Factors associated with adherence to the principles of responsible companion animal guardianship in a municipality in southeastern Brazil.

Responsible companion animal guardianship (RCAG) covers aspects that are relevant to both animal and human health. Understanding the factors associated with adherence to RCAG principles can guide accountability, animal welfare and disease control. The present observational study describes the level of adherence to RCAG by guardians living in a medium-sized Brazilian municipality and identifies the factors associated with inadequate guardianship. Questionnaires were administered to randomly selected guardians of 704 dogs. The binary outcome of the study, namely more and less adequate compliers to RCAG principles, was analyzed using a score list comprising eight variables relating to the care provided to the dogs, namely provision of veterinary services, vaccination against rabies, deworming, no access to the streets without supervision, walking with guardian, dog freedom at home, registration (microchipping) and satisfactory food supply. Factors possibly associated with less adequate adherence to RCAG by the guardian, such as socioeconomic and cultural features of the guardians, characteristics of the dogs and attributes of the guardian-animal interactions, were analyzed using multiple logistic regression models. The RCAG actions least adopted by guardians were animal registration, provision of veterinary care, walking with the dog and preventing access of the dogs to the streets without supervision. Individuals who cared for a single dog, a mixed breed dog or had previously lost a dog were less likely to show adequate adherence to RCAG. Conversely, guardians who owned cars, acquired dogs as puppies, lived in households with a maximum of four residents, cared for a sterilized dog or thought that caring for a dog was easier than (or as) expected, had a higher chance of showing adequate adherence to RCAG. The results verify that the socioeconomic and behavioral characteristic of guardians must be taken into consideration for understanding the adherence to RCAG. In addition, it is important to facilitate access to veterinary services and to raise awareness about the significance of a safe and healthy environment for companion animals.

Diagnostic Methods for Helicobacter pylori.

H. pylori infection is a significant global health concern. It cannot be diagnosed based solely on the patient's medical history and symptoms, and laboratory and imaging tests are often required to confirm the diagnosis. Both non-invasive and invasive methods are available for diagnosing H. pylori infection, including conventional and advanced detection techniques. It is not uncommon for patients to present with false-negative results due to the use of inadequate investigation methodologies, which prevents the adoption of appropriate clinical management. Thus, an analysis of the literature regarding the methods of diagnosis of H. pylori, with its advantages and disadvantages, is necessary. Publications in specialized scientific journals will undoubtedly contribute to facilitating access by professionals interested in the topic providing greater knowledge and potentially clinically useful guidance. In this review, the authors have sought to analyze and summarize the invasive and non-invasive methods, their applications, limitations, and the conditions that affect the sensitivity of the tests used for diagnosing H. pylori, an essential step for the successful treatment of this infection. It is essential to treat all patients infected with H. pylori. This represents a significant change in the approach, as previously treatment was recommended only for patients showing symptoms of infection. Therefore, it is crucial to understand the limitations of traditional diagnostic methods and help raise awareness among healthcare professionals about the latest advances in diagnosing this important bacterium.

Diagnosis of Canine Visceral Leishmaniasis by Flow Cytometry Serology using the rMELEISH Multiepitope Antigen Coupled in a Functional Bead.

BACKGROUND: Visceral leishmaniasis (VL) is a zoonotic disease, with dogs being the main reservoir of the Leishmania infantum parasite. OBJECTIVE: To develop a new flow cytometry test to diagnosis canine VL (CVL) diagnosis. METHODS: The current study addresses a new flow cytometry test using beads coupled to the multiepitope antigen rMELEISH. RESULTS: In the study set of samples a sensitivity (87.1%) and specificity (89.9%) was observed. Considering the dogs' clinical status, 20/20 (100.0%) of the symptomatic sera tested positive, while 19/22 (86.4%) of the oligosymptomatic and 16/20 (80.0%) of asymptomatic were positive. In the non-infected control, all samples (0/30) tested as negative. In the cross-reaction control, the test was more efficient in dogs infected with L. braziliensis (2/10) and Trypanosoma cruzi (0/10), than those with Babesia canis (4/10) and Ehrlichia canis (4/10). Dogs immunized with different vaccines (Leishmune, Leish-Tec®, or LBSap) did not present serological reactivity. CONCLUSION: The flow cytometry serology through coupling the antigen rMELEISH in functional beads showed high accuracy in diagnosing CVL.

