One Health Approach to Leptospirosis: Dogs as Environmental Sentinels for Identification and Monitoring of Human Risk Areas in Southern Brazil.

Leptospirosis has been a neglected, widespread and reemerging zoonotic disease of global importance. The One Health holistic approach combining human, animal and environmental health has been important for control and prevention of zoonotic disease. An urban municipality in Southern Brazil with a high prevalence of leptospirosis was selected by convenience, with asymptomatic human and canine individuals serologically tested using MAT against 30 Leptospira spp. serovars. Epidemiological questionnaires were assessed along with the historical national database, with associated risk factors to dog and human seropositivity analyzed using multiple logistic regression. The spatial distribution of retrospective human leptospirosis cases was analyzed using a Kernel map and overlapped dog seropositivity and historical flooding areas, demonstrating that domestic and asymptomatic dogs may be important environmental sentinels for leptospirosis in such urban areas, even in the absence of human seropositivity and low canine seropositivity. Associated risk factors for leptospirosis in dogs and humans according to multiple regression included healthy dogs (p = 0.02, odds ratio-OR = 0.17, confidence interval of 95%CI 0.03-0.71) with indoor access (p = 0.059, OR = 5.12, 95%CI 1.10-37.7) and human males (p = 0.042, OR = 2.44, 95%CI 1.08-6.11) with presenting calf pain (p = 0.005, OR = 3.14, 95%CI 1.46-7. 14), jaundice (p < 0.001, OR = 4.61, 95%CI 2.11-10.10) and renal failure (p = 0.008, OR = 4.49, 95%CI 1.49-13.76). The greater the rain precipitation (above 3 mm of average annual precipitation), the higher the number of leptospirosis cases in humans. In conclusion, dog active serosurvey and rain precipitation should be systematically reported and plotted altogether as a basis for the early detection and monitoring of human risk areas. Such findings may serve as a basis for public health policies in Brazil and other endemic countries worldwide.

Salmonella detection with LAMP and qPCR and identification of serovars of interest by multiplex qPCR in poultry carcasses.

Salmonella is present in the poultry production chain and is a major challenge in terms of food safety and animal health. The early Salmonella detection is one of the main tools to control and prevent the transmission of this pathogen. Microbiological isolation and serotyping to identify and differentiate Salmonella serovars are laborious processes, time-consuming, and expensive. Therefore, molecular diagnostic methods can be rapid and efficient alternatives to the detection of this pathogen. Thus, the aim herein was to standardize and evaluate the use of loop-mediated isothermal amplification (LAMP) in comparison with real-time PCR (qPCR) for detection of Salmonella associated with a multiplex qPCR for simultaneous identification and differentiation of S. Enteritidis, S. Typhimurium, S. Pullorum, and S. Gallinarum. The LAMP, qPCR, and multiplex qPCR assays were comparable in specificity. The three techniques were evaluated for specificity for 16 different serovars of Salmonella and for 37 strains of the serovars of interest. The limit of detection and the efficiency of the LAMP, qPCR, and multiplex qPCR reactions were determined. The techniques were applied to 33 samples of chicken carcasses and compared to the results of conventional microbiology for validation. As results, LAMP was specific in the detection of different Salmonella serovars but presented lower limit of detection ranging from 101 to 104 CFU/reaction. In comparison, qPCR could detect less cells (100 to 102 CFU/reaction), reaching equal specificity and better repeatability in the assays. The qPCR multiplexing for identification of the different serovars also showed good specificity, with the detection threshold between entre 101 and 102 CFU/reaction. The results obtained in the analyses on poultry carcasses suggested a correspondence between the results obtained in molecular methods and in conventional microbiology. Thus, the proposed assays are promising for the diagnosis of Salmonella in poultry carcasses, already proved to be faster and more efficient than conventional diagnostics techniques, being of great interest for poultry production, animal, and public health.

One Health Approach on Dog Bites: Demographic and Associated Socioeconomic Factors in Southern Brazil.

