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Made available in DSpace on 2022-04-29T08:37:20Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-01-01 The article presents an exploratory, descriptive study on Bad News Communication skills (CMN) of medical interns. The student’s perception was on learning for the CMN was investigated with a cross-sectional design and a mixed approach. A questionnaire was applied, 176 participants were obtained, and a focus group was held with 12 students. The data were analyzed through the calculation of means and standard deviations for quantitative variables and the focus group reports were subjected to Content Analysis. It was observed that in the group that had training, compared to the one who did not, ocurred twice frequency with more skill for CMN. In the CMN’s management, it was considered harder to be “honest without taking away hope” (69%) and “to deal with the patient’s emotion” (59%). 99.4% knew the SPIKES protocol, of which 41.5% considered the expression of emotions as its most difficult stage. Communication and handling of emotions were pointed out as the main difficulties in the doctorpatient relationship, with deficits in the teaching of CMN. It was found that CMN is not limited to the technical issue, but it involves attitudes that need to be addressed with different methodologies, as well as the implementation of educational policies in the medical field, especially given the demands that emerges with the covid-19 pandemic. Programa de Pós-graduação em Ciência Animal Universidade Estadual do Maranhão - UEMA, MA Curso de Medicina Veterinária Centro de Ciências Agrárias Universidade Estadual do Maranhão - UEMA, MA Programa de Pós-graduação Profissional em Defesa Sanitária Animal Universidade Estadual do Maranhão - UEMA, MA Departamento de Patologia Reprodução e Saúde Única Faculdade de Ciências Agrárias e Veterinárias - FCAV Universidade Estadual Paulista - UNESP, SP Programa de Pós-graduação Rede de Biodiversidade e Biotecnologia da Amazônia Legal Universidade Estadual do Maranhão - UEMA, MA Departamento de Patologia Reprodução e Saúde Única Faculdade de Ciências Agrárias e Veterinárias - FCAV Universidade Estadual Paulista - UNESP, SP

Made available in DSpace on 2020-12-12T02:30:49Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-10-01. Added 1 bitstream(s) on 2021-07-15T15:26:06Z : No. of bitstreams: 1 S1984-29612019000400685.pdf: 719893 bytes, checksum: fd5c07754236ed788ef5f2b434317dcb (MD5) Equine piroplasmosis, an economically important disease in horses, has so far not been reported in Pernambuco state, Brazil. This study aimed to evaluate the seroprevalence of anti-Babesia caballi and anti-Theileria equi antibodies based on the detection of these agents in equine blood and in ticks on horses in the municipality of Petrolina, Pernambuco, northeastern Brazil. Blood samples were drawn from 393 horses and sera were examined by ELISA. The presence of tick infestations was evaluated, and 101 ticks were subjected to DNA amplification for the detection of Babesia spp. by polymerase chain reaction (PCR). No parasites were detected in the blood smears. Anti-B. caballi and anti-T. equi antibodies were found in 27.2% (107/393) and 34.8% (137/393) horses, respectively. Infestation by Dermacentor nitens was detected in 4.3% (17/393) of the horses. There was no DNA amplification of the agents in ticks. The risk factors for the presence of anti-T. equi antibodies (P < 0.05) were: Purebred (P < 0.001), animals older than 156 months (P = 0.014), and the presence of ticks (P = 0.001). No risk factors for B. caballi were identified. This study confirmed the circulation of agents of equine piroplasmosis in the municipality of Petrolina, state of Pernambuco, Brazil. Laboratório de Doenças Parasitárias Universidade Federal do Vale do São Francisco – UNIVASF Universidade Federal de Minas Gerais – UFMG Faculdade de Ciências Agrárias e Veterinárias – FCAV Universidade Estadual Paulista Júlio de Mesquita Filho – UNESP Unidade Acadêmica de Medicina Veterinária Universidade Federal de Campina Grande – UFCG Faculdade de Ciências Agrárias e Veterinárias – FCAV Universidade Estadual Paulista Júlio de Mesquita Filho – UNESP

Made available in DSpace on 2018-12-11T17:16:34Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-10-01. Added 1 bitstream(s) on 2019-10-09T18:31:23Z : No. of bitstreams: 1 S1984-29612017000400525.pdf: 1552729 bytes, checksum: b43e4da3b617c8bd6a631e2f5192011d (MD5) The role of several feline vector-borne pathogens (FVBP) as a cause of disease in cats has not been clearly determined. In fact, with the exception of Bartonella spp. and hemoplasmas, FVBP in cats has not been clearly determined in Brazil yet. The present study aimed at identifying, by using molecular methods, the presence of FVBP in three cats showing non-specific clinical signs and inclusions suggestive of hemoparasites in blood smears. Cytauxzoon felis, ‘Candidatus Mycoplasma haemominutum’, Ehrlichia sp. closely related to Ehrlichia canis, and Anaplasma sp. closely related to Anaplasma phagocytophilum were detected in blood samples from two out of three sampled cats. Both cats positive for multiple FVBP did not show hematological and biochemical abnormalities. The present work emphasizes the need for molecular confirmation of co-infection by multiple FVBP in cats presenting non-specific clinical signs and inclusions resembling hemoparasites in blood smears. Laboratório de Imunoparasitologia Departamento de Patologia Veterinária Faculdade de Ciências Agrárias e Veterinárias Universidade Estadual Paulista Júlio de Mesquita Filho – UNESP Hospital Veterinário Jerônimo Dix-Huit Rosado Universidade Federal Rural do Semi-Árido – UFERSA Laboratório de Imunoparasitologia Departamento de Patologia Veterinária Faculdade de Ciências Agrárias e Veterinárias Universidade Estadual Paulista Júlio de Mesquita Filho – UNESP

Made available in DSpace on 2018-12-11T17:16:35Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-10-01. Added 1 bitstream(s) on 2021-07-15T15:21:56Z : No. of bitstreams: 1 S1984-29612017000400505.pdf: 810179 bytes, checksum: 880fd5f5b2da999610cca2592b2a3284 (MD5) Wild animals play an important role in carrying vectors that may potentially transmit pathogens. Several reports highlighted the participation of wild animals on the Anaplasma phagocytophilum cycle, including as hosts of the agent. The aim of this study was to report the molecular detection of an agent phylogenetically related to A. phagocytophilum isolated from a wild bird in the Midwest of the state of Paraná, Brazil. Fifteen blood samples were collected from eleven different bird species in the Guarapuava region. One sample collected from a Penelope obscura bird was positive in nested PCR targeting the 16S rRNA gene of Anaplasma spp. The phylogenetic tree based on the Maximum Likelihood analysis showed that the sequence obtained was placed in the same clade with A. phagocytophilum isolated from domestic cats in Brazil. The present study reports the first molecular detection of a phylogenetically related A. phagocytophilum bacterium in a bird from Paraná State. Laboratório de Doenças Infecciosas e Parasitárias Departamento de Medicina Veterinária Universidade Estadual do Centro-Oeste – UNICENTRO Laboratório de Imunoparasitologia Veterinária Departamento de Patologia Veterinária Faculdade de Ciências Agrárias e Veterinárias – FCAV Universidade Estadual Paulista “Júlio de Mesquita Filho” – UNESP Laboratório de Imunoparasitologia Veterinária Departamento de Patologia Veterinária Faculdade de Ciências Agrárias e Veterinárias – FCAV Universidade Estadual Paulista “Júlio de Mesquita Filho” – UNESP

Made available in DSpace on 2018-12-11T17:32:06Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-01-01. Added 1 bitstream(s) on 2019-10-09T18:32:52Z : No. of bitstreams: 1 S1984-29612017000100039.pdf: 1686498 bytes, checksum: bd23a1bfb172142b4943946c999cde47 (MD5) Dioctophyme renale is a nematode that can be found parasitizing the kidney, peritoneal cavity and, rarely, other organs of canids and mustelids. This disease has high occurrence in the municipality of Três Barras, state of Santa Catarina, thus making this an interesting area to study the epidemiological aspects of infection by D. renale in dogs. Among 197 dogs, 14.2% showed the parasite eggs in urine and 16.4% showed IgG antibodies anti-D. renale in serum samples according to the indirect ELISA method; among seropositive dogs, 15 (37.5%) animals did not show any parasite eggs in their urine. Parasitism was more frequent in females, and there was no finding of interference from age on parasitism. Factors such as water potential and presence of paratenic hosts in the studied region were reported by the owners of dogs and may have contributed to the occurrence of parasitism. Departamento de Medicina Veterinária Universidade do Contestado – UnC Departamento de Medicina Veterinária Preventiva e Reprodução Animal Universidade Estadual Paulista – UNESP Departamento de Patologia Veterinária Universidade Estadual Paulista – UNESP Departamento de Medicina Veterinária Preventiva e Reprodução Animal Universidade Estadual Paulista – UNESP Departamento de Patologia Veterinária Universidade Estadual Paulista – UNESP

Made available in DSpace on 2018-12-11T17:30:42Z (GMT). No. of bitstreams: 0 Previous issue date: 2016-01-01. Added 1 bitstream(s) on 2021-07-15T15:22:22Z : No. of bitstreams: 1 S1984-29612016000400516.pdf: 1212577 bytes, checksum: f8530cf20486ecacfd5efc14e1a0a5a4 (MD5) The transplacental transmission is the primary route of Neospora caninum infection in bovine herds around the world. This study aimed to determine the frequency of transplacental transmission of the parasite in dairy cattle of Agreste region of Pernambuco through serological tests (IFAT and ELISA). Three hundred sixteen serum samples from cows and heifers and their offspring were analyzed. The transplacental transmission rate was 72.22% (13/18) for cows and 69.23% (9/13) for heifers by IFAT. ELISA test showed transplacental transmission rate of 43.58% (17/39) for cows and 50% (9/18) for heifers. The transplacental transmission rates were similar, in both groups in test, but a higher seropositivity was found in cows by IFAT. Data were statistically analyzed using the chi-square and Fisher’s exact test. A significant relationship of dependence between seropositivity of mothers and their offspring was found. The more frequent IFAT antibody titers and ELISA levels for N. caninum were, respectively, 200 and between four (cows) and five (heifers and offspring). In the Spearman correlation, no association was found between the magnitude of antibody titers for N. caninum between mothers and their offspring. The kappa test showed an index of 0.35, indicating a mild correlation between the serological tests used. The study suggests that cows and heifers are the main transmitters of N. caninum in the studied region and that vertical transmission is the major form of transmission in dairy herds of the Agreste region of Pernambuco. Universidade Federal Rural de Pernambuco-UFRPE Clínica de Bovinos Universidade Federal Rural de Pernambuco-UFRPE Pós-graduação em Medicina Veterinária Universidade Estadual Paulista “Júlio de Mesquita Filho”-UNESP Departamento de Medicina Veterinária Universidade Federal Rural de Pernambuco-UFRPE Departamento de Patologia Animal Universidade Estadual Paulista “Júlio de Mesquita Filho”-UNESP Pós-graduação em Medicina Veterinária Universidade Estadual Paulista “Júlio de Mesquita Filho”-UNESP Departamento de Patologia Animal Universidade Estadual Paulista “Júlio de Mesquita Filho”-UNESP

Made available in DSpace on 2022-04-29T07:50:08Z (GMT). No. of bitstreams: 0 Previous issue date: 2016-01-01 Little is known about the occurrence of feline upper respiratory tract disease agents, namely Feline Herpesvirus type 1 (FHV-1) and Chlamydophila felis, and co-infection of these agents with Feline Immunodeficiency virus (FIV) and Feline Leukemia Virus (FeLV) in non-domestic felids in Brazil. Between 2009 and 2010, 72 conjunctival swab and serum samples were collected from eight non-domestic felid species [Leopardus pardalis, Leopardus tigrinus, Panthera leo, Panthera tigris, Puma concolor, Puma yagouaroundi, Oncifelis colocolo, and Panthera onca) maintained in captivity in Brazilian zoos. DNA extracted from conjunctival swabs were used in PCR assays for the detection of Chlamydophila sp, FHV-1, and retrovirus DNA, respectively. Antibodies to FIV and FeLV antigen were detected in non-domestic felid serum samples using a commercial ELISA kit. Antibodies to FIV were found only in five (6.9%) felids. No sampled non-domestic felid was positive for FeLV antigen detection. One (1.3%) out of 72 non-domestic felid conjunctival swab samples was positive for Chlamydophila_sp. and Feline Herpesvirus-1 in PCR. This felid was an ocelot and was negative for FIV and FeLV. The results of this survey showed the occurrence of co-infection with C. felis and FHV-1 in an ocelot (Leopardus pardalis) in Brazil. Universidade Estadual Paulista Faculdade de Ciências Agrárias e Veterinarias Departamento de Patologia Veterinária Universidade Estadual do Centro Oeste Departamento de Medicina Veterinária Universidade Estadual Paulista Faculdade de Ciências Agrárias e Veterinarias Departamento de Patologia Veterinária

Made available in DSpace on 2018-12-11T17:30:42Z (GMT). No. of bitstreams: 0 Previous issue date: 2016-01-01. Added 1 bitstream(s) on 2021-07-15T15:22:21Z : No. of bitstreams: 1 S1984-29612016000400414.pdf: 675364 bytes, checksum: a7a6bfb76a94de4c4bbd4b578bd2d8a0 (MD5) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Mycoplasma suis, the etiological agent of swine hemoplasmosis, has been neglected in swine herds around the world. Swine hemoplasmosis is frequently associated with hemolytic anemia, disgalacty, infertility and immunosuppression, and it results in significant economic losses. This study investigates the occurrence of M. suis in non-technified swine herds in the northeastern region of Brazil using quantitative PCR (qPCR) based on the 16S rRNA gene. Between March and August 2013, blood samples from 147 swine were collected during slaughter in the city of Mossoró, state of Rio Grande do Norte, northeastern Brazil. One hundred and twelve samples (76.19%) were positive for M. suis by qPCR assays. The range of Cqs and quantification (copies of a M. suis-16S rRNAl gene fragment/μL) was 20.86-37.89 and 1.64×101-6.64×107, respectively. One can conclude that M. suis infection have high occurrence (76,19%) in non‑technified swine-rearing systems in Mossoró in the state of Rio Grande do Norte, Brazil. Faculdade de Ciências Agrárias e Veterinárias Universidade Estadual Paulista “Júlio de Mesquita Filho”-UNESP Departamento de Ciência Animal Universidade Federal Rural do Semi-Árido-UFERSA Faculdade de Ciências Agrárias e Veterinárias Universidade Estadual Paulista “Júlio de Mesquita Filho”-UNESP