First report of putative Leishmania RNA virus 2 (LRV2) in Leishmania infantum strains from canine and human visceral leishmaniasis cases in the southeast of Brazil.

BACKGROUND: Leishmania RNA virus 1 (LRV1) is commonly found in South American Leishmania parasites belonging to the subgenus Viannia, whereas Leishmania RNA virus 2 (LRV2) was previously thought to be restricted to the Old-World pathogens of the subgenus Leishmania. OBJECTIVES: In this study, we investigated the presence of LRV2 in strains of Leishmania (L.) infantum, the causative agent of visceral leishmaniasis (VL), originating from different hosts, clinical forms, and geographical regions. METHODS: A total of seventy-one isolates were screened for LRV2 using semi-nested reverse transcription-polymerase chain reaction (RT-PCR) targeting the RNA-dependent RNA polymerase (RdRp) gene. FINDINGS: We detected LRV2 in two L. infantum isolates (CUR268 and HP-EMO) from canine and human cases, respectively. MAIN CONCLUSIONS: To the best of our knowledge, this is the first detection of LRV2 in the New World.

The Influence of Community Feeders and Commercial Food Outlets on the Spatial Distribution of Free-Roaming Dogs-A Photographic Capture and Recapture Study.

Understanding the distribution of dogs in the environment is relevant for establishing human and animal health actions. In the present study, we analyzed the influence of community feeders and commercial food outlets on the spatial distribution of free-roaming dogs in an urban area of a municipality in Southeast Brazil. The dogs were identified via photographic capture and recapture performed over five sampling efforts. The spatial densities of dogs were determined using the Kernel method. Spatial correlations between the distribution of free-roaming dogs and the locations of community feeders and commercial food outlets were analyzed using the K function. During the study, 1207 captures/recaptures were performed encompassing 554 dogs, the majority (62.6%) of which were males. Agglomerations of male and female dogs were observed in the areas where food was present. Positive spatial autocorrelations were detected between the distribution of dogs and food sources. The median distances between dogs and community feeders or commercial food outlets were 1.2 and 1.4 km, respectively, and the difference between these two was statistically significant. The presence of community feeders and food outlets demonstrates the influence of human activity, on the spatial distribution of free-roaming dogs. These results will be useful for developing strategies aimed at the improvement of animal welfare and the prevention of zoonoses.

rMELEISH: A Novel Recombinant Multiepitope-Based Protein Applied to the Serodiagnosis of Both Canine and Human Visceral Leishmaniasis.

BACKGROUND: visceral leishmaniasis (VL) is a critical public health problem in over ninety countries. The control measures adopted in Brazil have been insufficient when it comes to preventing the spread of this overlooked disease. In this context, a precise diagnosis of VL in dogs and humans could help to reduce the number of cases of this disease. Distinct studies for the diagnosis of VL have used single recombinant proteins in serological assays; however, the results have been variable, mainly in relation to the sensitivity of the antigens. In this context, the development of multiepitope-based proteins could be relevant to solving such problem. METHODS: a chimeric protein (rMELEISH) was constructed based on amino acid sequences from kinesin 39 (k39), alpha-tubulin, and heat-shock proteins HSP70 and HSP 83.1, and tested in enzyme-linked immunosorbent (ELISA) for the detection of L. infantum infection using canine (n = 140) and human (n = 145) sera samples. RESULTS: in the trials, rMELEISH was able to discriminate between VL cases and cross-reactive diseases and healthy samples, with sensitivity and specificity values of 100%, as compared to the use of a soluble Leishmania antigenic extract (SLA). CONCLUSIONS: the preliminary data suggest that rMELEISH has the potential to be tested in future studies against a larger serological panel and in field conditions for the diagnosis of canine and human VL.

Development of quantitative PCR and digital PCR for the quantification of Leishmania infantum in dogs.

