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      Prevalence and Factors Associated with Leishmania infantum Infection of Dogs from an Urban Area of Brazil as Identified by Molecular Methods

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          Abstract

          Background

          Various factors contribute to the urbanization of the visceral leishmaniasis (VL), including the difficulties of implementing control measures relating to the domestic reservoir. The aim of this study was to determine the prevalence of canine visceral leishmaniasis in an urban endemic area in Brazil and the factors associated with Leishmania infantum infection among seronegative and PCR-positive dogs.

          Methodology

          A cross-sectional study was conducted in Belo Horizonte, Minas Gerais, Brazil. Blood samples were collected from 1,443 dogs. Serology was carried out by using two enzyme-linked immunosorbent assays (Biomanguinhos/FIOCRUZ/RJ and “in house”), and molecular methods were developed, including PCR-RFLP. To identify the factors associated with early stages of infection, only seronegative (n = 1,213) animals were evaluated. These animals were divided into two groups: PCR-positive (n = 296) and PCR-negative (n = 917) for L. infantum DNA. A comparison of these two groups of dogs taking into consideration the characteristics of the animals and their owners was performed. A mixed logistic regression model was used to identify factors associated with L. infantum infection.

          Principal Findings

          Of the 1,443 dogs examined, 230 (15.9%) were seropositive in at least one ELISA, whereas PCR-RFLP revealed that 356 animals (24.7%) were positive for L. infantum DNA. Results indicated that the associated factors with infection were family income<twice the Brazilian minimum salary (OR 2.3; 95%CI 1.4–3.8), knowledge of the owner regarding the vector (OR 1.9; 95%CI 1.1–3.4), the dog staying predominantly in the backyard (OR 2.2; 95%CI 1.1–4.1), and a lack of previous serological examination for VL (OR 1.5; 95%CI 1.1–2.3).

          Conclusions

          PCR detected a high prevalence of L. infantum infection in dogs in an area under the Control Program of VL intervention. Socioeconomic variables, dog behavior and the knowledge of the owner regarding the vector were factors associated with canine visceral leishmaniasis (CVL). The absence of previous serological examination conducted by the control program was also associated with L. infantum infection. It is necessary to identify the risk factors associated with CVL to understand the expansion and urbanization of VL.

          Author Summary

          Visceral leishmaniasis (VL) is a disease caused by the parasite Leishmania infantum, and dogs are the most important domestic reservoirs of the agent. During recent decades, VL has expanded to large Brazilian urban centers. In the present work, we have demonstrated by using molecular techniques that the rate of canine infection as detected by serology has been considerably underestimated. Two groups of seronegative dogs (infected and non-infected according to molecular methods) were further evaluated from data obtained through interviews with owners of the animals. The factors associated with Leishmania infection in dogs were a family income of less than two minimum salaries, the knowledge of the owner regarding the vector, the dog spending most of its time in the backyard and the dog never having had a previous serological examination. Awareness regarding the factors associated with canine infection will improve health services and the understanding of the disease's expansion in urban areas.

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          Most cited references57

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          The role of conceptual frameworks in epidemiological analysis: a hierarchical approach.

          This paper discusses appropriate strategies for multivariate data analysis in epidemiological studies. In studies where determinants of disease are sought, it is suggested that the complex hierarchical inter-relationships between these determinants are best managed through the use of conceptual frameworks. Failure to take these aspects into consideration is common in the epidemiological literature and leads to underestimation of the effects of distal determinants. An example of this analytical approach, which is not based purely on statistical associations, is given for assessing determinants of mortality due to diarrhoea in children. Conceptual frameworks provide guidance for the use of multivariate techniques and aid the interpretation of their results in the light of social and biological knowledge.
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            Directions for the diagnosis, clinical staging, treatment and prevention of canine leishmaniosis.