Made available in DSpace on 2018-12-11T17:26:33Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-10-01. Added 1 bitstream(s) on 2021-07-15T15:22:16Z : No. of bitstreams: 1 S1984-29612015000400438.pdf: 1067227 bytes, checksum: bcba13d49a83d68be4c5b4ea8808ee31 (MD5) The present study reports the genetic diversity of Anaplasma marginale during anaplasmosis outbreaks in rural properties of the states of Goiás and São Paulo, Brazil. Mortality rates of 3.5% (37/1,050) in calves, 4.7% (45/954) in heifers and 1.1% (25/2,200) in lactating cows were observed in a cattle herd of the municipality of Mambaí, state of Goiás, central-western Brazil. In a cattle herd from the municipality of Lins, state of São Paulo, in southeastern Brazil, none of the animals died, despite presenting clinical signs suggestive of bovine anaplasmosis and exhibiting a drastic decrease in milk production. Thus, blood samples were collected from 100 animals with clinical signs suggestive of bovine anaplasmosis in the municipalities of Mambaí and Lins. Based on the microsatellite structure of the MSP1a of A. marginale, the genotypes E and H were observed in Lins, and the C, D and E genotypes were found in Mambaí. The analysis of the tandem repeat structures of the MSP1a showed nine different strains (τ-10 -15, α-β2, α-β3-13, α-β2 192, τ-β-100, α-β2-Γ, 193-β-100, 191-13-Γ and 191-13-18) in Lins and two (α-β3-Γ and E-F-φ2-F2) in Mambaí. Three new tandem repeats of MSP1a (191, 192 and 193) were described. The τ-10-15 and α-β3-Γ strains were predominantly associated with the occurrence of clinical anaplasmosis and mortality in calves, heifers and lactating cows. Departamento de Patologia Veterinária Universidade Estadual Paulista “Júlio de Mesquita Filho” – UNESP Direcção de Ciências Animais Universidad Agraria de La Habana – UNAH Departamento de Patologia Veterinária Universidade Estadual Paulista “Júlio de Mesquita Filho” – UNESP

Made available in DSpace on 2018-12-11T17:26:34Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-10-01. Added 1 bitstream(s) on 2019-10-09T18:32:21Z : No. of bitstreams: 1 S1984-29612015000400475.pdf: 767349 bytes, checksum: e612768493aaca7bc808f8a1dc356f73 (MD5) This study investigated occurrences of anti-Neospora caninum and anti-Toxoplasma gondii IgG antibodies by means of the enzyme-linked immunosorbent assay (ELISA) and indirect immunofluorescence assay (IFAT), along with risk factors associated with toxoplasmosis and neosporosis, in 204 dogs from urban and rural areas of the municipality of Araguaína, state of Tocantins, Brazil. One hundred and thirty samples (63.7%) were positive for T. gondii using ELISA: 57.1% and 70.7% in the urban and rural areas, respectively. The seropositivity frequency for T. gondii observed through IFAT was 57.4%, distributed between rural and urban areas as 62.6% and 52.4%, respectively. The factors associated with canine toxoplasmosis were age and breed (p

Made available in DSpace on 2022-04-29T07:12:55Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-10-01 The aim of this study was to evaluate parasitism kinetics and tissue lesions in the first week of infection by Neospora caninum in dogs fed Gallus gallus chorioallantoic membranes (CMs) previously infected in ovo. Five two-month-old pups were used. Each dog was given five CMs that were previously infected with N. caninum via the oral route. Four animals were euthanized in the first week of infection. All four dogs had their stools examined one week prior to and up to the day they were euthanized. The stools of the uneuthanized dog were collected for 30 days. After euthanasia, organ sections were utilized for histopathology, immunohistochemistry, indirect immunofluorescent tissue reactions, PCR and real-time PCR to detect parasites. Necropsy revealed that the small and large intestines, spleen, and lungs were affected. No oocysts or N. caninum DNA were identified in the stool samples. Real-time PCR was the most sensitive technique used to detect the protozoa in tissues, which were identified in 41% of the analyzed samples. Our results indicate that an experimental model using previously infected CMs appears to be a useful model for the study of the host-parasite relationship during the infection's acute phase. Departamento de Ciências Agrárias e Ambientais Universidade Estadual de Santa Cruz-UESC Campus Soane Nazaré de Andrade, Salobrinho, Ilhéus, BA Instituto de Ciências Biomédicas Universidade Federal de Uberlândia-UFU Campus Umuarama, Uberlândia, MG Departamento de Patologia Veterinária Universidade Estadual Julio Mesquita Filho-UNESP, Jaboticabal, SP Departamento de Parasitologia Animal Universidade Federal Rural do Rio de Janeiro-UFRRJ, Seropédica, RJ Departamento de Patologia Veterinária Universidade Estadual Julio Mesquita Filho-UNESP, Jaboticabal, SP

Made available in DSpace on 2022-04-29T05:36:23Z (GMT). No. of bitstreams: 0 Previous issue date: 2012-07-01 Hemotrophic mycoplasmas and Bartonella species are important pathogens that circulate between cats and invertebrate hosts, occasionally causing diseases in humans. Nevertheless, there are few reports on occurrences of these agents in cats in Brazil. The present study aimed to detect the presence of hemoplasma and Bartonella DNA by means of PCR and sequencing. FIV antigens and anti-FeLV antibodies, were studied by using a commercial kit on blood and serum samples, respectively, among 46 cats that were sampled during a spaying/neutering campaign conducted in Jaboticabal, SP. Three (6.5%) cats were positive for hemoplasmas: two (4.3%) for 'Candidatus M. haemominutum' and one (2.2%) for both M. haemofelis and 'Candidatus M. turicensis'. One of the two 'Candidatus M. haemominutum'-infected cats was also positive for FeLV antigens and showed antibodies for FIV. Two cats (4.3%) were positive for B. henselae. One of them was also positive for FeLV antigens. Eight cats (17.4%) were positive for FeLV, and just one (2.2%) showed anti-FIV antibodies. Bartonella species and hemoplasmas associated with infection due to retroviruses can circulate among apparently healthy cats. Laboratório de Imunoparasitologia, Departamento de Patologia Veterinária Faculdade de Ciências Agrárias e Veterinárias Universidade Estadual Paulista Júlio de Mesquita Filho - UNESP, Campus de Jaboticabal, Via de Acesso Prof. Paulo Donato Castellane s/n Zona Rural, CEP 14884-900, Jaboticabal, SP Faculdade de Ciências Agrárias e Veterinárias - FCAV Universidade Estadual Paulista - UNESP, Campus Araçatuba, Araçatuba, SP Laboratório de Imunoparasitologia, Departamento de Patologia Veterinária Faculdade de Ciências Agrárias e Veterinárias Universidade Estadual Paulista Júlio de Mesquita Filho - UNESP, Campus de Jaboticabal, Via de Acesso Prof. Paulo Donato Castellane s/n Zona Rural, CEP 14884-900, Jaboticabal, SP Faculdade de Ciências Agrárias e Veterinárias - FCAV Universidade Estadual Paulista - UNESP, Campus Araçatuba, Araçatuba, SP

Made available in DSpace on 2022-04-29T01:26:52Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-10-19 Serum samples from 714 equids of Itaguaí and Serrana microregions, Rio de Janeiro, southeastern Brazil, were examined by indirect fluorescent antibody test (titer 1:80) for Theileria equi. The prevalence in the microregions and factors associated with seropositivity were evaluated and the prevalence ratio (PR) calculated. The overall prevalence of T. equi infection was 81.09% (n = 579), with higher prevalence (p < 0.05) in the Itaguaí (85.43%) when compared to Serrana microregion (76.92%). The geographic area, altitude, farming condition and area of origin of equids were associated (p < 0.05) with seropositivity for T. equi. Equids reared in the Itaguaí microregion (PR = 1.11, p = 0.003) and at altitudes below 500 m (PR = 1.10; p = 0,014) were more likely to be seropositive for T. equi. Furthermore, when equids were born in the farm (PR = 1.10, p = 0.008) and reared with poor farming conditions (PR = 1.13, p = 0.018) they were more likely to be exposed to T. equi. The main ticks found on equids were Amblyomma cajennense and Dermacentor (Anocentor) nitens. The microregions studied are endemic areas for equine theileriosis and there exists enzootic stability for T. equi. Only factors related to the collection area of serum samples influenced the seropositivity of equids for T. equi in that region. Universidade Federal Rural do Rio de Janeiro UFRRJ Departamento de Medicina e Cirurgia Veterinária Instituto de Veterinária Universidade Federal Rural do Rio de Janeiro - UFRRJ Departamento de Patologia Veterinária Faculdade de Ciências Agrárias e Veterinárias Universidade Estadual Paulista - UNESP Departamento de Parasitologia Animal Instituto de Veterinária Universidade Federal Rural do Rio de Janeiro - UFRRJ Departamento de Patologia Veterinária Faculdade de Ciências Agrárias e Veterinárias Universidade Estadual Paulista - UNESP

Made available in DSpace on 2022-04-29T02:17:14Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-10-01 Neosporosis and toxoplasmosis are parasitic diseases which can cause reproductive problems in goats and sheep. The current study aimed to determine the occurrence of anti-Neospora caninum and anti-Toxoplasma gondii IgG antibodies in goats and sheep from the districts of Amarante do Maranhão and Buritirana, Imperatriz microregion, western area of Maranhão State, northeastern Brazil, and to assess factors associated to infection by these etiologic agents. Blood samples from 110 animals (46 goats and 64 sheep) from five herds were collected, and indirect immunofluorescence assay was used for serological testing. Of 46 goat samples, 17.39% (n = 8) showed anti-N. caninum antibodies and 4.35% (n = 2) anti-T. gondii, while of 64 sheep samples 4.69% (n = 3) and 18.75% (n = 12) showed anti-N. caninum and anti-T. gondii antibodies, respectively. No significant difference regarding the presence of domestic cats and/or dogs on the property and veterinary care was seen for both etiologic agents studied. However, food supplementation and animal reproductive failure were significantly (p < 0.05) for N. caninum among sheep and goats, respectively. The current study showed that goats and sheep in western Maranhão are exposed to N. caninum and T. gondii. It is the first evidence of these agents in small ruminants in this region. Departamento de Medicina e Cirurgia Veterinária - DMCV Instituto de Veterinária Universidade Federal Rural do Rio de Janeiro - UFRRJ Departamento de Parasitologia Animal - DPA Instituto de Veterinária Universidade Federal Rural do Rio de Janeiro - UFRRJ Escola de Medicina Veterinária e Zootecnia Universidade Federal do Tocantins - UFT Departamento de Patologia Veterinária Faculdade de Ciências Agrárias e Veterinárias - FCAV Universidade Estadual Paulista - UNESP Departamento de Patologia Veterinária Faculdade de Ciências Agrárias e Veterinárias - FCAV Universidade Estadual Paulista - UNESP

Made available in DSpace on 2022-04-29T08:48:40Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-04-01 Hemograms and acute-phase proteins in adult male New Zealand White rabbits that had been experimentally infected orally with sporulated oocysts of Eimeria stiedai were evaluated over a 28-day period. Fifty animals were used, divided into two groups: group A infected with 1 × 10 4 sporulated oocysts of E. stiedai and group B inoculated with distilled water. On the seventh day after infection, the infected animals presented anemia and leukocytosis with neutrophilia and monocytosis. Protein fractionation by means of electrophoresis identified 19 acute-phase proteins with molecular weights ranging from 24 to 238 kD. Ceruloplasmin, transferrin and haptoglobin showed high levels on the seventh day after infection, with gradual increases in their concentrations until the end of the experimental period. Thus, from the data of the present study, E. stiedai is considered to be a pyogenic etiological agent for which the infection level can be monitored through the leukocyte count and serum concentrations of ceruloplasmin, transferrin and haptoglobin, and these can be recommended as complementary tests. School of Veterinary Medicine and Zootechnics Federal University of Tocantins - UFT, BR 153, Km 112, Zona Rural, CP 132, CEP 77804-970, Araguaína - TO Postgraduate Program on Experimental Epidemiology Applied to Zoonoses University of São Paulo - USP Postgraduate Program on Industrial Biotechnology University of São Paulo - USP School of Agrarian and Veterinary Sciences São Paulo State University - UNESP School of Agrarian and Veterinary Sciences São Paulo State University - UNESP

