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Leishmaniasis is a zoonotic disease caused by over 20 species of protozoan parasites of the genus Leishmania. Infection is commonly spread by sandflies and produces a wide spectrum of clinical signs and symptoms. Therefore, from an epidemiological and therapeutic standpoint, it is important to detect and differentiate Leishmania spp. The objective of this study was to combinate in silico and in vitro strategies to evaluate the analytical specificity of primers previously described in the literature. According to electronic PCR (e-PCR) analysis, 23 out of 141 pairs of primers selected through literature search matched their previously reported analytical specificity. In vitro evaluation of nine of these primer pairs by quantitative PCR (qPCR) confirmed the analytical specificity of five of them at the level of Leishmania spp., L. mexicana complex or Leishmania and Viannia subgenera. Based on these findings, the combination of e-PCR and qPCR is suggested to be a valuable approach to maximize the specificity of new primer pairs for the laboratory diagnosis of infections with Leishmania spp.

Made available in DSpace on 2021-06-25T12:30:02Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-12-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Hospital das Clinicas, Faculdade de Medicina da Universidade de Sao Paulo (HC-FMUSP), Brazil Visceral leishmaniasis (VL) is marked by hyperactivation of a humoral response secreting high quantity of immunoglobulins (Igs) that are inaccessible to intracellular parasites. Here we investigated the contributions of the antibody response to the canine leishmaniasis pathogenesis. Using correlation and genome-wide association analysis, we investigated the relationship of anti-Leishmania infantum immunoglobulin classes levels with parasite burden, clinical response, renal/hepatic biochemical, and oxidative stress markers in dogs from endemic areas of VL. Immunoglobulin G (IgG) and IgA were positively correlated with parasite burden on lymph node and blood. Increased IgG, IgA and IgE levels were associated with severe canine leishmaniasis (CanL) whereas IgM was elevated in uninfected exposed dogs. Correlations of IgM, IgG and IgA with creatinine, urea, AST and ALT levels in the serum were suggested an involvement of those Igs with renal and hepatic changes. The correlogram of oxidative radicals and antioxidants revealed a likely relationship of IgM, IgG and IgA with oxidative stress and lipid pemxidation in the blood, suggested as mechanisms mediating tissue damage and CanL worsening. The gene mapping on chromosomal segments associated with the quantitative variation of immunoglobulin classes identified genetic signatures involved with reactive oxygen species generation, phagolysosome maturation and rupture, free iron availability, Thl/Th2 differenciation and, immunoglobulin clearance. The findings demonstrated the roles of the antibody response as resistance or susceptibility markers and mediators of CanL pathogenesis. In addition we pinpointed candidate genes as potential targets for the therapy against the damage caused by exacerbated antibody response and parasitism in VL. Univ Sao Paulo, Fac Med, Lab Patol Doencas Infecciosas, BR-01246903 Sao Paulo, Brazil Univ Estadual Paulista, Fac Ciencias Agr & Vet, Dept Med Vet Prevent & Reprod Anim, BR-14884900 Jaboticabal, SP, Brazil Univ Salvador, Escola Saude, BR-41720200 Salvador, BA, Brazil Univ Estadual Paulista, Fac Med Vet Aracatuba, Dept Apoio Prod & Saude Anim, BR-16015050 Aracatuba, SP, Brazil Univ Estadual Paulista, Fac Med Vet, Dept Clin Cirurgia & Reprod Anim, BR-16015050 Aracatuba, SP, Brazil Univ Sao Paulo, Fac Med Vet & Zootecnia, Dept Clin, BR-05508270 Sao Paulo, Brazil Univ Estadual Paulista, Fac Ciencias Agr & Vet, Dept Med Vet Prevent & Reprod Anim, BR-14884900 Jaboticabal, SP, Brazil Univ Estadual Paulista, Fac Med Vet Aracatuba, Dept Apoio Prod & Saude Anim, BR-16015050 Aracatuba, SP, Brazil Univ Estadual Paulista, Fac Med Vet, Dept Clin Cirurgia & Reprod Anim, BR-16015050 Aracatuba, SP, Brazil FAPESP: 2012/50285-9 FAPESP: 2012/05847-9 FAPESP: 2014/01095-8 CNPq: 476479/2012-6

Made available in DSpace on 2020-12-12T02:43:21Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-06-05 Background: Visceral leishmaniasis is an important but neglected disease that is spreading and is highly lethal when left untreated. This study sought to measure the Leishmania infantum seroprevalence in dogs, the coverage of its control activities (identification of the canine reservoir by serological survey, dog culling and insecticide spraying) and to evaluate its relationship with the occurrence of the disease in humans in the municipalities of Araçatuba and Birigui, state of São Paulo, Brazil. Methods: Information from 2006 to 2015 was georeferenced for each municipality and modeling was performed for the two municipalities together. To do this, latent Gaussian Bayesian models with the incorporation of a spatio-temporal structure and Poisson distribution were used. The Besag-York-Mollie models were applied for random spatial effects, as also were autoregressive models of order 1 for random temporal effects. The modeling was performed using the INLA (Integrated Nested Laplace Approximations) deterministic approach, considering both the numbers of cases as well as the coverage paired year by year and lagged at one and two years. Results: Control activity coverage was observed to be generally low. The behavior of the temporal tendency in the human disease presented distinct patterns in the two municipalities, however, in both the tendency was to decline. The canine serological survey presented as a protective factor only in the two-year lag model. Conclusions: The canine serological coverage, even at low intensity, carried out jointly with the culling of the positive dogs, suggested a decreasing effect on the occurrence of the disease in humans, whose effects would be seen two years after it was carried out. Department of Epidemiology School of Public Health Universidade de São Paulo (USP), Avenida Doutor Arnaldo 715 Department of Animal Health and Production School of Veterinary Medicine Unesp Center for Parasitology and Mycology Instituto Adolfo Lutz Regional Service 9 Superintendência de Controle de Endemias Zoonosis Control Center MRC-PHE Centre for Environment and Health Department of Epidemiology and Biostatistics Imperial College, Norfolk Place Department of Animal Health and Production School of Veterinary Medicine Unesp

A leishmaniose visceral é uma doença emergente e negligenciada em processo de expansão para áreas urbanas. A incidência da doença humana está relacionada com a infecção canina. Araçatuba e Birigui são municípios do Estado de São Paulo, Brasil, com soroprevalência da infecção canina de 8 a 10%, que empregam estratégias de controle voltadas ao reservatório canino baseado em inquérito sorológico e eutanásia dos cães soropositivos. Usando dados desses programas de controle para parametrizar modelos matemáticos, este estudo avaliou a eficácia dessas atividades. Estimamos que o controle atualmente empregado é capaz de reduzir em cerca de 20% a incidência de casos de leishmaniose visceral canina (LVC). Considerando-se um controle contínuo e um esforço das atividades de inquérito sorológico igual ao triplo da média do observado em Araçatuba e Birigui, a atividade de eutanásia de cães com diagnóstico positivo seria efetiva para o controle da infecção canina. Embora teoricamente possível, na prática, o controle da LVC com as estratégias preconizadas atualmente é insuficiente, pois exigiria superpor dificuldades enfrentadas por estas atividades como falta de recursos materiais, humanos e financeiros, além das questões éticas e jurídicas associadas. Visceral leishmaniasis is an emerging and neglected disease that is currently expanding to urban areas. The incidence of human disease is related to canine infection. Araçatuba and Birigui are municipalities (counties) in the state of São Paulo, Brazil, with 8-10% seroprevalence of canine infection and that employ control strategies targeting the canine reservoir, based on serological survey and culling of seropositive dogs. Using data from these control programs to parameterize mathematical models, this study assessed the efficacy of these activities. We estimated that current control is capable of reducing the incidence of canine visceral leishmaniasis (CVL) by approximately 20%. Assuming continuous control and three times the current serological survey activities in Araçatuba and Birigui, culling dogs with a positive CVL diagnosis would be effective for the control of canine infection. Although theoretically possible, in practice the control of CVL with the currently recommended strategies is insufficient, since it would require overcoming the difficulties in these activities, such as lack of material, human, and financial resources, besides associated ethical and legal issues. La leishmaniasis visceral es una enfermedad emergente y pasada por alto en proceso de expansión hacia áreas urbanas. La incidencia de la enfermedad humana está relacionada con la infección canina. Araçatuba y Birigui son municipios del estado de São Paulo, Brasil, con una seroprevalencia de infección canina de un 8 a un 10% que emplean estrategias de control dirigidas al reservorio canino, basado en una encuesta serológica y eutanasia de los perros seropositivos. Usando datos de esos programas de control para proporcionar parámetros en modelos matemáticos, este estudio evaluó la eficacia de esas actividades. Estimamos que el control actualmente empleado es capaz de reducir cerca de un 20% la incidencia de casos de leishmaniasis visceral canina (LVC). Si se considera un control continuo y un esfuerzo de las actividades de encuesta serológica igual al triple de la media de lo observado en Araçatuba y Birigui, la actividad de eutanasia de perros con diagnóstico positivo sería efectiva para el control de la infección canina. A pesar de que teóricamente es posible, en la práctica el control de la LVC con las estrategias preconizadas actualmente es insuficiente, puesto que exigiría superar dificultades a las que se enfrentan estas actividades como la falta de recursos materiales, humanos y financieros, además de las cuestiones éticas y jurídicas asociadas.

