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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)

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 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 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

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

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.

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.

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.

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.

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

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.

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.

Toxoplasmosis, caused by an obligate intracellular protozoan parasite, Toxoplasma gondii, is an worldwide parasitic disease, with significant importance for animal production and considerable impact to the public health. This study was aimed to evaluate the dynamic of the distribution of T. gondii in tissues of female Wistar rats and their puppies tissues, 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 follow: 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 the rats were sacrificed and the tissues were researched for the DNA of T. gondii by polymerase chain reaction (PCR) and the 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 most organs analyzed, although not prevent the establishment of infection with the parasite. And also, the immunization showed a favorable effect on the birth rate and litter size.

The objective of this study was to isolate and identify the main staphylococcal species causing bovine mastitis in 10 Brazilian dairy herds and study their capability to produce enterotoxins. Herds were selected based on size and use of milking technology, and farms were visited once during the study. 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. Identification of Staphylococcus spp. was performed using conventional microbiology, and PCR was used to determine the presence of enterotoxin-encoding genes ( sea , seb , sec , and sed ). Of the 1,318 CMT-positive milk samples, Staphylococcus spp. were isolated from 263 (19.9%). Of these isolates, 135 (51%) were coagulase-positive staphylococci (CPS) and 128 (49%) were coagulase-negative staphylococci (CNS). Eighteen different species of CNS were isolated, among which S. warneri , S. epidermidis and S. hyicus were the most frequent. The distribution of Staphylococcus species was different among herds: S. epidermidis was found in 8 herds, S. warneri was found in 7 herds, and S. hyicus in 6 herds. Some of the CNS species ( S. saprophyticus ssp. saprophyticus , S. auricularis , S. capitis , and S. chromogenes ) were isolated in only one of the farms. Genes related to production of enterotoxins were found in 66% (n=85) of all CNS and in 35% of the CPS isolates. For both CNS and CPS isolates, the most frequently identified enterotoxin genes were sea , seb , and sec ; the prevalence of sea differed between CPS (9.5%) and CNS (35.1%) isolates. Staphylococcus warneri isolates showed a greater percentage of sea than seb , sec , or sed , whereas S. hyicus isolates showed a greater percentage of sea than sec . Over 60% of CNS belonged to 3 major species, which carried 62.2 to 81.3% of the enterotoxin genes. The high prevalence highlights the potential for food poisoning caused by these species. For possible high-risk situations for food poisoning, such as milk produced with total bacterial counts greater than regulatory levels and stored under inappropriate temperatures, monitoring contamination with CNS could be important to protect human health. Because the prevalence of CNS intramammary infections in dairy herds is usually high, and these species can be found in great numbers in bulk milk, identification of risk factors for production of staphylococcal enterotoxins should be considered in future studies.