Submitted by PRISCILA IKEDA (priscila.ikeda@unesp.br) on 2021-10-10T17:54:41Z No. of bitstreams: 2 IkedaP - Tese completa - repositório.pdf: 8786902 bytes, checksum: e7241104156b26d98bbfd23569d1c98a (MD5) Tese-Parcial - IkedaP.pdf: 193638 bytes, checksum: 59ae6304e8d273447cdbe434da959039 (MD5) Approved for entry into archive by Alexandra Maria Donadon Lusser Segali null (alexmar@fcav.unesp.br) on 2021-10-15T14:44:36Z (GMT) No. of bitstreams: 1 ikeda_p_dr_jabo_par.pdf: 3605994 bytes, checksum: 441892a242e75dd5e411dd4a2ea6635c (MD5) Made available in DSpace on 2021-10-15T14:44:36Z (GMT). No. of bitstreams: 1 ikeda_p_dr_jabo_par.pdf: 3605994 bytes, checksum: 441892a242e75dd5e411dd4a2ea6635c (MD5) Previous issue date: 2021-08-05 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Estima-se que 75% das doenças emergentes compreendam zoonoses, cuja maioria tem como fontes de infecção animais selvagens; destas, cerca de 22,8% são veiculadas por vetores artrópodes. Desta forma, monitorar a presença de patógenos em animais selvagens em nichos compartilhados com seres humanos se torna um importante método preventivo de infecções zoonóticas. As famílias Bartonellaceae e Anaplasmataceae englobam Alphaproteobactérias Gram-negativas, intracelulares facultativas e obrigatórias, respectivamente, que vêm sendo identificadas em uma ampla variedade de mamíferos, incluindo seres humanos. Já os micoplasmas hemotróficos são bactérias Gram-negativas sem parede celular, de localização epieritrocitária, que podem causar desde infecções assintomáticas até anemia hemolítica severa, tanto em animais como em humanos. Coxiella burnetii é o principal representante da família Coxiellaceae (Gammaproteobactérias) e é conhecida por causar a Febre Q em humanos. Apesar de ser transmitido por aerossóis, esse agente vem sendo detectado em diversos ectoparasitos que talvez possam atuar em seu ciclo de vida. Já os piroplasmas são protozoários da ordem Piroplasmida e causam principalmente anemia hemolítica em animais de produção, domésticos e, eventualmente, em seres humanos. Embora esses agentes supracitados vêm sendo cada vez mais estudados em quirópteros, ainda pouco se sabe a respeito de sua ocorrência e diversidade genética em morcegos no Brasil. Os principais ectoparasitos encontrados nesses animais são representados por moscas das famílias Streblidae e Nycteribiidae, carrapatos, e ácaros das famílias Spinturnicidae e Macronyssidae. Considerando os patógenos de interesse em Saúde Pública supracitados e a expressiva representatividade de animais da ordem Chiroptera no Brasil, objetivou-se obter informações sobre a ocorrência e diversidade genética de agentes Anaplasmataceae, Bartonellaceae, Mycoplasmataceae, Coxiellaceae e protozoários da ordem Piroplasmida em quirópteros e seus respectivos ectoparasitos coletados em área periurbana da cidade de Campo Grande, estado do Mato Grosso do Sul, no centro-oeste brasileiro. Um total de 418 amostras (135 amostras de baço, 133 de sangue e 150 ectoparasitos) foram coletados de 135 animais das famílias Phyllostomidae, Vespertillionidae e Mollossidae. Por meio de técnicas moleculares verificou-se positividade de 18,13% (34/418) para Bartonella spp. dentre todas as amostras coletadas. Dessas, 17 amostras foram clonadas para o gene gltA a fim de obter 3 clones de cada uma para acessar a diversidade genotípica dentro da mesma amostra biológica. Com os 51 gltA-clones obtidos foi possível observar 13 genótipos diferentes, sendo que pelo menos dois ocorreram dentro de uma mesma amostra. Análises filogenéticas baseadas nos genes ftsZ, rpoB e nuoG confirmaram a alta diversidade de genótipos de Bartonella spp. em quirópteros e seus ectoparasitos. Para agentes Anaplasmataceae, foi observada positividade de 1,67% (7/418), 11,96% (50/418) e 13,63% (57/418) das amostras testadas para Anaplasma spp. (16S rRNA), Ehrlichia spp. (dsb) e Neorickettsia spp. (16S rRNA), respectivamente. As sequências obtidas mostraram-se similares a Anaplasma phagocytophilum, Ehrlichia minasensis, Neorickettsia risticii, e Neorickettsia findlayensis por análise pelo nBLAST, e filogeneticamente relacionado a Ehrlichia ruminantium baseado no gene gltA. Trata-se da primeira evidência molecular de agentes Anaplasmataceae em quirópteros e ectoparasitos associados no Brasil. Já para hemoplasmas, positividade de 13,6% (57/418) foi observada. Dentre as 24 sequências obtidas pelo gene 16S rRNA foi possível encontrar 12 genótipos diferentes que se distribuíram pelo macroclado do “grupo haemofelis”. As duas sequências 23S rRNA obtidas se posicionaram próximas a “Candidatus Mycoplasma haematohydrochaerus”, M. haemofelis e M. haemocanis. Todas as amostras de sangue e baço dos morcegos amostrados mostraram-se negativas na qPCR para C. burnetii baseada no gene IS1111. Para piroplasmídeos, 17 dos 135 (12,6%) morcegos amostrados mostraram-se positivos para agentes da ordem Piroplasmida (gene 18S rRNA). Trata-se do primeiro relato molecular de piroplasmídeos em quirópteros no Brasil. A análise filogenética mostrou a possível ocorrência de duas novas espécies: Piroplasmid n. sp. Artibeus spp., posicionado filogeneticamente próximo a Babesia canis e B. vogeli, e Piroplasmid n. sp. P. discolor, que formou um clado monofilético. Infelizmente, as amostras de ectoparasitos não puderam ser utilizadas na detecção de piroplasmídeos e C. burnetii. Enquanto moscas Streblidae mostraram positividade nos ensaios moleculares para Bartonella spp. (11/64), Ehrlichia spp. (7/64) e Mycoplasma spp. (1/64), carrapatos Ornithodoros hasei mostraram-se positivos para Ehrlichia spp. (3/19), Anaplasma spp. (1/19) e Neorickettsia spp. (1/19); por fim, ácaros Spinturniciidae e Macronyssidae apresentaram positividade para Bartonella spp. (4/21 Macronyssidae) e Ehrlichia spp. (18/21 Macronyssidae e 6/46 Spinturnicidae). Ainda, co-positividade para os diversos agentes pesquisados foi observada no presente estudo. Como conclusão, morcegos não-hematófagos de área periurbana do centro-oeste brasileiro atuam como hospedeiros para diversos genótipos de Bartonella spp. e hemoplasmas, para Ehrlichia spp. filogeneticamente associadas a E. ruminantium e para duas possíveis novas espécies de piroplasmídeos. It is estimated that 75% of emerging diseases comprise zoonoses, most of which have wild animals as sources of infection; of these, about 22.8% are carried by arthropod vectors. Thus, monitoring the presence of pathogens in wild animals in niches shared with humans becomes an important preventive method for zoonotic infections. Bartonellaceae and Anaplasmataceae families comprise Gram-negative, facultative and obligate intracellular Alphaproteobacteria, respectively, that have been identified in a wide variety of mammals, including humans. On the other hand, hemotrophic mycoplasmas are epierythrocytic Gram-negative bacteria lacking cell wall, which can cause from asymptomatic infections to severe hemolytic anemia, both in animals and in humans. Coxiella burnetii is the main representative of the Coxiellaceae family (Gammaproteobacteria) and is known to cause Q fever in humans. Despite being transmitted by aerosols, this agent has been detected in several ectoparasites that may act in its life cycle. Piroplasmids, on the other hand, are protozoa of the order Piroplasmida, which can cause hemolytic anemia in farm and domestic animals and, occasionally, in humans. Although the abovementioned agents have been increasingly studied in bats, little is known about their occurrence and genetic diversity in bats from Brazil. The main ectoparasites found in these animals are Streblidae and Nycteribiidae flies, soft ticks, and Spinturnicidae and Macronyssidae mites. Considering the abovementioned pathogens of interest in Public Health and the expressive representation of animals of the order Chiroptera in Brazil, this study aimed to investigate the occurrence and genetic diversity of Anaplasmataceae, Bartonellaceae, Mycoplasmataceae, Coxiellaceae agents and protozoa of the order Piroplasmida in chiropterans and their respective ectoparasites collected in a periurban area of the city of Campo Grande, state of Mato Grosso do Sul, in central-western Brazil. A total of 418 samples (135 spleen, 133 blood and 150 ectoparasites) were collected from 135 animals of the Phyllostomidae, Vespertillionidae and Mollossidae families. Based on molecular assays, positivity of 18.13% (34/418) for Bartonella spp. was found among all collected samples. Out of these, 17 samples had their amplified Bartonella gltA amplicons cloned in order to obtain 3 clones of each one aiming at assessing the genotypic diversity within the same sample. Among the 51 Bartonella gltA-clones obtained, 13 different genotypes were found, with at least two occurring within individual samples. Phylogenetic analyses based on the ftsZ, rpoB and nuoG genes confirmed the high genotype diversity of Bartonella spp. in bats and their ectoparasites. For Anaplasmataceae agents, positivity of 1.67% (7/418), 11.96% (50/418) and 13.63% (57/418) was observed for Anaplasma spp. (16S rRNA), Ehrlichia spp. (dsb) and Neorickettsia spp. (16S rRNA), respectively. The sequences obtained were closely related to Anaplasma phagocytophilum, Ehrlichia minasensis, Neorickettsia risticii, and Neorickettsia findlayensis by BLASTn analyses, and phylogenetically related to Ehrlichia ruminantium based on gltA gene. This is the first molecular evidence of Anaplasmataceae agents in bats and associated ectoparasites from Brazil. Positivity of 13.6% (57/418) for hemoplasmas was found. Among the 24 sequences obtained by the 16S rRNA gene, it was possible to find 12 different hemoplasma genotypes that were distributed into “Haemofelis group”. The two obtained 23S rRNA hemoplasma sequences were closely related to “Candidatus Mycoplasma haematohydrochaerus”, M. haemofelis and M. haemocanis. All bats’ blood and spleen samples were negative in the qPCR for C. burnetii based on the IS1111 gene. Seventeen of the 135 (12.6%) bats were positive for Piroplasmida (18S rRNA gene). This is the first molecular report of piroplasmids in bats from Brazil. Phylogenetic analysis showed the possible occurrence of two new species: Piroplasmid n. sp. Artibeus spp., which was closely related to Babesia canis and B. vogeli, and Piroplasmid n. sp. P. discolor, which formed a monophyletic clade. Unfortunately, the ectoparasite samples could not be used in the PCR assays for piroplasmids and C. burnetii. While Streblidae flies showed positivity in molecular assays for Bartonella spp. (11/64), Ehrlichia spp. (7/64) and Mycoplasma spp. (1/64), Ornithodoros hasei ticks were positive for Ehrlichia spp. (3/19), Anaplasma spp. (1/19) and Neorickettsia spp. (1/19). Spinturnicidae and Macronyssidae mites were positive for Bartonella spp. (4/21 Macronyssidae) and Ehrlichia spp. (18/21 Macronyssidae and 6/46 Spinturnicidae). Also, co-positivity for the different agents studied was observed. In conclusion, non-hematophagous bats from a periurban area of central-western Brazil act as hosts for several genotypes of Bartonella spp. and hemoplasms, Ehrlichia spp. closely related to E. ruminantium, and for two possible new species of piroplasmids. FAPESP: 17/14124-4 FAPESP: 19/13232-3 CAPES: 001

