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The performance of a commercial immunofluorescence assay (IFA commercial), an in-house immunofluorescence assay (IFA in-house) and an indirect enzyme-linked immunosorbent assay (ELISA) were evaluated in the detection of antibodies anti-C. burnetii in the serum of Q fever patients and persons without the disease. For the study, seropositive and seronegative samples for Q fever (n = 200) from a serum bank of the Instituto Adolfo Lutz in Brazil were used. Commercial IFA was considered in this study as the gold standard for diagnosing Q fever. The in-house IFA demonstrated good agreement with the commercial test, showing high sensitivity (91%) and specificity (97%) compared to the gold standard, with a Kappa coefficient of 0.8954. The indirect ELISA test showed lower agreement with the gold standard, showing low sensitivity (67%), although the specificity of the technique was high (97%) and the Kappa coefficient was moderate (0.6631). In-house IFA is an excellent alternative for diagnosing Q fever.

Trichosporonosis corresponds to a systemic fungal disease that leads to high mortality rates and is frequently associated with medical devices. It affects immunosuppressed patients in particular and is strongly linked to acquired human immunodeficiency, organ and tissue transplants, and malignant hematologic diseases such as leukemia and lymphomas. Trichosporon infections have been increasingly reported worldwide; however, little information is available either about their characteristics or the causative microorganism. Thus, the aims of the present study were: to investigate 59 yeasts of the genus Trichosporon by verifying the biofilm formation capacity of isolates; to analyze the susceptibility patterns of planktonic cells against the antifungals fluconazole, itraconazole, amphotericin-B, voriconazole, and caspofungin by comparing European Committee for Antimicrobial Susceptibility Testing (EUCAST) broth microdilution technique with the commercial method Etest; and to assess the susceptibility patterns of biofilm cells (sessile) against the same antifungals through broth microdilution. The ability to form biofilm on the surface of polystyrene plates was noted for all isolates, and 54.3% of samples were considered strong producers. Comparison between the antifungal susceptibility techniques evidenced that Etest showed higher and discordant minimum inhibitory concentrations (MICs) from those obtained by the microdilution method, especially for fluconazole, itraconazole, and caspofungin. Considering the susceptibility of biofilms, most species had high MIC50 and MIC90 against the tested antifungals, showing 4-to-66-fold higher concentrations for amphotericin B and 2-to-33-fold greater concentrations for caspofungin. These results highlight the importance of further studies with Trichosporon spp. for comparison between laboratory findings and in vivo response, considering both the susceptibility tests and the behavior of biofilm cells against drugs.

Made available in DSpace on 2022-04-28T19:49:05Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-12-01 Trichosporon spp. are widely distributed in the nature, comprising species that inhabit different ecological niches and can be found in the water, soil, and body surface of animals and humans. Such microorganisms have been classically associated with superficial infections; however, in the last decades, they have also been related to disseminated infections in immunocompromised patients, behaving as opportunistic agents, which demands rapid and accurate species identification for efficient therapy. Concordance level between the traditional phenotypic method and the molecular technique (gold standard) in the identification of all 59 Trichosporon samples was 59.3%. Identification concordance between MALDI-TOF spectrometry and the molecular technique was 71.2%. No isolate of environmental origin was identifiable by MALDI-TOF mass spectrometry (MS), and 100% of such environmental isolates were discordant for IGS region sequencing and phenotypic characterization. Both comparisons evidenced greatest concordance in the identification of T. asahii. The species T. debeurmannianum, T. dermatis, T. venhuisii and T. insectorum were not properly identified by both MALDI-TOF MS and the phenotypic technique. MALDI-TOF MS, in particular, seems to be appropriate to investigate yeasts of the genus Trichosporon; however, database updates are still necessary, especially for species that are not common in the clinical routine. With the aim of helping understand the aspects involved in early and accurate diagnosis of infections caused by this opportunistic agent, the present study compared the phenotypic, molecular (IGS region) and mass-spectrometry (MALDI-TOF) identification of 59 yeasts of the genus Trichosporon which had clinical and environmental origin and were kept in a mycology collection. São Paulo State University (Unesp) Institute of Biosciences, São Paulo (SP) Adolfo Lutz Institute (IAL) CLR II Biomedical Sciences Center, SP University of São Paulo (USP) School of Dentistry, SP Faculty of Medicine Federal University of Mato Grosso (UFMT), Av. Fernando Corrêa da Costa, 2.367, MT Irmandade da Santa Casa de Misericórdia de São Paulo Hospital, SP Darcy Vargas Children's Hospital, SP Bauru State Hospital (HEB), SP University of São Paulo (USP) Institute of Tropical Medicine, SP Adolfo Lutz Central Institute (IAL) Mycology Sector, SP São Paulo State University (Unesp) Institute of Biosciences, São Paulo (SP)

The aim of this study was to investigate the occurrence of fungi in dialysis water and dialysate, in addition to evaluating the susceptibility to antifungals and the biofilm production capacity of isolated microorganisms. The samples were collected in three hemodialysis units in Bauru (Brazil), every 15 days (July 2017–June 2018) at post-reverse osmosis, reuse, and dialysate points. The fungi were isolated by spread plate on Sabouraud dextrose agar. Filamentous fungi were phenotypically identified and yeasts were subjected to molecular evaluation of the ITS region. Susceptibility test to antifungals was carried out by the broth microdilution method and biofilm production capacity was evaluated in microtiter plates using crystal violet staining. Fungi were isolated in 52/216 (24.1%) samples, with an average count of 16.3 (10–40) CFU/mL. Overall, 61 microorganisms were identified, with 54 (88.5%) filamentous fungi and 7 (11.5%) yeasts. The main genera included were Penicillium, Cladosporium, Scedosporium, Rhinocladiella, Fusarium, and Emmonsia. Most isolates showed high values of minimum inhibitory concentration for 5-flucytosine and fluconazole and 35/45 (77.8%) isolates were classified as strong producers of biofilm. In order to increase the safety of the dialysis process, the adoption of control measures and monitoring of fungi in hemodialysis fluids is suggested. HIGHLIGHTS Fungi with pathogenic potential in hemodialysis fluids.; Fungi with low susceptibility to antifungals.; Fungi capable of producing biofilm.

Previous studies demonstrated that insecticide collars are highly effective in reducing canine visceral leishmaniasis (CVL); however, it is unclear if the efficacy differs by socioeconomic conditions across diverse communities. This study aimed fourfold: (i) to evaluate the protection of 4% impregnated deltamethrin collared (DMC) dogs in different areas of an endemic city for visceral leishmaniasis (VL); (ii) to analyze socioeconomic variables with the seroconversion rates; (iii) to analyze the indirect effect of DMC on untreated dogs in areas of intervention; and, (iv) to evaluate the potential transmission to other dogs in the same household when one positive dog is present. The study employed the municipality of Bauru, São Paulo, Brazil, as the area of interest and used Geographic Information System tools to fit binary logistic regression models. Dogs were divided into three cohort studies: intervention with DMC (I), indirect effect of DMC (IE), and control (C). Pre-intervention, lower mean income was associated with higher rates of CVL and a 142% increase in the odds of transmission (OR = 1.42, p-value = 0.001, CI 1.14,1.77). Post-intervention, lower-income areas depicted greater efficacy (76%) than higher-income areas (45%). The overall efficacy of DMC in preventing CVL was 63%; however, seroconversion rates were higher for IE (6.02%) than C (3.78%), revealing the failure of the indirect protection of DMC to manage the spread of the disease among the general non-wearing DMC canine population living in the same area. The protected dogs may repel the vectors, and non-protected dogs attract them, creating a higher transmission rate for non-protected dogs. Greater seroconversion was observed for living with an infected dog (10.20% in IE and 8.75% in C) than for the indirect effect of DMC, demonstrating the social burden of CVL. Overall, uncollared dogs have three times higher odds of being infected with CVL than DMC dogs (p 0.005), and uncollared dogs living with (an) infected dog(s) in the same household can reach 3.5 times higher odds than those living with negative ones (p 0.005). The results may assist in enhancing public policies and minimizing inequality in low and middle-income countries that suffer from neglected diseases such as VL.