Visceral leishmaniosis (VL) is a public health problem and its occurrence depends primarily on the presence of the vector and susceptible hosts; in the urban environment, the dog is the main reservoir. This study aimed to analyze the distribution of canine visceral leishmaniosis (CVL) and factors associated with it in an urban area endemic for VL. Analysis of the variables was based on 2755 epidemiological records of dogs positive and negative for CVL over a three-year period (2009, 2010 and 2011). A dog was considered positive when it presented amastigotes in the cytological examination of lymph node biopsy and/or was seropositive by immunoenzymatic and indirect immunofluorescence assays. CVL positive dogs were observed throughout the town, but significant differences were observed between the sectors analyzed (P0.0001), with two sectors showing higher positivity. CVL prevalence was 35.9% and was significantly associated with age and breed (P0.0001). Concerning symptoms, 44.3% of symptomatic dogs were positive for LV (P0.0001) in an urban area endemic for this zoonosis.

Made available in DSpace on 2018-11-26T15:45:41Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-09-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Euthanasia of infected dogs is one of the measures adopted in Brazil to control visceral leishmaniasis (VL) in endemic areas. To detect infected dogs, animals are screened with the rapid test DPP (R) Visceral Canine Leishmaniasis for detection of antibodies against K26/K39 fusion antigens of amastigotes (DPP). DPP-positives are confirmed with an immunoenzymatic assay probing soluble antigens of promastigotes (ELISA), while DPP-negatives are considered free of infection. Here, 975 dogs from an endemic region were surveyed by using DPP, ELISA and real-time PCR (qPCR) for the diagnosis of VL. When DPP-negative dogs were tested by qPCR applied in blood and lymph node aspirates, 174/887 (19.6%) were positive in at least one sample. In a second sampling using 115 cases, the DPP-negative dogs were tested by qPCR in blood, lymph node and conjunctival swab samples, and 36/79 (45.6%) were positive in at least one sample. Low-to-moderate pairwise agreement was observed between all possible pair of tests. In conclusion, the official diagnosis of VL in dogs in Brazilian endemic areas failed to accuse an expressive number of infected animals and the impact of the low accuracy of serological tests in the success of euthanasia-based measure for VL control need to be assessed. Univ Sao Paulo, Dept Med Vet Prevent & Saude Anim, Fac Med Vet & Zootecnia, Av Prof Dr Orlando Marques de Paiva 87, BR-05508270 Sao Paulo, SP, Brazil Univ Estadual Paulista, Fac Med Vet, Dept Med Vet, Campus Aracatuba, Aracatuba, SP, Brazil Univ Sao Paulo, Dept Med Vet, Fac Zootecnia & Engn Alimentos, Pirassununga, SP, Brazil Adolfo Lutz Inst, Ctr Parasitol & Micol, Sao Paulo, SP, Brazil Univ Sao Paulo, Fac Saude Publ, Dept Epidemiol, Sao Paulo, SP, Brazil Univ Estadual Paulista, Fac Med Vet, Dept Med Vet, Campus Aracatuba, Aracatuba, SP, Brazil FAPESP: 2011/21796-2 FAPESP: 2011/14892-5

ObjectiveTo evaluate the efficacy of a synthetic sex-aggregation pheromone of the sand fly vector Lu. longipalpis, co-located with residual insecticide, to reduce the infection incidence of Leishmania infantum in the canine reservoir, and to reduce sand fly vector abundance. To compare the outcomes to those resulting from fitting deltamethrin-impregnated collars to the canine reservoir.MethodsA stratified cluster-randomised trial was designed to detect a 50% reduction in canine incident infection after 24 months in 42 recruited clusters, randomly assigned to one of three intervention arms (14 cluster each): pheromone + insecticide, insecticide-impregnated dog collars, or placebo control. Infection incidence was measured by seroconversion to anti-Leishmania antibody, Leishmania parasite detection and canine tissue parasite loads. Changes in relative Lu. longipalpis abundance within households were measured by setting three CDC light traps per household.ResultsA total 1,454 seronegative dogs were follow-up for a median 15.2 (95% C.I.s: 14.6, 16.2) months per cluster. The pheromone + insecticide intervention provided 13% (95% C.I. 0%, 44.0%) protection against anti-Leishmania antibody seroconversion, 52% (95% C.I. 6.2%, 74·9%) against parasite infection, reduced tissue parasite loads by 53% (95% C.I. 5.4%, 76.7%), and reduced household female sand fly abundance by 49% (95% C.I. 8.2%, 71.3%). Variation in the efficacy against seroconversion varied between trial strata. Equivalent protection attributed to the impregnated-collars were 36% (95% C.I. 14.4%, 51.8%), 23% (95% C.I. 0%, 57·5%), 48% (95% C.I. 0%, 73.4%) and 43% (95% C.I. 0%, 67.9%), respectively. Comparison of the outcomes of the two interventions showed no statistically consistent differences in their efficacies; however, the errors were broad for all outcomes. Reductions in sand fly numbers were predominant where insecticide was located (chicken and dog sleeping sites), with no evidence of insecticide-induced repellency onto humans or dogs.ConclusionThe synthetic pheromone lure-and-kill approach provides protection particularly against L. infantum parasite transmission and sand fly vector abundance. The effect estimates are not dissimilar to those of the insecticide-impregnated collars, which are documented to reduce canine infection incidence, and human infection and clinical VL disease incidence, in different global regions. As a low-cost alternative or complimentary vector control tool, optimisation of best community deployment of the pheromone + insecticide are now underway.Author summaryThe sand fly vector of the intracellular parasite Leishmania infantum causing human and canine visceral leishmaniasis in the Americas is Lutzomyia longipalpis. Dogs are the proven reservoir. Vector control tools to reduce transmission suited to this predominantly exophilic vector are lacking. Insecticide-impregnated dog collars protect dogs against infectious bites from sand fly vectors, resulting in reductions of new infections in both dogs and humans. However, collars are costly particularly for endemic communities, and alternative approaches are needed. Recent bulk synthesis of a sex-aggregation pheromone produced by male Lu. longipalpis was shown to attract large numbers of conspecific females to lethal pyrethroid insecticides. This study, conducted in Brazil, evaluated the efficacy of this novel lure-and-kill approach to reduce seroconversion and infection incidence with L. infantum in the canine reservoir, in addition to measuring its impact on household abundance of Lu. longipalpis. Deployed in 14 stratified clusters, the outcomes were compared to those resulting attributed to the collars fitted to dogs in another 14 clusters; each intervention was compared to the 14 clusters that received placebo treatments. The beneficial effects of the lure-and-kill method were most noticeable on confirmed infection incidence and clinical parasite loads, and in reducing sand fly abundance. The overall effect of the two interventions were not statistically dissimilar, though the confidence intervals were broad. We conclude that the novel low-cost lure-and-kill approach should be added to the vector control toolbox against visceral leishmaniasis in the Americas.

Made available in DSpace on 2018-12-11T17:06:15Z (GMT). No. of bitstreams: 0 Previous issue date: 2016-09-15 Intensity of peripheral parasite infection has an important role in the transmission of Leishmania spp. from one host to another. As parasite load quantification is still an expensive procedure to be used routinely in epidemiological surveillance, the use of surrogate predictors may be an important asset in the identification of dogs with high transmitting ability. The present study examined whether common clinical and laboratory alterations can serve as predictors of peripheral parasitism in dogs naturally infected with Leishmania spp. Thirty-seven dogs were examined in order to establish correlations between parasite load (PL) in multiple peripheral tissues and common clinical and laboratory findings in canine visceral leishmaniasis (CVL). Quantitative polymerase chain reaction was employed to determine PL in conjunctival swabs, ear skin, peripheral blood and buffy coat. Additionally, a series of hematological, biochemical and oxidative stress markers were quantified. Correlations between net peripheral infection and severity of clinical alterations and variation in laboratory parameters were assessed through a new analytical approach, namely Compressed Parasite Load Data (CPLD), which uses dimension reduction techniques from multivariate statistics to summarize PL across tissues into a single variable. The analysis revealed that elevation in PL is positively correlated with severity of clinical sings commonly observed in CVL, such as skin lesions, ophthalmic alterations, onycogriphosis, popliteal lymphadenomegaly and low body mass. Furthermore, increase in PL was found to be followed by intensification of non-regenerative anemia, neutrophilia, eosinopenia, hepatic injury and oxidative imbalance. These results suggest that routinely used clinical and laboratory exams can be predictive of intensity of peripheral parasite infection, which has an important implication in the identification of dogs with high transmitting ability. Departamento de Clínica Cirurgia e Reprodução Animal Faculdade de Medicina Veterinária de Araçatuba UNESP- Univ Estadual Paulista, Rua Clóvis Pestana 793, Dona Amélia Departamento de Medicina Veterinária Preventiva e Reprodução Animal Faculdade de Ciências Agrárias e Veterinárias UNESP-Univ Estadual Paulista, Via de Acesso Prof. Paulo Donato Castellane s/n Departamento de Apoio Produção e Saúde Animal Faculdade de Medicina Veterinária de Araçatuba UNESP- Univ Estadual Paulista, Rua Clóvis Pestana 793, Dona Amélia School of Life Sciences and Warwick Infectious Disease and Epidemiology Research Group (WIDER), Gibbett Hill Road Departamento de Clínica Cirurgia e Reprodução Animal Faculdade de Medicina Veterinária de Araçatuba UNESP- Univ Estadual Paulista, Rua Clóvis Pestana 793, Dona Amélia Departamento de Medicina Veterinária Preventiva e Reprodução Animal Faculdade de Ciências Agrárias e Veterinárias UNESP-Univ Estadual Paulista, Via de Acesso Prof. Paulo Donato Castellane s/n Departamento de Apoio Produção e Saúde Animal Faculdade de Medicina Veterinária de Araçatuba UNESP- Univ Estadual Paulista, Rua Clóvis Pestana 793, Dona Amélia