Submitted by Amanda Barbosa Garcia (amandabarbosag@hotmail.com) on 2020-05-16T13:51:55Z No. of bitstreams: 1 Dissertação FINAL = capa + ficha + certificado.pdf: 1919232 bytes, checksum: 1ff84c3a76cfb25a3e290fef808c3aa9 (MD5) Submitted by Amanda Barbosa Garcia (amandabarbosag@hotmail.com) on 2020-05-16T13:51:55Z No. of bitstreams: 1 Dissertação FINAL = capa + ficha + certificado.pdf: 1919232 bytes, checksum: 1ff84c3a76cfb25a3e290fef808c3aa9 (MD5) Approved for entry into archive by Laudicélia Martins Arantes (lm.arantes@unesp.br) on 2020-05-18T16:57:36Z (GMT) No. of bitstreams: 1 garcia_ab_me_jabo.pdf: 1977988 bytes, checksum: 917cee9b89f275a12f13e9000eb6769e (MD5) Approved for entry into archive by Laudicélia Martins Arantes (lm.arantes@unesp.br) on 2020-05-18T16:57:36Z (GMT) No. of bitstreams: 1 garcia_ab_me_jabo.pdf: 1977988 bytes, checksum: 917cee9b89f275a12f13e9000eb6769e (MD5) Submitted by Amanda Barbosa Garcia (amandabarbosag@hotmail.com) on 2020-05-16T13:51:55Z No. of bitstreams: 1 Dissertação FINAL = capa + ficha + certificado.pdf: 1919232 bytes, checksum: 1ff84c3a76cfb25a3e290fef808c3aa9 (MD5) Submitted by Amanda Barbosa Garcia (amandabarbosag@hotmail.com) on 2020-05-16T13:51:55Z No. of bitstreams: 1 Dissertação FINAL = capa + ficha + certificado.pdf: 1919232 bytes, checksum: 1ff84c3a76cfb25a3e290fef808c3aa9 (MD5) Made available in DSpace on 2020-05-18T16:57:36Z (GMT). No. of bitstreams: 1 garcia_ab_me_jabo.pdf: 1977988 bytes, checksum: 917cee9b89f275a12f13e9000eb6769e (MD5) Previous issue date: 2020-04-08 Approved for entry into archive by Laudicélia Martins Arantes (lm.arantes@unesp.br) on 2020-05-18T16:57:36Z (GMT) No. of bitstreams: 1 garcia_ab_me_jabo.pdf: 1977988 bytes, checksum: 917cee9b89f275a12f13e9000eb6769e (MD5) Approved for entry into archive by Laudicélia Martins Arantes (lm.arantes@unesp.br) on 2020-05-18T16:57:36Z (GMT) No. of bitstreams: 1 garcia_ab_me_jabo.pdf: 1977988 bytes, checksum: 917cee9b89f275a12f13e9000eb6769e (MD5) Made available in DSpace on 2020-05-18T16:57:36Z (GMT). No. of bitstreams: 1 garcia_ab_me_jabo.pdf: 1977988 bytes, checksum: 917cee9b89f275a12f13e9000eb6769e (MD5) Previous issue date: 2020-04-08 Approved for entry into archive by Laudicélia Martins Arantes (lm.arantes@unesp.br) on 2020-05-18T16:57:36Z (GMT) No. of bitstreams: 1 garcia_ab_me_jabo.pdf: 1977988 bytes, checksum: 917cee9b89f275a12f13e9000eb6769e (MD5) Submitted by Amanda Barbosa Garcia (amandabarbosag@hotmail.com) on 2020-05-16T13:51:55Z No. of bitstreams: 1 Dissertação FINAL = capa + ficha + certificado.pdf: 1919232 bytes, checksum: 1ff84c3a76cfb25a3e290fef808c3aa9 (MD5) Approved for entry into archive by Laudicélia Martins Arantes (lm.arantes@unesp.br) on 2020-05-18T16:57:36Z (GMT) No. of bitstreams: 1 garcia_ab_me_jabo.pdf: 1977988 bytes, checksum: 917cee9b89f275a12f13e9000eb6769e (MD5) Approved for entry into archive by Laudicélia Martins Arantes (lm.arantes@unesp.br) on 2020-05-18T16:57:36Z (GMT) No. of bitstreams: 1 garcia_ab_me_jabo.pdf: 1977988 bytes, checksum: 917cee9b89f275a12f13e9000eb6769e (MD5) Made available in DSpace on 2020-05-18T16:57:36Z (GMT). No. of bitstreams: 1 garcia_ab_me_jabo.pdf: 1977988 bytes, checksum: 917cee9b89f275a12f13e9000eb6769e (MD5) Previous issue date: 2020-04-08 Submitted by Amanda Barbosa Garcia (amandabarbosag@hotmail.com) on 2020-05-16T13:51:55Z No. of bitstreams: 1 Dissertação FINAL = capa + ficha + certificado.pdf: 1919232 bytes, checksum: 1ff84c3a76cfb25a3e290fef808c3aa9 (MD5) Made available in DSpace on 2020-05-18T16:57:36Z (GMT). No. of bitstreams: 1 garcia_ab_me_jabo.pdf: 1977988 bytes, checksum: 917cee9b89f275a12f13e9000eb6769e (MD5) Previous issue date: 2020-04-08 Approved for entry into archive by Laudicélia Martins Arantes (lm.arantes@unesp.br) on 2020-05-18T16:57:36Z (GMT) No. of bitstreams: 1 garcia_ab_me_jabo.pdf: 1977988 bytes, checksum: 917cee9b89f275a12f13e9000eb6769e (MD5) Made available in DSpace on 2020-05-18T16:57:36Z (GMT). No. of bitstreams: 1 garcia_ab_me_jabo.pdf: 1977988 bytes, checksum: 917cee9b89f275a12f13e9000eb6769e (MD5) Previous issue date: 2020-04-08 Made available in DSpace on 2020-05-18T16:57:36Z (GMT). No. of bitstreams: 1 garcia_ab_me_jabo.pdf: 1977988 bytes, checksum: 917cee9b89f275a12f13e9000eb6769e (MD5) Previous issue date: 2020-04-08 Approved for entry into archive by Laudicélia Martins Arantes (lm.arantes@unesp.br) on 2020-05-18T16:57:36Z (GMT) No. of bitstreams: 1 garcia_ab_me_jabo.pdf: 1977988 bytes, checksum: 917cee9b89f275a12f13e9000eb6769e (MD5) Approved for entry into archive by Laudicélia Martins Arantes (lm.arantes@unesp.br) on 2020-05-18T16:57:36Z (GMT) No. of bitstreams: 1 garcia_ab_me_jabo.pdf: 1977988 bytes, checksum: 917cee9b89f275a12f13e9000eb6769e (MD5) Made available in DSpace on 2020-05-18T16:57:36Z (GMT). No. of bitstreams: 1 garcia_ab_me_jabo.pdf: 1977988 bytes, checksum: 917cee9b89f275a12f13e9000eb6769e (MD5) Previous issue date: 2020-04-08 Made available in DSpace on 2020-05-18T16:57:36Z (GMT). No. of bitstreams: 1 garcia_ab_me_jabo.pdf: 1977988 bytes, checksum: 917cee9b89f275a12f13e9000eb6769e (MD5) Previous issue date: 2020-04-08 Submitted by Amanda Barbosa Garcia (amandabarbosag@hotmail.com) on 2020-05-16T13:51:55Z No. of bitstreams: 1 Dissertação FINAL = capa + ficha + certificado.pdf: 1919232 bytes, checksum: 1ff84c3a76cfb25a3e290fef808c3aa9 (MD5) Approved for entry into archive by Laudicélia Martins Arantes (lm.arantes@unesp.br) on 2020-05-18T16:57:36Z (GMT) No. of bitstreams: 1 garcia_ab_me_jabo.pdf: 1977988 bytes, checksum: 917cee9b89f275a12f13e9000eb6769e (MD5) Made available in DSpace on 2020-05-18T16:57:36Z (GMT). No. of bitstreams: 1 garcia_ab_me_jabo.pdf: 1977988 bytes, checksum: 917cee9b89f275a12f13e9000eb6769e (MD5) Previous issue date: 2020-04-08 Approved for entry into archive by Laudicélia Martins Arantes (lm.arantes@unesp.br) on 2020-05-18T16:57:36Z (GMT) No. of bitstreams: 1 garcia_ab_me_jabo.pdf: 1977988 bytes, checksum: 917cee9b89f275a12f13e9000eb6769e (MD5) Made available in DSpace on 2020-05-18T16:57:36Z (GMT). No. of bitstreams: 1 garcia_ab_me_jabo.pdf: 1977988 bytes, checksum: 917cee9b89f275a12f13e9000eb6769e (MD5) Previous issue date: 2020-04-08 Made available in DSpace on 2020-05-18T16:57:36Z (GMT). No. of bitstreams: 1 garcia_ab_me_jabo.pdf: 1977988 bytes, checksum: 917cee9b89f275a12f13e9000eb6769e (MD5) Previous issue date: 2020-04-08 Approved for entry into archive by Laudicélia Martins Arantes (lm.arantes@unesp.br) on 2020-05-18T16:57:36Z (GMT) No. of bitstreams: 1 garcia_ab_me_jabo.pdf: 1977988 bytes, checksum: 917cee9b89f275a12f13e9000eb6769e (MD5) Made available in DSpace on 2020-05-18T16:57:36Z (GMT). No. of bitstreams: 1 garcia_ab_me_jabo.pdf: 1977988 bytes, checksum: 917cee9b89f275a12f13e9000eb6769e (MD5) Previous issue date: 2020-04-08 Approved for entry into archive by Laudicélia Martins Arantes (lm.arantes@unesp.br) on 2020-05-18T16:57:36Z (GMT) No. of bitstreams: 1 garcia_ab_me_jabo.pdf: 1977988 bytes, checksum: 917cee9b89f275a12f13e9000eb6769e (MD5) Made available in DSpace on 2020-05-18T16:57:36Z (GMT). No. of bitstreams: 1 garcia_ab_me_jabo.pdf: 1977988 bytes, checksum: 917cee9b89f275a12f13e9000eb6769e (MD5) Previous issue date: 2020-04-08 Made available in DSpace on 2020-05-18T16:57:36Z (GMT). No. of bitstreams: 1 garcia_ab_me_jabo.pdf: 1977988 bytes, checksum: 917cee9b89f275a12f13e9000eb6769e (MD5) Previous issue date: 2020-04-08 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Anaplasma marginale é uma bactéria, Gram negativa, intracelular obrigatória parasita de eritrócitos e o principal agente da Anaplasmose bovina. Esta doença causa anemia severa, provocando a redução do ganho de peso e da produção de leite e gerando grandes perdas econômicas na pecuária mundial. A diversidade genética desta bactéria vem sendo caracterizada com base na sequência das proteínas de superfície (MSPs), principalmente, na proteína MSP1α, sendo possível identificar as diferentes estirpes geográficas de acordo com as diferenças nas sequências de aminoácidos. O presente estudo teve como objetivo investigar a diversidade genética de A. marginale em bovinos de corte da raça Angus naturalmente infectados durante um surto da enfermidade. Vinte bovinos, com idades entre 6 e 9 meses, oriundos de uma fazenda no município de Itú, estado de São Paulo foram acompanhados por quatro meses e quatro colheitas de sangue foram realizadas. Amostras de soro foram submetidas à Ensaio Imunoenzimático Indireto (iELISA) para detecção de anticorpos IgG anti-A. marginale. As oitenta amostras de sangue total obtidas, foram submetidas à extração de DNA, PCR em tempo real quantitativa (qPCR) para o gene msp1β, semi-nested PCR (snPCR) para o gene msp1α, clonagem do fragmento alvo e sequenciamento pelo método de Sanger. As sequências obtidas foram submetidas à analises de diversidade genética pelo software RepeatAnalyzer. O ensaio Imunoenzimático Indireto (iELISA), utilizando Ag total, revelou baixa soroprevalência (22,5%) nos animais amostrados apesar de 100% de positividade na qPCR, com quantificação entre 103 e 107 número de cópias de DNA/μL, mostrando uma possível imunossupressão causada pela presença do patógeno. Na snPCR, baseada no gene msp1α, 57,5% (46/80) das amostras mostraram-se positivas. A análise de microssatélites de 36 sequências obtidas mostrou a presença do genótipo H (58,3%), F (25%), E (19,4%), C (2,7%) e G (2,7%). Através do programa Repeat Analyzer foram identificadas várias estirpes na regiao estudada, algumas delas nunca antes descritas na literatura, como 13 27 13 27 13 F; 16 F F; τ 27; 63 29 104 29; LJ1 13 LJ1 13; 16 F 17; 16 F 91 entre outras. Foi encontrada alta diversidade genética da bactéria A. marginale circulando nos animais estudados, identificando mais de uma estirpe circulante em um mesmo animal. Anaplasma marginale is a Gram-negative, obligatory intracellular erythrocyte parasite and the main agent of bovine Anaplasmosis. This disease causes severe anemia, causing a reduction in weight gain and milk production and generating major economic losses in livestock worldwide. The genetic diversity of this bacterium has been characterized based on the sequence of surface proteins (MSPs), mainly in the protein MSP1α, making it possible to identify the different geographical strains according to the differences in the amino acid sequences. The present study aimed to investigate the genetic diversity of A. marginale in Angus beef cattle naturally infected during an outbreak of the disease. Twenty cattle, aged 6 to 9 months, from a farm in the city of Itú, state of São Paulo, were followed for four months and four blood samples were taken. Serum samples were subjected to the Indirect Immunoenzymatic Assay (iELISA) to detect antibodies IgG anti-A. marginale. The eighty whole blood samples obtained were subjected to DNA extraction, quantitative real-time PCR (qPCR) for the msp1β gene, semi-nested PCR (snPCR) for the msp1α gene, cloning of the target fragment and sequencing by the Sanger method. The obtained sequences were subjected to genetic diversity analysis using the RepeatAnalyzer software. The Indirect Immunoenzymatic assay (iELISA) revealed low seroprevalence in the sampled animals despite 100% positivity in the qPCR, with quantification between 103 and 107 number of DNA copies/μL, showing a possible immunosuppression caused by the presence of the pathogen. In snPCR, based on the msp1α gene, 57.5% (46/80) of the samples were positive. Microsatellite analysis of 36 sequences obtained showed the presence of genotype H (58.3%), F (25%), E (19.4%), C (2.7%) and G (2.7%). Through the RepeatAnalyzer software, several strains were identified in the studied region, some of them never before described in the literature, such as 13 27 13 27 13 F; 16 F F; τ 27; 63 29 104 29; LJ1 13 LJ1 13; 16 F 17; 16 F 91 among others. High genetic diversity of A. marginale bacteria was found circulating in the animals studied, identifying more than one circulating strain in the same animal. 2018/05366-7 001

Submitted by CARLOS ANTÓNIO MATOS null (cmatos62@yahoo.com.br) on 2017-05-23T17:44:16Z No. of bitstreams: 1 Tese_Carlos_Antonio_Mato_2017.pdf: 2145169 bytes, checksum: a2aa0c6111ab05dde2597a7a07b2c0a7 (MD5) Approved for entry into archive by Luiz Galeffi (luizgaleffi@gmail.com) on 2017-05-23T18:34:48Z (GMT) No. of bitstreams: 1 matos_ca_me_jabo.pdf: 2145169 bytes, checksum: a2aa0c6111ab05dde2597a7a07b2c0a7 (MD5) Made available in DSpace on 2017-05-23T18:34:48Z (GMT). No. of bitstreams: 1 matos_ca_me_jabo.pdf: 2145169 bytes, checksum: a2aa0c6111ab05dde2597a7a07b2c0a7 (MD5) Previous issue date: 2017-04-19 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) A babesiose é uma doença infecciosa economicamente importante que afeta o gado bovino em todo o mundo, endêmica e importante causa de morbidade e mortalidade em bovinos no Brasil. A doença é causada por Babesia bigemina e B. bovis, protozoários parasitas intraeritrocíticos do filo Apicomplexa, agentes de enorme importância econômica em regiões tropicais e subtropicais. Merozoitos de B. bovis possuem em sua superfície, pelo menos, cinco glico-proteínas, que pertencem à família de antígenos variáveis de superfície do merozoíto (VMSA). A família VMSA de B. bovis inclui os genes msa-1, msa-2a1, msa-2a2, msa-2b e msa-2c. Estes antígenos são altamente imunogênicos e contêm epítopos sensíveis à neutralização e, por conseguinte, têm sido considerados como antígenos candidatos para o desenvolvimento de vacinas de subunidades contra B. bovis. No entanto, estes antígenos de superfície são geneticamente diversificados entre diferentes isolados de B. bovis, O que resulta em diferenças antigénicas entre vários isolados de B. bovis. A fim de avaliar a resposta imune humoral contra B. bovis e a diversidade genética de antígenos de superfície de merozoítos de B. bovis, amostras de soro e DNA de sangue de 30 bezerras, sendo 15 da Fazenda Pesagro, Seropédica, Rio de Janeiro, e outras 15 da Fazenda Germânia, Taiaçu, São Paulo, foram obtidas trimestralmente, desde o nascimento até aos 12 meses de idade. Os Anticorpos IgG para B. bovis foram detectados pelos testes de Imunofluorescência Indireta (RIFI) e Ensaio de Imunoadsorção Enzimático Indireto (IELISA). A Reação em Cadeia da Polimerase (PCR) foi utilizada com o objetivo de avaliar a diversidade genética de B. bovis, com base em genes que codificam antígenos de superfície de merozoítos (MSAs). Os resultados da sorologia demonstram que, a partir dos seis meses de idade, todas as bezerras foram soropositivas para B. bovis. Entre as 150 amostras de DNA testadas para a presença de três fragmentos de genes de msa de B. bovis, amplicons positivos foram obtidos em 14 amostras para msa-1, 24 para msa-2b e 34 para msa-2c de bezerras da Fazenda Pesagro e em 9 amostras para msa-1, 28 para msa-2b e 34 para msa-2c em amostras de bezerras da Fazenda Germânia. O sequenciamento foi realizado pelo método de Sanger para os fragmentos de genes msa-2b (n = 3), msa- 2c (n = 5) de Seropédica e msa-1 (n = 2), msa-2b (n = 5), msa-2c (n = 5) de Taiaçu. A análise filogenética foi realizada pelo método de Máxima verossimilhança e modelos GTR + G, GTR + G + I e GTR + G+ I para MSA-1, MSA-2b e MSA-2c, respectivamente. As sequências de MSAs dos genes msa-2b e msa-2c de Seropédica posicionaram-se em dois clados, enquanto que as sequências de MSAs dos genes msa-1 e msa-2b de Taiaçu posicionaram-se em dois clados, e sequências do gene msa-2c agruparam-se em um único clado. Os resultados da sorologia mostram que foi atingida a estabilidade endêmica em ambas as fazendas. Com base nas sequências do gene msa-2b, amplificadas no presente estudo, e a análise filogenética, pode-se afirmar que as amostras de B. bovis das fazendas Pesagro e Germânia dividem-se em dois grupos genotipicos, havendo um genótipo compartilhado por ambas as fazendas. Babesiosis, an economically important infectious disease that affects cattle worldwide, is endemic and important cause of morbidity and mortality of cattle in Brazil. The disease is caused by Babesia bigemina and B. bovis, apicomplexan intraerythrocytic protozoa parasites, which are agents of huge economic importance in tropical and subtropical regions. B. bovis merozoites present at least five (glyco-) proteins on their surfaces, which belong to a family of variable merozoite surface antigens (VMSA). The members of the VMSA family consist of merozoite surface antigen msa-1, msa-2a1, msa-2a2, msa-2b, and msa-2c. These antigens are highly immunogenic and contain neutralization-sensitive epitopes, and therefore have been considered as candidate antigens for developing subunit vaccines against B. bovis. However, these surface antigens are genetically diverse among different isolates of B. bovis, which results in antigenic differences among various B. bovis isolates. In order to evaluate the humoral immune response against B. bovis and genetic diversity of merozoite surface antigens of B. bovis, serum and DNA blood samples of 30 dairy calves, 15 from Seropedica, state of Rio de Janeiro, and the other 15 from a herd located in Taiaçu, state of São Paulo were obtained quarterly, since the birth up to 12 months of age. IgG antibodies to B. bovis were detected by Indirect Fluorescent Antibody Test (IFAT) and Enzyme-Linked Immunoadsorbent Assay (ELISA) tests. Polymerase Chain Reaction (PCR) was used aiming to assess the genetic diversity of B. bovis, based on genes that encode merozoite surface antigens (MSAs). The serology results demonstrate that up to six months of age all calves were seropositive to B. bovis. Among the 150 DNA blood samples tested for the presence of three B. bovis-msa gene fragments, positive amplicons were obtained in 14 samples for msa-1, 24 samples for msa-2b and 34 for msa-2c from calves of Pesagro herd, 9 samples for msa-1, 28 for msa-2b and 34 for msa-2c from calves of Germânia herd. Sequencing, by Sanger method, was performed for msa-2b (n=3), and msa-2c (n=5), genes fragments of Seropedica and msa-1(n=2), msa-2b (n=5), and msa-2c (n=5), genes fragments of Taiaçu. The phylogenetic analysis was performed by Maximum Likelihood method and models GTR+G, GTR+G+I and GTR+G+I for MSA-1, MSA-2b and MSA-2c, respectively. The Seropedica MSAs sequences namely msa-2b and msa-2c were positioned in two clades, and a Taiaçu MSAs sequence, that is msa-1 and msa-2b, were positioned in two clades, while msa-2c sequences were all grouped in only one clade. Serology results show that endemic stability has been achieved in both farms. Based on the msa-2b gene sequences amplified in the present study, and the phylogenetic analysis, it can be stated that the B. bovis samples from the Pesagro and Germânia farms are classified into two genotypic groups, having one genotype shared by both farms.

