin), ibe10 (invasiri), arid ecpA (adhesin) were investigated for the first time among cows with mastitis, where scores of clinical severity were assessed. Results of this study contribute to the characterization of virulence mechanisms and antimicrobial resistance profile of ExPEC variants that affect dairy cows with different scores of clinical mastitis. Sao Paulo State Univ, Sch Vet Med & Anim Sci, Dept Vet Hyg & Publ Hlth, BR-18618681 Botucatu, SP, Brazil Sao Paulo State Univ, Dept Microbiol & Immunol, BR-18618689 Botucatu, SP, Brazil Univ Estadual Campinas, Dept Genet Evolut Microbiol & Immunol, BR-13083970 Campinas, SP, Brazil Paulista Agcy Agribusiness Technol, BR-17030000 Bauru, SP, Brazil Sao Paulo State Univ, Sch Vet Med & Anim Sci, Dept Vet Hyg & Publ Hlth, BR-18618681 Botucatu, SP, Brazil Sao Paulo State Univ, Dept Microbiol & Immunol, BR-18618689 Botucatu, SP, Brazil FAPESP: 2015/19688-8 FAPESP: 2017/08102-8

Made available in DSpace on 2020-12-12T02:36:52Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-04-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG) Pró-Reitoria de Pesquisa, Universidade Federal de Minas Gerais The occurrence and characteristics of Clostridioides (previously Clostridium) difficile and Clostridium perfringens in the feces of diarrheic and non-diarrheic cats was investigated. Apparently healthy animals were more likely to be positive for C. perfringens type A (p = 0.009). Two isolates (0.7%), one each from a diarrheic and an apparently healthy cat, were positive for the enterotoxin-encoding gene but negative for the NetF-encoding gene. Six toxigenic C. difficile isolates were isolated, all RT106 and ST42, which is commonly reported in humans with C. difficile infection. Veterinary School Universidade Federal de Minas Gerais (UFMG), Av. Antônio Carlos, 6627 UNESP-São Paulo State University - Department of Veterinary Hygiene and Public Health UNESP-São Paulo State University - Department of Veterinary Hygiene and Public Health

Made available in DSpace on 2021-06-25T10:44:24Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-03-01 Caseous lymphadenitis (CL) in sheep is a chronic contagious disease caused by Corynebacterium pseudotuberculosis, commonly characterized by abscess formation in peripheral lymph nodes and disseminated infections. Nonetheless, other microorganisms, including with zoonotic relevance, can be isolated from CL-resembling lymph nodes. Currently, mycobacteria have been reported in visceral granulomatous lesions in small ruminants, a fact that poses a public health issue, particularly in slaughtered sheep intended for human consumption. Cytology using fine needle aspiration and microbiological culturing are suitable tests for routine diagnostic, whereas present drawbacks and molecular methods have been confirmatory. Data about the occurrence of mycobacteria in both lymph nodes with aspect of CL and apparently healthy visceral nodes of sheep slaughtered for human consumption are scarce. In this study, 197 visceral lymph nodes of sheep showed lymphadenitis and 202 healthy visceral lymph nodes of slaughtered sheep intended for human consumption were submitted to conventional bacteriological diagnosis, mycobacteria culturing, and cytological evaluation. Compatible Corynebacterium isolates were subjected to multiplex PCR targeting 16S rRNA, rpoB, and pld genes to detect C. pseudotuberculosis. Based on microbiological identification, C. pseudotuberculosis (86/197; 43.7%), streptococci γ-hemolytic (17/197; 8.6%), and Trueperella pyogenes (12/197; 6.1%) were prevalent in lymph nodes with abscesses, as opposed to staphylococci (53/202; 26.2%) in apparently healthy lymph nodes. No mycobacteria were isolated. Cytology identified 49.2% (97/197) Gram-positive pleomorphic organisms (coryneform aspect). Multiplex PCR confirmed genetic material of C. pseudotuberculosis in 74.4% (64/86) of the samples with C. pseudotuberculosis isolation and 66% (64/97) samples with cytological coryneform aspect (κ = 86.78%; 95% CI = 79.87–93.68%). These findings emphasize the prevalence of C. pseudotuberculosis in abscess formation among peripheral lymph nodes of sheep. Other bacteria were also identified in lymph nodes sampled that resembling C. pseudotuberculosis–induced infections that may difficult the diagnosis. Multiplex PCR revealed a valuable assay to detect C. pseudotuberculosis, in addition to routine methods applied to CL-diagnosis. No mycobacteria were identified in lymph nodes sampled, with and without apparent lesions. Nonetheless, due to public health impacts, this pathogen should be considered as a differential diagnosis of C. pseudotuberculosis–induced infections during inspection procedures of slaughtered sheep intended for human consumption. Department of Animal Production and Preventive Veterinary Medicine School of Veterinary Medicine and Animal Science São Paulo State University (UNESP) Laboratory of Cellular and Molecular Genetics Institute of Biologic Sciences Federal University of Minas Gerais (UFMG) School of Agrarian Sciences University of Western São Paulo (UNOESTE) Technology Faculty FATEC Zoonosis and Public Health Research Group Department of Biologial Sciences Feira de Santana State University (UEFS) Department of Animal Production and Preventive Veterinary Medicine School of Veterinary Medicine and Animal Science São Paulo State University (UNESP)

Made available in DSpace on 2022-05-01T09:00:54Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-11-01 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Rhodococcus equi is a well-known intracellular facultative bacterium that is opportunistic in nature, and a contagious disease-causing agent of pyogranulomatous infections in humans and multihost animals. Feline rhodococcosis is an uncommon or unnoticed clinical condition, in which the organism is usually refractory to conventional antimicrobial therapy. The pathogenicity of the agent is intimately associated with plasmid-governed infectivity, which is attributed to the presence of plasmid-encoded virulence-associated proteins (Vap). Three host-adapted virulence plasmid types (VAPs) have been distinguished to date: pVAPA, pVAPB, and pVAPN, whose infections are related to equine, pig, and bovine or caprine origin, respectively, while humans are infected by all three VAP types. Most virulence studies with R. equi plasmid types in animals involve livestock species. Conversely, data on the pathogenicity and human relevance of the virulence plasmid profile of R. equi isolated from cats remains unclear. This report describes a case of cellulitis-related R. equi that harbors the pVAPA-type in a cat with cutaneous lesion. Long-term therapy of the cat using marbofloxacin, a broad-spectrum third-generation fluoroquinolone, resulted effectiveness. pVAPA is a host-adapted virulent type that has been associated predominantly with pulmonary foal infections. Our cat had a history of contact with other cats, livestock (including horses), and farm environment that could have favored the transmission of the pathogen. Besides no clear evidence of cat-to-humans transmission of the pathogen, the identification of R. equi harboring pVAPA-type in a cat with cutaneous abscessed lesion represent relevance in human health because this virulent type has been described in people worldwide with clinical rhodococcal disorders. Veterinary Clinic of Companion Animals Pet Center Cariri UNESP-São Paulo State University Department of Animal Production and Preventive Veterinary Medicine Animal Vetlab Graduate Program in Animal Science School of Life Sciences Pontifícia Universidade Católica do Paraná – PUCPR Kitasato University Department of Animal Hygiene UNESP-São Paulo State University Department of Animal Production and Preventive Veterinary Medicine CNPq: 310345/2020-0

Rhodococcus equi is a well-recognized Gram-positive intracellular facultative bacterium that is opportunistic in nature, which causes pyogranulomatous infections in humans and multiple host animals. The pathogenicity of the microorganism has been attributed to the presence of plasmid-encoded virulence-associated proteins (Vap). To date, three host-associated virulence plasmid types of R. equi have been identified as follows: the circular pVAPA and pVAPB, related, respectively, to equine and porcine isolates, and a recently described linear pVAPN plasmid associated with bovine strains, although these three types are found in human isolates. Recent phylogenomic studies support the evidence that human R. equi infection is zoonotically acquired. Nevertheless, data regarding distribution and prevalence of the host-adapted virulence plasmid types of R. equi isolated from meat animals are scarce or unnoticed. Here, the three host-associated virulence plasmid types (pVAPA, pVAPB, and pVAPN) were investigated in 154 R. equi isolates recovered from lymph nodes of cattle with lymphadenitis (n = 31), faeces of cattle without enteric signs (n = 49), as well as different clinical specimens from human patients (n = 74). The analysis of virulence profile of 74 R. equi from humans revealed six (8.1%) isolates pVAPB (type 8), two (2.7%) pVAPN, and one (1.3%) pVAPB (type 11), all of which were from lung samples from people living with HIV/AIDS. From the lymph node samples of cattle, 41.9% (13 of 31) isolates revealed pVAPN type, whereas all isolates from faecal samples were negative for three host-associated types. Here, recently described bovine-associated pVAPN type was detected in R. equi isolates recovered from the lungs of people living with HIV/AIDS and lymph nodes from slaughtered cattle intended for human consumption; a finding that represents a public health concern, mainly in countries where undercooked or raw meat are traditionally consumed.