Made available in DSpace on 2013-08-12T17:59:20Z (GMT). No. of bitstreams: 0 Previous issue date: 2012-05-01 Made available in DSpace on 2013-09-30T18:26:21Z (GMT). No. of bitstreams: 0 Previous issue date: 2012-05-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:39:57Z No. of bitstreams: 0 Made available in DSpace on 2014-05-20T13:39:57Z (GMT). No. of bitstreams: 0 Previous issue date: 2012-05-01 INTRODUÇÃO: Toxoplasma gondii e Neospora caninum são parasitas Apicomplexa responsáveis por doenças sistêmicas em muitas espécies de animais, incluindo cães, o que representa grande importância em animais de estimação. MÉTODOS: Este estudo teve como objetivo determinar a prevalência da infecção de T. gondii e N. caninum em 50 cães com sinais neurológicos internados no Hospital Veterinário da Universidade Estadual Paulista (UNESP) na Cidade de Botucatu, Brasil. Todos os animais foram examinados para detecção de anticorpos por IFAT para ambos os parasitas. Tecidos de animais positivos foram analisados por bioensaio em camundongos (T. gondii) e gerbilos (N. caninum) e o DNA foi pesquisado por PCR. Amostras positivas para T. gondii por PCR foram analisadas por meio de análise de restrição de fragmentos polimórficos (restriction fragment length polymorphism-polymerase chain reaction - RFLP-PCR), utilizando-se 11 marcadores: SAG1, SAG2 (5'-3'SAG2 e, alt.SAG2), SAG3, Btub, GRA6, L358, c22 -8, C29-6, PK1 e Apico e o marcador CS3 para análise de virulência. RESULTADOS: Os anticorpos específicos foram detectados em 11/50 animais (22%; IC95% 12,8-35,3%) para T. gondii, e 7/50 (14%; IC95% 7,0-26,3%) para N. caninum. No bioensaio e PCR, 7/11 (63,6%; IC95% 34,9-84,8%) das amostras foram positivas para T. gondii, e 3/7 (42,9%; IC95% 15,7-75,5%) para N. caninum. Três diferentes genótipos foram identificados. Apenas um foi único. CONCLUSÕES: Estes dados confirmam a presença de T. gondii e N. caninum em cães do Brasil, e demonstra a importância do T. gondii como sentinela para a infecção e a variação genotípica deste parasita no Brasil. Introduction: Toxoplasma gondii and Neospora caninum are related Apicomplexa parasites responsible for systemic diseases in many species of animals, including dogs. Methods: This study aimed to determine the occurrence of T. gondii and N. caninum infections in 50 dogs with neurological signs that were admitted to the Veterinary Hospital of Universidade Estadual Paulista, City of Botucatu, Brazil. All animals were screened for antibodies using an immunofluorescent antibody test for both parasites. Tissues of positive animals were bioassayed in mice (T. gondii) and gerbils (N. caninum), and DNA was analyzed using the polymerase chain reaction (PCR). Positive samples for T. gondii by PCR were typed using restriction fragment length polymorphism-PCR for 11 markers: SAG1, SAG2 (5'-3'-SAG2 and alt.SAG2), SAG3, Btub, GRA6, L358, c22-8, c29-6, PK1 and Apico, and CS3 marker for virulence analysis. Results: Specific antibodies were detected in 11/50 (22%; 95% confidence interval (CI95%), 12.8-35.3%) animals for T. gondii and 7/50 (14%; CI95%, 7.02-26.3%) for N. caninum. In the bioassay and PCR, 7/11 (63.6%; CI95%, 34.9-84.8%) samples were positive for T. gondii and 3/7 (42.9%; CI95% I, 15.7-75.5%) samples were positive for N. caninum. Three different genotypes were identified, but only 1 was unique. Conclusions: These data confirm the presence of T. gondii and N. caninum in dogs from Brazil, indicating the importance of this host as a sentinel of T. gondii for human beings, and the genotypic variation of this parasite in Brazil. Univ Estadual Paulista, Fac Med Vet & Zootecnia, Dept Higiene Vet & Saúde Publica, Botucatu, SP, Brazil Univ Estadual Paulista, Fac Med Vet & Zootecnia, Dept Higiene Vet & Saúde Publica, Botucatu, SP, Brazil