Visceral leishmaniasis is an emerging and neglected disease that is currently expanding to urban areas. The incidence of human disease is related to canine infection. Araçatuba and Birigui are municipalities (counties) in the state of São Paulo, Brazil, with 8-10% seroprevalence of canine infection and that employ control strategies targeting the canine reservoir, based on serological survey and culling of seropositive dogs. Using data from these control programs to parameterize mathematical models, this study assessed the efficacy of these activities. We estimated that current control is capable of reducing the incidence of canine visceral leishmaniasis (CVL) by approximately 20%. Assuming continuous control and three times the current serological survey activities in Araçatuba and Birigui, culling dogs with a positive CVL diagnosis would be effective for the control of canine infection. Although theoretically possible, in practice the control of CVL with the currently recommended strategies is insufficient, since it would require overcoming the difficulties in these activities, such as lack of material, human, and financial resources, besides associated ethical and legal issues.A leishmaniose visceral é uma doença emergente e negligenciada em processo de expansão para áreas urbanas. A incidência da doença humana está relacionada com a infecção canina. Araçatuba e Birigui são municípios do Estado de São Paulo, Brasil, com soroprevalência da infecção canina de 8 a 10%, que empregam estratégias de controle voltadas ao reservatório canino baseado em inquérito sorológico e eutanásia dos cães soropositivos. Usando dados desses programas de controle para parametrizar modelos matemáticos, este estudo avaliou a eficácia dessas atividades. Estimamos que o controle atualmente empregado é capaz de reduzir em cerca de 20% a incidência de casos de leishmaniose visceral canina (LVC). Considerando-se um controle contínuo e um esforço das atividades de inquérito sorológico igual ao triplo da média do observado em Araçatuba e Birigui, a atividade de eutanásia de cães com diagnóstico positivo seria efetiva para o controle da infecção canina. Embora teoricamente possível, na prática, o controle da LVC com as estratégias preconizadas atualmente é insuficiente, pois exigiria superpor dificuldades enfrentadas por estas atividades como falta de recursos materiais, humanos e financeiros, além das questões éticas e jurídicas associadas.La leishmaniasis visceral es una enfermedad emergente y pasada por alto en proceso de expansión hacia áreas urbanas. La incidencia de la enfermedad humana está relacionada con la infección canina. Araçatuba y Birigui son municipios del estado de São Paulo, Brasil, con una seroprevalencia de infección canina de un 8 a un 10% que emplean estrategias de control dirigidas al reservorio canino, basado en una encuesta serológica y eutanasia de los perros seropositivos. Usando datos de esos programas de control para proporcionar parámetros en modelos matemáticos, este estudio evaluó la eficacia de esas actividades. Estimamos que el control actualmente empleado es capaz de reducir cerca de un 20% la incidencia de casos de leishmaniasis visceral canina (LVC). Si se considera un control continuo y un esfuerzo de las actividades de encuesta serológica igual al triple de la media de lo observado en Araçatuba y Birigui, la actividad de eutanasia de perros con diagnóstico positivo sería efectiva para el control de la infección canina. A pesar de que teóricamente es posible, en la práctica el control de la LVC con las estrategias preconizadas actualmente es insuficiente, puesto que exigiría superar dificultades a las que se enfrentan estas actividades como la falta de recursos materiales, humanos y financieros, además de las cuestiones éticas y jurídicas asociadas.

Made available in DSpace on 2019-10-04T12:32:45Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-01-01. Added 1 bitstream(s) on 2019-10-09T18:33:43Z : No. of bitstreams: 1 S0034-89102018000100275.pdf: 1044980 bytes, checksum: bb3c17f918e3d91041429010d5e6adf8 (MD5) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) OBJECTIVE: Estimate the coverage of control measures of visceral leishmaniasis and relate them with the occurrence of human visceral leishmaniasis in endemic urban area. METHODS: Cases of human and canine visceral leishmaniasis were considered as study population and evaluated by a serological survey conducted in Aracatuba, state Sao Paulo, from 2007 to 2015. The cases of human visceral leishmaniasis were geocoded by the address of the patients and the canine disease by the address of the dogs' owners. The coverage of serological survey, euthanasia, and insecticide spraying was calculated, as well as the canine seroprevalence and the incidence rates of human visceral leishmaniasis. The relationship between human visceral leishmaniasis and control measures was evaluated, as well as the seroprevalence by comparing maps and by linear regression. The relationship between the canine and the human disease was also evaluated by the Ripley's K function. RESULTS: The incidence rates of human visceral leishmaniasis showed a period of decline (2007 to 2009) and a period of stability (2010 to 2015), a behavior similar to that of canine seroprevalence. In general, the coverage of control measures was low, and the non-association with the incidence of human visceral leishmaniasis can be a result of the period analyzed and of the small number of analyzed units (sectors of the Superintendence for the Control of Endemic Diseases). The distribution of human cases showed spatial dependence with the distribution of seropositive dogs from 2007 to 2009. CONCLUSIONS: This study reaffirmed the relationship between the occurrence of the disease in humans and dogs, it verified a decrease in the rates of visceral leishmaniasis in Aracatuba over time, even at low coverage of control activities. However, further studies are needed to determine if factors beyond monitoring and control measures are involved in the reduction of incidences. Univ Sao Paulo, Fac Saude Publ, Programa Posgrad Saude Publ, Sao Paulo, SP, Brazil Superintendencia Controle Endemias, Serv Reg 9, Aracatuba, SP, Brazil Univ Estadual Paulista, Fac Med Vet Aracatuba, Dept Apoio Prod & Saude Anim, Lab Bioquim & Biol Mol, Aracatuba, SP, Brazil Adolfo Lutz Inst, Nucleo Parasitoses Sistem, Sao Paulo, SP, Brazil Prefeitura Municipal Aracatuba, Secretaria Municipal Saude, Ctr Controle Zoonoses, Aracatuba, SP, Brazil Fundacao Oswaldo Cruz, Programa Comp Cient, Rio De Janeiro, RJ, Brazil Univ Sao Paulo, Fac Saude Publ, Dept Epidemiol, Ave Doutor Arnaldo 715, BR-01246904 Sao Paulo, SP, Brazil Univ Estadual Paulista, Fac Med Vet Aracatuba, Dept Apoio Prod & Saude Anim, Lab Bioquim & Biol Mol, Aracatuba, SP, Brazil FAPESP: 2014/06518-4 FAPESP: 2014/177646

Made available in DSpace on 2018-12-11T17:22:17Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-07-31 Background: The incidence of visceral leishmaniasis (VL), one of the most important neglected diseases worldwide, is increasing in Brazil. The objectives of this study were to determine the canine VL (CanL) seroprevalence in an urban area of Araçatuba municipality and to evaluate its relationship with the characteristics of dogs and their owners. Results: The CanL seroprevalence in the study area was 0.081 (95% credible interval [CI]: 0.068-0.096). The following covariates/categories were positively associated with the occurrence of a seropositive dog: more than 10 dogs that had lived in the house (odds ratio [OR]=2.36; 95% CI: 1.03-5.43) (baseline: 0-10 dogs); house with dogs that previously died of VL (OR=4.85; 95% CI: 2.65-8.86) or died of causes other than old age (OR=2.26; 95% CI: 1.12-4.46) (baseline: natural or no deaths); dogs that spent the day in a sheltered backyard (OR=2.14; 95% CI: 1.05-4.40); dogs that spent the day in an unsheltered backyard or the street (OR=2.67; 95% CI: 1.28-5.57) (baseline: inside home). Spatial dependence among observations occurred within about 45.7m. Conclusions: The number of dogs that had lived in the house, previous deaths by VL or other cause, and the place the dog stayed during the day were associated with the occurrence of a VL seropositive dog. The short-distance spatial dependence could be related to the vector characteristics, producing a local neighbourhood VL transmission pattern. The geostatistical approach in a Bayesian context using integrated nested Laplace approximation (INLA) allowed to identify the covariates associated with VL, including its spatially dependent transmission pattern. USP Programa de Pós-graduação em Saúde Pública Faculdade de Saúde Pública, Avenida Doutor Arnaldo 715 Imperial College MRC-PHE Centre for Environment and Health Department of Epidemiology and Biostatistics, St. Mary's Campus, Norfolk Place Serviço Regional 9 Superintendência de Controle de Endemias, Rua Minas Gerais, 135 Unesp Laboratório de Bioquimica e Biologia Molecular Departamento de Apoio Producao e Saude Animal Faculdade de Medicina Veterinária de Araçatuba, Rua Clóvis Pestana 793 Núcleo de Parasitoses Sistêmicas Instituto Adolfo Lutz, Av. Doutor Arnaldo 355, 8o. Andar Cento de Laboratório Regional I Araçatuba Instituto Adolfo Lutz, R. Minas Gerais 135 Centro de Controle de Zoonozes, Rua Doutor Luiz de Almeida 145 Fundação Oswaldo Cruz Programa de Computação Científica, Avenida Brasil 4365, Antiga Residência Oficial Universidade de São Paulo (USP) Departamento de Epidemiologia Faculdade de Saúde Pública, Avenida Doutor Arnaldo 715 Unesp Laboratório de Bioquimica e Biologia Molecular Departamento de Apoio Producao e Saude Animal Faculdade de Medicina Veterinária de Araçatuba, Rua Clóvis Pestana 793