Made available in DSpace on 2018-12-11T17:34:33Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-10-01 A strain of an achlorophyllic alga, named PR24T, was isolated from cow milk samples from the state of Minas Gerais, Brazil. Based on 18S rDNA, 28S rRNA, D1/D2 region of the LSU rDNA and SSU rRNA gene sequence similarities, this strain was found to be a member of the genus Prototheca and closely related to Prototheca blaschkeae SAG2064T. However, the novel strain could easily be distinguished from recognized Prototheca species by internal transcribed spacer, species-specific PCR, single-strand conformation polymorphism-PCR analysis and phenotypic characteristics. The inability to grow in Sabouraud broth at pH 4.0 and the different cellular fatty acid composition clearly distinguished PR24T from the reference strain of P. blaschkeae. The combination of genotypic and phenotypic data indicates that strain PR24T represents a subspecies of P. blaschkeae, for which the name Prototheca blaschkeae subsp. brasiliensis subsp. nov. is proposed. The respective type strain is PR24T (=DSM 103592T=IHEM 26958T). Institute of Sciences of Food Production Italian National Research Council (CNR ISPA), Via Celoria 2 Institute of Agricultural Biology and Biotechnology Italian National Research Council (CNR IBBA), Via Einstein Council for Agricultural Research and Economics – Research Centre for Animal Production and Aquaculture (CREA-ZA), Via Lombardo 11 Department of Veterinary Hygiene and Public Health Universidade Estadual Paulista (UNESP) Department of Veterinary Medicine Federal University of Lavras Istituto Zooprofilattico Sperimentale della Lombardia e dell’Emilia Romagna (IZSLER), Via Einstein Department of Veterinary Hygiene and Public Health Universidade Estadual Paulista (UNESP)

Os patógenos de eliminação fecal figuram como uma das principais classes de microorganismos patogênicos para os humanos e animais, causando graves doenças entéricas e,ocasionalmente, extra-entéricas. Este grupo de organismos caracteriza–se por manterem–seviáveis e com capacidade infectante em condições hostis do ambiente, após a eliminaçãopelas fezes dos animais com e sem sinais entéricos. Devido ao contato cada vez mais estreitocom os humanos, os animais de companhia são elos importantes na cadeia de transmissão depatógenos entéricos. Os gatos domésticos representam um grupo importante de animaiscarreadores de patógenos entéricos com potencial zoonótico, uma vez que possuem hábitos decaça, de estabelecer territorialidade e de auto-limpeza. Dentre os patógenos de veiculaçãofecal de gatos domésticos de potencial zoonótico para humanos, merecem destaque:Escherichia coli, Salmonella spp., Campylobacter spp., Clostridium spp. Rhodococcus equi,Giardia spp. e Cryptosporidium spp. O presente estudo revisou os principais patógenosbacterianos e parasitários de gatos domésticos com ênfase aos aspectos de virulência,epidemiológicos, sinais clínicos, controle e reflexos em Saúde Pública

Trueperella pyogenes is an opportunistic pathogen that causes diverse pyogenic infections in livestock. The genes that encode the exotoxin pyolysin (plo) and other putative factors that promote adhesion of pathogen to host cells (fimbriae fimA, fimC, fimE, fimG, neuraminidases nanH, nanP, and collagen-binding protein cbpA) have been associated with virulence, particularly in mastitis and uterus infections of dairy cows. However, the role of these virulence markers in the pathogenicity of the agent in domestic animals infections still is incompletely understood. The genes plo, fimA, fimC, fimE, fimG, nanH, nanP, and cbpA were investigated in 71 T. pyogenes strains recovered from cattle, sheep, goats, dogs, equines, and a pig, recovered from mastitis (n = 35), and non-mastitis (n = 36) cases (abscesses, reproductive tract diseases, pneumonia, lymphadenitis, encephalitis). The most common genes harboured by the isolates were: plo (71/71 = 100·0%), fimA (70/71 = 98·6%), nanP (56/71 = 78·9%), fimE (53/71 = 74·6%), fimC (46/71 = 64·8%) and nanH (45/71 = 63·4%), whereas cbpA (6/71 = 8·4%) and fimG (4/71 = 5·6%) were uncommon. The most frequent genotypes were plo/fimA/fimE/fimC/nanH/nanP (17/71 = 23·9%), plo/fimA/fimE/nanH/nanP (13/71 = 18·3%), and plo/fimA/fimE/fimC/nanP (11/71 = 15·5%). No association was observed between the presence of genes vs clinical signs or host species. To the best of our knowledge, this is the first report on aforementioned virulence factors of pathogen detected in diseased horses and dogs. Significance and impact of the study The role of particular virulence factors of Trueperella pyogenes that determine different pyogenic infections among domestic animals is poorly understood. Eight putative virulence genes and genotype profiles of 71 isolates were investigated among different clinical manifestations in domestic animals. The most common genes were plo (71/71 = 100·0%), fimA (70/71 = 98·6%), nanP (56/71 = 78·9%), fimE (53/71 = 74·6%), fimC (46/71 = 64·8%) and nanH (45/71 = 63·4%), whereas plo/fimA/fimE/fimC/nanH/nanP (17/71 = 23·9%), plo/fimA/fimE/nanH/nanP (13/71 = 18·3%), and plo/fimA/fimE/fimC/nanP (11/71 = 15·5%) were the most frequent genotypes. Studies involving virulence factors are critical in the investigation of molecular epidemiology, pathogenicity, and hypothetical differences in the virulence among T. pyogenes strains from different geographical areas.

Made available in DSpace on 2018-12-11T17:10:23Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-05-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) In developing nations, 10–20% of the human cases of tuberculosis are caused by Mycobacterium bovis. However, this percentage may be underestimated because most laboratories in developing countries do not routinely perform mycobacterial cultures, and only a few have the systems in place to identify M. bovis. There are few studies investigating genotypic diversity and drug resistance in M. bovis from animal and/or human infections. The genotypic diversity of M. bovis strains obtained from bovine lymph nodes were investigated by spacer oligonucleotide typing (spoligotyping) and mycobacterial interspersed repetitive unit–variable-number tandem repeat typing (MIRU-VNTR). The phenotypic resistance to isoniazid and rifampicin and MIC values of the isolates were determined using the resazurin microtiter assay plate method (REMA). The evaluation of the possible genetic basis for such resistance was performed with GenoType MTBDRplus. Sixty-seven isolates were obtained, of which 11 (16%) were MDR-TB, 8 (12%) were isoniazid-resistant, and 2 (3%) were rifampicin-resistant. Mutations associated with drug resistance were not found. Genotyping techniques enabled the grouping of the strains into 12 clusters and 21 isolates with unique profiles. The high frequency of M. bovis reinforces the impact of the pathogen as a major causal agent of bovine tuberculosis in the study area. The resistance of the strains to drugs used for first-line treatment of human tuberculosis raises public health concerns. Further studies are required to elucidate the basis of drug resistance and genotypic diversity in M. bovis. Department of Veterinary Hygiene and Public Health School of Veterinary Medicine and Animal Sciences UNESP-Univ. Estadual Paulista, P.O. Box: 560, Prof. Dr. Walter Mauricio Correa Street, s/n, Rubiao Junior district Laboratory of Tuberculosis Research School of Pharmaceutical Sciences UNESP- Univ. Estadual Paulista, Araraquara-Jau, km 01 Department of Preventive Veterinary Medicine and Animal Health USP-Univ. de Sao Paulo, Prof. Dr. Orlando Marques de Paiva Avenue, 87 Department of Clinical Analysis and Biomedicine UEM-Univ. Estadual de Maringá, Colombo Avenue, 5790 Federal Veterinary Inspection Service State of Sao Paulo, Dr. Gabriel de Oliveira Rocha Street, 704 Department of Pathophysiology and Host Defense JATA-Japan Anti-tuberculosis Association RIT-Research Institute of Tuberculosis, Zip Code: 204-8533, 3-1-24 Matsuyama Department of Veterinary Hygiene and Public Health School of Veterinary Medicine and Animal Sciences UNESP-Univ. Estadual Paulista, P.O. Box: 560, Prof. Dr. Walter Mauricio Correa Street, s/n, Rubiao Junior district Laboratory of Tuberculosis Research School of Pharmaceutical Sciences UNESP- Univ. Estadual Paulista, Araraquara-Jau, km 01 FAPESP: #2014/04929-7 FAPESP: 2014/04929-7