Submitted by Luiz Ricardo Gonçalves null (tatinho_tva@hotmail.com) on 2016-07-27T18:25:27Z No. of bitstreams: 1 dissertação LRG.pdf: 2432897 bytes, checksum: 0ce4cc1f4bd2a278c07c6602aa1453d0 (MD5) Approved for entry into archive by Felipe Augusto Arakaki (arakaki@reitoria.unesp.br) on 2016-07-29T13:34:29Z (GMT) No. of bitstreams: 1 golcalves_lr_me_jabo.pdf: 2432897 bytes, checksum: 0ce4cc1f4bd2a278c07c6602aa1453d0 (MD5) Made available in DSpace on 2016-07-29T13:34:29Z (GMT). No. of bitstreams: 1 golcalves_lr_me_jabo.pdf: 2432897 bytes, checksum: 0ce4cc1f4bd2a278c07c6602aa1453d0 (MD5) Previous issue date: 2016-06-27 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Patógenos transmitidos por vetores artrópodes são mundialmente importantes, podendo causar enfermidades tanto no homen quanto nos animais. Recentemente, diversos estudos têm sido realizados a fim de elucidar o papel dos animais selvagens na epidemiologia desses patógenos. A identificação desses microrganismos, assim como dos seus vetores e hospedeiros, permite mapear áreas de ocorrência desses patógenos, auxiliando os órgãos competentes na elaboração de medidas de controle. Sendo assim, o presente estudo teve como objetivo detectar e caracterizar, por métodos moleculares, micoplasmas hemotróficos e bartonelas em amostras de baço de roedores selvagens e sinantrópicos distribuídos em cinco biomas brasileiros. Para tal, foram analisadas 500 amostras de roedores pertencentes a 51 espécies distribuídas em 13 estados. Dentre as 457 amostras de DNA positivas nos ensaios de PCR baseados nos genes Interphotoreceptor retinoid binding protein [IRBP] ou Glyceraldehyde-3-phosphate dehydrogenase [GAPDH], 100 (21,9%) mostraram-se positivas para Mycoplasma spp. por meio da cPCR baseada no gene 16S rRNA. A análise filogenética de 24 sequências do gene 16S rRNA mostrou a presença de dez diferentes clusters, todos agrupados dentro do grupo Mycoplasma haemofelis. O posicionamento filogenético associado com a baixa porcentagem de identidade entre as sequências amplificadas no presente estudo e aquelas previamente depositadas no Genbank sugere a circulação de novas espécies de hemoplasmas em roedores no Brasil. Por meio da qPCR, 117 (25,6%) amostras mostraram-se positivas para Bartonella sp. A análise de diversidade das sequências do gene gltA mostrou a presença de 15 haplótipos. A relação filogenética entre as sequências do gene gltA mostrou que a espécie de Bartonella detectada em roedores no Brasil é intimamente relacionada com Bartonella sp. denominada como grupo filogenético A detectada em roedores da Família Cricetidae na América do Norte, provavelmente formando uma única espécie ou um complexo de espécies. Por outro lado, as sequências amplificadas por meio de ensaios de cPCR adicionais, baseados nos genes ftsZ, groEL e pap-31, mostraram-se filogeneticamente próximas ao complexo Bartonella vinsonii. Por fim, as sequências de Bartonella amplificadas no presente estudo formaram um grupo monofilético e amplamente difundidas em sete diferentes espécies de roedores amostrados em três biomas. Tais achados sugerem uma provável dominância desse genótipo entre os roedores silvestres no Brasil. Copositividade para hemoplasmas e Bartonella sp. foi observada em 41 (9%) dos roedores amostrados. O presente estudo demonstrou, por meio de métodos moleculares, a ocorrência de micoplasmas hemotróficos e Bartonella sp. em roedores selvagens e sinantrópicos no Brasil. Arthropod-borne pathogens are wordwide important, causing diseases in humans and animals. Recently, several studies have been performed in order to elucidate the role of wild animals in the epidemiology of these pathogens. The identification of these microorganisms, as well as their vectors and hosts, allow to map the areas of occurrence of these pathogens, helping competent agencies in the development of control measures. Therefore, the present study aimed to detect and characterize, using molecular methods, hemotrophic mycoplasmas and Bartonella in wild and sinantropic rodent spleen samples distributed in five Brazilian biomes. For this purpose, 500 rodent samples belonging to 51 species distributed in 13 states were analyzed. Among 457 DNA samples positive to PCR assays based on the Interphotoreceptor retinoid binding protein [IRBP] or Glyceraldehyde-3-phosphate dehydrogenase [GAPDH]) genes, 100 (21.9%) were positive to Mycoplasma spp. by cPCR based on 16S rRNA gene. The phylogenetic analysis of 24 sequences of the 16S rRNA gene showed the presence of ten different clusters, which grouped within the Mycoplasma haemofelis group. The phylogenetic position associated with the low percentage of identity between the sequences amplified in the present study and those previously deposited in Genbank suggest the circulation of new hemoplasmas species in rodents in Brazil. Additionally, 117 (25.6%) samples were positive to Bartonella sp. by qPCR. The diversity analysis of the gltA gene sequences showed the presence of 15 haplotypes. The phylogenetic relationships between the sequences of gltA gene showed that Bartonella species detected in rodents in Brazil is closely related to Bartonella sp. namely phylogenetic group A detected in the rodents belonging to Cricetidae family in North America, probably constituting only one species, or a complex of species. On the other hand, the amplified sequences by additional cPCR assays based on ftsZ, groEL and pap-31 genes were phylogenetically positioned to Bartonella vinsonii complex. Finally, the Bartonella sequences amplified in the present study formed a monophyletic and widespread group in seven different species of rodents sampled in three biomes. These findings suggest a probable dominance of this genotype among wild rodents in Brazil. Co-positivity was observed in 41 (9%) of rodents sampled. The presente study demonstrated using molecular methods, the occurrence of hemotrophic mycoplasmas and Bartonella sp. in wild and synanthropic rodents in Brazil.

Made available in DSpace on 2016-04-01T17:55:01Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-10-15. Added 1 bitstream(s) on 2016-04-01T18:00:50Z : No. of bitstreams: 1 000860445.pdf: 1297564 bytes, checksum: 7fa2ff28bbc7d5672a1c49b6aaceef4a (MD5) Anaplasma marginale é o mais prevalente patógeno transmitido por carrapatos em bovinos nas regiões tropicais e subtropicais do mundo. A Proteína Principal de Superfície 1 alpha (MSP1a) do A. marginale contém um número variável de sequências repetidas na região animo-terminal e tem sido utilizada para a caracterização da diversidade genética desse patógeno. Nós realizamos um estudo longitudinal para averiguar a diversidade genética do A. marginale em um rebanho bovino leiteiro de Seropédica no estado do Rio de Janeiro e Taiaçu no estado de São Paulo, Brasil. Vinte bezerras foram avaliadas a cada três meses durante o primeiro ano de vida por esfregaço sanguíneo, Ensaio de Imunoadsorção Enzimático Indireto (ELISA), Reação de Imunofluorescência Indireta (RIFI) e Reação em Cadeia da Polimerase (nPCR/qPCR). Adicionalmente, as amostras positivas para o gene msp1a usando nPCR foram sequenciadas. A frequência do A. marginale variou de 10 - 90% no esfregaço sanguíneo, 20 - 80% no ELISA/RIFI e 15 - 100% na qPCR. O número de copias da msp1a por mL de sangue variou de 1,04 x 101 a 6.76 x 1012 (Seropédica RJ) e 1.20 x 101 a 5.17 x 106 (Taiaçu SP). Os resultados mostraram que a diversidade genética do A. marginale, em um grupo de bezerras até 1 ano de idade em Taiaçu (SP) foi baixa, com apenas três diferentes estirpes identificadas, mostrando o genótipo E. No entanto, os resultados mostraram que o diversidade genética do A. marginale, em um grupo de bezerras até 1 ano de idade em Seropédica (RJ) foi elevada, com dezenove diferentes estirpes identificadas, mostrando o genótipo E e G. As estirpes 4-63-27 (27.4%) e α-β3-Γ (77.8%) foram as mais comumente observadas em bezerras do estado do Rio de Janeiro e São Paulo, respectivamente. O novo conjunto de repetição da MSP1a 190 foi descrito em bezerras de Taiaçu e vinte e uma repetições em tandem da MSP1a resultou em novas sequências com alterações de... Anaplasma marginale is the most prevalent tick-borne pathogen in cattle in tropical and subtropical regions of the world. The A. marginale major surface protein 1 alpha (MSP1a) contains a variable number of tandem repeats in the amino terminal region and has been used for the characterization of pathogen genetic diversity. We conducted a longitudinal study to ascertain the genetic diversity of A. marginale in in a dairy cattle herd in Seropédica state of Rio de Janeiro and Taiaçu state of São Paulo, Brazil. Twenty calves were evaluated every three months during the first year of life by blood smear, Enzyme-linked Immunosorbent Assay (ELISA), Indirect Fluorescent Antibody Test (IFAT) and Polymerase Chain Reaction (nPCR/qPCR). Additionally, samples positive for the msp1a gene using nPCR were sequenced. The prevalence of A. marginale ranged from 10 to 90% according to blood smears, 20-80% using ELISA/IFAT and 15-100% using qPCR. The number of msp1a copies per mL of blood ranged from 1,04x101 to 6.76x1012 (Seropédica RJ) and 1.20 x 101 to 5.17 x 106 (Taiaçu SP). The results showed that the genetic diversity of A. marginale in a group of calves up to 1 year of age from Taiaçu (SP) was low, with only three different strains identified, showing the microsatellite genotype E. However, the results showed that the genetic diversity of A. marginale in a group of calves up to 1 year of age from Seropédica (RJ) was high, with nineteen different strains identified, showing the microsatellite genotype E and G. The strains 4-63-27 (27.4%) and α-β3-Γ (77.8%) were the most commonly observed in calves of Seropédica RJ and Taiaçu SP, respectively. The new MSP1a tandem repeat 190 was described in calves from Taiaçu and twenty-two MSP1a tandem repeats resulted in new sequences with amino acid changes, which were labeled as 165-186 in calves from Seropédica. In Seropedica, three animals were born infected, with strains 4-63-27, 78-242-25-31 and ...

Made available in DSpace on 2015-10-06T13:03:03Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-06-22. Added 1 bitstream(s) on 2015-10-06T13:19:02Z : No. of bitstreams: 1 000848356.pdf: 1331001 bytes, checksum: d6e3659c184a0dc66b218b791f493b49 (MD5) A malária é a endemia de maior impacto na Saúde Pública de países tropicais, devido à alta morbidade e mortalidade. Considerando a malária como uma zoonose, nos quais primatas podem funcionar como reservatórios de espécies de Plasmodium que podem infectar seres humanos nos levam a realizar estudos de malária em primatas com inquestionável relevância. Sabendo-se que geralmente a distribuição da malária humana e de primatas segue a mesma distribuição dos anofelinos, mosquitos vetores, neste trabalho investigaram-se a presença de Plasmodium spp. em primatas neotropicais e em anofelinos na Ilha de São Luís, Estado do Maranhão, Brasil. Colheram-se amostras de sangue de primatas neotropicais do CETAS-São Luís (n=141) e de vida livre (n=20) da Reserva Particular Sítio Aguahy, município de São José de Ribamar. Mosquitos anofelinos foram capturados nesta mesma reserva (n=380) e no Sítio Mangalho (n=36), localizado na Área de Proteção Ambiental do Maracanã, município de São Luís, totalizando em 54 ―pools‖. As amostras de sangue de primatas foram submetidas a testes morfológicos, sorológicos (RIFI e ELISA-teste) e moleculares (qPCR e PCR convencional) para identificação de Plasmodium. Os ―pools‖ de mosquitos foram ensaiados por testes moleculares para identificação de Plasmodium. Cinco primatas tiveram lâminas positivas (3,10%) com observação de formas trofozoíticas. Na RIFI quatro amostras de soro de primatas sororreagiram frente ao antígeno de P. malariae. Amplificaram para Plasmodium sp. na qPCR 34,16% (55/161) das amostras de primatas. Na PCR convencional 30,43% (49/161) foram positivas, sendo 47 (47/49) para P. brasilianum/P. malariae e 2 (2/49) para P. simium/P. vivax. Foram sequenciadas quatro amostras de DNA de primatas, que apresentaram identidade com P. malariae (n=2),com Plasmodium ZOOBH (n=1) e com P. falciparum (n=1). Três ―pools‖ de anofelinos foram positivos para Plasmodium na... Malaria is a disease of greater impact on Public Health in tropical countries because of the high morbidity and mortality. Considering malaria as a zoonosis in which primates can act as reservoirs of species of Plasmodium that can infect humans lead us to conduct malaria studies in primates with unquestionable relevance. It is generally know that the distribution of human and primate malaria following the same distribution of Anopheles mosquitoes vectors, in this study we investigated the presence of Plasmodium spp. in neotropical primates and Anopheles in São Luís Island, Maranhão State, Brazil. Samples were blood neotropical primates CETAS-São Luís (n = 141) and wild life (n = 20) Private Reserve Sítio Aguahy, São José de Ribamar. Anopheles mosquitoes were captured in the same reserve (n = 380) and Mangalho site (n = 36), located in the Environmental Protection Area of the Maracanã, São Luís, totaling 54 pools. The primate blood samples were subjected to morphological, serological (IFA and ELISA-test) and molecular (qPCR and conventional PCR) to identify Plasmodium. The pools of mosquitoes were assayed by testing for molecular identification of Plasmodium. Five primates had positive slides (3.10%) with observation trofozoíticas forms. In IFA four primate serum samples sororreagiram against the antigen of P. malariae. Amplified Plasmodium sp. qPCR at 34.16% (55/161) of samples of primates. In conventional PCR 30.43% (49/161) were positive, 47 (47/49) for P. brasilianum/P. malariae and 2 (2/49) to P.simium/P. vivax. Were sequenced DNA samples from four primates which showed identity with P. malariae (n = 2), Plasmodium ZOOBH (n = 1) and P. falciparum (n = 1). Three pools of Anopheles were positive for Plasmodium in qPCR (3/54) and conventional PCR. The sequencing samples showed identity to P. falciparum (n = 1), P. vivax (n = 1) and Plasmodium ZOOBH (EF090276). Due to the continued and increasing encroachment to forests by ...