Funda??o de Amparo ? Pesquisa do Estado de S?o Paulo, Brazil (FAPESP) grants no. 2012/50285-9 (M.D.L.), no. 2012/05847-9 (L.F.S.B.), and no. 2014/01095-8 (Y.T.U.); Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico (CNPq), Brazil grant no. 476479/2012-6 (M.D.L.); and Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (Capes). Universidade de S?o Paulo. Faculdade de Medicina Veterin?ria e Zootecnia. Departamento de Patologia Veterin?ria. S?o Paulo, SP, Brazil / Universidade Salvador. Escola de Sa?de. Salvador, BA, Brazil. Universidade Estadual Paulista. Faculdade de Ci?ncias Agr?rias e Veterin?rias. Departamento de Medicina Veterin?ria Preventiva e Reprodu??o Animal. Jaboticabal, SP, Brazil. Universidade de S?o Paulo. Faculdade de Medicina. Laborat?rio de Patologia de Doen?as Infecciosas. S?o Paulo, SP, Brazil. Universidade Salvador. Escola de Sa?de. Salvador, BA, Brazil. Universidade Salvador. Escola de Sa?de. Salvador, BA, Brazil. Universidade Salvador. Escola de Sa?de. Salvador, BA, Brazil. Universidade de S?o Paulo. Faculdade de Medicina. Laborat?rio de Patologia de Doen?as Infecciosas. S?o Paulo, SP, Brazil. Universidade de S?o Paulo. Faculdade de Medicina Veterin?ria e Zootecnia. Departmento de Cl?nica. S?o Paulo, SP, Brazil. Universidade Estadual Paulista. Faculdade de Medicina Veterin?ria. Departamento de Cl?nica, Cirurgia e Reprodu??o Animal. Ara?atuba, SP, Brazil. Universidade Estadual Paulista. Faculdade de Ci?ncias Agr?rias e Veterin?rias. Departamento de Medicina Veterin?ria Preventiva e Reprodu??o Animal. Jaboticabal, SP, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Universidade Estadual Paulista. Faculdade de Medicina Veterin?ria. Departamento de Cl?nica, Cirurgia e Reprodu??o Animal. Ara?atuba, SP, Brazil. Universidade Estadual Paulista. Faculdade de Medicina Veterin?ria. Departmento de Sa?de Animal e Produ??o. Ara?atuba, SP, Brazil. Universidade de S?o Paulo. Faculdade de Medicina. Laborat?rio de Patologia de Doen?as Infecciosas. S?o Paulo, SP, Brazil. The anti-inflammatory properties of sand fly saliva favor the establishment of the Leishmania infantum infection. In contrast, an antibody response against Lutzomyia longipalpis saliva is often associated with a protective cell-mediated response against canine visceral leishmaniasis. Genetic studies may demonstrate to what extent the ability to secrete anti-saliva antibodies depends on genetic or environmental factors. However, the genetic basis of canine antibody response against sand fly saliva has not been assessed. The aim of this study was to identify chromosomal regions associated with the anti-Lu. longipalpis salivary IgG response in 189 dogs resident in endemic areas in order to provide information for prophylactic strategies. Dogs were classified into five groups based on serological and parasitological diagnosis and clinical evaluation. Anti-salivary gland homogenate (SGH) IgG levels were assessed by Enzyme-Linked Immunosorbent Assay (ELISA). Genomic DNA was isolated from blood samples and genotyped using a SNP chip with 173,662 single nucleotide polymorphism (SNP) markers. The following linear regression model was fitted: IgG level = mean + origin + sex + age + use of a repellent collar, and the residuals were assumed as pseudo-phenotypes for the association test between phenotypes and genotypes (GWA). A component of variance model that takes into account polygenic and sample structure effects (EMMAX) was employed for GWA. Phenotypic findings indicated that anti-SGH IgG levels remained higher in exposed and subclinically infected dogs than in severely diseased dogs even in regression model residuals. Five associated markers were identified on chromosomes 2, 20 and 31. The mapped genes included CD180 (RP105) and MITF related to the rapid activation of B lymphocytes and differentiation into antibody-secreting plasma cells. The findings pointed to chromosomal segments useful for functional confirmation studies and a search for adjuvant molecules of the anti-saliva response.

Made available in DSpace on 2018-11-27T11:27:10Z (GMT). No. of bitstreams: 0 Previous issue date: 2016-04-01 To present a retrospective study from patients with spinal cysticercosis (SC), diagnosed within the last 30 years in Mexican and Indian neurological referral centers. This is a retrospective and comparative study of the clinical and radiological profile between Mexican and Indian patients with spinal neurocysticercosis during a 30-year period and a review of the literature during the same period. Twenty-seven SC patients were included: 19 from Mexico and 8 from India. SC presented predominantly with motor symptoms (21/27 patients): paraparesis and paraplegia were the most common signs; one-third of patients presented sphincter dysfunction. Imaging studies showed that parasites in vesicular stage were more frequent in patients from Mexico, while degenerative stages predominated in India. Association of subarachnoid cysticerci and hydrocephalus was observed only in Mexican patients. Despite the limitations of this study, the collected information supports the existence of differences in the clinical and radiological traits of SC patients between Asian and Latin-American hospitals. The possible biological factors that may underlie these differences are discussed. Inst Nacl Neurol & Neurocirug, Dept Neuroinfectol, Mexico City, DF, Mexico Hop La Pitie Salpetriere, Dept Neurol, Paris, France Univ Nacl Autonoma Mexico, Dept Immunol, Inst Invest Biomed, Mexico City 04510, DF, Mexico Univ Estadual Paulista, Arcatuba, Brazil Natl Inst Mental Hlth & Neurosci, Bangalore 560029, Karnataka, India Inst Nacl Neurol & Neurocirugia Manuel Velasco Su, Dept Infect Dis, Insurgentes 3877, Mexico City 14269, DF, Mexico Univ Estadual Paulista, Arcatuba, Brazil

Made available in DSpace on 2018-11-26T17:55:48Z (GMT). No. of bitstreams: 0 Previous issue date: 2016-04-01 Aims To detect the presence of Epstein-Barr virus (EBV) DNA in different sources of materials from a matched group of patients with oral lichen planus (OLP) and a group of people without OLP lesions, and to correlate the presence of virus with epidemiological variables of the groups studied. Methods Fresh tissue samples, saliva, exfoliated cells and plasma of 24 patients with OLP lesions (cases) and 17 patients without OLP lesions (controls) were collected. EBV was detected by nested PCR. Results Viral positivity was obtained in 62.5% of tissue samples; in 70.8% of exfoliated cell samples; in 33.3% of blood plasma samples and in 75% of saliva samples in the cases; and in 35.3% of tissue samples; 82.4% of exfoliated cell samples; in 47.1% of blood plasma samples and in 64.7% of saliva samples in the controls. There was a predominance of women in both groups. Variables not atrophic-erosive were most affected by EBV. Conclusions No relationship between EBV and OLP was found. However, all sources tested in this study were considered suitable for the detection of viruses. Aracatuba Sch Dent, Oral Oncol Ctr, Aracatuba, SP, Brazil Aracatuba Sch Dent, Dept Pathol & Clin Propaedeut, Aracatuba, SP, Brazil Aracatuba Sch Vet Med, Anim Hlth & Prod, Aracatuba, SP, Brazil

Leishmania spp. is a protozoan that maintains its life cycle in domestic and wild animals and it may include bats, a population that has increased in urban environments. This study aimed to investigate the presence of Leishmania spp. in bats captured strictly in urban areas that are endemic for visceral leishmaniasis. The spleen and skin samples of 488 bats from 21 endemic cities in northwestern São Paulo State, Brazil, were tested for the presence of Leishmania kDNA using real-time PCR. Differentiation from Trypanosoma spp. was achieved by amplifying a DNA fragment of the ribosomal RNA gene. The presence of Leishmania spp. kDNA was verified in 23.9% of bats and Trypanosoma spp. DNA was identified in 3.9%. Leishmania species differentiation revealed the presence of Leishmania amazonensis in 78.3% of the bats; L. infantum in 17.4%, and 1 sample (4.3%) showed a mix pattern of L. infantum and L. amazonensis. We also detected, for the first time, L. infantum and L. amazonensis DNA in Desmodus rotundus, the hematophagous bat. The presence of Leishmania spp. DNA in bats strictly from urban areas endemic for visceral leishmaniasis in the State of São Paulo, Brazil indicates that these wild and abundant animals are capable of harboring Leishmania spp. in this new scenario. Due to their longevity, high dispersion capacity and adaptability to synanthropic environments, they may play a role in the maintenance of the life cycle of Leishmania parasites.