Proteus spp. are opportunistic multidrug resistant enterobacteria associated with diverse clinical diseases in domestic animals. However, Proteus infections in domestic animals are often misdiagnosed or considered contaminants in microbiological cultures rather than a primary agent of disease. Descriptions of Proteus infections in domestic animals are typically restricted to case reports, retrospective studies, or surveillance of other microorganisms. The present study investigated multiple antibiotic resistance indices, minimum inhibitory concentrations (MICs), and ESBL production in 73 strains of Proteus mirabilis (n = 69) and Proteus vulgaris (n = 4) isolated from domestic animals with various clinical manifestations. In dogs, the pathogen was most commonly associated with cystitis (48.21), enteritis (21.42%), otitis (14.29%), and conjunctivitis (3.57%). In bovines, the microorganism was predominant in cases of enteritis (22.22%), abscess (11.11%), otitis (11.11%), omphalitis (11.11%), and peritonitis (11.11%), and in organ fragments (11.11%). In equines (50.0%) and cats (100.0%), diarrhea was the main clinical sign. In vitro standard disk diffusion assay showed that the most effective antimicrobials against the isolates were imipenem (98.63), norfloxacin (95.89), amikacin (95.89), levofloxacin (90.41), ceftriaxone (87.64), and florfenicol (87.67). In contrast, the isolates commonly showed resistance to novobiocin (95.89), azithromycin (57.53), and trimethropim/sulfamethoxazole (39.73). Among the 73 isolates, the efficacy of amoxicillin/clavulanic acid, gentamicin, ceftriaxone, and ciprofloxacin according to MICs was 87.67%, 86.30%, 84.93%, and 82.19%, respectively. The MIC50 values of amoxicillin/clavulanic acid, ceftriaxone, ciprofloxacin, and gentamicin were, respectively, 1.0, 0.004, 0.03, and 1.0 µg/mL. Thirty-three strains (45.21%) showed a antimicrobial multiple resistance index of ? 0.3. Multidrug resistance profiles of isolates were observed most frequently in dogs (n = 25; 75.76%), particularly in those with cystitis (n = 13; 52.0%), followed by bovines (n = 4; 12.12%), equines (n = 2; 6.06%), and cats (n = 2; 6.06%). Two (2.7%) strains, obtained from canine skin and feces, were diagnosed phenotypically as ESBL-producers. Here, we observed the diversity of Proteus infections in domestic animals. The presence of multidrug-resistant isolates and ESBL-producers reinforces the need for appropriate antimicrobial use and in vitro antimicrobial tests to support therapy.

Escherichia coli is a normal inhabitant of the enteric microflora of human and animal. Intestinal and extra-intestinal infections caused by E. coli in mammals are characterized by the presence of diversity of virulence factors. In addition it can be isolated from environment surrounding human and animal farms. E. coli is the main environmental pathogen causing clinical mastitis in dairy cattle. It causes a wide range of disease severity, from changes seen exclusively in milk to severe systemic signs. The severity of clinical mastitis has been conventionally classified into three levels: mild (grade 1), moderate (score 2), and severe (score 3). Recently, reports of cases of bovine mastitis caused by environmental agents have been on the rise, in particular in countries that have succeeded in controlling contagious microorganisms. Unlike enteric and certain extra-enteric conditions in domestic animals and humans, the impact of virulence factors on the occurrence of bovine mastitis due to E. coli, as well as the clinical severity of the cases, is not fully understood. In this regard, the present study reviewed the most relevant virulence factors of E. coli in human and animals, with emphasis in bovine mastitis. Escherichia coli (E. coli) é um micro-organismo que pertence a microbiota entérica de humanos e animais. As afecções entéricas e extra-entéricas causadas por E.coli em mamíferos são caracterizadas pela presença de ampla diversidade de fatores de virulência. Além disso, este micro-organismo também pode ser isolado em ambientes de produção animal. E. coli é o principal agente ambiental que causa mastite clínica em vacas leiteiras, provocando sinais clínicos que podem variar desde mudanças observadas exclusivamente no leite até sinais sistêmicos graves. A gravidade da mastite clínica é convencionalmente classificada em três níveis: leve (grau 1), moderada (grau 2), e grave (grau 3). Atualmente, os relatos de casos de mastite bovina causada por agentes ambientais têm aumentando, principalmente nos países que conseguiram controlar os micro-organismos contagiosos. Diferentemente de infecções entéricas e certas extra-entéricas em humanos e animais, o impacto dos fatores de virulência de E. coli na ocorrência de mastite bovina não é totalmente compreendido, assim como na gravidade clínica dos casos. Neste contexto, o presente estudo revisou os fatores de virulência de E. coli mais relevantes para humanos e animais, com ênfase na mastite bovina.

Made available in DSpace on 2019-10-06T16:49:27Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-06-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Prototheca spp. are pathogenic algae with important zoonotic potential. Most importantly, these algae often infect dairy cattle. Since there is no effective therapy against the algae, the standard recommendation is the disposal or culling of infected cows to avoid outbreaks. This study investigated the ability of blue light to inactivate pathogenic Prototheca species. Blue LED light (λ = 410 nm) was used to inactivate in vitro suspensions of P. zopfii genotypes 1 and 2, and P. blaschkeae. Our results showed that blue light irradiation induced a strain-specific dose-dependent algicidal effect against all tested strains. P. zopfii genotype 1, was more sensitive than genotype 2 and P. blaschkeae was the most tolerant. Even though we observed different inactivation kinetics, all strains presented significant photoinactivation levels within feasible procedure periods. Therefore, we conclude that blue light irradiation offers promising potential for the development of novel technologies that control contaminations and infections caused by Prototheca spp. Department of Internal Medicine School of Veterinary Medicine and Animal Science University of Sao Paulo Department of Microbiology Institute for Biomedical Sciences University of Sao Paulo Department of Clinical and Toxicological Analysis School of Pharmaceutical Sciences University of Sao Paulo Department of Veterinary Hygiene and Public Health Sao Paulo State University (UNESP) Department of Applied Microbiology Institute of Microbiology Faculty of Biology University of Warsaw BioLambda Scientific and Commercial LTD Department of Veterinary Hygiene and Public Health Sao Paulo State University (UNESP) FAPESP: 2016/25095-2

Made available in DSpace on 2019-10-06T16:16:57Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-04-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Canine morbillivirus (previously, canine distemper virus, CDV) is a highly contagious infectious disease-causing agent that produces immunosuppressive infections and multiple clinical signs. Canine toxoplasmosis is an opportunistic disease characterized by enteric, pulmonary, and neuromuscular signs that might be confused with CDV-induced infections. Rhodococcus equi is a Gram-positive intracellular facultative bacterium that is also opportunistic in nature, and causes pyogranulomatous infections in humans and multiple host animals, although canine rhodococcosis is rare or unrecognized. The pathogenicity of R. equi is intimately related to the presence of plasmid-encoded virulence-associated proteins (Vap). Three host-adapted virulence plasmid types of R. equi have been recognized: the circular pVAPA and pVAPB are associated with equine and porcine strains, respectively, and the recently detected linear pVAPN virulence plasmid is related to bovine isolates. Nevertheless, data regarding the detection of host-adapted virulence plasmid types of R. equi isolated from companion animals are scarce. This report describes a case of an uncommon coinfection due to R. equi, T. gondii and CDV, which was diagnosed in a pet dog with respiratory distress. In this case, CDV most likely induced immunosuppression, which facilitated opportunistic infections by R. equi and T. gondii. The analysis of the virulence profile of R. equi revealed the novel pVAPN plasmid type, initially related to bovine strains. This is the second report of the bovine-associated pVAPN type in a pet dog, with an unusual coinfection with T. gondii and CDV. These findings represent a public health concern due to the close contact between pet animals and their owners, particularly because the pVAPN plasmid type was recently detected in people with HIV/AIDS from the same geographical region. UNESP-São Paulo State University Department of Veterinary Hygiene and Public Health School of Veterinary Medicine and Animal Sciences UNESP Department of Veterinary Clinical Sciences VetMol Laboratory UEL-Londrina State University Department of Preventive Veterinary Medicine Kitasato University Department of Animal Hygiene UNESP-São Paulo State University Department of Veterinary Hygiene and Public Health School of Veterinary Medicine and Animal Sciences UNESP Department of Veterinary Clinical Sciences FAPESP: 2015/20585-9 CNPq: 306116/2016-2

EnglishPrototheca spp. have been reported as an emergent environmental mastitis pathogen in several countries. Biofilm formation is a significant factor associated with different degrees of virulence developed by many microorganisms, including Prototheca spp. The present study aimed to compare two growth conditions and two staining dyes to determine which combination was more appropriate to evaluate qualitatively and quantitatively the production of biofilm by P. zopfii. Biofilm formation was evaluated in polystyrene microplates under static and dynamic growth conditions and staining with crystal violet or cotton blue dye. All P. zopfii isolates from cows with mastitis were classified as biofilm-producers in all growth conditions and staining. The cotton blue dye proved to be more appropriate method to classify the intensity of P. zopfii biofilm production. Key words: Prototheca zopfii, dairy cattle; bovine mastitis, intramammary infections, microplate biofilm assay, virulence factor portuguesPrototheca spp. tem sido relatado como um patogeno ambiental causador de mastite bovina em varios paises. A formacao de biofilme e um fator associado a diferentes graus de virulencia desenvolvidos por muitos microrganismos, incluindo Prototheca zopfii. O presente estudo teve como objetivo comparar duas condicoes de crescimento e dois corantes para determinar a combinacao mais adequada para avaliar qualitativa e quantitativamente a producao de biofilme por P. zopfii. A formacao de biofilme foi avaliada em microplacas de poliestireno sob condicoes estaticas e dinâmicas de crescimento e coloracao com cristal violeta ou azul de algodao. Todos os isolados de P. zopfii de vacas com mastite foram caracterizados como produtores de biofilme, independentemente das condicoes de crescimento e coloracao. O corante azul de algodao demonstrou ser o metodo mais adequado para classificar a intensidade de producao de biofilme de P. zopfii. Palavras-chave: Prototheca zopfii; bovino de leite; mastite bovina; infeccoes intramamarias; ensaio de biofilme em microplaca; fator de virulencia