            Canine leishmaniosis (CanL) due to Leishmania infantum is a life threatening zoonotic disease with a wide distribution in four continents and importance also in non-endemic regions. The purpose of this report is to present a consensus of opinions on the diagnosis, treatment, prognosis and prevention of CanL in order to standardize the management of this infection. CanL is a disease in which infection does not equal clinical illness due to the high prevalence of subclinical infection among endemic canine populations. The most useful diagnostic approaches include serology by quantitative techniques and PCR. High antibody levels are associated with severe parasitism and disease and are diagnostic of clinical leishmaniosis. However, the presence of lower antibody levels is not necessarily indicative of disease and further work-up is necessary to confirm CanL by other diagnostic methods such as cytology, histopathology and PCR. We propose a system of four clinical stages, based on clinical signs, clinicopathological abnormalities and serological status. Suitable therapy and expected prognosis are presented for each of the stages. The combination of meglumine antimoniate and allopurinol constitutes the first line pharmaceutical protocol. However, although most dogs recover clinically after therapy, complete elimination of the parasite is usually not achieved and infected dogs may eventually relapse. Follow-up of treated dogs with blood counts, serum biochemistry, urinalysis, serology and PCR is essential for prevention of relapses. Protection against sand fly bites by topical insecticides is effective in reducing infection, and recent development of vaccines has indicated that prevention by vaccination is feasible.
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              Canine leishmaniosis - new concepts and insights on an expanding zoonosis: part one.

              Recent research has provided new insights on the epidemiology, pathology and immunology of canine leishmaniosis (CanL) and its genetic basis. The prevalence of infection in endemic areas is considerably higher than that of apparent clinical illness. In addition, infection spreads rapidly among dogs in the presence of optimal conditions for transmission. Infection involves a variety of granulomatous and harmful immune-mediated responses, and susceptibility to the disease is influenced by a complex genetic basis. These concepts will be instrumental for devising control programs. This review, the first in a series of two articles on CanL, presents an updated view on progress in elucidating the epidemiology and pathogenesis of this challenging disease, and the second part focuses on advances in diagnosis, treatment and prevention.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS Negl Trop Dis
                plos
                plosntds
                PLoS Neglected Tropical Diseases
                Public Library of Science (San Francisco, USA )
                1935-2727
                1935-2735
                August 2011
                16 August 2011
                : 5
                : 8
                : e1291
                Affiliations
                [1 ]Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil
                [2 ]Núcleo de Pesquisas em Ciências Biológicas, Instituto de Ciências Exatas e Biológicas, Universidade Federal de Ouro Preto, Ouro Preto, Minas Gerais, Brazil
                [3 ]Departamento de Ciências Biológicas, Universidade Federal de Alfenas, Alfenas, Minas Gerais, Brazil
                [4 ]Departamento de Farmácia, Escola de Farmácia, Universidade Federal de Ouro Preto, Ouro Preto, Minas Gerais, Brazil
                [5 ]Gerência Regional de Controle de Zoonoses, Secretaria Municipal de Saúde, Prefeitura de Belo Horizonte, Belo Horizonte, Minas Gerais, Brazil
                [6 ]Pós-Graduação em Infectologia e Medicina Tropical, Faculdade de Medicina, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil
                Institute of Tropical Medicine, Belgium
                Author notes

                Conceived and designed the experiments: ABR MJM MC. Performed the experiments: WC-V VMV BMR RDdOA-S LESR SLB MHFM. Analyzed the data: WC-V ABR MC. Contributed reagents/materials/analysis tools: ABR MC. Wrote the paper: WC-V ABR MJM MC. Responsible for the molecular methods: MJM VMV. Responsible for serological methods: ABR. Responsible for analysis: MC.

                Article
                PNTD-D-11-00047
                10.1371/journal.pntd.0001291
                3156685
                21858243
                4d607d46-42f6-4476-96a4-d6b5687fe166
                Coura-Vital et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
                History
                : 21 January 2011
                : 13 July 2011
                Page count
                Pages: 10
                Categories
                Research Article
                Medicine
                Epidemiology
                Infectious Disease Epidemiology
                Molecular Epidemiology
                Public Health
                Veterinary Science
                Veterinary Diseases
                Zoonotic Diseases
                Leishmaniasis
                Veterinary Parasitology
                Veterinary Epidemiology
                Veterinary Medicine
                Veterinary Diagnostics

                Infectious disease & Microbiology
                Infectious disease & Microbiology

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