Q fever and brucellosis are zoonoses that cause fever and other systemic clinical signs in humans; their occurrences are neglected and the differential diagnosis for some diseases is disregarded. This study aimed to investigate the seropositivity for Coxiella burnetii and Brucella spp. antibodies in patients suspected of dengue from 38 municipalities in the state of São Paulo, Brazil. The samples (n = 604) were obtained by convenience from the Adolfo Lutz Institute serum bank. Sera were subjected to an indirect immunofluorescence assay (IFA) using in-house and commercial diagnostic protocols to evaluate C. burnetii positivity. For Brucella spp., sera were subjected to rapid plate serum agglutination with buffered acidified antigen (AAT), slow tube serum agglutination (SAL), and 2-mercaptoethanol (2-ME) techniques. Associations and statistical inferences of the results were performed by logistic regression according to the clinical and demographic variables collected from the patients. Statistical analyses were performed using Statistical Analysis Software (SAS) and associations were considered when p value was

Made available in DSpace on 2022-04-29T08:31:24Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-08-06 Captive animals, despite the constant care provided, are susceptible to infections from different sources. We herein report the natural trypanosome infection of 11 (28.2% positive) out of 39 non-human primates from 13 different species, in a Brazilian zoological park. Immunofluorescent antibody test (IFAT) and conventional polymerase chain reaction (cPCR) ruled out Trypanosoma cruzi, the etiological agent of Chagas disease. However, sequencing performed with positive samples employing hsp70 primers revealed similarities from 86% to 88% to diverse trypanosomes, including T. cruzi, Trypanosoma grayi, Trypanosoma lewisi, Trypanosoma rangeli and Trypanosoma vivax. We believe that the low similarity values obtained by sequencing reflect the difficulties in the molecular identification of trypanosomes, which share a large portion of their genetic material; this similarity may also preclude the diagnosis of co-infection by more than one trypanosome species. Thus, our study demonstrates the presence of diverse trypanosomes in primates, which are susceptible to infection by these parasites. Mechanical devices such as windows and bed nets, etc., are required to avoid vector insects in these environments, in addition to preventive quarantining of animals recently introduced into zoos. Therefore, investigation of the parasites in both the animals already residing in the zoo and those being introduced is of paramount importance, although no easy task. Medical School São Paulo State University (UNESP) Graduate Course in Tropical Diseases, Botucatu Department of Preventive Veterinary Medicine and Animal Health Veterinary Medical School University of São Paulo (USP) Department of Animal Production and Preventive Veterinary Medicine School of Veterinary Medicine and Animal Science Unesp, Botucatu Regional Laboratories Center Adolfo Lutz Institute Department of Pediatrics Medical School Unesp, Botucatu Paulista Agency of Agribusiness Technology (APTA), Bauru Medical School São Paulo State University (UNESP) Graduate Course in Tropical Diseases, Botucatu Department of Animal Production and Preventive Veterinary Medicine School of Veterinary Medicine and Animal Science Unesp, Botucatu Department of Pediatrics Medical School Unesp, Botucatu

Made available in DSpace on 2021-07-14T10:54:05Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-06-28. Added 1 bitstream(s) on 2021-07-14T11:40:50Z : No. of bitstreams: 1 S1984-29612021000200319.pdf: 256672 bytes, checksum: 1da7ef90f5654d4c0380c4c41b98a3cd (MD5) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) A toxoplasmose está mundialmente distribuída e causa perdas na produção animal e problemas de saúde pública. Objetivou-se detectar Toxoplasma gondii e Sarcocystis spp. em 141 produtos cárneos de origem suína (49), bovina (48) e de quibe cru (44), comercializados em mercados de Botucatu, SP, Brazil. Realizou-se bioensaio das amostras em camundongos para isolamento do parasita, e detecção do DNA pela reação em cadeia pela polimerase, tendo como alvo a região do elemento repetitivo de 529 pares de bases (PCR-529-bp). Todas as amostras foram negativas ao bioensaio e 9 (6,38%) positivas à PCR, sendo 5/48 bovinas, 3/49 suínas e 1/44 quibe. Determinou-se a genotipagem das amostras positivas pela multiplex-, nested- e RFLP-PCR com 11 marcadores (SAG1, 5’-3’SAG2, alt.SAG2, SAG3, B-TUB, GRA6, L358, c22-8, c29-6, PK1, Apico). Obteve-se genótipo completo em uma amostra, semelhante a outros já identificados (MAS, TgCkBr89 e TgCkBr147). Nested- e RFLP-PCR do gene 18S rRNA das amostras positivas à PCR foram realizadas, e os produtos da nested-PCR, sequenciados e alinhados com dados do GenBank no NCBI. Quatro apresentaram 100% de homologia com T. gondii (L37415.1), duas Sarcocystis hominis (AF006471.1), duas Sarcocystis cruzi (AF176934.1), uma Sarcocystis hirsuta (AF006469.1), indicando a circulação de T. gondii e Sarcocystis spp. Toxoplasmosis occurs worldwide causing economic losses to the animal production and problems to the public health. The study aimed to detect Toxoplasma gondii and Sarcocystis spp.in 141 meat products from commercial meat cuts of pork, beef, and kibbeh sold in commercial markets from Botucatu, SP, Brazil. Samples were bioassayed in mice to isolate the parasite, and the parasite DNA detected by PCR targeting the 529 base pairs repeat element region (PCR-529-bp). All samples resulted negative on bioassay, whereas PCR positive for 9 (6,38%), distributed as 5/48 beef, 3/49 pork, and 1/44 kibbeh. PCR-positive were investigated for the the parasite genotype using multiplex-, nested-, and RFLP-PCR for 11 markers (SAG1, 5’-3’SAG2, alt.SAG2, SAG3, B-TUB, GRA6, L358, c22-8, c29-6, PK1, Apico). Complete genotype was determined on just one PCR-positive sample that matched MAS, TgCkBr89 and TgCkBr147 isolates already identified. In addition, nested- and RFLP-PCR targeting 18S rRNA was run for all PCR-positive samples and, the products, sequenced and aligned to the GenBank at NCBI website. Four samples showed 100% homology with T. gondii (GenBank #L37415.1), three with Sarcocystis hominis (GenBank #AF006471.1), two Sarcocystis cruzi (GenBank #AF176934.1), and one Sarcocystis hirsuta (GenBank #AF006469.1), indicating the circulation of T. gondii and Sarcocystis spp. Universidade Estadual Paulista, Faculdade de Medicina Veterinária e Zootecnia Instituto Adolfo Lutz, Laboratório Regional de Bauru Universidade do Oeste Paulista Universidade Estadual Paulista, Faculdade de Medicina Veterinária e Zootecnia CNPq: 559471/2010-6

Made available in DSpace on 2020-12-12T01:42:20Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-09-01 Water is indispensable for life and its quality is directly related to public health. The objective of this study was to investigate the presence of Pseudomonas aeruginosa in public water supply in municipalities in São Paulo State, Brazil. Analyses were carried out at the Adolfo Lutz Institute between February and December 2016, and included physicochemical (chlorine, pH, color, turbidity and nitrate), and microbiological parameters (total coliforms and Escherichia coli), as well as P. aeruginosa, with susceptibility tests to antimicrobial agents and biofilm production capacity by the strains isolated. In total, 251 water samples were evaluated and 19 (7.6%) presented P. aeruginosa. No significant differences were observed between the physicochemical parameters found in the positive and negative samples for this bacterium, but the samples containing total coliforms were also those with the highest positivity for P. aeruginosa. All samples with P. aeruginosa reported chlorine concentrations between 0.2 and 2.0 mg/L, as required by Brazilian legislation, demonstrating this bacterium’s resistance to conventional water treatment processes. Although not resistant to the antimicrobials tested, most strains isolated were classified as strong biofilm producers, emphasizing the need for further studies involving water supply quality. Instituto Adolfo Lutz Centro de Laboratório Regional de Bauru, Rua Rubens Arruda, Quadra 6, Centro Instituto Federal de Educação Ciência e Tecnologia do Triângulo Mineiro, Rua João Batista Ribeiro, 4000, Mercês Departamento de Microbiologia e Imunologia Universidade Estadual Paulista ‘Júlio de Mesquita Filho’ (UNESP) Instituto de Biociências, Caixa Postal 510, Distrito de Rubião Júnior Departamento de Microbiologia e Imunologia Universidade Estadual Paulista ‘Júlio de Mesquita Filho’ (UNESP) Instituto de Biociências, Caixa Postal 510, Distrito de Rubião Júnior