Despite being an important public health issue, particularly due to rabies, dog bites and associated risk factors have rarely been assessed by health services from a One Health perspective. Accordingly, the present study aimed to assess dog biting and associated demographic and socioeconomic risk factors in Curitiba, the eighth-largest Brazilian city with approximately 1.87 million people, based on the post-exposure prophylaxis (PEP) rabies reports between January/2010 and December/2015. The total of 45,392 PEP reports corresponded to an average annual incidence of 4.17/1000 habitants, mainly affecting white (79.9%, 4.38/1000 population), males (53.1%, 4.81/1000 population), and children aged 0-9 years (20.1%, 6.9/1000 population), with severe accidents associated with older victims (p < 0.001) and mainly caused by dogs known to the victims. An increase of USD 100.00 in the median neighborhood income was associated with a 4.9% (95% CI: 3.8-6.1; p < 0.001) reduction in dog bites. In summary, dog biting occurrence was associated with victims' low income, gender, race/color, and age; severe accidents were associated with elderly victims. As dog bites have been described as multifactorial events involving human, animal, and environmental factors, the characteristics presented herein should be used as a basis to define mitigation, control, and prevention strategies from a One Health perspective.

Spatial Owner-Dog Seroprevalence of Leptospira spp. Antibodies in Oceanic Islands and Costal Mainland of Southern Brazil.

Leptospirosis has been described as a disease neglected worldwide. Affecting humans and animals, the disease is often related to poor environmental conditions such as lack of sanitation and presence of synanthropic rodents. Despite being considered as a One Health issue, no study has focused on comparing owner-dog seroprevalence between islands and seashore mainland. Accordingly, the present study assessed anti-Leptospira spp. antibodies by applying microscopic agglutination test (MAT) methods to Leptospira and assessing associated risk factors via univariate and multivariate logistic regression analysis of owners and their dogs in islands and seashore mainland of southern Brazil. No anti-Leptospira spp. Seropositivity was found in 330 owner serum samples, while dogs presented an overall seroprevalence of 5.9%. All seropositive dogs reacted to serogroups of Leptospira interrogans, including 66.7% of Pyrogenes, 44.4% Canicola, 22.2% Icterohaemorrhagiae, 16.7% Australis; six reacted to more than one serogroup. No association was found among seropositivity and epidemiological variables, except that neighborhood dogs were less likely to be seropositive. Although no seropositivity was observed in owners, seropositivity in dogs had the potential to indicate such species as being sentinels for environmental exposure and potential human risk of infection.

Seropositivity of Anti-Toxoplasma gondii Antibodies in Owners and Their Dogs Living on Island and Mainland Seashore Areas of Southern Brazil.

Although toxoplasmosis has been considered among the most neglected zoonoses worldwide, no study has focused on the frequency and associated risk factors of owners and their dogs living on an island and mainland seashore areas. Accordingly, anti-Toxoplasma gondii IgG antibodies were screened by indirect fluorescent antibody test (IFAT) in owners and dogs from three oceanic islands and two nearby mainland harbor areas, with associated risk factors for toxoplasmosis assessed by univariable and multivariable logistic regression models. Overall, anti-T. gondii seropositivity was observed in 59/328 (18.0%) owners and 66/283 (23.3%) dogs, with no statistical difference between owners (p = 0.360) and dogs (p = 0.655) from islands and mainland areas. Consumption of local water springs (p = 0.016; OR = 2.11) was an associated risk factor for T. gondii seropositivity, and owners with the habit of spring water intake were twice more likely seropositive (p = 0.014; OR = 2.14). Presence of anti-T. gondii antibodies in dogs was associated with seropositive owners (p = 0.008; OR = 2.81), household consumption of beef meat (p = 0.042; OR = 1.7) and chicken (p = 0.026; OR = 2.9). Despite being lower than the worldwide prevalence, toxoplasmosis seropositivity in owners and their dogs in southern Brazil was influenced by the positive owner, water source, and meat consumption, and not by inhabiting islands or seashore mainland areas, presence of dogs, cats, or both. In addition, drinking water quality should always be considered a critical risk factor for toxoplasmosis on islands.

One health approach on serosurvey of anti-Leptospira spp. in homeless persons and their dogs in South Brazil.