Made available in DSpace on 2018-11-26T15:47:26Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-02-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Although a national programme for control of visceral leishmaniosis (VL) is being run in Brazil, the disease continues to spread. This programme is essentially based on culling infected dogs from endemic regions. Thus, there is an urgent need to develop other control measures against VL to deter its advance. Here, a subunit vaccine, a recombinant vaccine, an insecticide-impregnated collar and the associations between these measures were evaluated for reducing the incidence of Leishmania infection in dogs. This was through a cohort study conducted in an endemic region of Brazil, considering the incidence and time of total exposure over a period of 1 year. The incidence of VL was estimated by means of serological and molecular diagnostic tests, 180 and 360 days after the application of the control measures. The estimates of the effectiveness (EF) were not significant in any cohort. The EF of the subunit vaccine, the recombinant vaccine and the collar were 26.4%, 32.8% and 57.7% and the upper limit of the 95% confidence interval for EF were 63.7%, 67.9% and 82.5%, respectively. In conclusion, under the conditions of this study, none of the immunogens for VL control was sufficiently effective to protect dogs against infection. On the other hand, use of collars impregnated with insecticide seems to constitute a method with better prognosis, corroborating other studies in this field. Univ Sao Paulo, Fac Med Vet & Zootecnia, Dept Med Vet Prevent & Saude Anim, Sao Paulo, SP, Brazil Univ Estadual Paulista, Fac Med Vet, Dept Med Vet, Campus Aracatuba, Aracatuba, SP, Brazil Univ Sao Paulo, Fac Zootecnia & Engn Alimentos, Dept Med Vet, Pirassununga, SP, Brazil Adolfo Lutz Inst, Ctr Parasitol & Micol, Sao Paulo, SP, Brazil Univ Sao Paulo, Fac Saude Publ, Dept Epidemiol, Sao Paulo, SP, Brazil Univ Estadual Paulista, Fac Med Vet, Dept Med Vet, Campus Aracatuba, Aracatuba, SP, Brazil FAPESP: 2011/21796-2 FAPESP: 2011/14892-5

Made available in DSpace on 2018-12-11T16:44:44Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-10-02 Animal experimentation is a controversial topic, especially among the general public and the scientific community. Thirty-eight undergraduate students attending the College of Veterinary Medicine–São Paulo State University in the municipality of Araçatuba, São Paulo, Brazil, were followed up between 2008 and 2011 and were asked to complete an annual questionnaire focused on different aspects of animal experimentation, including the animal species involved, the objectives of the research, ethics, animal welfare and euthanasia. Most students agreed that animal testing is not morally incorrect, and the dynamics of students’ attitudes were notable: undergraduates tended to change their opinion over time, with junior students opposing animal experimentation more than seniors do, indicating that the more scientific knowledge the students acquire, the more favourable to animal experimentation they become. Nevertheless, they agreed that research must consider the basic principles of animal welfare. Department of Animal Clinics Surgery and Reproduction College of Veterinary Medicine UNESP–Univ Estadual Paulista Department of Production and Animal Health College of Veterinary Medicine; UNESP–Univ Estadual Paulista Laboratoire des Processus Infectieux à Trypanosomatidés Département Infection et Epidemiologie Institut Pasteur Department of Animal Clinics Surgery and Reproduction College of Veterinary Medicine UNESP–Univ Estadual Paulista Department of Production and Animal Health College of Veterinary Medicine; UNESP–Univ Estadual Paulista

Made available in DSpace on 2018-12-11T17:12:17Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-07-01 Background Oral leukoplakia is considered a potentially malignant lesion for the development of squamous cell carcinoma, and various risk factors may be related to its development and malignant transformation, including the human papillomavirus (HPV). The aim of this case-control study was to detect the presence of HPV in fresh tissue, plasma and saliva samples obtained from patients with and without oral leukoplakia, and verify the correlation of the presence of DNA of HPV between different sources of materials. Methods In this study, 32 patients with oral leukoplakia and 24 patients selected in a case-control manner were included. DNA extraction from the samples was performed, and afterwards it was amplified by nested polymerase chain reaction (nPCR) for the detection of HPV (nPCR: MY09-MY11/GP05+-GP06+). Results The DNA of HPV was found in 68.75% of the fresh tissue samples; in 50% of plasma, and in 62.5% of saliva samples in the group of patients with leukoplakia; in comparison with 45.8%, 54%, and 45.8%, in the fresh tissue, plasma and saliva samples, respectively, in the control group. Conclusion Based on the present study, there was no difference in the rate of HPV detection in patients with or without oral leukoplakia. However all sources tested in this study were considered suitable for HPV detection, especially plasma samples, which showed be an important non-invasive source of HPV detection in leukoplakia patients. Oral Oncology Center and Department of Pathology and Clinical Propaedeutics Araçatuba School of Dentistry São Paulo State University (UNESP – Univ Estadual Paulista) Department of Animal Health and Production School of Veterinary Medicine São Paulo State University (UNESP – Univ Estadual Paulista) Oral Oncology Center and Department of Pathology and Clinical Propaedeutics Araçatuba School of Dentistry São Paulo State University (UNESP – Univ Estadual Paulista) Department of Animal Health and Production School of Veterinary Medicine São Paulo State University (UNESP – Univ Estadual Paulista)

Bovine cysticercosis is a problem distributed worldwide that result in economic losses mainly due to the condemnation of infected carcasses. One of the difficulties in applying control measures is the identification of the source of infection, especially because cattle are typically acquired from multiple farms. Here, we tested the utility of an animal movement network constructed with data from a farm that acquires cattle from several other different farms to map the major contributors of cysticercosis propagation. Additionally, based on the results of the network analysis, we deployed a sanitary management and drug treatment scheme to decrease cysticercosis’ occurrence in the farm. Six farms that had commercial trades were identified by the animal movement network and characterized as the main contributors to the occurrence of cysticercosis in the studied farm. The identification of farms with a putative risk of Taenia saginata infection using the animal movement network along with the proper sanitary management and drug treatment resulted in a gradual decrease in cysticercosis prevalence, from 25% in 2010 to 3.7% in 2011 and 1.8% in 2012. These results suggest that the animal movement network can contribute towards controlling bovine cysticercosis, thus minimizing economic losses and preventing human taeniasis. RESUMO: A cisticercose bovina é um problema distribuído mundialmente e que resulta em perdas econômicas, principalmente devido à condenação de carcaças infectadas. Uma das dificuldades em se controlar esta zoonose no Brasil é a prática de aquisição de bovinos de múltiplas fazendas, tornando quase impossível a identificação da fazenda de origem do gado infectado, onde as medidas de controle devem ser aplicadas. Objetivou-se avaliar uma rede de movimentação animal construída com dados de uma fazenda de gado de corte que adquire animais de diferentes locais, com o objetivo de mapear as fazendas que mais contribuíam para a propagação da cisticercose. Adicionalmente, com base na análise da rede de movimentação, manejo sanitário e protocolo de tratamento adequados foram aplicados para diminuir a ocorrência de cisticercose na fazenda em estudo. Seis propriedades que tinham trocas comerciais foram identificadas pela rede de movimentação de bovinos e caracterizadas como as principais contribuintes para a ocorrência da cisticercose na fazenda em estudo. A identificação de fazendas com risco de infecção por Taenia saginata por meio da rede de movimentação animal juntamente com o manejo sanitário e protocolo de tratamento adequados resultaram em diminuição gradual da prevalência da cisticercose de 25%, em 2010, para 3,7% em 2011 e 1,8% em 2012. Estes resultados sugerem que a estratégia de análise da rede de movimentação do gado, associadas ao manejo sanitário e tratamento adequados podem contribuir para o controle da cisticercose bovina, minimizando assim as perdas econômicas e prevenindo a teníase humana.

Visceral leishmaniosis is a zoonotic disease that is transmitted by Lutzomyia longipalpis sandflies. Dogs are the main peri-urban reservoir of the disease, and progression of canine leishmaniosis is dependent on the type of immune response elaborated against the parasite. Type 1 immunity is characterized by effective cellular response, with production of pro-inflammatory cytokines such as tumour necrosis factor alpha (TNF-α). In contrast, Type 2 immunity is predominantly humoral, associated with progression of the disease and mediated by anti-inflammatory cytokines such as interleukin 10 (IL-10). Although seemly important in the dynamics of leishmaniosis, other gene products such as toll-like receptor 2 (TRL-2) and inducible nitric oxide synthase (iNOS) exert unclear roles in the determination of the type of immune response. Given that the dog skin serves as a micro-environment for the multiplication of Leishmania spp., we investigated the parasite load and the expression of TLR-2, iNOS, IL-10 and TNF-α in the skin of 29 infected and 8 control dogs. We found that increased parasite load leads to upregulation of TLR-2, IL-10 and TNF-α, indicating that abundance of these transcripts is associated with infection. We also performed a xenodiagnosis to demonstrate that increased parasitism is a risk factor for infectiousness to sandflies.

Made available in DSpace on 2018-12-11T17:08:34Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-01-30 Quantification of Leishmania infantum load via real-time quantitative polymerase chain reaction (qPCR) in lymph node aspirates is an accurate tool for diagnostics, surveillance and therapeutics follow-up in dogs with leishmaniasis. However, qPCR requires infrastructure and technical training that is not always available commercially or in public services. Here, we used a machine learning technique, namely Radial Basis Artificial Neural Network, to assess whether parasite load could be learned from clinical data (serological test, biochemical markers and physical signs). By comparing 18 different combinations of input clinical data, we found that parasite load can be accurately predicted using a relatively small reference set of 35 naturally infected dogs and 20 controls. In the best case scenario (use of all clinical data), predictions presented no bias or inflation and an accuracy (i.e., correlation between true and predicted values) of 0.869, corresponding to an average error of ±38.2 parasites per unit of volume. We conclude that reasonable estimates of L. infantum load from lymph node aspirates can be obtained from clinical records when qPCR services are not available. São Paulo State University (Unesp). School of Veterinary Medicine Araçatuba. Department of Clinics Surgery and Animal Reproduction São Paulo State University (Unesp) School of Agricultural and Veterinarian Sciences Jaboticabal Department of Preventative Veterinary Medicine and Animal Reproduction USP − Universidade de São Paulo Departamento de Patologia Veterinária Faculdade de Medicina Veterinária e Zootecnia São Paulo State University (Unesp) School of Veterinary Medicine Araçatuba Department of Support Production and Animal Health São Paulo State University (Unesp). School of Veterinary Medicine Araçatuba. Department of Clinics Surgery and Animal Reproduction São Paulo State University (Unesp) School of Agricultural and Veterinarian Sciences Jaboticabal Department of Preventative Veterinary Medicine and Animal Reproduction São Paulo State University (Unesp) School of Veterinary Medicine Araçatuba Department of Support Production and Animal Health