Made available in DSpace on 2019-10-04T12:31:28Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-10-18 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Leishmaniasis is a vector-borne parasitic protozoan infection that affects mammals and involves a complex epidemiology. Although dogs are considered the main reservoir in zoonotic visceral leishmaniasis (VL), the possible presence of other mammalian species acting as reservoirs has been associated as a possible cause of lack of success in the control of human VL in many endemic areas. The knowledge about natural infections of some species is still scarce, such as nonhuman primates (NHP), especially from the genus Callithrix (marmosets). We investigated the infection by Leishmania (Leishmania) infantum, the agent of VL in the Americas, in 26 marmosets captured monthly, from April 2014 to March 2015, in an environmentally protected area (EPA) in Southeastern Brazil. The EPA has undergone significant environmental changes and has a transmission focus of canine VL since 2009. Serology was performed through the direct agglutination test, which detected low antibody titers in seven marmosets (7/26; 26.9%, 95% confidence interval 9.9-44.0), being five Callithrix penicillata (black-tufted-ear marmoset) and two Callithrix jacchus (white-tufted-ear marmoset). The presence of the DNA of Leishmania was investigated in blood and skin samples by PCR and genetic sequencing. This is the first report of the detection of L. (L.) infantum in the skin of a marmoset, which was verified in a sample from one C. penicillata. The results demonstrate the natural infection of marmosets by L. (L.) infantum and may suggest the participation of these animals as hosts in the parasite's transmission cycle in the EPA. However, more comprehensive studies are needed to elucidate their role on the VL epidemiology in this area and also in different endemic areas, especially because these NHP are increasingly in contact with humans and domestic animals, particularly due to environmental changes. Univ Estadual Campinas, Dept Publ Hlth, Fac Med Sci, Campinas, SP, Brazil Adolfo Lutz Inst, Ctr Syst Parasit Dis, Sao Paulo, Brazil Adolfo Lutz Inst, Bauru Ctr Reg Labs 2, Bauru, Brazil Sao Paulo State Univ, Sch Vet Med & Anim Sci, Dept Vet Hyg & Publ Hlth, Botucatu, SP, Brazil Sao Paulo State Univ, Sch Vet Med & Anim Sci, Dept Vet Hyg & Publ Hlth, Botucatu, SP, Brazil FAPESP: 2014/27212-0 FAPESP: 2014/13049-0 FAPESP: 2016/02572-0

A leptospirose é uma zoonose de grande importância em saúde pública. Cento e quatro (104) amostras de urina de bovinos abatidos na região centro-oeste do Estado de São Paulo, Brasil, foram avaliadas para detectar Leptospira spp. As amostras foram mantidas em PBS e em culturas de EMJH e Fletcher. Observamos crescimento de microrganismos em 13 amostras do meio Fletcher, embora não tenha sido observado no meio EMJH. Por PCR, 25 animais (amostras) foram positivos com os primers LEP1 / LEP2, sendo que 16 amostras foram mantidas em PBS, seis em meio Fletcher e três em meio EMJH. As 25 amostras foram submetidas a PCR com o gene LipL32 para verificação da patogenicidade. Uma amostra amplificada e por sequenciamento resultou em 99,77% de similaridade com Leptospira kirschneri sorovar Canicola e Leptospira interrogans sorovar Pyrogenes. Este é o primeiro relato de isolamento de Leptospira kirschneri sorovar Canicola e Leptospira interrogans sorovar Pyrogenes em amostra de urina bovina na Região Centro-Oeste do estado de São Paulo, Brasil. Concluímos a importância da leptospirose no rebanho avaliado e a atenção para o risco ocupacional, devido à possibilidade de urina contaminada nas pastagens e disseminação da infecção para outros animais e pessoas contactantes.

Made available in DSpace on 2021-06-25T12:42:01Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-03-26. Added 1 bitstream(s) on 2021-07-15T15:24:36Z : No. of bitstreams: 1 S1678-91992021000100502.pdf: 1071475 bytes, checksum: 5ffc20edee55763e04682223a0062bd3 (MD5) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Background: The early symptoms of leptospirosis and dengue fever are difficult to distinguish and can cause diagnostic confusion. Due to the large dengue epidemics that has occurred in Brazil in recent years, it is possible that cases of leptospirosis were unreported. Therefore, we performed a retrospective study to detect leptospirosis in patients who were tested for dengue, but whose laboratory diagnoses were negative. Methods: Sera samples from 2,017 patients from 48 cities located in the central region of Sao Paulo state, Brazil, were studied. All samples were subjected to the microscopic agglutination test (MAT), 305 of which were taken from patients five days or less since the onset of symptoms, and were additionally subjected to real-time polymerase chain reaction (PCR). Results: The overall prevalence of leptospirosis cases was 21 (1.04%), with 20 through MAT (18 for Icterohaemorrhagiae and two for the Cynopteri serogroup) and one through PCR (amplicon sequencing compatible with Leptospira interrogans). According to previously established criteria, eight cases of leptospirosis were classified as confirmed and 13 as probable. The Brazilian notification system for health surveillance had no records for 16 patients positive for leptospirosis and, thus, they were considered unreported cases. Statistical analyses revealed that the prevalence of leptospirosis was higher in men (1.56%) than in women (0.56%), and the mean age was higher in positive patients (43.7 years) than in negative ones (32.3 years). Conclusion: The results indicated that patients suspected of dengue fever had evidence of leptospirosis or Leptospira infection, and most of these cases were unreported in the Brazilian notification system. The high burden of dengue may contribute to the misdiagnosis of leptospirosis, and health professionals should increase their awareness of leptospirosis as an important differential diagnosis of patients with suspicion of dengue. Sao Paulo State Univ UNESP, Sch Vet Med & Anim Husb, Dept Anim Prod & Prevent Vet Med, Botucatu, SP, Brazil Adolfo Lutz Inst, Ctr Reg Lab Bauru 2, Bauru, SP, Brazil Butantan Inst, Lab Appl Toxinol, Sao Paulo, SP, Brazil Sao Paulo State Univ UNESP, Sch Vet Med & Anim Husb, Dept Anim Prod & Prevent Vet Med, Botucatu, SP, Brazil FAPESP: 2015/06446-6

The development of new approaches that allow early assessment of which cases of COVID-19 will likely become critical and the discovery of new therapeutic targets are urgent demands. In this cohort study, we performed proteomic and laboratorial profiling of plasma from 163 patients admitted to Bauru State Hospital (Bauru, SP, Brazil) between May 4th and July 4th, 2020, who were diagnosed with COVID-19 by RT-PCR nasopharyngeal swab samples. Plasma samples were collected upon admission for routine laboratory analyses and shotgun quantitative label-free proteomics. Based on the course of the disease, the patients were further divided into 3 groups: a) mild symptoms, discharged without admission to an intensive care unit (ICU) (n=76); b) severe symptoms, discharged after admission to an ICU (n=56); c) critical, died after admission to an ICU (n=31). White cells and neutrophils were significantly higher in severe and critical patients compared to mild ones. Lymphocytes were significantly lower in critical patients compared to mild ones and platelets were significantly lower in critical patients compared to mild and severe ones. Ferritin, TGO, urea and creatinine were significantly higher in critical patients compared to mild and severe ones. Albumin, CPK, LDH and D-dimer were significantly higher in severe and critical patients compared to mild ones. PCR was significantly higher in severe patients compared to mild ones. Proteomic analysis revealed marked changes between the groups in plasma proteins related to complement activation, blood coagulation, antimicrobial humoral response, acute inflammatory response, and endopeptidase inhibitor activity. Higher levels of IREB2, GELS, POLR3D, PON1 and ULBP6 upon admission to hospital were found in patients with mild symptoms, while higher levels of Gal-10 were found in critical and severe patients. This needs to be validated in further studies. If confirmed, pathways involving these proteins might be potential new therapeutic targets for COVID-19.

Made available in DSpace on 2022-04-29T08:32:40Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-08-01 When it comes to visceral leishmaniasis (VL) in Brazil, one of the main targets of public health policies of surveillance is the control of domestic canine reservoirs of Leishmania infantum. This paper aims to evaluate the effect of the dog population and household environment for the maintenance of natural foci in the transmission to human and animal hosts in an endemic city for VL, Bauru, in Brazil. We collected 6, 578 blood samples of dogs living in 3, 916 households from Nov.2019 to Mar.2020 and applied geospatial models to predict the disease risk based on the canine population. We used Kernel density estimation, cluster analysis, geostatistics, and Generalized Additive Models (GAM). To validate our models, we used cross-validation and created a receiver operating characteristic (ROC) curve. We found an overall canine VL (CVL) seroprevalence of 5.6% for the sampled dogs, while for the households, the positivity rate was 8.7%. Odds ratios (OR) for CVL increased progressively according to the number of canines for >2 dogs (OR 2.70); households that already had CVL in the past increased the chances for CVL currently (OR 2.73); and the cases of CVL increase the chances for human VL cases (OR 1.16). Our models were statistically significant and demonstrated a spatial association between canine and human disease cases, mainly in VL foci that remain endemic. Although the Kernel density ratio map had the best performance (AUC = 82), all the models showed high risk in the city's northwest area. Canine population dynamics must be considered in public policies, and geospatial methods may help target priority areas and planning VL surveillance in low and middle-income countries. Parasitology and Mycology Center Adolfo Lutz Institute (IAL) Sao Paulo Adolfo Lutz Institute Regional Laboratories Center II Bauru, Bauru Center for Zoonoses Control of Bauru Health Secretariat of Bauru Bioterium Nucleos Adolfo Lutz Institute (IAL) Sao Paulo Institute of Geography Federal University of Uberlándia, Uberlándia Department of Geography Sao Paulo State University Faculty of Sciences and Technology (FCT/UNESP) Presidente Prudente Department of Geography Sao Paulo State University Faculty of Sciences and Technology (FCT/UNESP) Presidente Prudente