Made available in DSpace on 2019-10-06T16:42:41Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-01-01 Many efforts have been made to understand the pathogenesis of bovine mastitis to reduce losses and promote animal welfare. Staphylococcus aureus may cause bovine clinical mastitis, but it is mainly associated with subclinical infection, which is usually persistent and can easily reoccur. Here, we conducted a comparative genomic analysis between strains of S. aureus causing subclinical infection (Sau170, 302, 1269, 1364), previously sequenced by our group, and two well-characterized strains causing clinical mastitis (N305 and RF122) to find differences that could be linked to mastitis outcome. A total of 146 virulence-associated genes were compared and no appreciable differences were found between the bacteria. However, several nonsynonymous single nucleotide polymorphisms (SNPs) were identified in genes present in the subclinical strains when compared to RF122 and N305, especially in genes encoding host immune evasion and surface proteins. The secreted and surface proteins predicted by in silico tools were compared through multidimensional scaling analysis (MDS), revealing a high degree of similarity among the strains. The comparison of orthologous genes by OrthoMCL identified a membrane transporter and a lipoprotein as exclusive of bacteria belonging to the subclinical and clinical groups, respectively. No hit was found in RF122 and N305 for the membrane transporter using BLAST algorithm. For the lipoprotein, sequences of Sau170, 302, 1269, and 1364 with identities between 68–73% were found in the MDS dataset. A conserved region found only in the lipoprotein genes of RF122 and N305 was used for primer design. Although the polymerase chain reaction (PCR) on field isolates of S. aureus did not validate the findings for the transporter, the lipoprotein was able to separate the clinical from the subclinical isolates. These results show that sequence variation among bovine S. aureus, and not only the presence/absence of virulence factors, is an important aspect to consider when comparing isolates causing different mastitis outcomes. Departamento de Bioquímica e Biologia Molecular Universidade Federal de Viçosa Núcleo de Análise de Biomoléculas Universidade Federal de Viçosa Instituto Nacional de Ciência e Tecnologia em Interações Planta Praga/BIOAGRO Universidade Federal de Viçosa Departamento de Higiene Veterinária e Saúde Pública Faculdade de Medicina Veterinária e Zootecnia UNESP/Botucatu Departamento de Higiene Veterinária e Saúde Pública Faculdade de Medicina Veterinária e Zootecnia UNESP/Botucatu

Rhodococcus equiis responsible for infections in multiple-host animals. In humans, the prevalence of rhodococcus has increased worldwide and represents an emergent risk.R. equiis a soil-borne opportunistic bacterium isolated from feces of a wide variety of domestic species, except cats; thus, there is no known potential risk of its transmission from humans. Here, the mono- and cooccurrence ofRhodococcus equiand other bacteria and selected virulence markers were investigated in feces of nondiarrheic cats from urban (n=100) and rural (n=100) areas. Seven (7/200=3.5%)R. equiisolates were recovered in ceftazidime, novobiocin, and cycloheximide (CAZ-NB) selective media, exclusively of cats from three distinct farms (p=0.01), and these cats had a history of contact with horses and their environment (p=0.0002). None of theR. equiisolates harbored hosted-adapted plasmid types associated with virulence (pVAPA, pVAPB, and pVAPN). One hundred seventy-fiveE. coliisolates were identified, and 23 atypical enteropathogenicE. coli(aEPEC), 1 STEC (Shiga-toxin producingE. coli), and 1 EAEC (enteroaggregativeE. coli) were detected. Eighty-sixC. perfringenstype A isolates were identified, and beta-2 and enterotoxin were detected in 21 and 1 isolates, respectively. FiveC. difficileisolates were identified, one of which was toxigenic and ribotype 106. The main cooccurring isolates in cats from urban areas wereE. coliandC. perfringensA (26/100=26%),E. coliandC. perfringenstype Acpb2+(8/100=8%), and aEPEC (eae+/escN+) andC. perfringenstype A (5/100=5%). In cats from farms, the main cooccurring isolates wereE. coliandC. perfringenstype A (21/100=21%),E. coliandC. perfringenstype Acpb2+8/100=8%), andE. coliandR. equi(4/100=4%). We identified, for the first time,R. equiin nondiarrheic cats, a finding that represents a public health issue because rhodococcus has been reported in both immunosuppressed and immunocompetent humans, particularly people living with HIV/AIDS.

Made available in DSpace on 2020-12-12T02:29:51Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-07-01 Ehrlichia and Anaplasma species are the most common tick-borne disease (TBD) pathogens in dogs worldwide. Ehrlichia canis, the aetiological agent of the Canine Monocytic Ehrlichiosis (CME), is known to replicate within the cytoplasm of mononuclear cells into clusters of organisms called morulae. However, detection of morulae in neutrophils is commonly observed in dogs infected by Ehrlichia ewingii or Anaplasma phagocytophilum. We report uncommon clinical cases of canine ehrlichiosis presenting morulae compatible with E. ewingii and A. phagocytophilum in dogs from two distinct regions of Brazil. Eight dogs were admitted to two veterinary teaching hospitals from Brazil, showing clinical or haematological signs suggestive of TBD. Blood or peritoneal fluid was withdrawn for haematological and cytologic analysis. All samples were evaluated by PCR assays for Ehrlichia and Anaplasma using genus-specific primers for dsb, 16S rRNA and groEL genes, followed by sequencing. Samples were also evaluated by nested PCR assays for the 16S rRNA gene of E. ewingii and groEL gene of A. phagocytophilum and Anaplasma platys. Seven dogs revealed thrombocytopenia, six dogs had monocytosis and five presented lymphopenia and anaemia. All dogs showed morulae structures compatible with Ehrlichia spp. in neutrophils and were PCR-positive for the dsb and 16S rRNA gene fragments of Ehrlichia, with sequences showing 100% identity with multiple E. canis sequences deposited in the GenBank™. Sequencing of 16S rRNA and groEL gene fragments from one PCR-positive dog showed 100% identity with A. platys. Overall, our data suggest that in endemic regions for E. canis, that is Brazil, the presence of morulae in neutrophils may indicate infection by this bacterium. Herein, morulae were also found in neutrophils present in the peritoneal fluid of a dog. Also, this is the first report of E. canis and Hepatozoon canis co-infection in neutrophils from naturally infected dogs confirmed by DNA sequencing. Laboratory of Virology and Rickettsial Infections Veterinary Hospital Federal University of Mato Grosso (UFMT) Veterinary Clinical Laboratory Department of Veterinary Clinics São Paulo State University (UNESP) Department of Veterinary Hygiene and Public Health São Paulo State University (UNESP) Laboratory of Clinical Pathology Veterinary Medicine College University of Cuiabá (UNIC) Laboratory of Microbiology and Molecular Biology Veterinary Hospital of the Faculty of Veterinary Medicine Federal University of Mato Grosso (UFMT) Department of Veterinary Medicine Federal University of Paraná (UFPR) Veterinary Clinical Laboratory Department of Veterinary Clinics São Paulo State University (UNESP) Department of Veterinary Hygiene and Public Health São Paulo State University (UNESP)

Made available in DSpace on 2018-12-11T17:31:11Z (GMT). No. of bitstreams: 0 Previous issue date: 2016-01-01 Diarrhea is a major clinical problem affecting foals up to 3 months of age. The aim of this study was to identify enteric microorganisms involved in monoinfections and coinfections and the associated virulence factors in healthy and diarrheic foals. Diarrheic (D) (n=56) and nondiarrheic (ND) foals (n=60) up to three months of age were studied. Fecal samples were analyzed for identification of infectious agents (microbiological culturing, molecular techniques, and microscopic analyses). Escherichia coli fimH (30% versus 25%), Salmonella spp. (25% versus 7%), Strongyloides westeri (25% versus 25%), Clostridium perfringens type A (21% versus 10%), E. coli ag43 (20% versus 35%), Strongylus (11% versus 18%), and vapA-positive Rhodococcus equi (5% versus 2%) were the most frequent enteric pathogens detected in D and ND foals, respectively. The frequency of toxin A-positive C. perfringens was significantly increased in the D (p=0.033) compared with the ND animals. R. equi strains harboring virulent plasmids were also identified (VapA 85-kb type I and VapA 87-kb type I) in D and ND foals. Coinfections were observed in 46% of the D and 33% of the ND foals. Our results demonstrate the great diversity of enteric pathogens, virulence factors, and coinfections involved in enteric infections of foals. Department of Veterinary Clinical Sciences Universidade Estadual Paulista (UNESP) Department of Veterinary Hygiene and Public Health Universidade Estadual Paulista (UNESP) Department of Preventive Veterinary Medicine Federal University of Minas Gerais (UFMG) Department of Genetics Evolution and Bioagents Campinas State University (Unicamp) Department of Preventive Veterinary Medicine University of S�o Paulo Department of Animal Hygiene Kitasato University Department of Veterinary Clinical Sciences Universidade Estadual Paulista (UNESP) Department of Veterinary Hygiene and Public Health Universidade Estadual Paulista (UNESP)