Universidade de S?o Paulo. Faculdade de Medicina Veterin?ria e Zootecnia. Departamento de Patologia Veterin?ria. S?o Paulo, SP, Brazil / Universidade de S?o Paulo. Faculdade de Medicina. Laborat?rio de Patologia de Doen?as Infecciosas. S?o Paulo, SP, Brazil. Universidade Estadual Paulista. Faculdade de Ci?ncias Agr?rias e Veterin?rias. Departamento de Medicina Veterin?ria Preventiva e Reprodu??o Animal. Jaboticabal, SP, Brazil. Universidade de S?o Paulo. Faculdade de Medicina. Laborat?rio de Patologia de Doen?as Infecciosas. S?o Paulo, SP, Brazil. Universidade de S?o Paulo. Faculdade de Medicina. Laborat?rio de Patologia de Doen?as Infecciosas. S?o Paulo, SP, Brazil. Universidade de S?o Paulo. Faculdade de Medicina. Laborat?rio de Patologia de Doen?as Infecciosas. S?o Paulo, SP, Brazil. Universidade Estadual Paulista. Faculdade de Medicina Veterin?ria. Departamento de Cl?nica, Cirurgia e Reprodu??o Animal. Ara?atuba, SP, Brazil. Universidade de S?o Paulo. Faculdade de Medicina. Laborat?rio de Patologia de Doen?as Infecciosas. S?o Paulo, SP, Brazil. Universidade de S?o Paulo. Faculdade de Medicina. Laborat?rio de Patologia de Doen?as Infecciosas. S?o Paulo, SP, Brazil / Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Universidade Estadual Paulista. Faculdade de Medicina Veterin?ria. Departamento de Cl?nica, Cirurgia e Reprodu??o Animal. Ara?atuba, SP, Brazil Universidade Estadual Paulista. Faculdade de Medicina Veterin?ria. Departmento de Sa?de Animal e Produ??o. Ara?atuba, SP, Brazil Universidade de S?o Paulo. Faculdade de Medicina. Laborat?rio de Patologia de Doen?as Infecciosas. S?o Paulo, SP, Brazil. A genome-wide association study (GWAS) could unravel the complexity of the cell-mediated immunity (CMI) to canine leishmaniasis (CanL). Therefore, we scanned 110,165 single-nucleotide polymorphisms (SNPs), aiming to identify chromosomal regions associated with the leishmanin skin test (LST), lymphocyte proliferation assay (LPA), and cytokine responses to further understand the role played by CMI in the outcome of natural Leishmania infantum infection in 189 dogs. Based on LST and LPA, four CMI profiles were identified (LST?/LPA?, LST+/LPA?, LST?/LPA+, and LST+/LPA+), which were not associated with subclinically infected or diseased dogs. LST+/LPA+ dogs showed increased interferon gamma (IFN-?) and tumor necrosis factor alpha (TNF-?) levels and mild parasitism in the lymph nodes, whereas LST?/LPA+ dogs, in spite of increased IFN-?, also showed increased interleukin-10 (IL-10) and transforming growth factor ? (TGF-?) levels and the highest parasite load in lymph nodes. Low T cell proliferation under low parasite load suggested that L. infantum was not able to induce effective CMI in the early stage of infection. Altogether, genetic markers explained 87%, 16%, 15%, 11%, 0%, and 0% of phenotypic variance in TNF-?, TGF-?, LST, IL-10, IFN-?, and LPA, respectively. GWAS showed that regions associated with TNF-? include the following genes: IL12RB1, JAK3, CCRL2, CCR2, CCR3, and CXCR6, involved in cytokine and chemokine signaling; regions associated with LST, including COMMD5 and SHARPIN, involved in regulation of NF-?B signaling; and regions associated with IL-10, including LTBP1 and RASGRP3, involved in T regulatory lymphocytes differentiation. These findings pinpoint chromosomic regions related to the cell-mediated response that potentially affect the clinical complexity and the parasite replication in canine L. infantum infection.

The aim of this study was to detect cross infections by Leishmania spp. and Trypanosoma spp. using enzyme-linked immunosorbent assay (ELISA), indirect fluorescent antibody test (IFAT) and polymerase chain reaction (PCR). Thus, 408 blood samples were collected from dogs domiciled in Aracatuba Municipality, Sao Paulo State, Brazil; the dogs were of both sexes, of several breeds and aged 6 months. For Leishmania spp., 14.95 % (61 out of 408) of dogs were reactive using IFAT. Positivity was 20.10 % (82 out of 408) using ELISA and 29.66 % (121 out of 408) using PCR, with significant differences for the sex and age of these animals (p

Leishmaniasis is kept in nature by the participation of several animal species. This study evaluated the presence of Leishmania spp. in skin samples of free-ranging marsupials Micoureus paraguayanus (n = 95) and Didelphis albiventris (n = 191), captured in Morro do Diabo State Park and in sections of its surrounding forest, in the region of Pontal do Paranapanema, Sao Paulo State, Brazil. The samples were tested for the presence of kDNA of Leishmania spp. by polymerase chain reaction (PCR) and by real time PCR (qPCR). All samples from D. albiventris tested by PCR were negative for the presence of kDNA of Leishmania spp. However, when tested by qPCR, the positivity was 1.6%. A positivity of 7.4% by PCR and 11.6% by qPCR was observed for M. paraguayanus. Sixty-four per cent (9/14) of positive animals were limited to the same forest fragment. Presence of Leishmania (Leishmania) amazonensis and Leishmania (Viannia) braziliensis was detected in M. paraguayanus samples. While D. albiventris is the most studied marsupial species due to its urban habits, other marsupial species such as M. paraguayanus can be potential reservoirs of Leishmania spp. and should also be studied.

Cysticercosis is a major cause of economic loss in bovine production due to meat condemnation. Chemotherapy is being used in Brazilian cattle and a diagnostic test to improve the treatment program is desired. We produced monoclonal antibodies against crude (TAEB) and cyst fluid (TAEF) Taenia saginata metacestode antigens using immunized BALB/c mice. After cell fusion, 10 TAEB and nine TAEF hybrids were selected and cloned resulting in 18 IgG(1) and 32 IgM TAEB clones, and 9 IgG(1) and 9 IgM TAEF clones. Ascites was produced and Western blot testing was performed resulting in reactivity to protein fractions of low molecular weight (18kDa), 43, 55, 66 and 100kDa. The indirect immunofluorescence test, with one monoclonal antibody against crude and one against cyst fluid antigens, recognized antigenic fractions of both the scolex and the bladder wall of metacestodes from naturally infected bovine.

The Leishmune vaccine has been used in endemic areas to prevent canine visceral leishmaniasis in Brazil, but cytokine production induced by vaccination has rarely been investigated in dogs. This study aimed to evaluate the immune response of dogs vaccinated with Leishmune FML vaccine (Fort Dodge) against total antigen of Leishmania (Leishmania) chagasi (TAg) and FML. Twenty healthy dogs from Aracatuba, Sao Paulo, Brazil, an endemic leishmaniasis area, received three consecutive subcutaneous injection of Leishmune vaccine at 21-day intervals. PBMC were isolated before and 10 days after completing vaccination and lymphoproliferative response and antibody production against FML or total promastigote antigen were tested. Cytokines IFN-gamma, IL-4 and TNF-alpha were measured in culture supernatant and CD4+/CD25+ and CD8+/CD25+ T cell presence was determined. Analysis of the data indicated that the vaccine conferred humoral responses (100%) against both antigens and cellular immunity to FML (85%) and total antigen (80%), the supernatant of cultured cells stimulated with TAg and FML showed an increase in IFN-gamma (P

Made available in DSpace on 2016-07-19T13:51:17Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-04-01. Added 1 bitstream(s) on 2016-07-19T14:08:07Z : No. of bitstreams: 1 S1676-24442010000200003.pdf: 159026 bytes, checksum: c2652900208285190bbab98104c177df (MD5) Fundação para o Desenvolvimento da UNESP (FUNDUNESP) Introdução: Recentemente o papilomavírus humano (HPV) tem sido associado à carcinogênese oral. A metodologia empregada na detecção do vírus é uma das maiores causas observadas da grande variabilidade nas taxas de detecção do HPV. Objetivo: Este estudo comparou a sensibilidade de detecção do DNA do HPV em casos de carcinoma epidermoide de lábio utilizando a amplificação do DNA viral por reação em cadeia da polimerase (PCR) ou nPCR. Material e método: Foram utilizadas 33 amostras provenientes de casos de carcinoma epidermoide de lábio. Para as extrações do DNA utilizou-se o sistema QIAamp DNA Mini Kit. Como controle interno utilizou-se o gene da b-globina. Das 33 amostras iniciais, 30 foram positivas para o gene b-globina, sendo utilizadas para detectar o DNA viral. Comparou-se a amplificação do DNA viral pelos métodos da PCR com os oligonucleotídeos MY09/MY11 e nPCR, empregando-se os pares de oligonucleotídeos iniciadores MY09/MY11 e, na segunda etapa, o par GP5+/GP6+. O controle positivo para a presença do DNA do HPV utilizado foi a linhagem de células HeLa e, como controle negativo, a mistura de amplificação sem DNA. A análise dos produtos de PCR e nPCR para HPV foi realizada por eletroforese em gel de poliacrilamida a 8%. Resultados: Utilizando-se o método da PCR, a amplificação do DNA do HPV foi constatada em dois casos. Com a nPCR foi verificada presença de DNA viral em 13 das 30 amostras. Conclusão: Com a utilização da nPCR, a detecção do HPV nos casos estudados aumentou mais de seis vezes. Introduction: Human papillomavirus (HPV) has been currently associated with oral carcinogesis. The methodology applied in virus detection is one of the main reasons for the great variability observed in HPV detection. Objectives: This study compared HPV DNA detection efficiency in lip squamous cell carcinoma samples (SCC) using viral DNA amplification by PCR or nPCR. Methods: Thirty three samples of lip squamous cell carcinoma were selected. DNA extractions were performed with QIAamp DNA minikit. The internal control was b-globin gene. Thirty out of 33 initial samples were positive for b-globin gene, which were employed to detect viral DNA. The amplification of viral DNA was compared by PCR method through MY09/MY11 oligonucleotides and nPCR through MY09/MY11 oligonucleotides in the first stage and GP5+/GP6+ in the second stage. HeLa cells were used as positive control for HPV DNA presence and the amplification mixture without DNA as negative control. The analysis of PCR and nPCR products for HPV DNA was performed through gel electrophoresis in polyacrylamide at 8%. Results: HPV DNA amplification was verified in two cases by the use of PCR method. HPV DNA presence was verified in 13 out of 30 samples by the use of nPCR. Conclusion: HPV DNA detection of lip SCC increased sixfold in the studied cases with the employment of nPCR. Universidade Estadual Paulista Júlio de Mesquita Filho, Departamento de Apoio, Produção e Saúde Animal da Faculdade de Medicina Veterinária de Araçatuba da UNESP Universidade Estadual Paulista Júlio de Mesquita Filho, Departamento de Apoio, Produção e Saúde Animal da Faculdade de Medicina Veterinária de Araçatuba da UNESP. FUNDUNESP: 00149/07

