One Health approach to Coxiella burnetii in Brazilian indigenous communities.

Indigenous health has posted complex challenges worldwide, particularly due to historical economic, territorial, social and environmental processes, which may lead to emergence and reemergence of pathogens. In addition to few Coxiella burnetii serosurveys in vulnerable populations, especially in developing tropical countries, no comprehensive One Health approach has focused on human-animal infection along with potential environmental determinants. Accordingly, this study aimed to assess the seroprevalence of anti-C. burnetii antibodies in indigenous populations and their dogs from 10 indigenous communities distributed in southern and southeastern Brazil, along with the correspondent healthcare professionals. In overall, 8/893 (0.90%; 95% CI 0.45-1.76) indigenous and 1/406 (0.25%) dog samples were seropositive, with 7/343 (2.04%) individuals the 1/144 (0.69%) dog from the Ocoy community, located in the city of São Miguel do Iguaçu, bordering Argentina at south, and far 10 km at west from Paraguay. All 84 healthcare professionals tested seronegative.

Risk factors for toxocariasis during incarceration: the One Health intervention approach.

Despite potential exposure to soil-transmitted helminths, especially when stray dogs and cats are present, toxocariasis in inmate populations remains to be established. Accordingly, the present study assessed the seroprevalence and associated risk factors of toxocariasis at the Women's State Penitentiary of Parana, Brazil. A total of 234/370 (63.2%; 95% CI 58.2-68.0) women inmates and 28/87 (32.2%; 95% CI 23.3-42.6) correctional officers were seropositive for anti-Toxocara spp. IgG by ELISA, with inmates 2.62-fold more likely positive (p = 0.00000026). The univariate model has identified that non-white (OR = 1.58, p = 0.047) and older than 39 years (OR = 1.28, p = 0.032) inmates were associated with mild but significant odds for seropositivity. Elementary or higher educational level was considered a protective factor for seropositivity. The presence of Toxocara spp. eggs was observed in 10/15 (66.7%) collected soil samples by centrifuge-flotation in Zinc Sulfate, and molecular analysis by PCR identified only Toxocara cati in these eggs. An intervention program was established with regular trap-neuter-release, with gradual removal for adoption (donation campaigns), treatment, and euthanasia when necessary (particularly due to advanced sporotrichosis). In addition, an educational awareness agenda was proposed, aiming to reduce soil contamination and accidental intake by the incarcerated population. A total of 40 feral cats were trapped, 20 males and 20 females, mostly adults. After trapping, 36 cats were neutered, treated, and microchipped in the Veterinary Teaching Hospital (VTH) at the Federal University of Paraná. Five trapped feral cats were euthanized, four diagnosed with advanced sporotrichosis, and one already neutered cat (not herein) with complications due to feline immunodeficiency virus (FIV). Female inmates presented higher seroprevalence for Toxocara spp. antibodies when compared to correctional officers, significantly associated with age, self-declared ethnicity (non-white), and lack of formal education. Despite the non-natural scenario of a state penitentiary, the One Health approach of Toxocara spp. has highlighted the interdisciplinary nature of the study and its relevance in understanding the complex interactions between human, animal, and environmental factors, particularly impacting female inmates. Further studies should establish the rate of inmate infection over time while deprived of liberty.

Influence of Housing Temperature and Genetic Diversity on Allogeneic T Cell-Induced Tissue Damage in Mice.

The objective of this study was to determine how housing temperature and genetic diversity affect the onset and severity of allogeneic T cell-induced tissue damage in mice subjected to reduced intensity conditioning (RIC). We found that adoptive transfer of allogeneic CD4+ T cells from inbred donors into sub-lethally irradiated inbred recipients (I→I) housed at standard housing temperatures (ST; 22-24 °C) induced extensive BM and spleen damage in the absence of injury to any other tissue. Although engraftment of T cells in RIC-treated mice housed at their thermo-neutral temperature (TNT; 30-32 °C) also developed similar BM and spleen damage, their survival was markedly and significantly increased when compared to their ST counterparts. In contrast, the adoptive transfer of allogeneic T cells into RIC-treated outbred CD1 recipients failed to induce disease in any tissue at ST or TNT. The lack of tissue damage was not due to defects in donor T cell trafficking to BM or spleen but was associated with the presence of large numbers of B cells and myeloid cells within these tissues that are known to contain immunosuppressive regulatory B cells and myeloid-derived suppressor cells. These data demonstrate, for the first time, that housing temperature affects the survival of RIC-treated I→I mice and that RIC-conditioned outbred mice are resistant to allogeneic T cell-induced BM and spleen damage.

A review on microRNA detection and expression studies in dogs.