Made available in DSpace on 2020-12-10T20:01:08Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-05-29. Added 1 bitstream(s) on 2021-07-15T15:25:05Z : No. of bitstreams: 1 S1678-91992020000100314.pdf: 5541148 bytes, checksum: 0557e47fe298c3fd72e9ef0243fc3ca6 (MD5) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Background: Extracellular vesicles (EVs) are small membrane-bound vesicles of growing interest in vetetinary parasitology. The aim of the present report was to provide the first isolation, quantification and protein characterization of EVs from buffalo (Bubalus bubalis) sera infected with Theileria spp. Methods: Infected animals were identified through optical microscopy and PCR. EVs were isolated from buffalo sera by size-exclusion chromatography and characterized using western blotting analysis, nanoparticle tracking analysis and transmission electron microscopy. Subsequently, the proteins from isolated vesicles were characterized by mass spectrometry. Results: EVs from buffalo sera have shown sizes in the 124-140 nm range and 306 proteins were characterized. The protein-protein interaction analysis has evidenced biological processes and molecular function associated with signal transduction, binding, regulation of metabolic processes, transport, catalytic activity and response to acute stress. Five proteins have been shown to be differentially expressed between the control group and that infected with Theileria spp., all acting in the oxidative stress pathway. Conclusions: EVs from buffaloes infected with Theileria spp. were successfully isolated and characterized. This is an advance in the knowledge of host-parasite relationship that contributes to the understanding of host immune response and theileriosis evasion mechanisms. These findings may pave the way for searching new EVs candidate-markers for a better production of safe biological products derived from buffaloes. Sao Paulo State Univ UNESP, Botucatu Med Sch FMB, Grad Program Trop Dis, Botucatu, SP, Brazil Fed Univ Sao Carlos UFSCar, Dept Physiol Sci, Lab Biochem & Mol Biol, Sao Carlos, SP, Brazil Univ Zagreb, Clin Internal Dis, ERA Chair Team VetMedZg, Fac Vet Med, Zagreb, Croatia Univ Campinas UNICAMP, Inst Biol, Dept Biochem & Tissue Biol, Campinas, SP, Brazil Adolfo Lutz Inst, Ctr Reg Labs 2, Bauru, SP, Brazil Paulista Agcy Agribusiness Technol APTA, Bauru, SP, Brazil Univ Glasgow, Inst Biodivers Anim Hlth & Comparat Med, Glasgow, Lanark, Scotland Sao Paulo State Univ UNESP, Grad Program Clin Res, Botucatu Med Sch FMB, Botucatu, SP, Brazil Sao Paulo State Univ UNESP, Ctr Study Venoms & Venomous Anim CEVAP, Botucatu, SP, Brazil Sao Paulo State Univ UNESP, Botucatu Med Sch FMB, Grad Program Trop Dis, Botucatu, SP, Brazil Sao Paulo State Univ UNESP, Grad Program Clin Res, Botucatu Med Sch FMB, Botucatu, SP, Brazil Sao Paulo State Univ UNESP, Ctr Study Venoms & Venomous Anim CEVAP, Botucatu, SP, Brazil FAPESP: 2014/13299-7 CNPq: 458919/2014-4 CNPq: 563582/2010-3 CAPES: 23038.006285/2011-21 CAPES: 23038.008557/2010 CNPq: 441463/2019-3

Some animals have an important relationship with fungal infections, and searching for pathogens in animal samples may be an opportunity for eco-epidemiological research. Since studies involving wildlife are generally restricted, using samples from road kills is an alternative. The aim of this study was to verify whether pathogenic fungi of public health importance occur in wildlife road kills from Santa Catarina State, Brazil. Organ samples (n = 1063) from 297 animals were analysed according to Polymerase Chain Reaction (PCR) using universal primers to detect fungi in general and, subsequently, using primers specific to Paracoccidioides brasiliensis, Histoplasma capsulatum and Cryptococcus spp. There were 102 samples positive for fungal species. Eight samples were positive for P. brasiliensis, three samples were positive for Cryptococcus spp. and one sample had coinfection by these two fungi. No sample was positive for Histoplasma spp. according to the molecular detection. Genetic sequencing allowed the identification of Fungal sp. in 89 samples, Cryptococcus neoformans in two samples and Aspergillus penicillioides in three samples. This study shows the importance of wild animals in the epidemiology of fungal infections and assists in the mapping of pathogen occurrence in a region that was not previously evaluated.

Made available in DSpace on 2018-12-11T17:18:37Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-03-21 Fusarium species have emerged as responsible for a broad spectrum of infections, including superficial, locally invasive and disseminated ones, especially in the hospital environment. Since there are few reports of invasive and disseminated fusariosis in children, the aim of this study was to report four cases of nosocomial infection caused by this microorganism in children with cancer hospitalized in a public children’s hospital located in Brazil. Two of these patients were female and two were male. All patients presented febrile neutropenia, while three patients had acute lymphocytic leukemia and one patient had Wilms’ tumor as underlying disease. In two cases, fungi were isolated from blood and identified as Fusarium oxysporum species complex after phenotypic and genotypic studies, while in two other cases fungi were isolated from skin biopsies and identified as Fusarium solani species complex. One patient died 12 days after the onset of cutaneous lesions. All isolates, after susceptibility testing, presented high levels of minimum inhibitory concentration for itraconazole, voriconazole and amphotericin B. Considering the emergence of filamentous fungi as etiologic agents of nosocomial infections, health professionals should be aware of the problems these infections, especially fungal ones, may cause to debilitated patients. Hospital Infantil Darcy Vargas Faculdade de Odontologia Universidade de São Paulo (USP) Departamento de Farmácia Universidade Estadual do Centro-Oeste do Paraná (UNICENTRO) Secção de Micologia Instituto Adolfo Lutz (IAL) Núcleo de Ciências Biomédicas Instituto Adolfo Lutz (IAL) CLR II Departamento de Microbiologia e Imunologia Universidade Estadual Paulista (UNESP) Instituto Adolfo Lutz - Rua Rubens Arruda, s/n, quadra 06, Centro Departamento de Microbiologia e Imunologia Universidade Estadual Paulista (UNESP)

Hemoplasma infections are emerging and wild fauna can represent an important reservoir of these pathogens. However, there are very few epidemiological studies about the occurrence of hemoplasmas in wild cats around the world. The purpose of this study is twofold: (1) evaluate the occurrence and phylogeny of hemoplasmas in captive wild felines at a zoo in the state of Paraná, Brazil, and (2) verify the correlation between subpopulations of these bacteria and the hematological and biochemical parameters of the animals. PCR was used to detect hemoplasmas in the blood of three cougars (Puma concolor), a jaguar (Panthera onca), a tiger (Panthera tigris) and a lion (Panthera leo), followed by sequencing and phylogenetic analysis. The cougars and jaguar were found to be hemoplasma-positive by PCR. The phylogenetic analysis of the 16S rRNA gene sequences enabled the identification of genotypes of ‘Candidatus Mycoplasma haemominutum’ circulating in this zoo. The identified sequences were closely related to hemoplasma sequences originating from domestic cats and other wild cats, but the infected cougars and jaguar were healthy and showed no hematological or biochemical changes. It was concluded that P. concolor and P. onca are exposed to ‘Candidatus Mycoplasma haemominutum’ in Paraná, but further research is suggested to assess the resistance of wild cats to different hemoplasma subpopulations.