Leishmaniasis is a zoonotic disease with worldwide distribution. In the Americas, the causative agent of the visceral form is the protozoa Leishmania (Leishmania) infantum. Transmission to the host or vertebrate reservoir occurs through the bite of infected arthropod females like Lutzomyia longipalpis. The epidemiological connection between the infection in dogs and humans generate constant studies about the relationship between the parasite and the canine host, including the development of methods and tests for the detection and quantification ofLeishmania (L.) infantum. Both conventional PCR (cPCR) and quantitative PCR (qPCR) can be used in the diagnosis of the parasite. Dropet Digital PCR (ddPCR) is another useful tool. Knowing the parasite load and its relationship with the clinical signs of naturally infected dogs is useful in research development and for establishing treatments that reduce the transmission of the disease. In this study, thirty-nine clinical samples of spleen from dogs naturaly infected by L. infantum were collected after necropsy. Two molecular tools were used to quantify the parasite load (qPCR and ddPCR) and there was 100% agreement in the results of the them. The tools developed in this work are important for the detection of L. infantum in dogs and humans. Droplet Digital PCR does not require a standard curve and is easy to standardize. In such manner, this new tool can generate more in-depth information in the broad debate about parasitic loads and the pathogenesis of leishmaniasis.

First report of putative Leishmania RNA virus 2 (LRV2) in Leishmania infantum strains from canine and human visceral leishmaniasis cases in the southeast of Brazil

BACKGROUND Leishmania RNA virus 1 (LRV1) is commonly found in South American Leishmania parasites belonging to the subgenus Viannia, whereas Leishmania RNA virus 2 (LRV2) was previously thought to be restricted to the Old-World pathogens of the subgenus Leishmania. OBJECTIVES In this study, we investigated the presence of LRV2 in strains of Leishmania (L.) infantum, the causative agent of visceral leishmaniasis (VL), originating from different hosts, clinical forms, and geographical regions. METHODS A total of seventy-one isolates were screened for LRV2 using semi-nested reverse transcription-polymerase chain reaction (RT-PCR) targeting the RNA-dependent RNA polymerase (RdRp) gene. FINDINGS We detected LRV2 in two L. infantum isolates (CUR268 and HP-EMO) from canine and human cases, respectively. MAIN CONCLUSIONS To the best of our knowledge, this is the first detection of LRV2 in the New World.

Responsible companion animal guardianship is associated with canine visceral leishmaniasis: an analytical cross-sectional survey in an urban area of southeastern Brazil.

BACKGROUND: Responsible companion animal guardianship (RCAG) comprises a set of concepts involving activities, behavior and care that guardians must provide to ensure the welfare of their animals. When such principles are disregarded, the risk of animals developing zoonotic diseases, such as canine visceral leishmaniasis (CVL), increases. This disease is a public health problem in many urban settings in Brazil because dogs are the main reservoirs of Leishmania and are involved in the transmission of the parasites to humans. Our analytical cross-sectional epidemiological survey aimed to investigate the prevalence of CVL in a city in southeastern Brazil and to establish the association between the disease and a number of predictor variables including dog traits, socioeconomic status of guardians, ecological features of the domicile and RCAG. RESULTS: Our study showed that the global prevalence of CVL in the sample canine population was 6.7% (47/704). All variables related to better dog care were associated with lower chances of infection. Multiple regression analysis revealed that the chances of animals being seropositive for CVL were significantly (p < 0.05) higher when guardians had no formal education or possessed a university degree (vs. those with complete primary or secondary schooling) and when dogs were sheltered outside the house and had free access to the streets. An additional novel finding was that dogs that were acquired as puppies presented half of the chance of developing the disease in comparison with those acquired at the adult stage. Geographically weighted logistic regression coefficients showed that the strengths of the predictor/CVL associations varied depending on the studied geographical space. Both models demonstrated that the associations were always in the same directions. CONCLUSIONS: Our findings indicate that regardless of age and mode of acquisition, adult dogs should be submitted to clinical evaluation and tests for CVL. RCAG can exert positive effects on the control of CVL.