UNLABELLED The virulence-plasmid profile of Rhodococcus equi strains isolated from Suidae and humans is similar. Recent evidence suggests that the consumption of pork products contaminated with faeces might be a potential source of R. equi infections in humans, mainly to patients with rhodococcosis without history of contact with pigs or pig farms. This study investigated the virulence-associated genes (vapA and vapB) and plasmid profiles of R. equi among the 150 samples of small intestinal content obtained from slaughtered pigs. In addition, all samples were subjected to microbiological culture in conventional sheep blood agar and CAZ-NB, TCP and TVP selective media. A total of 40 (26·7%) of the samples recovered R. equi, with two samples recovering isolates harbouring the VapB type 8 plasmid. Among the 150 pigs sampled herein, CAZ-NB was considered the best selective medium for the isolation of R. equi from faeces. Our results provide evidence that the contamination of slaughtered pig carcasses with pathogenic R. equi might occur through faeces, representing a public health concern. Furthermore, this study is the first description of R. equi strains carrying the VapB plasmid in the gut of pigs. SIGNIFICANCE AND IMPACT OF THE STUDY Intermediately virulent (VapB) is a common plasmid-type harboured by R. equi isolated from pigs and humans with AIDS. Curiously, humans with rhodococcosis usually have no history of contact with pigs or pig farms. Virulence-plasmid profile of 40 R. equi isolated among 150 small intestine content samples from pigs revelled two carrying isolates with the VapB type-8 plasmids. Moreover, comparison of three selective culture media shows that CAZ-NB was the best. Our results provide evidence that contamination of slaughtered pig carcasses with pathogenic R. equi might occur through faeces, representing a public health concern. Furthermore, R. equi carrying VapB type-8 plasmids types are described for the first time in the gut of the pig.

Made available in DSpace on 2016-07-07T12:35:48Z (GMT). No. of bitstreams: 0 Previous issue date: 2015. Added 1 bitstream(s) on 2016-07-07T12:44:45Z : No. of bitstreams: 1 S0036-46652015000300251.pdf: 121351 bytes, checksum: 9340a4491c232605998380c2a55ea104 (MD5) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Nocardia é um microorganismo ubiquitário relacionado a infecções piogranulomatosas, com difícil resolução tecidual em humanos e animais. A doença é mundialmente emergente devido ao aumento de doenças e tratamentos imunossupressores. Este relato de casos ocorridos no Brasil visa apresentar a identificação molecular dos isolados e o padrão de sensibilidade a antimicrobianos por disco-difusão e concentração inibitória mínima (CIM) através de fitas E-test®. Os casos ocorreram em homens, em idade adulta. Três quadros foram cutâneos, dois pulmonares, um neurológico e um sistêmico. O quadro respiratório, o neurológico e um sistêmico estavam associados à doença ou terapia imunossupressoras. O sequenciamento do gene 16S rRNA (1491pb) possibilitou a identificação de quatro isolados de Nocardia farcinica, dois de Nocardia nova e um de Nocardia asiatica. N. farcinica foi observada em dois casos dermatológicos, um pulmonar e um quadro sistêmico, N. nova foi isolada de um caso neurológico e outro pulmonar; e N. asiatica em um caso dermatológico. O teste de disco-difusão mostrou que amicacina (100%), amoxicilina/clavulanato (100%), cefalexina (100%) e ceftiofur (100%) foram mais efetivos; enquanto gentamicina (43%), sulfametoxazol/trimetoprim (43%) e ampicilina (29%) foram menos efetivos. No entanto, no teste de concentração inibitória mínima (CIM), apenas um dos quatro isolados da espécie Nocardia farcinica mostrou-se resistente a sulfametoxazole-trimetropina. Nocardia is a ubiquitous microorganism related to pyogranulomatous infection, which is difficult to treat in humans and animals. The occurrence of the disease is on the rise in many countries due to an increase in immunosuppressive diseases and treatments. This report of cases from Brazil presents the genotypic characterization and the antimicrobial susceptibility pattern using the disk-diffusion method and inhibitory minimal concentration with E-test® strips. In summary, this report focuses on infections in young adult men, of which three cases were cutaneous, two pulmonary, one neurological and one systemic. The pulmonary, neurological and systemic cases were attributed to immunosuppressive diseases or treatments. Sequencing analysis of the 16S rRNA segments (1491 bp) identified four isolates of Nocardia farcinica, two isolates of Nocardia nova and one isolate of Nocardia asiatica. N. farcinica was involved in two cutaneous, one systemic and other pulmonary cases; N. nova was involved in one neurological and one pulmonary case; and Nocardia asiatica in one cutaneous case. The disk-diffusion antimicrobial susceptibility test showed that the most effective antimicrobials were amikacin (100%), amoxicillin/clavulanate (100%), cephalexin (100%) and ceftiofur (100%), while isolates had presented most resistance to gentamicin (43%), sulfamethoxazole/trimethoprim (43%) and ampicillin (29%). However, on the inhibitory minimal concentration test (MIC test), only one of the four isolates of Nocardia farcinica was resistant to sulfamethoxazole/trimethoprim. Universidade Federal de Santa Maria, Departamento de Medicina Veterinária Preventiva Universidade Federal de São Paulo, Departamento de Ciências Biológicas Universidade Estadual Paulista, Departamento de Higiene Veterinária e Saúde Pública, Faculdade de Medicina Veterinária e Zootecnia de Botucatu FAPESP: 2009/56037-1 FAPESP: 2010/53494-5

Brucellosis remains as a public health concern worldwide. In domestic animals, the disease is characterized by reproductive disorders in male and female. Besides extensive use of serological tests and recent development of molecular biology techniques, microbiological culture of Brucella species is yet considered a “gold standard” method for diagnosis. Here, semen of 335 bovine bulls was subjected simultaneously to microbiological culture in Brucella agar, Farrell media, and CITA media to evaluate comparatively the best selective media for isolation of Brucella sp. Among all 335 samples, B. abortus B19 strain was isolated from semen of five (1.49%) bulls using the three selective media. However, Farrell media was considered the best selective media for microbiological diagnosis, because of allowed isolation of B. abortus B19 strain from bull semen without bacterial commensal or fungal contamination of plates. Key words: Brucella agar, Farrell media, CITA media, bovine bulls, semen.

Made available in DSpace on 2018-11-12T17:28:18Z (GMT). No. of bitstreams: 0 Previous issue date: 2018. Added 1 bitstream(s) on 2018-11-12T17:36:25Z : No. of bitstreams: 1 S0036-46652018005000602.pdf: 432009 bytes, checksum: 1a143450dcdaac86bfa9abe9436f4f3d (MD5) ABSTRACT Although the tuberculin test represents the main in vivo diagnostic method used in the control and eradication of bovine tuberculosis, few studies have focused on the identification of mycobacteria in the milk from cows positive to the tuberculin test. The aim of this study was to identify Mycobacterium species in milk samples from cows positive to the comparative intradermal test. Milk samples from 142 cows positive to the comparative intradermal test carried out in 4,766 animals were aseptically collected, cultivated on Lowenstein-Jensen and Stonebrink media and incubated for up to 90 days. Colonies compatible with mycobacteria were stained by Ziehl-Neelsen to detect acid-fast bacilli, while to confirm the Mycobacterium genus, conventional PCR was performed. Fourteen mycobacterial strains were isolated from 12 cows (8.4%). The hsp65 gene sequencing identified M. engbaekii (n=5), M. arupense (n=4), M. nonchromogenicum (n=3), and M. heraklionense (n=2) species belong to the Mycobacterium terrae complex. Despite the absence of M. tuberculosis complex species in the milk samples, identification of these mycobacteria highlights the risk of pathogen transmission from bovines to humans throughout milk or dairy products, since many of mycobacterial species described here have been reported in pulmonary and extrapulmonary diseases both in immunocompetent and immunocompromised people. Universidade Estadual Paulista Faculdade de Medicina Veterinária e Zootecnia Departamento de Higiene Veterinária e Saúde Pública Universidade de São Paulo Faculdade de Medicina Veterinária e Zootecnia Departamento de Medicina Veterinária Preventiva e Saúde Animal Universidade de Nariño Departamento de Biología Universidade do Rio Verde Universidade Estadual Paulista Faculdade de Medicina Veterinária e Zootecnia Departamento de Higiene Veterinária e Saúde Pública

The aim of the present study was to compare the test performances of three commercial enzyme immunoassays (EIAs) against the toxigenic culture using the cytotoxicity assay as the gold standard. All EIAs showed >78% sensitivity, and the lowest specificity was 92.6%. These results suggest that EIAs could be useful for the diagnosis of Clostridium difficile infection in foals.