Leishmaniasis affect millions of people, causing morbidity and mortality, especially in developing tropical and subtropical countries. Unfortunately, the possibilities of treatment for these infections are still quite limited and most of the available drugs present serious side effects. The objective of this paper was to evaluate the therapeutic role of amiodarone and itraconazole in the treatment of cutaneous leishmaniasis caused by Leishmania (Leishmania) amazonensis . In order to perform this evaluation, hamsters were infected with 1 × 10 6 metaciclic promastigotes of the parasite in the hind footpad and, after the onset of the lesions, were treated with glucantime, amiodarone, itraconazole, glucantime and amiodarone, glucantime and itraconazole or amiodarone and itraconazole. The treatments' efficacy was evaluated per analysis of the size of the cutaneous lesions and by parasitic investigation of the infected foot (by histopathological examination and PCR) and possible side effects were analyzed taking into account the weight of the animals and some biochemical and metabolic parameters (glucose, urea, creatinine, AST, ALT and ALP). The results have shown that, in hamsters, amiodarone and itraconazole, either used isolated or in combination, are unable to stop the development of cutaneous lesions caused by L. (L.) amazonensis , but improve the activity of glucantime in the treatment of these lesions and seem to present no evident side effects. More studies are necessary in order to investigate the clinical potential of these combinations, so there can be the possibility of broadening the therapeutic options available, especially in resistant cases.

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

Made available in DSpace on 2018-12-11T17:07:44Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-11-01 Fundação Oswaldo Cruz Background: Canine visceral leishmaniasis (CVL) is a worldwide parasitic zoonosis caused by Leishmania (Leishmania) infantum around the world. Canids are the definitive hosts and sand flies the intermediate hosts. Objective: To test the hypothesis that a new species-specific primers (Lch14:Lch15, targeting a multiple alignment for L. infantum kDNA minicircle) is an efficient diagnostic tool for L. infantum. Methods: The presence of L. infantum DNA was assessed in blood samples of 69 stray dogs using the conventional PCR (cPCR) and quantitative PCR (qPCR). Additional 50 lymph nodes and 50 bone marrow samples (positive and negative samples for parasitological tests) from dogs from endemic and nonendemic areas for CVL were also used. Results: L. infantum strains, and all positive lymph node and bone marrow samples for parasitological test gave positive results for cPCR and qPCR, presenting analytical sensitivity of ∼10° parasite mL−1. For the blood samples, 40/69 (58%; CI 95%; 46%-69%) resulted positive for L. infantum in both tests. All positive samples were confirmed by sequencing. Conclusion: This study showed the importance of the specific detection of L. infantum based on species-specific primers by molecular techniques, highlighting the application as a confirmation method in epidemiological studies and to adopt the best control measures. Department of Pathobiology and Population Medicine College of Veterinary Medicine Mississippi State University Mississippi State Regional Laboratory of Bauru Adolfo Lutz Institute Institute of Collective Health Federal University of Bahia Department of Veterinary Hygiene and Public Health School of Veterinary Medicine and Animal Science UNESP Universidade Estadual Paulista Department of Veterinary Hygiene and Public Health School of Veterinary Medicine and Animal Science UNESP Universidade Estadual Paulista

Cows are probably the main source of contamination of raw milk with Staphylococcus aureus. Mammary glands with subclinical mastitis can shed large numbers of Staph. aureus in milk. Because of the risk of this pathogen to human health as well as animal health, the aim of this paper was to describe an outbreak of mastitis caused by methicillin-resistant Staph. aureus (MRSA), oxacillin-susceptible mecA-positive Staph. aureus (OS-MRSA), and methicillin-susceptible Staph. aureus (MSSA) on a dairy farm. Milk samples were obtained from all quarters, showing an elevated somatic cell count by the California Mastitis Test. The isolates were identified by phenotypic and genotypic methods. Staphylococcus spp. were isolated from 53% (61/115) of the milk samples, with 60 isolates identified as Staph. aureus (98.4%) and 1 isolate identified as Staphylococcus epidermidis (1.6%). The presence of the mecA gene was verified in 48.3% of Staph. aureus isolates. Of the Staph. aureus isolates, 23.3% were MRSA and 25.0% were OS-MRSA. The total of mastitis cases infected with MRSA was 12.2%. The detection of this large percentage of mastitis cases caused by MRSA and OS-MRSA is of great concern for the animals' health, because β-lactams are still the most important antimicrobials used to treat mastitis. In addition, Staph. aureus isolates causing bovine mastitis represent a public health risk.

We evaluated 345 wild animals from southern and south-eastern Brazil to understand their role in vaccinia virus (VACV) transmission cycle. VACV DNA was detected in rodents, marsupials, chiroptera and cingulate, expanding the knowledge of VACV host range in wildlife that could potentially act as source of infection in rural and urban areas.

Made available in DSpace on 2019-10-04T12:40:28Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-08-05 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) This is the first report of the yeast Apiotrichum veenhuisii (formerly Trichosporon veenhuisii) causing disease in humans; its virulence and in vitro behavior against antifungals were also studied. The sample was isolated from biopsy fragments of disseminated lesions on the skin of a pediatric patient with acute myeloid leukemia. The studied virulence factors evidenced that the strain tested negative for secretion of the enzymes proteinase, phospholipase, and hemolysin. The isolate was characterized as low biofilm producer. Except for amphotericin B and voriconazole, the sample presented high minimum inhibitory concentration values against azole and echinocandins. Adolfo Lutz Inst, CLR 2, Nucleo Ciencias Biomed, Bauru, SP, Brazil Univ Sao Paulo, Fac Odontol, Sao Paulo, SP, Brazil Hosp Infantil Darcy Vargas, Sao Paulo, SP, Brazil Univ Estadual Paulista, Dept Microbiol & Imunol, Botucatu, SP, Brazil Univ Sao Paulo, Fac Med, Sao Paulo, SP, Brazil Adolfo Lutz Inst, Nucleo Ciencias Quim & Bromatol, Bauru, SP, Brazil Univ Estadual Ctr Oeste Parana, Guarapuava, Parana, Brazil Univ Estadual Paulista, Dept Microbiol & Imunol, Botucatu, SP, Brazil

Made available in DSpace on 2020-12-12T02:30:13Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-03-01 Bats are essential to the global ecosystem, but their ability to harbour a range of pathogens has been widely discussed, as well as their role in the emergence and re-emergence of infectious diseases. This paper describes the first report of coinfection by two zoonotic agents, rabies virus (RABV) and the fungus Histoplasma suramericanum in a bat. The bat was from the Molossus molossus species, and it was found during the daytime in the hallway of a public psychiatric hospital in a municipality in São Paulo State, southeastern Brazil. RABV infection was diagnosed by the direct fluorescent antibody test and mouse inoculation test. The fungus was isolated by in vitro culture. Both diagnoses were confirmed by molecular techniques. Phylogenetic analysis showed that the fungus isolate had proximity to H. suramericanum in the Lam B clade, while the RABV isolate was characterized in the Lasiurus cinereus lineage. Since the M. molossus bat was found in a peri-urban transition area (urban/peri-urban), the possibility of cross-species transmission of this RABV lineage becomes more plausible, considering that this scenario may provide shelter for both M. molossus and L. cinereus. These are relevant findings since there has been an increase in bat populations in urban and peri-urban areas, particularly due to environmental modifications and anthropogenic impacts on their habitat. Thus, the detection of two zoonotic agents in a bat found in a public hospital should raise awareness regarding the importance of systematic surveillance actions directed towards bats in urban areas. São Paulo State University (UNESP) Municipal Health Department State University of Campinas (UNICAMP) Oswaldo Cruz Foundation Adolfo Lutz Institute Center of Regional Laboratories II São Paulo State University (UNESP)

Made available in DSpace on 2019-10-06T16:18:21Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-06-01 Leishmaniasis is considered a parasitic disease that still causes serious consequences for mankind, because it presents a high mortality rate worldwide. Considered multi-hosts, the parasites of the genus Leishmania are able of infecting a wide variety of animal species. The dog was considered the main source of infection of visceral leishmaniasis (VL), in the urban area. However, the role of other animal species in the epidemiological cycle of the disease, such as cattle, remains unclear. Therefore, the aim of the present study was to evaluate the occurrence of Leishmania spp. in 100 bovines (Bos taurus) from an area endemic for canine VL, using blood culture and molecular analysis. By the sequencing analysis, one sample showed 100% similarity with Leishmania infantum. The results provide the first case of L. infantum isolation in one bovine from the periurban areas of Bauru, state of São Paulo, Brazil. Department of Tropical Diseases Botucatu Medical School São Paulo State University - UNESP Department of Veterinary Hygiene and Public Health São Paulo State University - UNESP Department of Preventive Medicine and Animal Health University of Sao Paulo - USP Adolfo Lutz Institute Bauru II Regional Laboratory Center Paulista Agency of Agribusiness Technology APTA Department of Tropical Diseases Botucatu Medical School São Paulo State University - UNESP Department of Veterinary Hygiene and Public Health São Paulo State University - UNESP