Accuracy of diagnostic tests for American tegumentary leishmaniasis: a systematic literature review with meta-analyses.

OBJECTIVES: To analyse the accuracy of American tegumentary leishmaniasis (ATL) diagnostic methods and evaluate the quality of the existing publications by means of a systematic review. METHODS: Diagnostic tests evaluated in at least two studies with common reference standards were included in the sensitivity and/or specificity meta-analyses. Quality and susceptibility to bias were analysed using the QUADAS-2 and STARD tools. RESULTS: The title and abstract of 3387 publications were evaluated after deduplication resulting from database searches. Thirty-eight studies were included in the review, and 26 of them had results inserted in meta-analyses. The diagnostic methods with the highest pooled sensitivity values were ELISA, polymerase chain reaction (PCR), indirect immunofluorescence reaction and Montenegro's intradermal reaction. Cytometry was assessed in only two studies and presented 100% sensitivity in both. Smear slide microscopy and histopathology showed low pooled values of sensitivity. For specificity, the highest pooled values were identified for PCR. High values were also identified for ELISA, except for studies in which the reference standard for defining negative participants included individuals with Chagas' disease or paracoccidioidomycosis, which also occurred for cytometry. IFR had lower specificities than ELISA. There was a predominance of case-control designs of phase 1 or 2 and only four studies were strongly recommended as evidence generators. Several reference standards were adopted, and different methods were assessed in a small number of studies. CONCLUSION: PCR showed the highest accuracy for the diagnosis of ATL, and its use should be encouraged in clinical practice. ELISA is recommended for the screening of suspected individuals, but the possibility of cross-reactions should be considered. New validation studies for the tests evaluated in few publications and studies of phase 3 with appropriate methods are needed.

OBJECTIFS: Analyser l'exactitude des méthodes de diagnostic de la leishmaniose tégumentaire américaine (LTA) et évaluer la qualité des publications existantes au moyen d'une analyse systématique. MÉTHODES: Les tests diagnostiques évalués dans au moins deux études avec des étalons de référence communs ont été inclus dans les méta-analyses de sensibilité et/ou de spécificité. La qualité et la sensibilité au biais ont été analysées à l'aide des outils QUADAS-2 et STARD. RÉSULTATS: Le titre et le résumé de 3387 publications ont été évalués après déduplication résultant de recherches dans la base de données. 38 études ont été incluses dans la revue et 26 d'entre elles ont eu des résultats inclus dans des méta-analyses. Les méthodes de diagnostic avec les valeurs de sensibilité poolées les plus élevées étaient ELISA, la réaction en chaîne par polymérase (PCR), la réaction d'immunofluorescence indirecte et la réaction intradermique du Monténégro. La cytométrie a été évaluée dans seulement deux études et présentait une sensibilité de 100% dans les deux. La microscopie et l'histopathologie sur lame de frottis ont montré de faibles valeurs poolées de sensibilité. Pour la spécificité, les valeurs poolées les plus élevées ont été identifiées pour la PCR. Des valeurs élevées ont également été identifiées pour l'ELISA, à l'exception des études dans lesquelles la norme de référence pour définir les participants négatifs incluait des individus atteints de la maladie de Chagas ou de paracoccidioïdomycose, qui s'est également produite pour la cytométrie. L'IFR avait des spécificités plus faibles que l'ELISA. Il y avait une prédominance des essais cas-témoins de phases 1 ou 2 et seules quatre études étaient fortement recommandées comme génératrices de preuves. Plusieurs normes de référence ont été adoptées et différentes méthodes ont été évaluées dans un petit nombre d'études. CONCLUSION: la PCR a montré la plus grande exactitude pour le diagnostic de l'ATL et son utilisation doit être encouragée dans la pratique clinique. L'ELISA est recommandé pour le dépistage des personnes suspectées, mais la possibilité de réactions croisées doit être considérée. De nouvelles études de validation des tests évalués dans quelques publications et des études de phase 3 avec des méthodes appropriées sont nécessaires.

Effects of Gender, Sterilization, and Environment on the Spatial Distribution of Free-Roaming Dogs: An Intervention Study in an Urban Setting.