Made available in DSpace on 2014-10-01T13:08:38Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-02-01Bitstream added on 2014-10-01T14:03:01Z : No. of bitstreams: 1 S0100-736X2014000200007.pdf: 310675 bytes, checksum: d6ee1deeffd6997b27a18ae2f43e2bff (MD5) O sistema silvipastoril caracteriza-se por aumentar a produção de leite, com maior número de vacas por hectare devido ao maior aporte de proteína na dieta. Neste sistema as vacas são alimentadas, além do pasto, de pequenas árvores e arbustos. O objetivo do presente estudo foi determinar os principais indicadores de qualidade do leite e agentes causais de mastite em vacas criadas em sistema silvipastoril. Foram avaliadas a composição (teor de gordura, proteína total, lactose, extrato seco, extrato seco desengordurado e nitrogênio uréico), contagem de células somáticas (CCS), contagem bacteriana total (CBT), ocorrência de mastite clínica e subclínica, isolamento microbiológico, perfil de sensibilidade bacteriana in vitro e detecção de resíduos antimicrobianos no leite produzido por 100 vacas, bem como do tanque de expansão e latões em propriedades do Vale do Cauca, Colômbia. Os teores médios dos principais constituintes do leite foram 3,24% de gordura, 3,27% de proteína total, 4,40% de lactose, 10,62% de extrato seco, 8,57% de extrato seco desengordurado e 15,82mg/dL de nitrogênio uréico, enquanto do tanque de expansão e latões foi 3,51% de gordura, 3,20% de proteína total, 4,34% de lactose, 11,72% de extrato seco, 8,47% de extrato seco desengordurado e 14,57mg/dL de nitrogênio uréico. A celularidade média dos quartos mamários e do tanque de expansão foi 141.252,75 CS/mL e 363.078,05 CS/mL respectivamente. A CBT média dos quartos mamários e do tanque de expansão foi 4.466,84 UFC/mL e 24.547,01 UFC/mL. Os principais micro-organismos isolados dos quartos mamários foram Corynebacterium bovis, Staphylococcus aureus, Staphylococcus hyicus, Staphylococcus spp., Streptococcus dysgalatiae, enquanto do tanque de expansão foram identificados Streptococcus spp., Enterobacter cloacae, Hafnia alveii, Streptococcus hemolítico e Streptococcus spp., com maior frequência. A presença de resíduos de antimicrobianos em leite de vacas e do tanque ou latão foi detectada em 30% e 86% das amostras, respectivamente. O sistema silvipastoril mostrou ser uma boa alternativa para produção de leite em vacas. No entanto, são necessários cuidados no tratamento mamário para evitar resíduos no leite e a análise de todos os parâmetros de qualidade para garantir um produto diferenciado. The silvopastoral system is characterized by increasing the production of milk with a greater number of cows per hectare due to the higher amount of protein in the diet. In silvopastoral system cows are fed in addition to pasture, small trees and shrubs. The aim of this study was determinate the main indicators of milk quality and mastitis causal agents in cows bred on silvopastoral system. We evaluated the composition (fat, total protein, lactose, solids, dry extract, nonfat dry and urea nitrogen), somatic cell count (SCC), total bacterial count (TBC), occurrence of clinical and subclinical mastitis, microbiological isolation, in vitro antimicrobial sensitivity profile and detection of antimicrobial residues in milk produced by 100 cows raised in silvopastoral systems, as well as the bulk tank and churns in farms of Cauca Valley, Colombia. The concentration of the major constituents of milk were 3.24% fat, 3.27% total protein, 4.40% lactose, 10.62% dry extract, 8.57% nonfat dry and 15.82mg/dL urea nitrogen, while the bulk tank and churns was 3.51% fat, 3.20% total protein, 4.34% lactose, 11.72% dry extract, 8.47% nonfat dry and 14.57mg/dL urea nitrogen. The cell count of the cows and the bulk tank was 141,252.75 CS/mL and 363,078.05 CS/mL respectively. The TBC mean in cows and the bulk tank was 4,466.84 CFU/mL and 24,547.01 CFU/mL respectively. The main microorganisms isolated from the udder cows were Corynebacterium bovis, Staphylococcus aureus, Staphylococcus hyicus, Staphylococcus spp., Streptococcus dysgalatiae, while the bulk tank were identified more often Streptococcus spp., Enterobacter cloacae, Hafnia alveii, hemolytic Streptococcus and Streptococcus spp. Antimicrobial residues in cow milk and bulk or churn were detected in 30% and 86% respectively. The silvopastoral system showed to be good alternative to milk production in cow. However is important the care with antimicrobial residues in milk and the analysis of all quality parameters to ensure a differentiated product. Universidade Estadual Paulista Faculdade de Medicina Veterinária e Zootecnia Departamento de Higiene Veterinária e Saúde Pública Universidade Estadual Paulista Faculdade de Medicina Veterinária e Zootecnia Departamento de Higiene Veterinária e Saúde Pública

Nocardia spp. infections can cause severe damage to the mammary gland due to suppurative pyogranulomatous lesions and lack of clinical cure in response to conventional antimicrobial therapy. Although Nocardia infections are considered relatively uncommon in cows, there has been an apparent worldwide increase in the incidence of bovine mastitis caused by Nocardia spp, perhaps due to environmental transmission of this ubiquitous pathogen. The objectives of present study were to determine: (i) species distribution of 80 Nocardia isolates involved in bovine mastitis (based on molecular methods); and (ii) antimicrobial susceptibility pattern of all isolates from three geographical areas in Brazil. In this study, Nocardia nova (80%) was the most frequently isolated species, followed by Nocardia farcinica (9%). Additionally, Nocardia puris, Nocardia cyriacigeorgica, Nocardia veterana, Nocardia africana, and Nocardia arthritidis were detected using 16S rRNA sequencing. This is apparently the first report of N. puris, N. veterana, N. cyriacigeorgica, N. arthritidis and N. africana in association with bovine mastitis. Based on the disk diffusion test, isolates were most frequently resistant to cloxacillin (75%), ampicillin (55%) and cefoperazone (47%), whereas few Nocardia spp. were resistant to amikacin, cefuroxime or gentamicin.

Submitted by Guilherme Lemeszenski (guilherme@nead.unesp.br) on 2013-08-22T18:45:40Z No. of bitstreams: 0 Made available in DSpace on 2013-08-22T18:45:40Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-05-01 Made available in DSpace on 2013-09-30T19:34:45Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-05-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T15:09:26Z No. of bitstreams: 0 Made available in DSpace on 2014-05-20T15:09:26Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-05-01 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ) SR-2/UERJ PRONEX Corynebacterium diphtheriae, Corynebacterium ulcerans and Corynebacterium pseudotuberculosis constitute a group of potentially toxigenic microorganisms that are related to different infectious processes in animal and human hosts. Currently, there is a lack of information on the prevalence of disease caused by these pathogens, which is partially due to a reduction in the frequency of routine laboratory testing. In this study, a multiplex polymerase chain reaction (mPCR) assay that can simultaneously identify and determine the toxigenicity of these corynebacterial species with zoonotic potential was developed. This assay uses five primer pairs targeting the following genes: rpoB (Corynebacterium spp), 16S rRNA (C. ulcerans and C. pseudotuberculosis), pld (C. pseudotuberculosis), dtxR (C. diphtheriae) and tox [diphtheria toxin (DT) ]. In addition to describing this assay, we review the literature regarding the diseases caused by these pathogens. of the 213 coryneform strains tested, the mPCR results for all toxigenic and non-toxigenic strains of C . diphtheriae, C. ulcerans and C. pseudotuberculosis were in 100% agreement with the results of standard biochemical tests and PCR-DT. As an alternative to conventional methods, due to its advantages of specificity and speed, the mPCR assay used in this study may successfully be applied for the diagnosis of human and/or animal diseases caused by potentially toxigenic corynebacterial species. Universidade do Estado do Rio de Janeiro Faculdade de Ciências Médicas Laboratório de Difteria e Corinebactérias de Importância Clínica Universidade Federal de Minas Gerais Instituto de Ciências Biológicas Departamento de Biologia Universidade Federal da Bahia (UFBA) Instituto de Ciências da Saúde Departamento de Biointeração Universidade do Cairo Faculdade de Medicina Veterinária Fundação Universidade Federal do Vale do São Francisco Universidade Estadual Paulista Escola de Medicina Veterinária e Ciência Animal Departamento de Higiene Veterinária e Saúde Pública Universidade Estadual Paulista Escola de Medicina Veterinária e Ciência Animal Departamento de Higiene Veterinária e Saúde Pública

Summary Reasons for performing the study Toxin detection and screening could contribute to knowledge of the transmission patterns, risk factors and epidemiology of Clostridium difficile and Clostridium perfringens. Objective To isolate C. difficile and C. perfringens and to detect A/B toxins in faecal samples from diarrhoeic and nondiarrhoeic foals. Study design Cross-sectional observational study. Methods A total of 153 samples from foals were collected: 139 samples from farms and 14 samples from diarrhoeic foals admitted to a veterinary hospital. The A/B toxins were detected by cytotoxicity assay. All suspected colonies of C. perfringens were subjected to polymerase chain reaction for detection of the major toxin genes (α, β, e and ι) and for detection of β2-, NetB- and enterotoxin-encoding genes. Furthermore, C. difficile and C. perfringens isolates were evaluated for in vitro antimicrobial susceptibility. Results Seven of 153 (4.6%) samples, all from diarrhoeic foals, were positive for C. difficile A/B toxin. Of these, 5 of 14 (35.7%) were from hospitalised foals, and only 2 of 63 (3.2%) diarrhoeic foal samples were from farms (P = 0.002). Clostridium perfringens was isolated from 31 (20.3%) foals, of which 21 of 76 (27.6%) were diarrhoeic and 10 of 76 (13.2%) were nondiarrhoeic, demonstrating a difference between these 2 groups (P = 0.045). Only 4 strains were positive for the β2-encoding gene (cpb2). All C. difficile and C. perfringens isolates were susceptible to metronidazole and vancomycin. Conclusions The present report highlights the need for laboratory diagnostics to differentiate C. difficile-associated infection in foals from other causes of diarrhoea to facilitate adequate antimicrobial therapy. Potential relevance More studies are needed to clarify the role of C. perfringens as a primary agent of diarrhoea in foals.