MicroRNAs (miRNAs) are small non-coding RNAs that function by post-transcriptional regulation of gene expression. Their stability and abundance in tissue and body fluids makes them promising potential tools for both the diagnosis and prognosis of diseases and attractive therapeutic targets in humans and dogs. Studies of miRNA expression in normal and disease processes in dogs are scarce compared to studies published on miRNA expression in human disease. In this literature review, we identified 461 peer-reviewed papers from database searches using the terms "canine," "dog," "miRNA," and "microRNA"; we screened 244 for inclusion criteria and then included a total of 148 original research peer-reviewed publications relating to specific miRNA expression in canine samples. We found an overlap of miRNA expression changes between the four groups evaluated (normal processes, non-infectious and non-inflammatory conditions, infectious and/or inflammatory conditions, and neoplasia) in 39 miRNAs, 83 miRNAs in three of the four groups, 110 miRNAs in two of the three groups, where 158 miRNAs have only been reported in one of the groups. Additionally, the mechanism of action of these overlapping miRNAs varies depending on the disease process, elucidating a need for characterization of the mechanism of action of each miRNA in each disease process being evaluated. Herein we also draw attention to the lack of standardization of miRNA evaluation, consistency within a single evaluation method, and the need for standardized methods for a direct comparison.

One health approach to toxocariasis in Brazilian indigenous populations, their dogs, and soil contamination.

Introduction: Although socioeconomic vulnerability and lifestyle factors may contribute to the transmission of Toxocara spp., no study has investigated indigenous populations in Brazil using the One Health approach. Methods: Accordingly, this study assessed anti-Toxocara spp. antibodies in Brazilian indigenous people and healthcare professionals by enzyme-linked immunosorbent assay. Presence of Toxocara spp. eggs (feces and hair) in dogs as definitive hosts and in soil samples of the indigenous communities were also recovered and molecularly investigated. Results: Overall, 342/463 (73.9%) indigenous individuals and 46/147 (31.3%) non-indigenous healthcare professionals were seropositive for Toxocara spp. In addition, T. canis eggs were retrieved from 9/194 (4.6%) dog fecal samples and 4/204 (2.0%) dog hair samples, mainly from the Paraná State communities (3/42; 7.1%). Soil contamination was observed only in the Paraná State communities (36/90; 40.0%), with the molecular detection of T. canis. River water consumption was also associated with indigenous seropositivity (Odds ratio, 11.4). Discussion: Indigenous individuals in Paraná State communities were 2.72-fold more likely to be seropositive than those in São Paulo State, likely due to a lack of sanitary infrastructure. In this scenario, a primarily soil-transmitted disease may also have become waterborne, with embryonated eggs probably spread to water supplies by rain. Full-time healthcare professionals in daily contact with indigenous communities were 9.2-fold more likely to be seropositive than professionals who visited sporadically, suggesting exposure to Toxocara spp. during their work and raising health concerns. In addition, the findings herein showed a significantly higher seroprevalence in indigenous people than in healthcare workers (χ2 = 85.5; p < 0.0001), likely due to overtime exposure to Toxocara spp. In conclusion, Brazilian indigenous communities are highly exposed to toxocariasis, with poor infrastructure and contact with contaminated river water as associated risk factors and a higher risk of infection in healthcare professionals working full-time in these communities.

Molecular detection and characterization of SARS-CoV-2 in cats and dogs of positive owners during the first COVID-19 wave in Brazil.

Despite previous reports of SARS-CoV-2 infection in dogs and cats worldwide, the type of swab sample used for its detection through RT-qPCR needs to be better compared and described. Accordingly, as part of a multicenter study in Brazil, the aim of the present study was to assess which rectal or oropharyngeal swabs would be more appropriate for detecting SARS-CoV-2 in cats and dogs, through viral load comparison. Pets of owners diagnosed with COVID-19 in the last 7 days were eligible. A total of 148 animals from four of the five Brazilian geographical regions were analyzed, among which 10/48 cats (20.83%) and 11/100 dogs (11.00%) were positive. The results suggested that oropharyngeal swabs should be considered for SARS-CoV-2 detection, particularly in cats, due to the higher cDNA viral load. Also, the genomic results showed similarities between SARS-CoV-2 animal variants and human variants that were circulating at the time of sampling, thus corroborating the existence of zooanthroponotic transmission. In conclusion, the present study highlighted the importance of SARS-CoV-2 monitoring among cats and dogs, as virus modification may indicate the possibility of mutations in animals and spillover back to owners. Thus, positive individuals should always self-isolate from their pets during COVID-19, to prevent trans-species transmission and mutation.

Toxoplasma gondii exposure in Brazilian indigenous populations, their dogs, environment, and healthcare professionals.