Information concerning the factors affecting the circulation and distribution of free-roaming dogs is crucial in developing control actions and limiting the spread of zoonoses. The present study analyzes the influence of gender, sterilization, and environment on the spatial distribution of free-roaming dogs in urban settings. Animals were captured/recaptured in seven consecutive morning sampling efforts conducted at 2-monthly intervals in control and intervention areas in a medium-size town in southeastern Brazil. Capture locations were georeferenced and captured animals were microchipped before being released at their original capture sites. Dogs captured in the intervention area were subjected additionally to surgical sterilization prior to release. Home range (HR) areas were calculated by applying the minimum convex polygon method to dogs that had been captured at least three times. Land coverage zones were determined from satellite images and overlaid on maps of the study areas along with the locations of 22 commercial food outlets. HR areas showed a global mean of 448 m2 and a median of 28 m2, values that were smaller than those reported previously for dogs in rural regions. The median HR of females (64.m2) was higher than that of males (15 m2), while median HRs of animals in the control and intervention areas were similar (27 and 28.5 m2, respectively). Variability of HR was high, although animals with small HRs predominated. Free-roaming dogs grouped primarily in urbanized and transitional regions, and their spatial distribution was positively correlated with locations of commercial food outlets. While sterilization did not influence HR size, the search for food was a key factor in determining mobility and spatial aggregation of free-roaming dogs. Our findings are pertinent in understanding the ecology of free-roaming dogs in urban environments and will be applicable to strategies aimed at promoting animal welfare and preventing the dissemination of zoonoses.

Antileishmanial activity evaluation of thiazolidine-2,4-dione against Leishmania infantum and Leishmania braziliensis.

Leishmaniasis is responsible for approximately 65,000 annual deaths. Despite the mortality data, drugs available for the treatment of patients are insufficient and have moderate therapeutic efficacy in addition to serious adverse effects, which makes the development of new drugs urgent. To achieve this goal, the integration of kinetic and DSF assays against parasitic validated targets, along with phenotypic assays, can help the identification and optimization of bioactive compounds. Pteridine reductase 1 (PTR1), a validated target in Leishmania sp., is responsible for the reduction of folate and biopterin to tetrahydrofolate and tetrahydrobiopterin, respectively, both of which are essential for cell growth. In addition to the in vitro evaluation of 16 thiazolidine-2,4-dione derivatives against Leishmania major PTR1 (LmPTR1), using the differential scanning fluorimetry (ThermoFluor®), phenotypic assays were employed to evaluate the compound effect over Leishmania braziliensis (MHOM/BR/75/M2903) and Leishmania infantum (MHOM/BR/74/PP75) promastigotes viability. The ThermoFluor® results show that thiazolidine-2,4-dione derivatives have micromolar affinity to the target and equivalent activity on Leishmania cells. 2b is the most potent compound against L. infantum (EC50 = 23.45 ± 4.54 µM), whereas 2a is the most potent against L. braziliensis (EC50 = 44.16 ± 5.77 µM). This result suggests that lipophilic substituents on either-meta and/or-para positions of the benzylidene ring increase the potency against L. infantum. On the other hand, compound 2c (CE50 = 49.22 ± 7.71 µM) presented the highest selectivity index.

Toxicity and Anti-promastigote Activity of Benzoxazinoid Analogs Against Leishmania (Viannia) braziliensis and Leishmania (Leishmania) infantum.

Purpose: Here, we aim to evaluate the antileishmanial activity of compounds with a benzoxazinoid (BX) skeleton, previously synthesized by our group, against Leishmania (Viannia) braziliensis and Leishmania (Leishmania) infantum promastigotes. Methods: Anti-promastigote activity, as well as cytotoxicity, were determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetric assays. The selectivity index (SI) for each compound was calculated using a ratio of the cytotoxicity of compounds and the geometric mean (GM) of antileishmanial concentrations to each species tested. The comparisons between groups were carried out using a t test or analysis of variance (one-way ANOVA). A P value of less than 0.05 was considered significant. Results: All the compounds tested were active, with IC50 falling between 92±6.19 µg/mL and 238±6.57 µg/mL for L. braziliensis, and 89±6.43 µg/mL and 188±3.58 µg/mL against L. infantum. Bex2, Bex3, Pyr1, Pyr2, and Pyr4 were compounds that showed activity similar to the drug Glucantime®, exhibited low cytotoxicity against splenic hamster cells (CC50 raging between >400 and 105.7±2.26 µg/mL) and had favorable selectivity indices (SI 1.12 to 3.96). Conclusion: The analogs in question are promising prototypes for the pharmaceutical development of novel, safer and more effective leishmanicidal agents.