Submitted by Guilherme Lemeszenski (guilherme@nead.unesp.br) on 2013-08-22T18:45:01Z No. of bitstreams: 1 S1984-29612010000400013.pdf: 246150 bytes, checksum: 32423ac229af4fa3459ac8e95324d569 (MD5) Made available in DSpace on 2013-08-22T18:45:01Z (GMT). No. of bitstreams: 1 S1984-29612010000400013.pdf: 246150 bytes, checksum: 32423ac229af4fa3459ac8e95324d569 (MD5) Previous issue date: 2010-12-01 Made available in DSpace on 2013-09-30T18:24:10Z (GMT). No. of bitstreams: 2 S1984-29612010000400013.pdf: 246150 bytes, checksum: 32423ac229af4fa3459ac8e95324d569 (MD5) S1984-29612010000400013.pdf.txt: 11039 bytes, checksum: 30209624c3291ee387b4aa567eb0303f (MD5) Previous issue date: 2010-12-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:38:28Z No. of bitstreams: 2 S1984-29612010000400013.pdf: 246150 bytes, checksum: 32423ac229af4fa3459ac8e95324d569 (MD5) S1984-29612010000400013.pdf.txt: 11039 bytes, checksum: 30209624c3291ee387b4aa567eb0303f (MD5) Made available in DSpace on 2014-05-20T13:38:28Z (GMT). No. of bitstreams: 2 S1984-29612010000400013.pdf: 246150 bytes, checksum: 32423ac229af4fa3459ac8e95324d569 (MD5) S1984-29612010000400013.pdf.txt: 11039 bytes, checksum: 30209624c3291ee387b4aa567eb0303f (MD5) Previous issue date: 2010-12-01 Neste trabalho, é relatada a infecção natural por Leishmania em um gato doméstico no qual, formas amastigotas do parasito foram observadas em imprint de linfonodo poplíteo. Reações sorológicas positivas e negativas foram observadas pelo teste de imunoadsorção enzimática (ELISA) e reação de imunofluorescência indireta (RIFI), respectivamente. A reação em cadeia da polimerase (PCR) revelou que a sequência de nucleotídeos foi idêntica à Leishmania (L.) chagasi. Este é o primeiro relato da doença em felino da cidade de Andradina, Estado de São Paulo, Brasil, área considerada endêmica para leishmaniose visceral canina e humana. This work describes natural infection by Leishmania in a domestic cat where amastigote forms of the parasite were observed in the popliteal lymph node imprint. Positive and negative serological reactions were observed by enzyme-linked immunosorbent assay (ELISA) and indirect immunofluorescence assay (IFA), respectively. Polymerase chain reaction (PCR) revealed that the nucleotide sequence of the sample was identical to Leishmania (L.) chagasi. This is the first report of the disease in felines of the city of Andradina, SP, an area considered endemic for canine and human visceral leishmaniasis. Universidade Estadual Paulista Faculdade de Ciências Agrárias e Veterinárias Departamento de Medicina Veterinária Preventiva Universidade Estadual Paulista Faculdade de Odontologia Departamento de Clínica, Cirurgia e Reprodução Animal Universidade Estadual Paulista Instituto de Biociências Departamento de Parasitologia Universidade Estadual Paulista Faculdade de Medicina Veterinária e Zootecnia Departamento de Higiene Veterinária e Saúde Pública Fundação Educacional de Andradina Faculdade de Ciências Agrárias e Veterinárias Universidade Estadual Paulista Departamento de Apoio, Produção e Saúde Animal Universidade Estadual Paulista Faculdade de Ciências Agrárias e Veterinárias Departamento de Medicina Veterinária Preventiva Universidade Estadual Paulista Faculdade de Odontologia Departamento de Clínica, Cirurgia e Reprodução Animal Universidade Estadual Paulista Instituto de Biociências Departamento de Parasitologia Universidade Estadual Paulista Faculdade de Medicina Veterinária e Zootecnia Departamento de Higiene Veterinária e Saúde Pública Universidade Estadual Paulista Departamento de Apoio, Produção e Saúde Animal

The aim of this work was to molecularly detect Leishmania species in 52 cats from Andradina Municipality, Sao Paulo State, Brazil. The direct parasitological test was performed by using imprints of poplited lymph node, bone marrow and spleen to verify amastigote forms of Leishmania spp. The samples that were positive parasitological tests were subjected to molecular analysis (PCR) and sequencing. Infection was detected for 5.76% (3/52) of the examined cats and two had presence of amastigote forms of Leishmania spp. in lymph nodes. Polymerase chain reaction (PCR) of kinetoplast minicircle DNA, indicated positive amplification for samples of spleen and lymph nodes and the sequencing resulted in 97% similarity with Leishmania (L.) chagasi. This study proved the occurrence of infection with Leishmania (L.) chagasi in felines from Andradina municipality, Sao Paulo State.

Toxoplasmosis is the major parasitic disease affecting sheep. It is important for veterinary medicine, animal science and public health since it causes reproductive and economic losses in the herd, as well as damaging human health due to consumption of contaminated meat and milk, which can facilitate zoonotic transmission. Detection of Toxoplasma gondii in ovine milk and lack of data in the literature describing differentiation between acute and chronic disease for this species stimulated the elaboration of the present research project. To achieve the aim of this study, the animals were allocated to two groups of 20 ewes each, of which group 1 was composed of animals with positive serology and group 2 with negative serology. Acute and chronic stages of the disease were differentiated by modified direct agglutination test (MAT), in which antigens were fixed with formalin (MAT-AF) and methanol (MAT-AM). The parasite was detected in milk by polymerase chain reaction (PCR), and the molecular identity of the amplified products was confirmed by sequencing. The serological results indicated that sheep had a chronic infection profile. T. gondii DNA was detected in seven milk samples from five seropositive sheep, and twice in milk of two sheep. Sequences of species shared 97-100% identity with T. gondii. These findings allowed the hypothesis that the peripartum period may also lead to the resurgence of tissue T. gondii tachyzoites cysts which can circulate again and be excreted in the milk. This study used sheep naturally infected with T. gondii as a prerequisite for further investigations on the possible participation of this species in toxoplasmosis epidemiology and as a potential transmission route related to consumption of milk from infected sheep.