Made available in DSpace on 2015-10-06T13:03:26Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-06-15. Added 1 bitstream(s) on 2015-10-06T13:18:31Z : No. of bitstreams: 1 000848350.pdf: 2552487 bytes, checksum: 834f8a43bf7d63a12d8041a4a5022654 (MD5) Agentes das Ordens Rickettsiales, Haemosporida e Rhizobiales são causadores de enfermidades que acometem várias espécies animais, podendo representar um risco à saúde dos mesmos, inclusive de seres humanos. Aves carnívoras podem infestar-se e infectar-se com muitas espécies de parasitas, incluindose carrapatos e hemoparasitas, sendo os hábitos de predação fatores de favorecimento à transmissão e translocação de diversos agentes etiológicos. O objetivo deste estudo foi pesquisar, por meio de métodos indiretos (Reação de Imunofluorescência Indireta - RIFI) e diretos (esfregaços sanguíneos, PCR em Tempo Real e PCR Convencional) os agentes das Famílias Anaplasmataceae (Ehrlichia, Anaplasma, Neorickettsia), Bartonellaceae (Bartonella spp.) e Ordem Haemosporida (Plasmodium, Haemoproteus e Leucocytozoon) em ectoparasitas e amostras de sangue de aves carnívoras, realizando-se um estudo filogenético dos agentes encontrados. Para tanto, procedeu-se à colheita de 121 amostras de sangue total e 88 amostras de soro de aves pertencentes às Ordens Accipitriformes, Falconiformes, Strigiformes e Cathartiformes, capturadas nos estados de São Paulo e Rio de Janeiro. Em relação aos ectoparasitas, foram recolhidas três larvas e uma ninfa ingurgitada de carrapatos do gênero Amblyomma de um gavião carijó (Rupornis magnirostris) no Estado de São Paulo. Na RIFI para E. chaffeensis e A. phagocytophilum, 03 (3,41%) e 05 (5,68%) amostras apresentaram sororreatividade, respectivamente. Todas as amostras testadas na RIFI para E. canis mostraram-se negativas. Na avaliação de esfregaços sanguíneos corados pelo Giemsa, não foram encontradas estruturas compatíveis com hemoparasitas. Em relação a PCR em Tempo Real (qPCR), não foram encontrados animais PCR positivos para Ehrlichia chaffeensis (gene vlpt) e Anaplasma phagocytophilum (gene msp-2). Na qPCR Multiplex para Ehrlichia spp., Anaplasma spp. (gene groE) e... Rickettsiales, Haemosporida and Rhizobiales agents cause diseases that affect various animal species, including humans. Carnivorous birds become infested and infected with many species of parasites, including ticks and hemoparasites, and the predation habits favoring transmission and translocation of various etiological agents. The aim of this study was to investigate, using indirect methods (Immunofluorescence - IFAT) and direct methods (blood smears, Real-Time PCR and Conventional PCR) agents of Families Anaplasmataceae (Ehrlichia, Anaplasma, Neorickettsia), Bartonellaceae (Bartonella spp.) and Haemosporida Order (Plasmodium, Haemoproteus and Leucocytozoon) in ectoparasites and blood samples carnivorous birds, performing a phylogenetic study of these agents. Therefore, we collected 121 blood samples and 88 serum samples from birds belonging to the Accipitriformes, Falconiformes, Strigiformes and Cathartiformes Orders, captured in the states of São Paulo and Rio de Janeiro. Regarding the ectoparasites, we collected three larvae and one engorged nymphal Amblyomma ticks of a Rupornis magnirostris in São Paulo. At IFAT for E. chaffeensis and A. phagocytophilum, 03 (3.41%) and 05 (5.68%) samples showed seroreactivity, respectively. Blood smears stained by Giemsa were analyzed and hemoparasites were not found. Regarding the Real-Time PCR (qPCR), in multiplex qPCR for Ehrlichia spp., Anaplasma spp. (groE gene) and Bartonella spp. (nuoG gene) observed 12 samples positive for Anaplasma spp. (9.91%). In conventional PCR-based 16SrRNA gene for Anaplasmataceae and cytochrome B gene for hemosporidia, eight birds were PCR positive for Ehrlichia spp (6.61%, five in PCR for E. chaffeensis and three in PCR for E. canis), three for Haemoproteus spp (2.48%) and Plasmodium spp (0.83%). In phylogenetic analysis, four samples of Ehrlichia spp. positioned on the same branch Ehrlichia identified in wild animals in Brazil and the US human isolate. The ...

Made available in DSpace on 2015-08-20T17:09:40Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-02-12. Added 1 bitstream(s) on 2015-08-20T17:26:39Z : No. of bitstreams: 1 000837650_20170101.pdf: 129928 bytes, checksum: bcf92a65a2ae8f3fe651ecb9b7e44699 (MD5) Bitstreams deleted on 2017-01-02T15:03:50Z: 000837650_20170101.pdf,. Added 1 bitstream(s) on 2017-01-02T15:05:07Z : No. of bitstreams: 1 000837650.pdf: 3998019 bytes, checksum: 27c55eb98a37acdb1cdea68262f691ce (MD5) O agente etiológico da leishmaniose visceral no Brasil, é a Leishmania chagasi. Ela é transmitida pela picada do flebotomíneo Lutzomyia longipalpis, que adquire o parasito ao realizar o hematofagismo em animais infectados. O presente trabalho teve como objetivo estudar a resposta imune de camundongos BALB/c imunizados com a proteína recombinante produzido a partir do gene A2-L de L. chagasi, amostra isolada de um cão atendido no Hospital Veterinário da FCAV-UNESP, campus de Jaboticabal-SP, e avaliar a capacidade dessa proteína recombinante em induzir imunoproteção nos animais, após desafio com o parasito. Quarenta camundongos foram divididos em quatro grupos: G1, 10 animais inoculados com promastigotas de L. chagasi; G2, 10 animais inoculados com a proteína recombinante A2-L; G3, 10 animais imunizados com a proteína recombinante A2-L e desafiados com promastigotas de L. chagasi; G4, 10 animais inoculados com solução salina (controle negativo). Os parâmetros da resposta imune humoral avaliados foram anticorpos da classe IgG e das subclasses IgG1 e IgG2a, pelo ELISA indireto. Quanto à resposta imune celular, avaliou-se a produção de células CD4+ e CD8+, pela técnica de citometria de fluxo e, as imunomarcações de CD4+, CD8+, iNOS, macrófagos, MHC I, MHC II e Linfócitos B, pela técnica de imunohistoquímica. A produção de citocinas (IL-2, IFN-γ, TNF-α, IL-4 e IL-10), pela técnica de PCR em Tempo Real Quantitativo de Transcrição Reversa (RTqPCR) e, a carga parasitária dos baços dos camundongos infectados com o parasita por qPCR. O grupo G3 teve um padrão de resposta humoral do tipo Th1=Th2, enquanto o grupo G1, o padrão de resposta foi do tipo Th1

Made available in DSpace on 2014-06-11T19:27:16Z (GMT). No. of bitstreams: 0 Previous issue date: 2012-03-01Bitstream added on 2014-06-13T19:55:39Z : No. of bitstreams: 1 barros_ld_me_jabo.pdf: 316486 bytes, checksum: 47f0ed48430e97c60257700b27571deb (MD5) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) O presente trabalho teve como objetivos determinar a soro-ocorrência de anticorpos, isolar e caracterizar geneticamente T. gondii de pombos Zenaida auriculata de vida livre do município de Londrina, Paraná. Foram capturados 206 pombos da espécie Z. auriculata utilizando armadilhas tipo arapuca, em três regiões do município de Londrina, dentre elas o campus universitário, zona urbana e rural. Após a eutanásia em câmara de CO2, realizou-se a punção cardíaca de cada animal para a colheita de sangue e obtenção de soro para a pesquisa de IgG anti-T. gondii pelo Teste de Aglutinação Modificado (MAT). Em seguida foi realizada a necropsia para a colheita de tecidos (pulmão, coração, fígado, cérebro e músculo peitoral) para o isolamento do agente por meio do bioensaio em camundongos. Os isolados obtidos foram caracterizados por meio da PCR-RFLP utilizando 12 marcadores genéticos. Os resultados foram comparados e classificados de acordo com os genótipos presentes no ToxoDB (http://toxodb.org/toxo/). No MAT, 22,3% (46/206) das amostras foram consideradas soropositivas, com títulos variando de 16 até 4096. A soro-ocorrência de acordo com a região de captura foi de 56%, 12,1% e 6,2% para campus universitário, zona urbana e zona rural respectivamente (p

Made available in DSpace on 2014-06-11T19:27:59Z (GMT). No. of bitstreams: 0 Previous issue date: 2012-02-09Bitstream added on 2014-06-13T18:56:57Z : No. of bitstreams: 1 figueiredo_map_me_jabo.pdf: 822824 bytes, checksum: 5e062bcbd66ebcff34c0479b4b807b5d (MD5) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Os Estados que compõem a Amazônia Legal respondem por 99% dos casos de malária humana registrados no Brasil. O Estado do Maranhão, atualmente, é responsável por 1% dos casos dessa região. Acredita-se que, na Região Amazônica ocorra a transmissão de malária de primatas para humanos, devido a relatos atuais de resultados soroepidemiológicos de humanos e primatas. O presente estudo objetivou realizar o diagnóstico morfológico, sorológico e molecular de Plasmodium spp. em primatas neotropicais na Ilha de São Luís, Estado do Maranhão, Brasil. Para tal, foram utilizadas como técnicas de diagnóstico: a microscopia de luz, a reação em cadeia pela polimerase (PCR) e a imunofluorescência indireta (RIFI). No período de junho de 2009 a abril de 2010, foram amostrados 70 primatas, sendo 50 provenientes do Centro de Triagem Animais Silvestres (CETAS), localizado no município de São Luís, e 20 de vida livre, capturados em reserva particular localizada no município de São José de Ribamar, Estado do Maranhão. Foram avaliadas pela microscopia de luz 140 lâminas (duas de cada animal), das quais cinco (7,1%) foram positivas. Pela RIFI não se detectou anticorpos anti-Plasmodium spp. Pela PCR, dos 70 animais amostrados, foi possível observar produtos amplificados para Plasmodium spp. em 13 amostras, das quais oito (61,54%) eram de animais de vida livre e cinco (38,46%) de CETAS. Pelo pequeno tamanho dos fragmentos dos produtos amplificados, foi necessário cloná-los em vetor pGEM-T Easy para sequenciamento, no entanto, não foram obtidos resultados confirmatórios The states that comprise the Amazon region are incriminated for 99% of registered cases of human malaria in Brazil. Currently, the state of Maranhão is responsible for 1% of cases in Amazonic region. According to recent seroepidemiological studies, the transmission of malaria from primates to humans occurs in Amazon region. Our study aimed to detect Plasmodium spp. in neotropical primate from island of São Luis, Maranhão State, Brazil, using morphological (light microscopy), molecular (Polymerase Chain Reaction [PCR]) and serological (Indirect immunofluorescence Assay [IFA]) techniques. Between June/2009 and April/2010, we sampled 70 primates, 50 from the Wildlife Screening Center (CETA), located in the city of São Luís, and 20 captured in a private reserve located in São José de Ribamar, state of Maranhão. Based on blood smear examinations, five (7.1%) monkeys were positive to Plasmodium spp. All animals showed seronegative to Plasmodium spp. by IFA. Plasmodium spp.-amplicons were detected in 13 animals; eight out of positive monkeys (61.54%) were free-ranging animals and five (38.46%) maintained in capitivity in CETAS. Regarding the small size of amplified fragments, we cloned the amplicons into pGEM-T Easy vector aiming at obtaining a higher sequencing quality; however, we did not reached satisfactory results