Space-time analysis of the incidence of human visceral leishmaniasis (VL) and prevalence of canine VL in a municipality of southeastern Brazil: Identification of priority areas for surveillance and control.

Human and canine visceral leishmaniasis (HVL and CVL, respectively) represent serious public health issues in Brazil. The surveillance and control measures currently employed have had limited effect in impeding the territorial expansion of the disease and in reducing the number of cases. We have investigated the space-time distribution of HVL incidence rates and CVL prevalence in the coverage areas of the 148 primary healthcare units in Belo Horizonte (MG, Brazil) during a 6-year period in order to identify those that should be prioritized for disease control actions. Data were smoothed using the empirical Bayes method and analyzed by space-time scanning and application of univariate global Moran's I index and local indicators of spatial association (LISA) statistics to identify spatial autocorrelations. Point data of CVL were analyzed using the Kernel method. Bivariate global Moran's I and LISA techniques were employed to identify spatial correlations between HVL and CVL. Based on our results, we were able to formulate two proposals for establishing the prioritization of coverage areas, namely: (i) classification of maximum priority areas as identified by bivariate LISA for HVL and CVL, and (ii) combination of maximum priority areas with high priority areas as identified by univariate LISA for HVL. According to our proposals, 27 coverage areas in Belo Horizonte were categorized as maximum priority and a further 13 were classified as high priority. Our proposals, which are based on practical, feasible and inexpensive statistical tools, will contribute to a better understanding of VL distribution in urban settings and improving the efficiency of governmental control programs.

IgG avidity index and complete blood count as biomarkers of clinical disease in naturally infected dogs with Leishmania infantum.

Canine visceral leishmaniosis (CVL), a parasitic disease caused by Leishmania infantum, may evolve to a chronic condition and lead to death. Evaluation of infected dogs is important to establish the clinical and laboratory parameters involved in the evolution of the disease. The objectives of the present study were to discriminate a canine population (n = 52) into sub-clinical and clinically affected dogs based on signs and scores, to evaluate the hematological, biochemical, histopathological and parasitological parameters of the two dog groups, and to analyze the results by multivariate regression analysis with the aim of establishing biomarkers of CVL clinical disease. The most common signs observed in the clinically affected dogs (n = 29) were hyperkeratosis, weight loss, onychogryphosis, pale mucosa and lymphadenomegaly. In the multivariate analysis, animals presenting high IgG avidity index and low red blood, lymphocyte and eosinophil counts, and low serum urea concentration had an increased probability of being classified as clinically affected (p < 0.05). All five parameters were considered to be strong biomarkers for monitoring the clinical disease, while IgG avidity percentage was strongly correlated with the number of clinical signs and could function as an indicator of the duration of infection. This is the first report on the application of IgG avidity and of multivariate regression analysis in establishing associations between the clinical signs of CVL and host biomarkers. Since avidity index (AI) percentages were strongly correlated with the number of clinical signs, it could be useful in clinical practice for auxiliary diagnosis of CVL and monitoring disease progression. A limitation of this study is the lack of information on co-infections by Anaplasma platys, Babesia canis vogeli, Ehrlichia canis and Hepatozoon canis. Therefore future studies should evaluate the influence of such co-infections on the associations studied using multivariate methods with larger samples.

Molecular characterization of Leishmania infantum isolates in municipalities of the state of Minas Gerais, Brazil.