Prototheca species have increasingly been reported to be opportunistic pathogens that cause mastitis in dairy herds, and it poses an emergent problem because at present, there are no effective therapies for the treatment of protothecal mastitis. This study investigated the in vitro algicidal effect of guanidine on 75 Prototheca zopfii genotype 2 strains isolated from 75 cases of clinical and subclinical bovine mastitis. All strains were susceptible to guanidine in vitro with minimal algaecide concentrations ranging from 0·001 to 0·035%. Guanidine is known to have a high microbicidal effect and is considered to be a new generation microbicidal compound. It is not toxic to human mucous membranes and conjunctivas at low concentrations and has been used as a disinfectant in swimming pools and as an antiseptic for human wounds. The algicidal action of guanidine at low concentrations indicates that it could be an alternative disinfectant or antiseptic for cleaning of the dairy environment and milking equipment, in pre- and postdipping solutions, in the chemical dry therapy of bovine teats and even in the intramammary therapy of P. zopfii infections. This is the first report of the in vitro algicidal effect of guanidine on P. zopfii strains of animal origin. Significance and Impact of the Study: Prototheca zopfii genotype 2 is an opportunistic pathogen of bovine mastitis. To date, no effective therapies against protothecal mastitis have been developed. The in vitro algicidal effect of guanidine on 75 P. zopfii genotype 2 strains isolated from cows revealed that all of the isolates were susceptible to the compound at low concentrations, which indicates that guanidine may be used as an antiseptic/disinfectant for dairy milking equipment, in pre- and postdipping solutions, and as a chemical dry therapy or an intramammary therapy. This study describes the in vitro algicidal effect of guanidine on P. zopfii for the first time. © 2017 The Society for Applied Microbiology

Tuberculosis remains as the world’s biggest threat. In 2014, human tuberculosis ranked as a major infectious disease by the first time, overcoming HIV death rates. Bovine tuberculosis is a chronic disease of global distribution that affects animals and can be transmitted to humans by the consumption of raw milk, representing a serious public health concern. Despite the efforts of different countries to control and eradicate bovine tuberculosis, the high negative economic impact on meat and milk production chains remains, given the decreased production efficiency (approximately 25%), the high number of condemned carcasses, and increased animal culling rates. This scenario has motivated the establishment of official programs based on regulations and diagnostic procedures. Although Mycobacterium tuberculosis and Mycobacterium bovis are the major pathogenic species to humans and bovines, respectively, nontuberculous mycobacteria within the Mycobacterium genus have become increasingly important in recent decades due to human infections, including the ones that occur in immunocompetent people. Diagnosis of mycobacteria can be performed by microbiological culture from tissue samples (lymph nodes, lungs) and secretions (sputum, milk). In general, these pathogens demand special nutrient requirements for isolation/growth, and the use of selective and rich culture media. Indeed, within these genera, mycobacteria are classified as either fast- or slow-growth microorganisms. Regarding the latter ones, incubation times can vary from 45 to 90 days. Although microbiological culture is still considered the gold standard method for diagnosis, molecular approaches have been increasingly used. We describe here an overview of the diagnosis of Mycobacterium species in bovine milk.

Submitted by Guilherme Lemeszenski (guilherme@nead.unesp.br) on 2013-08-22T18:41:33Z No. of bitstreams: 1 S1517-83822012000300029.pdf: 150600 bytes, checksum: 304a83b70e504b99d1843f16702e9731 (MD5) Made available in DSpace on 2013-08-22T18:41:33Z (GMT). No. of bitstreams: 1 S1517-83822012000300029.pdf: 150600 bytes, checksum: 304a83b70e504b99d1843f16702e9731 (MD5) Previous issue date: 2012-09-01 Made available in DSpace on 2013-09-30T19:28:08Z (GMT). No. of bitstreams: 2 S1517-83822012000300029.pdf: 150600 bytes, checksum: 304a83b70e504b99d1843f16702e9731 (MD5) S1517-83822012000300029.pdf.txt: 17954 bytes, checksum: 6616562b0a141977fd7a804b8c4a7e67 (MD5) Previous issue date: 2012-09-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:39:52Z No. of bitstreams: 2 S1517-83822012000300029.pdf: 150600 bytes, checksum: 304a83b70e504b99d1843f16702e9731 (MD5) S1517-83822012000300029.pdf.txt: 17954 bytes, checksum: 6616562b0a141977fd7a804b8c4a7e67 (MD5) Made available in DSpace on 2014-05-20T13:39:52Z (GMT). No. of bitstreams: 2 S1517-83822012000300029.pdf: 150600 bytes, checksum: 304a83b70e504b99d1843f16702e9731 (MD5) S1517-83822012000300029.pdf.txt: 17954 bytes, checksum: 6616562b0a141977fd7a804b8c4a7e67 (MD5) Previous issue date: 2012-09-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Two strains of Nocardia spp. were isolated from bovine milk of two individual bulk tank. Molecular identification classified the strains as Nocardia farcinica and Nocardia cyriacigeorgica. The thermorresistance to boiling of the isolates was carried out and was observed bacterial growth after boiling. Our findings indicate the potential risk of pathogen transmission to humans through contaminated milk with Nocardia spp. Universidade Estadual Paulista Faculdade de Medicina Veterinária Departamento de Higiene Veterinária e Saúde Pública Chiba University Medical Mycology Research Center Universidade Estadual Paulista Faculdade de Medicina Veterinária Departamento de Higiene Veterinária e Saúde Pública FAPESP: 10/53494-5 FAPESP: 09/56037-1

Virulence of Rhodococcus equi strains from 20 humans in Brazil was investigated by using a polymerase chain reaction to characterize isolates as virulent (VapA), intermediately virulent (VapB), and avirulent. Nine isolates were obtained from human immunodeficiency virus (HIV)-positive patients, six from HIV-negative patients, and five from patients of unknown status. Five isolates were VapB positive, four were VapA positive, and eleven were avirulent. Among the nine isolates from HIV-positive patients, five contained VapB plasmids and two contained VapA plasmids. Five VapB-positive isolates had the type 8 virulence plasmid. Eleven of the patients had a history of contact with livestock and/or a farm environment, and none had contact with pigs.

Made available in DSpace on 2019-10-06T16:16:34Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-01-01 Escherichia coli is the most common contaminant of the bovine uterus in the first few weeks postpartum, and one of the most important pathogens involved in uterine infections. This bacterium is characterised by diverse virulence factors (VF) however, the profile of E. coli VF in physiologic postpartum uterine contamination is not well established. Therefore, the present study aimed to investigate the prevalence of intrauterine bacteria, a set of virulence factors and phylogroups of E. coli isolates, leukogram and uterine cytology in 75 Holstein cows at 24 h (Sampling time 1) and 14 days (Sampling time 2) postpartum. Escherichia coli, α-haemolytic Streptococcus, Trueperella pyogenes, and Pasteurella multocida were the most common microorganisms isolated in pure culture, whereas E. coli and Clostridium perfringens Type A, E. coli and α-haemolytic Streptococcus, and E. coli and Proteus mirabilis were the most frequent microorganisms in mixed colonies at both studied sampling times. In the 59 E. coli isolates after 24 h (n = 35) and 14 days (n = 24) postpartum, the genes detected for VF were fimH, iucD/aer, kps, hlyA, usp, vt1 and vt2. Most E. coli strains detected in both moments of study belonged to Commensal phylogenetic groups A and B1, whereas Pathogenic groups D and B2 were identified at 24 h and 14 days postpartum respectively. All cows presented suitable immune response against the presence of bacteria in uterine lumen, observed by leukocytosis, neutrophilia, lymphocytosis and monocytosis at leukogram and a high number of polymorphonuclear leukocytes in uterine cytology, in both studied moments. Thus, a complex diversity of aerobic and anaerobic bacteria is involved in uterine contamination in the early postpartum of cows, besides the predominance of E. coli. Moreover, the genes fimH, iucD/aer, hlyA, kps, usp, vt1 and vt2 play a key role in the virulence of E. coli in this period. Department of Animal Reproduction and Veterinary Radiology School of Veterinary Medicine and Animal Science (FMVZ) São Paulo State University (UNESP), Prof. Doutor Walter Mauricio Correa Street Department of Genetics Evolution and Bioagents Institute of Biology University of Campinas, 255 Monteiro Lobato Street, Postal Address 6109 Department of Veterinary Clinics School of Veterinary Medicine and Animal Science (FMVZ) São Paulo State University (UNESP), Prof. Doutor Walter Mauricio Correa Street Department of Veterinary Hygiene and Public Health School of Veterinary Medicine and Animal Sciences (FMVZ) São Paulo State University (UNESP) Prof. Doutor Walter Mauricio Correa Street Rubião Junior Postal Address 560 Department of Animal Reproduction and Veterinary Radiology School of Veterinary Medicine and Animal Science (FMVZ) São Paulo State University (UNESP), Prof. Doutor Walter Mauricio Correa Street Department of Veterinary Clinics School of Veterinary Medicine and Animal Science (FMVZ) São Paulo State University (UNESP), Prof. Doutor Walter Mauricio Correa Street Department of Veterinary Hygiene and Public Health School of Veterinary Medicine and Animal Sciences (FMVZ) São Paulo State University (UNESP) Prof. Doutor Walter Mauricio Correa Street Rubião Junior Postal Address 560