Made available in DSpace on 2014-06-11T19:32:51Z (GMT). No. of bitstreams: 0 Previous issue date: 2012-01-12Bitstream added on 2014-06-13T18:44:22Z : No. of bitstreams: 1 andre_mr_dr_jabo.pdf: 1327619 bytes, checksum: 69c97d185f41eb670b28bd9bfcbe3759 (MD5) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Doenças transmitidas por vetores artrópodes são mundialmente importantes para a saúde humana e animal. Recentemente, diversos estudos têm sido realizados a fim de investigar o possível papel dos animais selvagens na epidemiologia destas enfermidades. A identificação de reservatórios selvagens para estes hemoparasitas ajudaria no entendimento da epi enfermidades por eles causadas, principalmente aquelas de caráter zoonótico. O presente estudo teve como objetivo pesquisar em amostras de sangue de carnívoros selvagens mantidos em cativeiro a presença de infecção ou exposição a agentes Anaplasmataceae (Ehrlichia canis, E. chaffeensis, E. ewingii, Neorickettsia risticii, N. helminthoeca, Anaplasma platys e A. phagocytophilum), Rickettsiaceae (Orientia tsutsugamushi e Rickettsia sp. dos Grupos da Febre Maculosa e do Tifo), Babesia sp., Cytauxzoon sp. (somente para os felídeos), micoplasmas hemotróficos (Mycoplasma haemofelis, Candidatus M. haemominutum, Candidatus M. turicensis, M. haemocanis e Candidatus M. haematoparvum) e Hepatozoon sp., utilizando métodos sorológicos e moleculares. Para tal, foram amostrados 167 felídeos e 100 canídeos selvagens mantidos em cativeiro nos estados de São Paulo, Mato Grosso e Distrito Federal. Doze felídeos (7,2%) e três (3%) canídeos mostraram-se soropositivos frente ao antígeno de E. canis. Apenas um canídeo (1%) mostrou-se soropositivo para A. phagocytophilum. Nenhum animal mostrou-se soropositivo para E. chaffeensis ou N. risticii. A análise filogenética baseada em um fragmento de 350 pb do gene 16S rRNA não foi suficientemente robusta para diferenciar entre as espécies de Ehrlichia spp. Os fragmentos de DNA de Ehrlichia spp. encontrados em quatro felídeos foram posicionados em um ramo distinto daquela de E. canis e E. chaffeensis, com base... Vector-borne diseases are worldwide important to human and animal health. Recently, several studies have been done aiming to investigate the role of wild animals in the epidemiology of these diseases. The identification of wild reservoirs for these hemoparasites would help in a better understanding of the epidemiology of these diseases, mainly that with zoonotic character. The present work aimed to investigate the presence of infection or exposure to Anaplasmataceae (Ehrlichia canis, E. chaffeensis, E. ewingii, Neorickettsia risticii, N. helminthoeca, Anaplasma platys e A. phagocytophilum) and Rickettsiaceae (Orientia tsutsugamushi and Spotted Fever and Typhus Group Rickettsia sp.), Babesia sp., Cytauxzoon sp., hemotrophic hemoplasmas (Mycoplasma haemofelis, Candidatus M. haemominutum, Candidatus M. turicensis, M. haemocanis e Candidatus M. haematoparvum) and Hepatozoon sp. in captive wild carnivores blood samples by molecular and serological methods. Blood samples were collected from 167 wild felids and 100 wild canids, maintained in captivity in zoos from São Paulo and Mato Grosso states, and Distrito Federal. Twelve felids (7.2%) and three canids (3%) were seropositive to E. canis. Only one canid (1%) was seropositive to A. phagocytophilum. None was seropositive to E. chaffeensis neither N. risticii. The phylogenetic analysis of 350bp of 16S rRNA gene was not sufficiently robust to support the differentiation of Ehrlichia species. The Ehrlichia spp. DNA found in four felids was in a distinct clade from E. chaffeensis and E. canis by omp-1 phylogenetic analysis. The phylogenetic analysis of dsb gene showed the infection by E. canis in one sampled crab-eating fox. Seventeen felids (10%) and ten canids (10%) was positive to Anaplasma spp. PCR, which was in the same clade than A. phagocytophilum and A. platys by... (Complete abstract click electronic access below)

Made available in DSpace on 2014-06-11T19:27:22Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-02-25Bitstream added on 2014-06-13T19:14:41Z : No. of bitstreams: 1 jusi_mmg_me_jabo_parcial.pdf: 92332 bytes, checksum: e6a16d4b0f9ea4ba45ca5712eba954e3 (MD5) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) A leishmaniose visceral é uma zoonose, considerada como uma entre seis doenças tropicais mais importantes nos países em desenvolvimento. O agente etiológico da enfermidade no Brasil, a Leishmania chagasi, é transmitida pela picada do flebotomíneo Lutzomyia longipalpis, que adquire o parasito ao realizar o hematofagismo em animais infectados. No ambiente doméstico, o cão é considerado o principal reservatório da leishmaniose visceral e, em consequência disso, é também o principal alvo das campanhas de controle da forma humana da doença. Neste trabalho foi avaliado o potencial diagnóstico para a leishmaniose visceral canina de um antígeno recombinante, produzido a partir do gene de uma proteína, isolado em L. chagasi, amostra Jaboticabal. O gene isolado apresenta similaridade com a família A2 de L. chagasi e foi expresso utilizando o vetor pET28a em E. coli. A proteína de 11 KDa expressa, foi avaliada pelo ensaio imunoenzimático (ELISA). O ELISA com o antígeno recombinante His6_A2 detectou anticorpos anti-A2 em 52% dos cães infectados com L. chagasi. Enquanto que, 67% dos soros de animais vacinados e 65% dos animais de áreas-não endêmicas, foram negativos pelo ELISA_A2. Os testes de Western-blotting e Dot-ELISA realizado com soros de camundongos inoculados com essa proteína His_A2 demonstrou resultados positivos de antigenicidade e imunogenicidade. Todos os sequenciamentos de DNA mostraram homologia com genes da família A2. Visceral leishmaniasis is a zoonosis and one of six most important tropical diseases in developing countries. The causative agent of the disease in Brazil, Leishmania chagasi, is transmitted by the bite of the sandfly Lutzomyia longipalpis, who acquired the parasite biting infected animals. In domestic environment, the dog is considered the main reservoir of visceral leishmaniasis, and in consequence it is also the main target of campaigns to control the disease in human beings. This study evaluated the potential of a recombinant antigen produced from the A2 gene Jaboticabal strain L. chagasi for diagnosis of canine visceral leishmaniasis (LVC). The isolated gene showed similarity with L. chagasi family A2 gene, and was expressed using pET28a expression vector in E. coli. The 11-kDa protein expressed was evaluated by enzyme linked immunosorbent assay (ELISA) using known positive and negative sera, previously tested by Indirect Immunofluorescence Assay (IFA). The His-A2 recombinantantigen detected anti-A2 antibodies in 52% of L. chagasi infected dogs. Seventy-seven percent of vaccinated dogs and 65% of dogs from non-endemic areas were both negative by A2-ELISA. The antigenicity and immunogenicity of the expressed protein was confirmed by Western blot and Dot-ELISA assays, performed using sera from mice inoculated with this protein.

Made available in DSpace on 2014-06-11T19:27:58Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-12-11Bitstream added on 2014-06-13T20:36:41Z : No. of bitstreams: 1 sacchi_abv_me_jabo.pdf: 876901 bytes, checksum: bbd58ecdb2c29112b3d0b1112c51b549 (MD5) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) O presente trabalho teve por objetivo pesquisar no sangue de cervos-do-pantanal (B. dichotomus), a presença de DNA de agentes rickettsiais, tais como Ehrlichia chaffeensis, E. ewingiii, E. canis, Anaplasma phagocytophilum, A. marginale e Neorickettsia risticii, pela Reação em Cadeia pela Polimerase (PCR), além de estudar a prevalência de anticorpos anti-E. chaffeensis e anti-A. phagocytophilum utilizando-se a Reação de Imunofluorescência Indireta (RIFI). Os 143 animais utilizados foram capturados durante o desenvolvimento do Projeto Cervo-do-Pantanal (1998 a 2002) na área de inundação da Usina Hidrelétrica de Porto Primavera, no Rio Paraná, e divididos em quatro sub-populações de acordo com o local e momento da captura, em áreas denominadas MS01, MS02 (Mato Grosso do Sul, antes e após a inundação, respectivamente), PX (Rio do Peixe, depois da primeira cota de inundação) e AGUA (Rio Aguapeí, depois da inundação). Na avaliação sorológica, as freqüências de animais positivos para anticorpos anti-E. chaffeensis e anti-A. phagocytophilum foram de 76,76% e 20,20% em MS01, 88,88% e 22,22% em PX, 88,88% e 5,55% em MS02, e 94,12% e 5,88% em AGUA, respectivamente. Na pesquisa de DNA, dos 143 animais testados, 61 (42,65%) foram positivos para E. chaffeensis, sendo 38 (38,38%) de MS01, 4 (44,44%) de PX, 12 de MS02 (66,66%) e 7 (41,18%) de AGUA. Amostras positivas enviadas para seqüenciamento revelaram 100% de similaridade com amostras de Ehrlichia chaffeensis da Argentina e dos Estados Unidos (números de acesso no Genbank EU826516.2 e AF416764.1, respectivamente). Já para o genogrupo de A. phagocytophilum, 70 cervos (48,95%) foram considerados positivos. Destes, 51 (51,51%) são provenientes de MS01, 12 (66,66%) de MS02 e 7 (41,18%) de AGUA. Animais provenientes de PX foram todos negativos. Amostras positivas enviadas para seqüenciamento... The present work had as objective to research in the blood of marsh deer (B. dichotomus), the presence of rickettsial agents DNA such as Ehrlichia chaffeensis, E. ewingii, E. canis, Anaplasma phagocytophilum, A. marginale e Neorickettsia risticii, by the Polimerase Chain Reaction (PCR), as well as studying the prevalence of anti-E. chaffeensis and anti-A. phagocytophilum antibodies using Indirect Imunofluorescence Reaction (IFA). The 143 animals that were used were captured during the development of the Marsh Deer Project (1998 to 2002) in the flood area of the Porto Primavera Hydroelectric Power Station, in Parana River, and divided in four sub-populations according to the local and moment of the capture, in areas denominated MS01, MS02 (Mato Grosso do Sul, before and after flood respectively), PX (Peixe’s River, after the first flood quota) and AGUA (Aguapeí River, after flood). In the serological avaliation, the frequency of positive animals for anti-E. chaffeensis and anti-A. phagocytophilum antibodies were 76,76% and 20,20% in MS01, 88,88% and 22,22% in PX, 88,88% and 5,55% in MS02, and 94,12% and 5,88% in AGUA, respectively. In the DNA research, from the 143 tested animals, 61 (42,65%) were positive for E. chaffeensis, 38 (38,38%) from MS01, 4 (44,44%) from PX, 12 from MS02 (66,66% and 7 (41,18%) from AGUA. Positive samples send to sequency demonstrate 100% of similarity with samples of Ehrlichia chaffeensis from Argentina and United States (numbers of access in Genbank EU826516.2 and AF416764.1, respectively). For the A. phagocytophilum genogroup, 70 deers (48,95%) were considered positive. Of these, 51 (51,51%) come from MS01, 12 (66,66%) from MS02 and 7 (41,18%) from AGUA. The animals that come from PX were all negative. Positive samples send to sequencing demonstrated 99% of similarity with a sample of Anaplasma... (Complete abstract click electronic access below)

Made available in DSpace on 2014-06-11T19:33:26Z (GMT). No. of bitstreams: 0 Previous issue date: 2008-07-31Bitstream added on 2014-06-13T21:06:05Z : No. of bitstreams: 1 furuta_pi_dr_jabo.pdf: 1073038 bytes, checksum: 5e772861c92b1e24fce5e65feea5cf02 (MD5) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) A neosporose é uma doença causada pelo protozoário Neospora caninum, e causa um grande impacto econômico na pecuária mundial. Sabe-se que muitos mamíferos são considerados hospedeiros intermediários, e que recentemente, foi confirmado o papel das aves no ciclo de vida de N. caninum como hospedeiros intermediários. O presente trabalho teve como finalidade estudar o ciclo evolutivo e patogenia de N. caninum em cães alimentados com ovos embrionados de Gallus gallus domesticus previamente inoculados com taquizoítas. Realizou-se diariamente o exame de fezes dos animais, pela técnica de flutuação, constatando a presença de oocistos nas fezes a partir de 7 dia apósinfecção. Anticorpos anti- N. caninum foram pesquisados semanalmente nos cães pela RIFI, sendo observados anticorpos a partir dos 14dai. Semanalmente um cão foi eutanasiado e durante a necropsia observou-se em todos os cinco animais, aumento dos linfonodos mesentéricos e das tonsilas, presença de muco no duodeno e jejuno e hepatoesplenomegalia, com hiperplasia de polpa branca do baço. Pelo exame histopatológico detectou-se reatividade dos linfonodos, lesões no fígado, coração, pulmão, baço e rim. Pelas técnicas de imunoistoquímica, imunofluorescência indireta dos tecidos e no Real-time PCR, observou-se a presença do parasito. Os resultados obtidos apontam que, os ovos embrionados infectados pelo parasito mostraram eficiência como modelo experimental para infectar cães, que eliminaram oocistos nas fezes, mesmo não apresentando sinais clínicos da neosporose. Ademais, os cães infectados apresentaram prematura soroconversão pela prévia administração de bicarbonato de sódio e o uso das técnicas citadas tornou possível a detecção do parasito nos tecidos analisados. Neosporosis is a protozoan disease caused by Neospora caninum and affects the reproductive system in cattle worldwide. Infection by the parasite has been widely described in mammals, and recently the role of birds in its lifecycle was confirmed as intermediate hosts. This work aimed to study the life cycle of N. caninum and the pathogeny in dogs that were fed with embryonated eggs inoculated with tachyzoites. The tested dogs were followed up for oocyst shedding, by flotation technique, and it was detected at 7 days after infection. Antibodies anti- N. caninum were detected at 14dai. Once a week, one dog was euthanized and at the necropsy alterations was observed on the mesenteric lymph nodes and tonsils, presence of mucus at duodenum and jejunum portions, and hepatosplenomegaly. At the histopathological examination lymph nodes alterations were noticed ranging from moderate to intense. In addition, there were alterations in liver, heart, lung, spleen and kidney. The parasite was detected by immunochemistry and indirect immunofluorescense tests as well as by Real-time PCR. The results herein presented suggest that inoculated embryonated eggs showed to be an efficient model to infect dogs, because of shedding oocysts, and no clinical alterations. Moreover, the infected dogs presented earlier seroconvertion because of sodium bicarbonate solution administration and the parasite detection in the analyzed tissues was successful by the techniques used.

Made available in DSpace on 2014-06-11T19:31:11Z (GMT). No. of bitstreams: 0 Previous issue date: 2008-03-24Bitstream added on 2014-06-13T18:06:52Z : No. of bitstreams: 1 nakaghi_ach_dr_jabo.pdf: 627442 bytes, checksum: 53f339a01b25f98c06e48eed8f59cc13 (MD5) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) O presente estudo objetivou clonar e analisar a expressão do gene p28 da amostra Jaboticabal de Ehrlichia canis e avaliar sua utilização no diagnóstico molecular e sorológico da erliquiose canina. A PCR, baseada no gene p28 de E. canis, foi capaz de detectar o DNA de um único parasita entre um bilhão de células. Amostras sangüíneas de cães com suspeita clínica de erliquiose foram testadas pela PCR, baseada no gene p28 e pela nested PCR para detecção do gene 16S rRNA. O fragmento amplificado do gene p28 foi observado em 51,25% (n=41) das 80 amostras examinadas, e todas elas foram positivas para o gene 16S rRNA de E. canis, entretanto, em outros 21,25% (n=17) das amostras apenas o gene 16S rRNA foi detectado. A caracterização molecular do gene p28 mostrou similaridade com outras amostras de E. canis. Para a expressão da proteína recombinante P28, foram testados dois sistemas diferentes com linhagens de Escherichia coli BL21(DE3), BL21 Star(DE3)pLysS, BL21(DE3) pLysS e BL21-CodonPlus(DE3)-RIL. A análise pelo Western-blotting revelou reatividade de várias frações protéicas com anticorpo policlonal anti-E. canis. Porém, quando a membrana foi incubada com anticorpo monoclonal para detecção de moléculas de histidina, não houve reatividade, mostrando a ausência de expressão da proteína P28. The aim of this study was to clone and express the p28 gene of Ehrlichia canis Jaboticabal strain and evaluate its application in molecular and serological diagnosis of canine ehrlichiosis. The p28-based PCR was able to detect E. canis DNA of one parasite among one billion cells. Blood samples from dogs for which a diagnosis of canine ehrlichiosis was suspected were tested by p28-based PCR and by 16S rRNA-based nested PCR. Amplified fragment from p28 gene were observed in 51.25% (n=41) among 80 assayed samples and all of this positive samples were also positives for 16S rRNA gene of E. canis, however in 21.25% (n=17) only 16S rRNA gene was detected. The molecular characterization of p28 gene showed similarity with others strains of E. canis. Expression of P28 recombinant protein was tested with two different systems and in Escherichia coli BL21(DE3), BL21 Star(DE3)pLysS, BL21(DE3) pLysS and BL21-CodonPlus(DE3)-RIL strains. After expression induction, Westernblotting showed reactivity of some protein fractions against anti-E. canis antibodies. However, when samples were incubated with anti-histidine monoclonal antibodies, they did not react, demonstrating non-expression of P28 protein.