Background: Little is known about the genetic diversity of Leishmania infantum isolates in the state of Minas Gerais, Brazil. Methods: Molecular characterization of hsp70, mpi and ITS1 was carried out for 29 isolates of L. infantum from the bone marrow of naturally infected dogs from the cities of Divinópolis, Pará de Minas and Brumadinho, located in the central-west, central and central regions of the state of Minas Gerais, Brazil, respectively. Results: Analysis of the parasite nucleotide sequences demonstrated very high homogeneity of the studied samples. Conclusions: In the endemic regions studied, parasites are genotypically indistinguishable.

Abundance, survival, recruitment and effectiveness of sterilization of free-roaming dogs: A capture and recapture study in Brazil.

The existence of free-roaming dogs raises important issues in animal welfare and in public health. A proper understanding of these animals' ecology is useful as a necessary input to plan strategies to control these populations. The present study addresses the population dynamics and the effectiveness of the sterilization of unrestricted dogs using capture and recapture procedures suitable for open animal populations. Every two months, over a period of 14 months, we captured, tagged, released and recaptured dogs in two regions in a city in the southeast region of Brazil. In one of these regions the animals were also sterilized. Both regions had similar social, environmental and demographic features. We estimated the presence of 148 females and 227 males during the period of study. The average dog:man ratio was 1 dog for each 42 and 51 human beings, in the areas without and with sterilization, respectively. The animal population size increased in both regions, due mainly to the abandonment of domestic dogs. Mortality rate decreased throughout the study period. Survival probabilities did not differ between genders, but males entered the population in higher numbers. There were no differences in abundance, survival and recruitment between the regions, indicating that sterilization did not affect the population dynamics. Our findings indicate that the observed animal dynamics were influenced by density-independent factors, and that sterilization might not be a viable and effective strategy in regions where availability of resources is low and animal abandonment rates are high. Furthermore, the high demographic turnover rates observed render the canine free-roaming population younger, thus more susceptible to diseases, especially to rabies and leishmaniasis. We conclude by stressing the importance of implementing educational programs to promote responsible animal ownership and effective strategies against abandonment practices.

Reliability of techniques used in the diagnosis of canine visceral leishmaniasis by the national control program in Brazil: A survey in an area of recent transmission.

One of the key components of the Brazilian Program for the Control of Visceral Leishmaniasis (PCLV) is the euthanasia of Leishmania-infected canine reservoirs, the detection of which depends on a screening procedure involving a Dual Path Platform® (DPP) immunoassay and a confirmatory enzyme-linked immunosorbent assay (ELISA). The aims of the present study were to evaluate the reliability of these techniques in a region of recent transmission of canine VL, to follow up the seroconversion 3-4 months after the initial diagnosis of DPP reactive but ELISA indeterminate or non-reactive dogs, and to identify the species of Leishmania in circulation in the area. Each animal was submitted to DPP under field conditions, performed by municipal health workers using peripheral blood (DPP-field), to DPP under laboratory conditions using serum (DPP-lab) and to ELISA using serum. The agreements between the tests were determined using McNemar's χ2 test, Cohen's kappa coefficient (k) at the 95% confidence interval and prevalence-adjusted bias-adjusted kappa (PABAK). Of the 1130 dogs examined, 74.2% were non-reactive in all three tests applied. Based on the PCLV positive-infection criterion, seroprevalence was 8.9% (101/1130) with 83.2% (84/101) of infected animals showing reactivity in all three tests while 7.8% (8/101) were reactive in DPP-field and ELISA and 8.9% (9/101) in DPP-lab and ELISA. The proportions of disagreements were substantial in all comparisons. Inter-rater reliability between DPP-field and ELISA (k=0.55; PABAK=0.78) and DPP-lab and ELISA (k=0.59; PABAK=0.81) were considered moderate, while that between DPP-field and DPP-lab (k=0.61; PABAK=0.79) was classified as marginally good. The proportion of seroconversions in DPP reactive animals that were initially ELISA indeterminate was significantly higher than in those that were DPP reactive but initially ELISA non-reactive. Restriction fragment length polymorphism analysis revealed the presence of Leishmania infantum, the etiologic agent of VL, in bone marrow samples from VL-infected animals. Our data showed that the techniques and protocols currently employed in the PCLV screening approach are not entirely reliable. Further consideration should be given to monitoring dogs with undetermined results in ELISA and a better training should be provided for health workers responsible for performing DPP tests applied under field conditions.