Submitted by Guilherme Lemeszenski (guilherme@nead.unesp.br) on 2013-08-22T18:57:01Z No. of bitstreams: 1 S0102-311X2009001200021.pdf: 75958 bytes, checksum: 3f0821826d4d18d55a23f75ee02e9857 (MD5) Made available in DSpace on 2013-08-22T18:57:01Z (GMT). No. of bitstreams: 1 S0102-311X2009001200021.pdf: 75958 bytes, checksum: 3f0821826d4d18d55a23f75ee02e9857 (MD5) Previous issue date: 2009-12-01 Made available in DSpace on 2013-09-30T19:51:29Z (GMT). No. of bitstreams: 2 S0102-311X2009001200021.pdf: 75958 bytes, checksum: 3f0821826d4d18d55a23f75ee02e9857 (MD5) S0102-311X2009001200021.pdf.txt: 16679 bytes, checksum: 20e9bddd709fb0b8a2bcea6406b7ee51 (MD5) Previous issue date: 2009-12-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:42:22Z No. of bitstreams: 2 S0102-311X2009001200021.pdf: 75958 bytes, checksum: 3f0821826d4d18d55a23f75ee02e9857 (MD5) S0102-311X2009001200021.pdf.txt: 16679 bytes, checksum: 20e9bddd709fb0b8a2bcea6406b7ee51 (MD5) Made available in DSpace on 2014-05-20T13:42:22Z (GMT). No. of bitstreams: 2 S0102-311X2009001200021.pdf: 75958 bytes, checksum: 3f0821826d4d18d55a23f75ee02e9857 (MD5) S0102-311X2009001200021.pdf.txt: 16679 bytes, checksum: 20e9bddd709fb0b8a2bcea6406b7ee51 (MD5) Previous issue date: 2009-12-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Fundação para o Desenvolvimento da UNESP (FUNDUNESP) As pesquisas relativas a agressões por animais baseiam-se em dados de registros de pacientes atendidos nos serviços de vigilância epidemiológica da raiva. Analisaram-se as características de animais agressores, obtidas a partir de dados de 10.616 fichas de envio de amostras para diagnóstico da raiva na região noroeste do Estado de São Paulo, Brasil, entre 1993-2007. Desse total de fichas, 61,5% continham informações sobre agressão e, dentre os animais agressores (25%), a maioria foi de cães (67%), seguidos por gatos (21,8%) e quirópteros (8,1%). em 92,1% dos relatos as vítimas eram pessoas e 82,3% dos animais eram domiciliados. A maioria dos cães agressores tinha até um ano de idade. Houve associação significante (p < 0,0001) entre agressividade e sexo, sendo os machos mais agressores do que as fêmeas. Dos animais positivos para raiva, 75,9% (183/241) eram agressores. Os dados obtidos são importantes para a instituição de programas de prevenção de mordeduras, baseados no perfil não só da vítima, mas também do animal agressor, além de permitirem estabelecer fatores de risco para ocorrência de agressões. Epidemiological factors related to animal bites in humans and other animals were obtained from a database with 10,616 records of animal specimens sent for rabies diagnosis in northwest São Paulo State, Brazil, from 1993 to 2007. of this total, 61.5% contained information on the bites, and among the biting animals (25%), the majority were dogs (67%), followed by cats (21.8%), and bats (8.1%). In 92.1% of the reports the victims were humans, and 82.3% of the animals were home pets. The majority of the biting dogs were less than a year old. There was a significant association (p < 0.0001) between aggressiveness and gender (with male animals more aggressive than females). of the rabies-positive animals, 75.9% (183/241) were biters. The data are important for establishing bite prevention programs based on the profile of both victims and biting animals, besides identifying risk factors for animal bites. Universidade Estadual Paulista Júlio de Mesquita Filho Universidade Estadual Paulista Júlio de Mesquita Filho FAPESP: 04/06740-7 FAPESP: 06/02612-0 FUNDUNESP: 00829/2008-DFP

Submitted by Guilherme Lemeszenski (guilherme@nead.unesp.br) on 2013-08-22T18:44:56Z No. of bitstreams: 1 S1984-29612008000300001.pdf: 42329 bytes, checksum: c71076b90570012b2dfdffc4b532a856 (MD5) Made available in DSpace on 2013-08-22T18:44:56Z (GMT). No. of bitstreams: 1 S1984-29612008000300001.pdf: 42329 bytes, checksum: c71076b90570012b2dfdffc4b532a856 (MD5) Previous issue date: 2008-09-01 Made available in DSpace on 2013-09-30T18:25:11Z (GMT). No. of bitstreams: 2 S1984-29612008000300001.pdf: 42329 bytes, checksum: c71076b90570012b2dfdffc4b532a856 (MD5) S1984-29612008000300001.pdf.txt: 27264 bytes, checksum: 1745b5c726fb0990a1c3e8c62c843652 (MD5) Previous issue date: 2008-09-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:42:35Z No. of bitstreams: 2 S1984-29612008000300001.pdf: 42329 bytes, checksum: c71076b90570012b2dfdffc4b532a856 (MD5) S1984-29612008000300001.pdf.txt: 27264 bytes, checksum: 1745b5c726fb0990a1c3e8c62c843652 (MD5) Made available in DSpace on 2014-05-20T13:42:35Z (GMT). No. of bitstreams: 2 S1984-29612008000300001.pdf: 42329 bytes, checksum: c71076b90570012b2dfdffc4b532a856 (MD5) S1984-29612008000300001.pdf.txt: 27264 bytes, checksum: 1745b5c726fb0990a1c3e8c62c843652 (MD5) Previous issue date: 2008-09-01 O cão doméstico desempenha importante papel como reservatório na transmissão da leishmaniose visceral ao homem, zoonose de grande importância em saúde pública. Realizou-se avaliação epidemiológica da leishmaniose visceral em 1.112 cães domiciliados no município de Poxoréo, estado do Mato Grosso e observou-se prevalência de 7,8%. Observou-se ainda associação estatisticamente significativa entre a prevalência de leishmaniose visceral canina e as variáveis faixa etária, presença de sinais clínicos e presença de outra espécie animal co-habitando com os cães avaliados, tendo sido as galinhas mais freqüentemente observadas entre os animais soropositivos. O sexo, a coleta de lixo domiciliar bem como a renda familiar não apresentaram associação estatisticamente significativa com a prevalência da leishmaniose visceral canina. A análise dos resultados sugere que cães com idade superior a sete anos e a , presença de outra espécie animal co-habitando com os cães podem ser fatores de risco para a leishmaniose visceral canina. Dogs play an important role as reservoir in the domestic cycle of visceral leishmaniasis, a serious public health problem. An epidemiological survey in 1,112 dogs was conducted at the Municipality of Poxoréo State of Mato Grosso, Brazil, using indirect immunofluorescence antibody test where the prevalence was 7.8%. Significant association was found between prevalence of canine visceral leishmaniasis and age of the dogs. Clinical signs and presence of other animals in the backyard, like chicken being more likely associated with seropositivity. Gender, garbage collection in the residence and family financial income were not associated with visceral leishmaniasis prevalence. Analysis of the results suggests that dogs aging more than 7 years and presence of another animal species co-inhabiting with the dogs may be risk factors for canine visceral leishmaniasis. Universidade Estadual Paulista Júlio de Mesquita Filho Departamento de Higiene Veterinária e Saúde Pública Faculdade de Medicina Veterinária e Zootécnica Unesp-Botucatu Instituto de Biociências Unesp-Araçatuba Departamento de Apoio, Produção e Saúde Animal Universidade Estadual Paulista Júlio de Mesquita Filho Departamento de Higiene Veterinária e Saúde Pública Faculdade de Medicina Veterinária e Zootécnica Unesp-Botucatu Instituto de Biociências Unesp-Araçatuba Departamento de Apoio, Produção e Saúde Animal Univ Estadual Paulista, Fac Med Vet & Zootecn, Dept Higiene Vet & Saúde Publ, BR-18618000 Botucatu, SP, Brazil