Although leptospirosis has been described as a worldwide bacterial zoonosis primarily affecting vulnerable populations, to date no study has focused on concomitant serosurvey of homeless persons and their dogs. The aim of the present study was, to use a One Health approach to serologically assess homeless persons and their dogs in 3 major cities of south Brazil (São Paulo, Curitiba, and Foz do Iguaçu). Environmental information was obtained with an epidemiological questionnaire given to all participants. A total of 200 human and 75 dog samples were tested for anti-Leptospira spp. antibodies to thirty different serovars using the microscopic agglutination test. None of the homeless persons were positive while 5 of the 75 (6.7%) dogs were positive. Among homeless population, 89% (177 of 200) were male, 61% (122 of 200) self-declared Non-white, and 67% (134 of 200) were educated up to the 8th school grade. Lower exposure of homeless persons to Leptospira spp. in the present study when compared to other vulnerable populations (slum and low-income residents) may be result of less direct exposure as they are able to rapidly change locations in response to flooding events. In addition, these results may reflect the effectiveness of a specific healthcare service provided to people living in the streets in the 3 cities. While dogs may be used as environmental sentinels for leptospirosis, the low seropositivity results found in this report may indicate low transmission risk to homeless owners in direct daily contact with their dogs.

Serological survey of anti-Leptospira spp. antibodies in individuals with animal hoarding disorder and their dogs in a major city of Southern Brazil.

BACKGROUND: Individuals with hoarding disorder (HD) presented a persistent difficulty in detaching from objects and/or animals. Unhealthy conditions, frequently found in cases of animal HD (AHD), may favour environmental contamination and the spread of zoonotic pathogens. Despite that, only one study of zoonotic diseases in individuals with AHD and their companion animals has been conducted to date. OBJECTIVES: This study aimed to assess the seroprevalence of anti-Leptospira spp. antibodies in individuals with AHD and their dogs in a major city of Southern Brazil. METHODS: Blood samples were obtained from 264 dogs (21 households) and 19 individuals with AHD (11 households) and tested by microscopic agglutination test. RESULTS: All human samples were seronegative. Seropositivity was found in 16/264 (6.1%; CI 95% 3.3-9.6%) dogs from 11/21 (52.38%) households, with titres ranging from 100 up to 400, and Copenhageni (10/16; 62.5%) was the most frequent serovar. Surprisingly, seropositivity of hoarded dogs found herein was among the lowest reportedly observed in other dog populations of Brazil. Two epidemiological variables were significantly associated with seropositivity in dogs: the presence of cat hoarding (p = 0.004) and the report of flood occurrence in the household (p = 0.031). CONCLUSIONS: No individuals with AHD were seropositive, and besides the lower seroprevalence of dogs, they probably had contact with Leptospira spp. at some point in their life. Since dogs can be considered potential sentinels in leptospirosis, public health programs must become aware of the risk of leptospirosis cases in households of individuals with AHD and nearby communities.

A fast and cheap in-house magnetic bead RNA extraction method for COVID-19 diagnosis.

COVID-19 has posed a worldwide public health challenge affecting millions of people in different countries. Rapid and efficient detection of SARS-CoV-2 is essential for pandemic control. Reverse Transcription quantitative PCR (RT-qPCR) of nasopharyngeal swabs is the gold standard method for the virus detection, but the high demand for tests has substantially increased the costs and reduced the availability of reagents, including genetic material purification kits. Thus, the present study aimed to compare two bead-based RNA extraction methods (an in-house and a commercial kit) from nasopharyngeal swabs and RT-qPCR detection of SARS-CoV-2. Twenty-five positive and five negative nasopharyngeal swab samples were subjected to extraction of nucleic acids using both methods in an automated platform. Both protocols revealed a high correlation between Cycle Quantifications (Cqs) (r = 0.99, p < 0.0001). In addition, the in-house kit was 89.5 % cheaper when compared to the mean cost of commercial RNA extraction kits. The results show that the in-house protocol is an affordable and reliable option for RNA extraction for SARS-CoV-2 detection from nasopharyngeal swabs.