Made available in DSpace on 2019-10-06T15:57:08Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-11-01 Bats are considered to play a significant role in the epidemiology of histoplasmosis, worldwide. We investigated the occurrence of H. capsulatum in lung samples from 89 bats, from urban areas in Southeastern Brazil, using nested PCR based on ribosomal DNA. Fungal DNA was detected in 31/89 samples (34.8%), of which 13/31 were Molossids (41.9%), 4/31 Eumops spp. (12.9%), 2/31 Artibeus lituratus (6.5%), and 12/31 others (38.7%). This is the first report of natural infection by H. capsulatum in A. lituratus in Southeastern Brazil, which reinforces the importance of these synanthropic animals in the epidemiology of histoplasmosis in urban areas. School of Veterinary Medicine and Animal Science UNESP Univ Estadual Paulista Department of Veterinary Hygiene and Public Health Department of Pathobiology and Population Medicine College of Veterinary Medicine Mississippi State University Institute of Biosciences UNESP Univ Estadual Paulista Department of Microbiology and Immunology Department of Public Health School of Medical Sciences State University of Campinas - UNICAMP Adolfo Lutz Institute Regional Laboratory of Bauru School of Veterinary Medicine and Animal Science UNESP Univ Estadual Paulista Department of Veterinary Hygiene and Public Health Institute of Biosciences UNESP Univ Estadual Paulista Department of Microbiology and Immunology

There is a growing concern about the participation of wild hosts and reservoirs in the epidemiology of leishmaniasis, particularly within the context of increasingly frequent environmental changes and the expansion of the One Health concept. This work is a molecular research of infection by Leishmania spp. among the wildlife of an environmentally protected area located in the municipality of Campinas, São Paulo, Brazil. The studied area has a history of intense environmental changes, with notifications of human cases of cutaneous leishmaniasis in the 1990s, and a focus of canine visceral leishmaniasis since 2009. Eighty-two wild mammals were sampled by monthly captures in this region over a 1-year period. Blood samples were collected from each animal and subjected to DNA extraction and PCR using primers for the region of the internal transcribed spacer-1. The results of gene sequencing for the first time revealed the infection of opossums (Didelphis albiventris) by Leishmania spp., subgenera Leishmania and Viannia, in Campinas. These findings, in addition to environmental and historical characteristics of the studied area, indicate a possible role of wildlife in the introduction and/or maintenance of natural foci of leishmaniasis transmission.

Made available in DSpace on 2018-12-11T17:15:32Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-11-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Objective: To analyze the prevalence of feline leishmaniosis in 55 asymptomatic cats or assess clinical disease in a visceral leishmaniasis-endemic area using parasitological, serological and molecular techniques. Methods: Fifty-five adult domestic and non-purebred cats held at an animal shelter in a rural area of the municipality of Ilha Solteira, São Paulo, Brazil, a region endemic for canine visceral leishmaniosis, were evaluated. A total of 5 mL of blood was collected from each animal, for blood culture, serological (indirect fluorescent antibody test, soluble extract ELISA, ELISA recombinant K39 antigen) and molecular tests (PCR). At the time of blood collection, the cats underwent clinical evaluation for the presence of clinical signs. Results: Flagellate protozoa were found in nine blood cultures (16.4%). Seropositivity by indirect fluorescent antibody test was observed in 32 serum samples (62.7%). ELISA-SE showed seropositivity in 37 cats (72.5%), while ELISA-rK39 revealed positive results in 11 cats (21.6%). Leishmania spp. was detected by PCR in whole blood samples from five cats (9.1%). Based on clinical evaluation, it was possible to detect the presence of clinical signs (alopecia, emaciation, pinna lesions, nose lesions, skin lesions) in 30 animals (54.5%). Conclusions: To the knowledge of the authors, this is the first report of Leishmania spp. in domestic cats in the epidemiological cycle of leishmaniosis in the studied area, requiring more attention in the establishment of preventive steps to control this disease. Department of Tropical Diseases Botucatu Medical School São Paulo State University - UNESP Department of Preventive Medicine and Animal Health Faculty of Veterinary Medicine and Animal Husbandry USP-Sao Paulo Department of Microbiology and Immunology São Paulo State University UNESP - Botucatu Department of Biology and Animal Husbandry São Paulo State University UNESP Adolfo Lutz Institute Bauru II Regional Laboratory Center Paulista Agency of Agribusiness Technology APTA Department of Tropical Diseases Botucatu Medical School São Paulo State University - UNESP Department of Microbiology and Immunology São Paulo State University UNESP - Botucatu Department of Biology and Animal Husbandry São Paulo State University UNESP FAPESP: 2010/13602-0

Epidemiological studies on endemic mycosis can be improved using molecular biology techniques to elucidate the role of bats as reservoirs and vectors of pathogenic fungi for infection of other animals and humans. The objective of this study was to explore the presence of Histoplasma capsulatum, Cryptococcus spp. and Paracoccidioides brasiliensis in insectivorous, frugivorous and nectarivorous bats collected in urban areas. We analysed 172 bats collected by the Epidemiological Surveillance Agency in 12 municipalities of the Midwest region of the state of Sao Paulo, Brazil. Spleen, liver, intestine and lung samples were subjected to microbiological culture and nested PCR analyses. Prevalence of H. capsulatum infection was 8.1% (14/172), with one bat found to be positive by fungal culturing, 12 positive by nested PCR and one positive by both methods. Two insectivorous bats were found positive by nested PCR for Cryptococcus spp., one in the spleen and the other in the spleen and lung. Two insectivorous bats showed natural infection by P. brasiliensis, in the spleen of one bat and the spleen and liver of the other. Our results reinforce the importance of bats as fungal dispersers in urban environments and the importance of constant epidemiologic surveillance because these synanthropic animals are in close contact with humans and animals.

Made available in DSpace on 2018-12-11T17:09:20Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-05-01 Bats have aroused growing attention in the public health sphere because they are considered the main reservoir of rabies virus (RABV) in the Americas, in places where canine rabies is under control. Antigenic and genetic studies of RABV isolates have been used to describe the epidemiological profile of rabies and to identify possible hosts/reservoirs for different epidemiological cycles. This study describes the antigenic and genotypic characterization of 19 RABV isolates from central nervous system samples of non-hematophagous bats (Mammalia: Chiroptera). These bats were diagnosed as RABV positive by direct fluorescent antibody and mouse inoculation tests. Antigenic characterization using a panel of eight monoclonal antibodies revealed that 7 of 19 RABV isolates from these bats belonged to variant 3, for which the hematophagous bat species Desmodus rotundus is the main reservoir, and 1 of 19 RABV isolates from an insectivorous bat belonged to variant 4, which is characteristic of these bats. The remaining 11 RABV samples were divided into six non-compatible profiles. The isolates were subjected to reverse transcription polymerase chain reaction for the N gene and partially sequenced. Genetic characterization of these isolates was performed by grouping the sequences obtained with known RABV lineages. The sequences were grouped in clusters by the phylogenetic inference neighbor-joining method, together with another 89 homologous sequences obtained from GenBank. This analysis grouped the isolates into four known lineages: Nyctinomops Brazil, Myotis Brazil, Eptesicus Brazil and D. rotundus Brazil, as well as another cluster that may define a RABV lineage not yet characterized, here named Myotis Brazil II, for which bats of the genus Myotis apparently act as reservoirs. This assumption of a new lineage is also based on the observation of amino acid substitutions, with an average intraspecific identity of 99.8%, varying from 99.6 to 100.0% for nucleotides and 100.0% for amino acids. Department of Veterinary Medicine and Public Health School of Veterinary Medicine and Animal Science São Paulo State University (UNESP), Rubião Jr. District Pasteur Institute, 393 Paulista Ave. Department of Public Health School of Medical Sciences State University of Campinas, 126 Tessália Vieira de Camargo St. Adolfo Lutz Institute Bauru II Center of Regional Laboratories, 6000 Rubens Arruda St. Department of Veterinary Medicine and Public Health School of Veterinary Medicine and Animal Science São Paulo State University (UNESP), Rubião Jr. District

Hemoparasitic infections are tick-borne diseases, which affect animals and humans. Considering the importance of canine hemoparasitic infections in veterinary clinics, this study aimed to determine the occurrence of Anaplasma platys, Ehrlichia canis and Babesia vogeli in blood samples from 182 dogs not domiciled in the city of Pato Branco, southwestern region of Paraná State, Brazil, using polymerase chain reaction (PCR). The prevalence of A. platys and B. vogeli was 32.9% and 10.9% respectively, and A. platys infection prevailed (p