Spatial distribution of tuberculosis from 2002 to 2012 in a midsize city in Brazil.

BACKGROUND: Tuberculosis (TB) remains a major public health problem in many developing countries. Exploratory spatial analysis is a powerful instrument in spatial health research by virtue of its capacity to map disease distribution and associated risk factors at the population level. The aim of the present study was to describe the epidemiologic characteristics and spatial distribution of new cases of TB reported during the period 2002-2012 in Divinopolis, a midsized city located in the state of Minas Gerais, southeastern Brazil. METHODS: Sociodemographic and clinical data relating to the study cases were retrieved from the national Brazilian database and geocoded according to residential address. Choropleth and kernel density maps were constructed and a spatial-temporal analysis was performed. Tracts defined by the 2010 national census were classified as sectors with higher or lower densities of new TB cases based on the kernel density map. Multivariate logistic analysis was used to compare the two types of sectors according to income, level of literacy and population density. RESULTS: A total of 326 new cases of TB were reported during the study period. Residential addresses relating to 309 (94.8 %) of these were available in the SINAN database and the locations were geocoded and mapped. The average incidence of TB during the study period was 14.5/100,000 inhabitants. Pulmonary TB was the most predominant form (73.6 %) and 74.5 % of patients had been cured. The percentage of cases was highest in males (67.8 %) and individuals aged 25-44 years (41.1 %), and lowest in children aged less than 15 years (4.6 %). The disease was spatially distributed throughout the urban district. The incidence rate among urban census tracts ranged from 0.06 to 1.1 %, and the disease occurred predominantly in the downtown area (99.3 %). Higher population density was associated significantly with increased odds of living in a sector with a "higher density of cases", even after adjusting for income and education (odds ratio = 13.7). CONCLUSIONS: The highest density of cases was strongly associated with higher population density but not with lower income or level of literacy.

Canine visceral leishmaniasis in an urban setting of Southeastern Brazil: an ecological study involving spatial analysis.

BACKGROUND: The physical characteristics of the environment influence the composition, distribution and behavior of the vectors and mammalian hosts involved in the transmission of visceral leishmaniasis (VL), thereby affecting the epidemiology of the disease. In Brazil, urbanization of human VL is a recent phenomenon and represents an issue of particular concern to local health authorities. The present study aimed to establish the degree of spatial dependency between canine and human VL in the municipality of Divinópolis, Minas Gerais, Brazil, and to identify priority risk areas in which stricter control measures should be implemented. METHODS: The selected canine population comprised 3,652 dogs distributed within 11 strata and 1,247 urban blocks. Serum samples were collected between March 2013 and February 2014. Serodiagnosis of dogs was performed using the enzyme-linked immunosorbent assay and the indirect fluorescent-antibody test. The blocks sampled for canine VL and the addresses of the 16 confirmed cases of human VL notified in Divinópolis during the period 2007-2013 were georeferenced. Spatial analysis of the data was performed using Kernel density estimation, Ripley's bivariate K-function and directional distribution methods. RESULTS: The overall prevalence of seropositive animals was 4.63% (range 3.95-5.31) (n =169) and varied in different strata between 0.9 (range 0.0-1.91) and 8.73% (range 5.65-11.81). A positive spatial dependency was detected between human and canine VL in which the occurrence of human cases of the disease tended to concentrate in locations that were close to areas with a higher incidence of canine VL. The priority risk area could be clearly distinguished from Kernel density estimation and standard deviational ellipse plots in which the human VL ellipse was totally enclosed within the canine VL ellipse. CONCLUSIONS: The results presented herein will enable the Municipal Health Office of Divinópolis to devise a more effective management plan for human VL in which specific strategies would be applied to areas presenting different levels of risk. This spatial evaluation of leishmaniasis model could be applied in other urban areas of Brazil.