The aim of this study was to compare the prevalence of virulence genes in 158 Escherichia coli strains isolated from 51 clinical cases of UTIs, 52 of pyometra and from 55 fecal samples from healthy dogs by PCR. papC was found in 12 (23.5%) strains isolated from UTIs, 19 (36.5%) from pyometra and 10 (18.2%) from feces. papGII was observed in 3 (5.8%) strains from pyometra, and papGIII in 10 (19.6%) from UTIs, 15 (28.8%) from pyometra and 9 (16.4%) from feces. sfaS was detected in 22 (43.1%) strains from UTIs, 24 (46.1%) from pyometra and 19 (34.5%) from feces. hlyA was observed in 17 (33.3%) strains from UTIs, 18 (34.6%) from pyometra and 7 (12.7%) from feces, while cnf-1 was detected in 11 (21.6%) from UTIs, 21 (40.4%) from pyometra and 9 (16.4%) from feces. iucD was observed in 12 (23.5%) strains from UTIs, 9 (17.3%) from pyometra and 1 (1.8%) from feces. usp was found 17 (33.3%) isolates from UTIs and 36 (69.9%) from pyometra.

Submitted by Guilherme Lemeszenski (guilherme@nead.unesp.br) on 2013-08-22T18:56:05Z No. of bitstreams: 1 S0102-09352008000200031.pdf: 199936 bytes, checksum: 9dcfd9480c09ffecebb4f743fef16ea4 (MD5) Made available in DSpace on 2013-08-22T18:56:05Z (GMT). No. of bitstreams: 1 S0102-09352008000200031.pdf: 199936 bytes, checksum: 9dcfd9480c09ffecebb4f743fef16ea4 (MD5) Previous issue date: 2008-04-01 Made available in DSpace on 2013-09-30T19:49:36Z (GMT). No. of bitstreams: 2 S0102-09352008000200031.pdf: 199936 bytes, checksum: 9dcfd9480c09ffecebb4f743fef16ea4 (MD5) S0102-09352008000200031.pdf.txt: 12684 bytes, checksum: bb933daa6147470698b4d06f3f14ef5f (MD5) Previous issue date: 2008-04-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:39:49Z No. of bitstreams: 2 S0102-09352008000200031.pdf: 199936 bytes, checksum: 9dcfd9480c09ffecebb4f743fef16ea4 (MD5) S0102-09352008000200031.pdf.txt: 12684 bytes, checksum: bb933daa6147470698b4d06f3f14ef5f (MD5) Made available in DSpace on 2014-05-20T13:39:49Z (GMT). No. of bitstreams: 2 S0102-09352008000200031.pdf: 199936 bytes, checksum: 9dcfd9480c09ffecebb4f743fef16ea4 (MD5) S0102-09352008000200031.pdf.txt: 12684 bytes, checksum: bb933daa6147470698b4d06f3f14ef5f (MD5) Previous issue date: 2008-04-01 Relata-se a ocorrência de graves sintomas de mastite hiperaguda em vaca, causada por Klebsiella pneumoniae, na terceira semana de lactação. Descrevem-se aspectos epidemiológicos, sintomas clínicos, procedimentos de diagnóstico microbiológico, conduta terapêutica e medidas de controle. UNESP Faculdade de Medicina Veterinária e Zootecnia UNESP Faculdade de Medicina Veterinária e Zootecnia

Salmonellosis is a foodborne disease caused by bacteria of the genus Salmonella, being pigs and pork-products potentially important for its occurrence. In recent decades, some serovars of Salmonella have shown increase of resistance to conventional antimicrobials used in human and animal therapy, with serious risks for public health. The aim of this study was to evaluate feces (n=50), mediastinal (n=50), mesenteric (n=50) and mandibular (n=50) lymph nodes obtained from slaughter houses for Salmonella spp. Positive samples were serotyped and subjected to an in vitro antimicrobial susceptibility test, including the extended-spectrum beta-lactamase (ESBL) production. Salmonella species were identified in 10% (20/200) of total samples. From these, 20% (10/50) were identified in the submandibular lymph nodes, 18% (9/50) in the mesenteric lymph nodes, 2% (1/50) in feces and 0% (0/50) in the mediastinal lymph nodes. The serotypes found were Salonella Typhimurium (55%), S. enterica subsp. enterica 4,5,12: i: - (35%), S. Brandenburg and S. Derby with 5% (5% each). All strains showed resistance to at least one antimicrobial; 90% were resistant to four or more antimicrobials, and 15% were multidrug-resistant. Resistance to ciprofloxacin, tetracycline and nalidixic acid was particularly prevalent amongst the tested serovars. Here, we highlighted the impact of pigs in the epidemiological chain of salmonellosis in domestic animals and humans, as well as the high antimicrobial resistance rates of Salmonella strains, reinforcing the necessity for responsible use of antimicrobials for animals as an emergent One Health issue, and to keep these drugs for human therapy approaches. RESUMO: Nas últimas décadas, o aumento de cepas circulante de Salmonella concomitantemente a resistência microbiana tem despertado a preocupação dos órgãos de Saúde Pública. Deste modo, o objetivo do presente trabalho foi pesquisar a presença de Salmonella a partir de fezes (n=50), linfonodos mediastinos (n=50), mesentéricos (n=50) e submandibular (n=50) oriundos de um abatedouro suíno. As cepas isoladas foram sorotipadas e testadas quanto a resistência antimicrobiana. A presença de Salmonella isolada foram em 10% (20/200) do total de amostras, sendo 20% dos linfonodos submandibulares, 18% dos linfonodos mesentéricos e 2% das fezes. Os sorotipos encontrados foram S. Typhimurium (55%), S. enterica subsp. enterica 4,5,12: i: - (35%), S. Brandenburg (5%) e S. Derby (5%). Todas a cepas apresentaram resistência a pelo menos um antimicrobiano testado, sendo 90% resistente pelo menos quatro antimicrobianos. Destes, 15% foram classificadas como multidrogas resistentes. Os antimicrobianos mais resistentes entre os sorovares isolados foram a ciprofloxacina, tetraciclina e o ácido nalidixico. A presença de cepas de Salmonella resistente a antimicrobianos na espécie suína tem gerado um grande impacto epidemiológico entre homem e animal, reforçando cada vez mais a necessidade do uso adequado de drogas principalmente relacionado com o tema “One Health”.

Made available in DSpace on 2015-10-21T13:09:49Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-07-09 Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) SR-2/UERJ Oswaldo Cruz Institute grants Many new, emerging and re-emerging diseases of humans are caused by pathogens which originate from animals or products of animal origin. Cotynebacterium lactis, a recently described species of the genus anynebacterium, was first isolated from milk of asymptomatic cows. In the present study a cutaneous abscess caused by C. lactis in a dog was recognized by cytologic and histologic examination in addition to 16S rRNA gene analysis of the microorganism. Therefore, C lactis should be included among other bacterial species recognized as emerging pathogens for companion animals. (C) 2015 Elsevier B.V. All rights reserved. Universidade Federal Rural Semi Arido UFERSA, Mossoro, RN, Brasil Universidade Estadual Paulista UNESP, Departamento Veterinária, Higiene &Saude Publica, Botucatu, SP, Brasil Faculdade de Medicina Veterinária e Zootecnia FAMED, Garça, SP, Brasil Fundacao Oswaldo Cruz, Inst Oswaldo Cruz IOC Fiocruz, Lab Genet Mol Microrganismos, Rio De Janeiro, RJ, Brazil Fundacao Oswaldo Cruz, Inst Nacl Controle Qualidade Saude INCQS Fiocruz, Rio De Janeiro, RJ, Brazil Universidade do Estado Rio de Janeiro, Faculdade de Ciencias Médicas FCM UERJ, Lab Difteria &Corinebacterias Importancia Clin L, Rio De Janeiro, RJ, Brasil Universidade Estadual Paulista (UNESP), Departamento Veterinária, Higiene &Saude Publica, Botucatu, SP, Brasil

Introduction: Bovine vaccinia virus (VACV) is a well‐known zoonotic agent related to exanthemous lesions in skin and mucous membranes of dairy cattle and humans, characterized by the formation of vesicles, pustules and ulcers. Mastitis is one of the most common infectious diseases of dairy herds. Bovine mammary infections are caused mainly by bacterial micro‐organisms, especially staphylococci. To the best of our knowledge, intramammary coinfection with VACV and Staphylococcus aureus in cows has not been reported previously. Case presentation: During an outbreak of exanthematic bovine VACV infection with animals showing vesicles, pustules and haemorrhagic ulcers on the teats, milk samples were collected for mastitis detection. Conclusion: The present report describes a case of intramammary coinfection by VACV and S. aureus in a bovine VACV outbreak.