Made available in DSpace on 2018-11-26T17:10:37Z (GMT). No. of bitstreams: 0 Previous issue date: 2016-09-01. Added 1 bitstream(s) on 2019-10-09T18:26:23Z : No. of bitstreams: 1 S0037-86822016000500602.pdf: 908415 bytes, checksum: 4b877ac021b8f1efc0335a51d2e6c35a (MD5) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Introduction: Road-killed wild animals host zoonotic pathogens such as Toxoplasma gondii, offering a new opportunity for the epidemiological study of these infectious organisms. Methods: This investigation aimed to determine the presence of T. gondii and other apicomplexan parasites in tissue samples of 64 road-killed wild animals, using polymerase chain reaction (PCR). Positive samples were then typed by PCR-restriction fragment length polymorphism (RFLP) using 7 markers: SAG1, 5'-3' SAG2, SAG3, BTUB, c29-6, PK1, and Apico. PCR-RFLP targeting 18S ribosomal RNA (rRNA) genes was also performed on all samples to detect other apicomplexan parasites. Results: T. gondii DNA was detected in 16 tissue samples from 8 individual animals, as follows: 1 Cerdocyon thous (crab-eating fox), 1 Didelphis albiventris (white-eared opossum), 1 Lutreolina crassicaudata (lutrine opossum), 2 Myrmecophaga tridactyla (giant anteater), 1 Procyon cancrivorus (crab-eating raccoon), and 2 Sphiggurus spinosus (Paraguay hairy dwarf porcupine). Seven different T. gondii genotypes were identified, 6 of which were novel. Typing by 18S rRNA verified these 16 T. gondii-infected samples, and identified 1 Sarcocystis spp.-infected animal [Dasypus novemcinctus (nine-banded armadillo)]. The amplified T. gondii (GenBank accession No. L37415.1) and Sarcocystis spp. 18S rRNA products were confirmed by sequencing. Conclusions: Our results indicate that T. gondii is commonly present in wild mammals, which act as sources of infection for humans and animals, including other wild species. The approach employed herein proved useful for detecting T. gondii and Sarcocystis spp. in the environment and identifying their natural reservoirs, contributing to our understanding of host-parasite interactions. Adolfo Lutz Inst, Lab Reg Bauru, Sao Paulo, Brazil Univ Estadual Paulista, Fac Med Vet & Zootecnia, Dept Higiene Vet & Saude Publ, Botucatu, SP, Brazil Mississippi State Univ, Dept Pathobiol & Populat Med, Coll Vet Med, Mississippi State, MS 39762 USA Univ Estadual Paulista, Fac Med Vet & Zootecnia, Dept Higiene Vet & Saude Publ, Botucatu, SP, Brazil FAPESP: 08/09378-8 FAPESP: 08/08291-6

Leptospirosis is an infectious disease of worldwide importance. The development of diagnostic techniques allows sick animals to be identified, reservoirs to be eliminated and the disease prevented and controlled. The present study aimed to compare different techniques for diagnosing leptospirosis in sheep. Samples of kidney, liver and blood were collected from 465 animals that originated from a slaughterhouse. The sera were analyzed by the Microscopic Agglutination Test (MAT), and kidney and liver samples of seropositive animals were analyzed using four techniques: bacteriological culture, the Warthin Starry (WS) technique, conventional PCR (cPCR), and quantitative PCR (qPCR). With the MAT, 21 animals were positive (4.5%) to serovars Hardjo (n=12), Hebdomadis (n=5), Sentot (n=2), Wolfii (n=1) and Shermani (n=1). Titers were 100 (n=10), 200 (n=2), 400 (n=6) and 1600 (n=3). No animal was positive by bacteriological culture; four animals were positive by the WS technique in kidney samples; six animals were positive by cPCR in kidney samples; and 11 animals were positive by qPCR, eight of which in kidney samples and three in liver. The bacterial quantification revealed a median of 4.3 bacteria/μL in liver samples and 36.6 bacteria/μL in kidney samples. qPCR presented the highest sensitivity among the techniques, followed by cPCR, the WS technique and bacteriological culture. These results indicate that sheep can carry leptospires of the Sejroe serogroup, and demonstrate the efficiency of quantitative PCR to detect Leptospira spp. in tissue samples.

Synanthropic rodents, mainly rats and mice, become ecologically associated with men due to changes in their ecosystems caused by human activities. These animals may take part in the epidemiological cycles of several diseases, including toxoplasmosis. The presence of serum antibodies to Toxoplasma gondii in 43 rodents captured in the urban area of Umuarama, PR, Brazil, was verified by modified agglutination test (MAT). Brain and heart samples were also collected and bioassayed in mice for the isolation of the parasite. Isolated samples were analyzed by 12 multilocus genotyping. Although all rodents were seronegative, the parasite was isolated in one mouse (Mus musculus) and one rat (Rattus rattus). Genotyping showed that these samples were similar to those previously isolated from cats in the state of Parana, Brazil.

Paracoccidioides brasiliensis infections have been little studied in wild and/or domestic animals, which may represent an important indicator of the presence of the pathogen in nature. Road-killed wild animals have been used for surveillance of vectors of zoonotic pathogens and may offer new opportunities for eco-epidemiological studies of paracoccidiodomycosis (PCM). The presence of P. brasiliensis infection was evaluated by Nested-PCR in tissue samples collected from 19 road-killed animals; 3 Cavia aperea (guinea pig), 5 Cerdocyon thous (crab-eating-fox), 1 Dasypus novemcinctus (nine-banded armadillo), 1 Dasypus septemcinctus (seven-banded armadillo), 2 Didelphis albiventris (white-eared opossum), 1 Eira barbara (tayra), 2 Gallictis vittata (grison), 2 Procyon cancrivorus (raccoon) and 2 Sphiggurus spinosus (porcupine). Specific P. brasiliensis amplicons were detected in (a) several organs of the two armadillos and one guinea pig, (b) the lung and liver of the porcupine, and (c) the lungs of raccoons and grisons. P. brasiliensis infection in wild animals from endemic areas might be more common than initially postulated. Molecular techniques can be used for detecting new hosts and mapping 'hot spot' areas of PCM.

Toxoplasma gondii is one of the most significant parasite, due its importance in veterinary medicine and in public health, considered a food-borne pathogens, there is no available drug treatments to eliminate it from animal tissue, this reinforce the search for a vaccine against this parasite. This study was aimed to evaluate the dynamic of the distribution of T. gondii in tissues of female Wistar rats and their milk, after the immunization by oral rote with irradiated tachyzoites. One week after pregnancy confirmation, rats was challenged by gavage with T. gondii bradyzoites, oocysts or tachyzoites of T. gondii. Forty-eight pregnant rats were grouped as follows: immunized and challenged with bradyzoites (BZ*); non-immunized and challenged with bradyzoites (BZ); immunized and challenged with oocysts (OC*); non-immunized and challenged with oocysts (OC); immunized and challenged with tachyzoites (TZ*); non-immunized and challenged with tachyzoites (TZ); only immunized (I); control group (C). After parturition, milk samples were collected for 3 weeks and then rats were sacrificed and the tissues and milk samples were researched for T. gondii parasite load determined by the quantitative PCR (qPCR). It was verified that the immunization with irradiated tachyzoites of T. gondii induced the reduction of parasitic load in muscle samples in rats challenged by bradyzoites and oocysts, although not enabled the development of sterile immunity. The detection of parasite DNA in milk was found throughout the lactation period, from immunized and non-immunized rats, however no differences were found in the parasite load caused by immunization.