Submitted by Guilherme Lemeszenski (guilherme@nead.unesp.br) on 2013-08-22T18:57:00Z No. of bitstreams: 1 S0102-311X2008000400024.pdf: 277207 bytes, checksum: a9e9f7c133c347b124ef5d64e3418123 (MD5) Made available in DSpace on 2013-08-22T18:57:00Z (GMT). No. of bitstreams: 1 S0102-311X2008000400024.pdf: 277207 bytes, checksum: a9e9f7c133c347b124ef5d64e3418123 (MD5) Previous issue date: 2008-04-01 Made available in DSpace on 2013-09-30T19:51:24Z (GMT). No. of bitstreams: 2 S0102-311X2008000400024.pdf: 277207 bytes, checksum: a9e9f7c133c347b124ef5d64e3418123 (MD5) S0102-311X2008000400024.pdf.txt: 16746 bytes, checksum: 8c0b3d0771b82277d8a17362a67185e7 (MD5) Previous issue date: 2008-04-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T15:13:21Z No. of bitstreams: 2 S0102-311X2008000400024.pdf: 277207 bytes, checksum: a9e9f7c133c347b124ef5d64e3418123 (MD5) S0102-311X2008000400024.pdf.txt: 16746 bytes, checksum: 8c0b3d0771b82277d8a17362a67185e7 (MD5) Made available in DSpace on 2014-05-20T15:13:21Z (GMT). No. of bitstreams: 2 S0102-311X2008000400024.pdf: 277207 bytes, checksum: a9e9f7c133c347b124ef5d64e3418123 (MD5) S0102-311X2008000400024.pdf.txt: 16746 bytes, checksum: 8c0b3d0771b82277d8a17362a67185e7 (MD5) Previous issue date: 2008-04-01 No período de 1994 a 2004, a população canina de Araçatuba, São Paulo, Brasil, registrou duas importantes zoonoses: a raiva e a leishmaniose visceral. Analisaram-se as mudanças ocorridas nessa população durante esse período, utilizando resultados de censos caninos e de coletas censitárias de sangue realizados em 1994, 1999 e 2004. A relação cão/10 habitantes variou significativamente, passando de 1,7 em 1994 para 2,0 em 1999 e para 1,8 em 2004. A porcentagem de cães com até um ano de idade passou de 20% para 32,5% e o número de eutanásias realizadas também aumentou após 1999, com a introdução da leishmaniose visceral. O número de cães e a estrutura etária variaram nos diversos setores do município e aqueles com maior porcentagem de animais com até dois anos de idade apresentaram maior ocorrência de casos de leishmaniose visceral humana e canina. Tais resultados decorrem de ações de controle adotadas nos setores com casos humanos de leishmaniose visceral, porém, o aumento da população canina mais jovem pode resultar em aumento da susceptibilidade destes cães à doença, favorecendo a manutenção da mesma na área. From 1994 to 2004, the canine population in Araçatuba, São Paulo State, Brazil, suffered two major canine zoonoses: rabies and visceral leishmaniasis. Changes in the dog population during this period were evaluated using canine census data from 1994 and 2004 and the results of blood samples for diagnosis of canine visceral leishmaniasis in 1999. The ratio of dogs per 10 inhabitants varied from 1.7 in 1994 to 2.0 in 1999 and 1.8 in 2004. The percentage of puppies less than 1 year of age increased from 20% to 32.5%, and the number of euthanized dogs also increased after 1999, when visceral leishmaniasis began to appear. The number of dogs and percentage of puppies varied between different areas of the city, and neighborhoods with a higher percentage of young animals showed more cases of both human and canine leishmaniasis. This result may be due to control measures applied in these areas in response to cases of human and canine visceral leishmaniasis, but the increase in the younger canine population can be accompanied by increased susceptibility in these animals, thus favoring maintenance of the disease in the area. Agência Nacional de Vigilância Sanitária Universidade Estadual Paulista Departamento de Apoio, Produção e Saúde Animal Universidade Estadual Paulista Departamento de Apoio, Produção e Saúde Animal

The age structure of the dog population is essential for planning and evaluating control programs for zoonotic diseases. We analyzed data of an owned-dog census in order to characterize, for the first time, the structure of a dog population under compulsory culling in a visceral leishmaniasis endemic area (Panorama, São Paulo State, Brazil) that recorded a dog-culling rate of 28% in the year of the study. Data on 1,329 households and 1,671 owned dogs revealed an owned dog:human ratio of 1:7. The mean age of dogs was estimated at 1.73 years; the age pyramid indicated high birth and mortality rates at the first year of age with an estimated cumulative mortality of 78% at the third year of age and expected life span of 2.75 years. In spite of the high mortality, a growth projection simulation suggested that the population has potential to grow in a logarithmic scale over the years. The estimated parameters can be further applied in models to maximize the impact and minimize financial inputs of visceral leishmaniasis control measures. Resumo: É importante conhecer a estrutura etária da população canina para melhor planejar e avaliar os programas de controle de zoonoses. Os autores analisaram os dados de um censo de cães com donos para caracterizar, pela primeira vez, a estrutura etária de uma população canina sujeita à eliminação compulsória numa área endêmica de leishmaniose visceral (Panorama, Estado de São Paulo, Brasil) que registrou uma taxa de eliminação canina de 28% no ano do estudo. Os dados para 1.329 domicílios e 1.671 cães com donos mostraram uma razão de cães para humanos de 1:7. A média de idade dos cães foi estimada em 1,73 anos; a pirâmide etária indicou altas taxas de natalidade e de mortalidade no primeiro ano de vida, com uma estimativa de mortalidade cumulativa de 78% aos três anos de idade, e uma expectativa de vida de 2,75 anos. Apesar da alta mortalidade, uma simulação de projeção de crescimento sugeriu que a população tem o potencial de crescer numa escala logarítmica ao longo dos anos. Os parâmetros estimados podem ser aplicados também a modelos para maximizar o impacto e minimizar os insumos financeiros de medidas de controle da leishmaniose visceral. Resumen: El conocimiento de la estructura etaria de una población de perros es esencial para la planificación de programas de control de zoonosis. Se analizaron datos de un censo de población canina domiciliada, con el objetivo de caracterizar, por primera vez, la estructura de una población de perros domiciliados en un área donde la eutanasia de perros positivos en leishmaniasis visceral es obligatoria (Panorama, São Paulo, Brasil), y que registró un 28% de casos de eutanasia en el año en que el censo fue realizado. Los datos de 1.329 domicilios y 1.671 perros resultaron en una razón perro:hombre de 1:7. La edad media de los perros fue estimada en 1,73 años; la pirámide de edad indica altas tasas de nacimiento y mortalidad hasta 1 año de vida, con tasa de mortalidad acumulada de un 78% en el tercer año de vida, y expectativa de vida de 2,75 años. A pesar de la alta tasa de mortalidad, la simulación de crecimiento poblacional sugiere que esta población tiene potencial de crecimiento en escala logarítmica a lo largo de los años. Los parámetros estimados pueden ser utilizados en modelos para maximizar el impacto y minimizar los costes de las medidas de control de la enfermedad.

International audience; Abstract One of the measures to control visceral leishmaniosis (VL) in Brazil is the identification and culling of the canine reservoir. There is much controversy concerning this strategy, including the proper identification of positive dogs and the fact that the host-parasite relationship changes over time make it more challenging. A dynamic cohort of 62 dogs was followed every three months using serological and parasitological examinations and PCR. Positivity by PCR was higher than by serology and by parasitological examinations and showed a tendency to decrease over time, while serology tended to increase after six months. Concomitant positivity in all tests was observed in 10.4% of the samples, and negativity in 29.1%. Overall sensitivity ranged from 43.6 to 64.1%, and was not uniform over time. The proportion of dogs with or without clinical signs was not different by cytology or PCR but PCR was able to identify a larger number of asymptomatic dogs compared to ELISA and immunochromatography. PCR can be useful for surveillance of areas where cases of canine VL have not yet been detected and in which control strategies can be implemented to limit the spread of the disease. Despite the advance in diagnostic tools CVL diagnosis remains a challenge.; Resumo Uma das medidas de controle da leishmaniose visceral (LV) no Brasil se baseia na identificação e eliminação do reservatório canino. Existe considerável controvérsia relativa a esta estratégia incluindo a correta identificação dos cães positivos e a variação temporal da relação hospedeiro-parasita, o que torna esta medida ainda mais desafiadora. Uma coorte dinâmica de 62 cães foi acompanhada trimestralmente utilizando-se métodos sorológicos, parasitológicos e a PCR. A taxa de positividade por PCR foi maior em comparação à dos métodos sorológicos e parasitológicos, e mostrou tendência à diminuição com o passar do tempo, enquanto que a positividade sorológica apresentou tendência a aumento, após seis meses. Observou-se positividade concomitante em todos os testes em 10,4% das amostras e, negatividade concomitante, em 29,1%. A sensibilidade geral variou de 43,6% a 64,1%, não sendo uniforme ao longo do estudo. A proporção de cães com e sem sinais clínicos que foram positivos ao exame parasitológico ou à PCR não foi estatisticamente diferente. Contudo, foi possível identificar como positivos um maior número de animais assintomáticos por meio da técnica de PCR, em comparação aos testes ELISA e imunocromatográfico. A PCR pode ser bastante útil para a vigilância epidemiológica de áreas onde casos de LV canina ainda não tenham sido descritos e onde estratégias de controle podem ser implantadas para limitar a disseminação da doença. Não obstante o avanço nas ferramentas diagnósticas, diagnosticar a LVC continua um desafio.