Made available in DSpace on 2014-06-11T19:27:58Z (GMT). No. of bitstreams: 0 Previous issue date: 2008-02-25Bitstream added on 2014-06-13T18:56:53Z : No. of bitstreams: 1 andre_mr_me_jabo.pdf: 427316 bytes, checksum: 366b68e3f9cae8ab0938b9ab1032ffb9 (MD5) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Poucos relatos têm sido feitos sobre o diagnóstico da erliquiose e babesiose em felinos domésticos e selvagens brasileiros, os quais são baseados diretamente pela presença de mórulas em leucócitos e piroplasmas em eritrócitos, e indiretamente pela detecção de anticorpos anti-Ehrlichia canis. O presente estudo teve como objetivo realizar a detecção molecular de E. canis e B. canis e a presença de anticorpos da classe IgG contra esses hemoparasitas em amostras de sangue e soro, respectivamente. Neste utilizamos 72 felídeos selvagens brasileiros mantidos em cativeiro em algumas instituições e zoológicos. Pela Reação de Imunofluorescência Indireta (RIFI), dezoito (25%) e cinqüenta e três (73,6%) dos 72 animais amostrados foram sororeagentes frente aos antígenos de E. canis e B. canis, respectivamente. Na PCR para E. canis, onze (15,3%) dos 72 animais amostrados foram positivos. Os amplicons foram confirmados por seqüenciamento e o DNA de E. canis encontrado mostrou grande similaridade genética com amostras de E. canis isoladas no Brasil, México, Portugal, Grécia e Taiwan, com 98% de similaridade. Nenhuma das amostras foi positiva na PCR para B. canis. Destaca-se a importância e a primeira detecção molecular de E. canis e presença de anticorpos anti-E. canis e anti-B. canis em felídeos selvagens brasileiros mantidos em cativeiro. Few are the reports that have been carried out on ehrlichiosis and babesiosis diagnostic in Brazilian domestic and wild felids, which are based directly on the presence of morulae in leucocytes and piroplasms in erythrocytes, and indirectly by detection of antibodies against E. canis. The aim of this study was to detect molecularly E. canis and B. canis and the presence of anti-E. canis and B. canis IgG antibodies in the blood and sera samples, respectively, from 72 Brazilian wild captive felids maintained in some instituitions and zoos. Eighteen (25.0%) and fifty-three (73.6%) out of 72 animals were seroreagent for E. canis and B. canis antigen, respectively, by IFA (Indirect Immunofluorescent Assay). Eleven (15.3%) of the 72 samples were positive for nPCR E. canis. The amplicons were confirmed by sequencing and the E. canis DNA found appeared to be closely related to E. canis samples from Brazil, Mexico, Portugal, Greece and Taiwan with 98% percent identity. None of the 72 samples were positive for B. canis by PCR. This is the first molecular detection of E. canis and presence of seroreactivity for both B. canis and E. canis in Brazilian wild captive felids.

Made available in DSpace on 2014-06-11T19:23:42Z (GMT). No. of bitstreams: 0 Previous issue date: 2007-08-10Bitstream added on 2014-06-13T20:11:24Z : No. of bitstreams: 1 galhardo_a_me_jabo.pdf: 395101 bytes, checksum: e772cc6cb2c6cd3e0d11abdb749af7ff (MD5) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) A cetamina, um anestésico dissociativo, é amplamente empregado para a indução e manutenção da anestesia em cães. Possui dois enantinômeros, sendo o S(+), de atividade anestésica superior. Com este estudo objetivou-se correlacionar os parâmetros clínicos com os aspectos farmacocinéticos da cetamina S(+), bem como avaliar, o método de HPLC para quantificação deste fármaco e, as alterações farmacodinâmicas e farmacoinéticas observadas na associação deste com o midazolam. Para tanto, a cetamina S(+) foi administrada em oito cães, sendo quatro fêmeas da raça Beagle e quatro machos sem raça definida, pela via intravenosa na dose de 10mg/kg, associada ou não ao midazolam (0,2mg/kg), em duas fases distintas. A análise farmacocinética foi realizada através da cromatografia líquida de alta performance (HPLC) em amostras plasmáticas de quatros fêmeas e um macho. Os parâmetros clínicos foram obtidos por meio da biotelemetria nas quatro fêmeas e, semiotecnicamente, nos quatro machos, por um período de 12 horas. Observouse que a concentração máxima, cinco minutos após a aplicação, foi de 35,1 e 18,7 æg/mL, a meia vida de distribuição de 0,142 e 0,234 horas, a meia vida de eliminação de 3,70 e 4,06 horas, o clearance de 2,40 e 4,36 L/kg.h e o volume de distribuição de 9,70 e 16,47 L/kg, para a cetamina S(+) isolada e associada ao midazolam, respectivamente. Não foram detectadas diferenças estatísticas para os parâmetros farmacocinéticos analisados, porém foram detectadas em 15 alguns momentos para os parâmetros clínicos. Pode-se concluir que a presença do midazolam não altera os parâmetros clínicos e farmacocinéticos da cetamina S(+) em cães e, que a biodisponibilidade foi coincidente com as alterações clínicas monitoradas por meio da biotelemetria. Ketamine, a dissociative anesthetic agent, is widely used for induction and maintenance of anesthesia in dogs. It has two enantiomers, from which the S(+) has a greater anesthetic activity. The aim of this study was to correlate clinical parameters with pharmacokinetics aspects of ketamine S(+) as well as evaluating, the HPLC method for quantification this drug and, observed the pharmacodinamics and pharmacokinetics alterations in the association of this drug with midazolam it. For this, the ketamine was administered intravenously in eight dogs (four females beagles and four crossbreed males) at a dose of 10mg/kg, associated of not to midazolam (0.2mg/kg), in two distinct phases. Pharmacokinetic analysis was accomplished through High Performance Liquid Chromatography (HPLC) in serum samples of four females and a male. Clinical parameters were obtained through biotelemetry in the four females and semiotecnically in the four males, for 12 hour period. It was noticed that five mintues after the administration the maximum concentration was 35.1 and 18.7 æg/mL, Half-life of distribution was 0.142 and 0.234 hours, and half life of elimination was 3.7 and 4.06 hours, clearance was 2.40 and 4.36 L/kg.h and the volume of distribution was 9.70 and 16.47 L/kg, for ketamine S(+) isolated and associated to midazolam, respectively. No statistical difference was noticed in 17 the pharmacokinetic parameters, however there were differences in clinical parameters in some periods. In conclusion, the presence of midazolam does not alter clinical and pharmacokinetics parameters of ketamine S(+) in dogs and the biodisponibility of the drug was in accordance with clinical alterations monitored through biotelemetry.

Made available in DSpace on 2014-06-11T19:33:26Z (GMT). No. of bitstreams: 0 Previous issue date: 2007-03-15Bitstream added on 2014-06-13T19:23:20Z : No. of bitstreams: 1 mineo_twp_dr_jabo.pdf: 3433348 bytes, checksum: fac62f81f1b33434ac64c0e811ae8d5a (MD5) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Neospora caninum é um protozoário do Filo Apicomplexa, que foi primeiramente descrito como causa de encefalomielite em filhotes caninos sorologicamente negativos para Toxoplasma gondii. Estudos anteriores neste importante hospedeiro da cadeia epidemiológica de N. caninum demonstram que as respostas de anticorpos IgG são tardiamente detectadas e que a infecção clínica é de difícil indução. Desta forma, este trabalho objetivou o estudo da imunidade de cães frente à infecção oral por N. caninum. Os resultados obtidos a partir de análises de diversos animais experimentalmente infectados indicam que os cães apresentam uma prolongada fase aguda da infecção, com eliminação de oocistos associado à queda nos níveis de linfócitos T CD4+ e CD8+ e diminuição de MHC de classe II por células apresentadoras de antígeno. Adicionalmente, os animais apresentam soroconversão instável durante o mesmo período, sendo que somente IgG1 e IgG3 foram detectados em adultos e filhotes, respectivamente, entre o 2o e 3o mês de infecção. De forma concomitante, observa-se uma predominância da expressão de citocinas imunomoduladoras como TGF 1, IL-4 e IL-10. Após dois meses de infecção, o perfil da resposta se inverte, sendo observado picos de produção dos marcadores CD4 e CD8 de linfócitos T e citocinas próinflamatórias como IFN , IL-6 e IL-12, além do aumento nos títulos de anticorpos, principalmente IgG1 e IgG4 nos cães jovens. Com base nestes resultados, conclui-se que os cães apresentam uma relação de equilíbrio com N. caninum, a qual induz nesta espécie uma modulação da resposta imunológica durante a fase de merogonia. Neospora caninum is an Apicomplexan parasite firstly described as the cause of encephalomyelitis in puppies serologically negative to Toxoplasma gondii. Previous reports on the parasite s definitive host indicate a late IgG antibody response and that clinical disease is difficult to be induced. The aim of this study was to investigate canine immunity during N. caninum oral infection. The results obtained from the analysis of infected animal s samples indicate that dogs present a protracted acute phase, with oocyst shedding correlated to a drop in CD4+ and CD8+ T cell levels, and low MHC class II expression by antigen presenting cells. Additionally, the dogs presented an unstable seroconversion pattern in the same period, with only IgG1 and IgG3 being detected in adult dogs and puppies, respectively, between the second and third months of infection. Concomitantly, dominant Th2 cytokine expression was observed, with peak expression levels of TGF 1, IL-4 and IL-10. After 2 months of infection, the immunity profile shifts towards a Th1 response, with high levels of CD4 and CD8 lymphocytary marker production and pro-inflammatory cytokine expression (IFN , IL-6 and IL-12), besides of the raise in antibody levels, especially IgG1 and IgG4 in puppies. Based in the results presented herein, we may conclude that dogs present a balanced host-parasite relationship, modulating the host immune response during N. caninum merogony.

Made available in DSpace on 2014-06-11T19:33:26Z (GMT). No. of bitstreams: 0 Previous issue date: 2006-09-21Bitstream added on 2014-06-13T20:44:55Z : No. of bitstreams: 1 dagnone_as_dr_jabo.pdf: 2655933 bytes, checksum: 8f6122da8b67a36f40a31689501140e2 (MD5) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Este estudo foi realizado objetivando-se detectar e identificar a presença de material genômico de agentes pertencentes à Família Anaplasmataceae em 55 cães com inclusões citoplasmáticas encontrados em células leucocitárias granulocíticas, monocíticas e plaquetas, compatíveis com infecção por Ehrlichia canis, E. chaffeensis, E. ewingii, A. phagocytophilum, A. platys ou Neorickettsia risticii. A ocorrência natural de Babesia canis também foi avaliada pois vetor Rhipicephalus sanguineus, é o principal carrapato encontrado em cães no Brasil. Também objetivou-se realizar, através da utilização de técnicas de Seqüenciamento de um fragmento do gene 16S rRNA, a caracterização parcial dos fragmentos das amostras positivas encontradas neste estudo, e comparar as seqüências dos fragmentos obtidos com as seqüências apresentadas previamente no GenBank, iniciando a realização de estudos filogenéticos das seqüências encontradas. Amostras de sangue foram colhidas de 25 e 30 cães atendidos nos Hospitais Veterinários da Universidade Estadual Paulista-Unesp, Câmpus de Jaboticabal-SP e Universidade para o Desenvolvimento do Estado e da região do Pantanal-UNIDERP, Campo Grande- MS, respectivamente. Dentre as inclusões citoplasmáticas encontradas neste estudo foi observado uma grande diversidade na morfologia, tamanho, coloração e localização das inclusões. DNA de agentes Família Anaplasmataceae foram detectados em 45/55 das amostras sangüíneas examinadas (81,8%), sendo destas 32 (58,18%) e 13 (23,63%) positivas para Ehrlichia canis e Anaplasma platys, respectivamente. Dez animais que foram incluídos neste estudo por apresentarem inclusões citoplasmáticas foram negativos quando uma segunda confecção de esfregaço de papa leucocitária corada pelo Giemsa foi realizada, e também... The aim of this study was to evaluate the natural occurrence of Anaplasmataceae agents (Ehrlichia canis, E. chaffeensis, E. ewingii, A. phagocytophilum, A. platys and Neorickettsia risticii) using molecular techniques. For this purpose a group of fifty-five supposedly infected dogs from two different Brazilian states (São Paulo and Mato Grosso do Sul) that showed suggestive Anaplasmataceae agent intracitoplasmic inclusions in white blood cells and platelets were analysed. The natural occurrence of Babesia canis was also evaluated, because its vector is also Brazilian dog tick, Rhipicephalus sanguineus. Sequencing of a partial phragment of 16S rRNA gene from the positive samples were performed to conduct phylogenetic study. The evaluated blood cells of the studied dog population displayed a wide variety in shapes, sizes, stain pattern and localization of the inclusions. Anaplasmataceae DNA were amplified in 45/55 (81.8%) examined blood samples. Thirty-two samples were positive for E. canis (58.18%) and thirteen for Anaplasma platys (23.63%). Ten animals were negative by PCR and an analyses of a second Buffy coat smear. The obtained nucleotide sequences were analysed for similarity of the 16S rRNA gene sequence using Blast with other sequences available in the GenBank database. The sequences obtained from E. canis positive were closely related to E. canis from Spain (Acession number AY 394465) presenting an identity percentage of between 94.82% and 99.67%. Sequences from Anaplasma sp and Anaplasma platys positive samples were closely related to A. platys from Venezuela (Genbank Acession number AF 399917) presenting an identity betwwen 96.52% and 99.69%, and A. platys from Spain (Genbank Acession number AY 530 806\0 presenting... (Complete abstract, click electronic access below)