Made available in DSpace on 2015-10-21T13:09:47Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-01-01. Added 1 bitstream(s) on 2015-10-22T09:53:10Z : No. of bitstreams: 1 S0100-736X2015000100009.pdf: 316526 bytes, checksum: 43771ebadd5f763631679efb3929cffe (MD5) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Klebsiella pneumoniae é um agente ambiental comum de mastite clínica e subclínica que afetam vacas leiteiras e pode estar presente no produto final, reduzindo a sua qualidade. Mastite causada por K. pneumoniae é ainda mais grave devido à sua má resposta à antibioticoterapia, rápida evolução para choque tóxico e morte do animal. Este trabalho teve como objetivo estudar a prevalência deste patógeno entre os rebanhos leiteiros em dez fazendas localizadas em diferentes municípios do Estado de São Paulo com base no tamanho do rebanho e uso de tecnologia de ordenha. Todas as glândulas mamárias das vacas em lactação foram examinadas usando o California Mastitis Test (CMT) e caneca de fundo telado. Foram colhidas amostras de leite (20mL) de todos os quartos CMT- positivos e dos tanques de expansão, também foram colhidos swab de fezes, membros posteriores dos animais, cama dos animais e sala de ordenha. O isolamento e identificação de K. pneumoniae foi realizada através de cultura microbiológica convencional, ensaio bioquímico e Reação em Cadeia da Polimerase, utilizando primers desenhados e testados no Laboratório de Biologia Molecular aplicada à Zoonoses (FMVZ, Unesp-Botucatu) com base na região do gene de 16S rRNA. Este estudo incluiu 1067 animais. Foram detectados seis casos de infecção intramamária por K. pneumoniae em seis diferentes animais em duas fazendas. Ainda, K. pneumoniae foi isolada em 77 swabs (34 de camas em 9 propriedades, 7 de salas de pré-ordenha em 5 propriedades, 6 de salas de ordenha em 4 propriedades, 11 do reto de animais em 6 propriedades e 19 de membros posteriores em 7 propriedades. A análise molecular confirmou o agente K. pneumoniae. K. pneumoniae foi isolada pelo menos em uma localização em todas as propriedades leiteiras., salientando a necessidade de manter a higiene nas fazendas leiteiras a fim de controlar a mastite por esse patógeno. Klebsiella pneumoniae is a common environmental agent of clinical and subclinical mastitis affecting dairy herds, and may be present in the final product decreasing its quality. Mastitis caused by K. pneumoniae is even more severe due to its poor response to antibiotic therapy, rapid evolution to toxic shock and death of the animal. This paper aimed to study the prevalence of this pathogen among dairy herds in ten farms located in different municipalities of Sao Paulo State based on size and use of milking technology. All mammary glands of all lactating cows were screened using the California Mastitis Test (CMT) and a strip cup. A single aseptic milk sample (20mL) was collected from all CMT-positive quarters and bulk tanks, whereas swab samples were collected from feces, hind limbs of the animals, bedding and milking parlor. Identification of K. pneumoniae was performed using conventional microbiology culture, biochemical assay and Polimerase Chain Reaction. The primers were designed and tested at the Laboratory of Molecular Biology applied to Zoonoses (FMVZ, Unesp-Botucatu) targeting the 16S rRNA gene. This study included 1067 animals. Six cases of intramammary infection by K. pneumoniae were detected in six different cows in two farms. Moreover, K. pneumoniae was isolated in 77 swabs (34 from bedding in 9 farms, 7 from waiting rooms in 5 farms, 6 from milking parlors in 4 farms, 11 from rectums in six farms, and 19 from hindlimbs in 7 farms. Molecular analysis confirmed the agent was K. pneumoniae. At least one strain of the agent was identified in a certain site in all farms, showing the need of maintaining the hygiene in dairy farms. Sao Paulo State Univ, UNESP, Dept Vet Hyg &Publ Hlth, BR-18618970 Botucatu, SP, Brazil Sao Paulo State Univ, UNESP, Dept Vet Hyg & Publ Hlth, BR-18618970 Botucatu, SP, Brazil FAPESP: 2009/10864-7

The present research aimed to carry out an epidemiological study, recording the prevalence of Candida spp. causing infection, as well as the colonization, distribution of the different species in clinical specimens from different sectors of a university hospital, risk factors and their susceptibility to antifungal drugs. Over a one-year period, 100 samples of Candida from 67 patients were isolated and identified, which 74% were characterized as colonization and 26% as nosocomial infections. C. albicans was the most frequent (40%), followed by C. tropicalis (25%), C. parapsilosis (21%), C. glabrata (9%), C. rugosa (2%), C. novergensis (1%), C. krusei (1%) and C. guilliermondii (1%). Non-albicans Candida species represented 71.4% of the colonization cases and 52.1% of infection. C. albicans was the most common species found in secretions and blood, C. parapsilosis was the most isolated from venous catheter, while C. tropicalis and C. glabrata were the most frequently isolated species in probes. The hospital wards with the largest number of yeasts were the Intensive Care Units (45%). Renal failure and multiple traumas were the most frequent underlying diseases and the mainly risk factors for colonization or infection were antibiotic therapy and invasive procedures. Most of the samples showed high susceptibility to the antifungal agents studied. Epidemiological investigations of these agents in the hospital environment are very important, especially at the studied hospital, so that preventive measures may be taken against infections.

Background Leishmaniasis is a rapidly expanding zoonosis that shows increasing urbanization. Concern exists regarding the role of wildlife in visceral leishmaniasis (VL) transmission, due to frequent natural or anthropogenic environmental changes that facilitate contact between wildlife, humans and their pets. The municipality of Campinas, in southeastern Brazil, initially recorded VL in 2009, when the first autochthonous case was confirmed in a dog living in an upscale residential condominium, located inside an environmentally protected area (EPA). Since then, disease transmission remains restricted to dogs inhabiting two geographically contiguous condominiums within the EPA. Methodology/Principal findings We conducted a cross-sectional study of the VL focus to investigate Leishmania spp. infection in domestic dogs, wild mammals and sand flies using molecular tools and recommended serological techniques. Canine seroprevalences of 1.5% and 1.2% were observed in 2013 and 2015, respectively. Six insect species, confirmed or suspected vectors or potential transmitters of Leishmania, were identified. Two specimens of the main L. (L.) infantum vector in Brazil, Lutzomyia longipalpis, were captured in the EPA. Natural infection by L. (L.) infantum was recorded in one Expapillata firmatoi specimen and two Pintomyia monticola. Natural infection by L. (L.) infantum and Leishmania subgenus Viannia was also detected in two white-eared opossums (Didelphis albiventris), a known reservoir of VL. Geographical coordinates of each sampling of infected animals were plotted on a map of the EPA, demonstrating proximity between these animals, human residences, including the dogs positive for VL, and forest areas. Conclusions/Significance The EPA, which is inhabited by humans, has an active VL focus. The risk of establishing and maintaining disease transmission foci in similar scenarios, i.e. wild areas that undergo environmental modifications, is evident. Moreover, different epidemiological profiles of VL must be included to elaborate prevention and control measures that consider the particularities of each transmission area., Author summary Leishmaniasis are neglected tropical diseases that represent a major public health problem in the world. New outbreaks have been recorded in the outskirts of some cities and phlebotomine vectors show geographic expansion, potentially associated with environmental and climate changes. We investigated a recent focus of visceral leishmaniasis (VL) in dogs from two residential condominiums located in an environmentally protected area in the municipality of Campinas, São Paulo, southeastern Brazil. The purpose of the study was to investigate Leishmania (L.) infantum infection, the etiological agent of the disease, in vector insects (phlebotominae), domestic (dog) and wild hosts. Antibody prevalence was determined in 1.5% and 1.2% of dogs examined in 2013 and 2015, respectively. In addition, natural infection by the VL agent was confirmed in two species of sand flies and two white-eared opossums. All positive animals were captured near human residences and in forest areas. Scenarios like these, with changes in ecosystems and the proximity of wild and domestic animals and humans, can promote the occurrence of zoonoses. The participation of wildlife in the transmission cycle of zoonoses has assumed greater importance in recent years as a result of the consolidation of the One Health concept.

Made available in DSpace on 2015-02-02T12:39:35Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-08-05Bitstream added on 2015-02-02T13:07:51Z : No. of bitstreams: 1 S1678-91992014000200331.pdf: 799385 bytes, checksum: 4bfde4303e714913f79c5fa140c54f6c (MD5) Item merged in doublecheck by Felipe Arakaki (fe.arakaki@gmail.com) on 2015-11-17T10:41:22Z Item was identical to item(s): 111450, 113598 at handle(s): http://hdl.handle.net/11449/112350, http://hdl.handle.net/11449/114490 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Background: Road-killed wild animals have been classified as sentinels for detecting such zoonotic pathogens as Leishmania spp., offering new opportunities for epidemiological studies of this infection.Methods: This study aimed to evaluate the presence of Leishmania spp. and Leishmania chagasi DNA by PCR in tissue samples (lung, liver, spleen, kidney, heart, mesenteric lymph node and adrenal gland) from 70 road-killed wild animals.Results: DNA was detected in tissues of one Cavia aperea (Brazilian guinea pig), five Cerdocyon thous (crab-eating fox), one Dasypus septemcinctus (seven-banded armadillo), two Didelphis albiventris (white-eared opossum), one Hydrochoerus hydrochoeris (capybara), two Myrmecophaga tridactyla (giant anteater), one Procyon cancrivorus (crab-eating raccoon), two Sphiggurus spinosus (porcupine) and one Tamandua tetradactyla (lesser anteater) from different locations in the Central Western part of Sao Paulo state. The Leishmania chagasi DNA were confirmed in mesenteric lymph node of one Cerdocyon thous. Results indicated common infection in wild animals.Conclusions: The approach employed herein proved useful for detecting the environmental occurrence of Leishmania spp. and L. chagasi, as well as determining natural wild reservoirs and contributing to understand the host-parasite interaction. 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 FAPESP: 2008/09378-8 FAPESP: 2008/08291-6