Although Toxoplasma gondii exposure has been reported in indigenous populations worldwide, a One Health approach has not been applied to date. This study concurrently assessed T. gondii exposure in indigenous populations, and their dogs, environment, and indigenous or non-indigenous healthcare professionals (HPs). Human and dog serum samples from 9 indigenous communities in Brazil were assessed by indirect immunofluorescence antibody test for anti-T. gondii antibodies. Soil samples (30 per community) were processed with PCR to amplify T. gondii DNA. Associated risk factors and seroprevalence were analyzed using logistic regression models. Human seropositivity and type of water source were assessed by generalized linear mixed model (GLMM) with binomial error distribution, and game meat consumption with chi-squared test. Overall, 225/463 (49%) indigenous persons were seropositive for anti-T. gondii antibodies. Of all the HPs, 67/168 (40%) were positive, and included 54/147 (37%) positive non-indigenous HPs. Indigenous persons more likely to be seropositive compared with non-indigenous HPs (OR: 1.63; 95% CI: 1.11-2.39). A total of 97/253 (38%) dogs were seropositive and highly associated with seropositive owners (p < 0.001). Based on univariate analysis for indigenous individuals, state location of community (p < 0.001), ethnicity (p < 0.001), consumption of game meat (p < 0.001), type of water source (p < 0.001), and educational level (p = 0.026) were associated with seropositivity. Logistic regression showed that indigenous seropositivity was associated with eating game meat (p = 0.002), drinking water from rivers (p < 0.001), and inversely proportional to the educational level. According to univariate analysis for non-indigenous HP, age (p = 0.005), frequency of visits to the indigenous populations (p < 0.001), consumption of water at the indigenous communities (p < 0.001), and ingestion of raw meat (p = 0.023) were associated with T. gondii seropositivity. Logistic regression revealed living outdoors (p = 0.042), habit of hunting (p = 0.008), and drinking river water (p = 0.007) as risk factors associated to seropositivity in dogs. In addition, indigenous communities lacking water treatment had higher seroprevalence for all groups including indigenous persons (GLMM; z = -7.153; p < 0.001), their dogs (GLMM; z = -2.405; p = 0.0162), and all HPs (GLMM; z = -2.420; p = 0.0155). Human seropositivity was associated with that of their dogs (p < 0.001). A single soil sample, out of 270 (0.37%), was positive for T. gondii by PCR. Our results indicate water source is a risk for human and dog toxoplasmosis in indigenous communities; both share similar exposure. Moreover, quality water access was shown to be crucial to prevent toxoplasmosis in both total and non-indigenous HPs who work in these indigenous communities.

Molecular detection of feline hemoplasmas and retroviruses in free-roaming and shelter cats within a university campus.

Objectives: The aim of the present study was to assess the frequency of hemoplasma, feline immunodeficiency virus (FIV) and feline leukemia virus (FeLV) infections in cats living in an on-campus shelter and free-roaming cats within a university campus in Brazil. Methods: Blood samples were tested using quantitative PCR for hemoplasma, FIV and FeLV. Positive hemoplasma samples were sequenced. Associations between hemoplasma detection and living situation, sex, flea and/or tick parasitism, and coinfection with FIV and FeLV, were assessed using Fisher's exact test and the respective odds ratios were calculated. Results: Overall, 6/45 (13.3%) cats tested positive: four (8.9%) were infected with 'Candidatus Mycoplasma haemominutum' and two (4.4%) with Mycoplasma haemofelis. All positive samples were from free-roaming cats (6/15; 40.0%) and had statistically significantly lower packed cell volumes (P = 0.037). Although 5/23 (21.7%) males and 1/22 (4.6%) females were positive, no statistically significant association between sex and hemoplasma infection was found (P = 0.19). Viral quantitative PCR (qPCR) was performed on 43/45 samples, among which 2/43 (4.7%) were positive for FIV and none for FeLV. Only one cat (2.3%) was coinfected with hemoplasma and FIV (P = 0.26). In addition, 4/6 (66.7%) cats that tested positive for hemoplasmas were infested by fleas (P = 0.0014) and/or ticks (P = 0.25). Conclusions and relevance: These results show that even if the free-roaming cat population is clinically healthy and has adequate access to food, it may present flea infestation and hemoplasma infection with lower packed cell volume values.

Seroprevalence and associated risk factors of strongyloidiasis in indigenous communities and healthcare professionals from Brazil.

Strongyloides stercoralis, a pathogenic roundworm, is considered endemic in several tropical and subtropical areas worldwide. Indigenous populations have the highest soil-transmitted helminthiases-related mortality rates, but the prevalence and risk factors associated with S. stercoralis in Brazilian indigenous populations have not been established. Thus, the present study aimed to assess the seroprevalence and associated risk factors for S. stercoralis in indigenous communities and the healthcare professionals serving them in Brazil. Indigenous populations living in nine communities and healthcare professionals were tested for anti- S. stercoralis antibodies by ELISA. A questionnaire was used to assess socio-epidemiological information. Associated risk factors for seropositivity were tested by chi-square or Fisher's exact tests, using univariate analyses and multivariate logistic regression. Overall, 174/463 (37.6%; CI 95%: 33.3-42.1) indigenous persons and 77/147 (52.4%; 95% CI: 44.3-60.3) healthcare professionals were seropositive for anti- S. stercoralis antibodies. Seropositivity among the two groups was statistically significant (p = 0.0016; OR = 0.547; 95% CI: 0.376-0.796) and revealed that healthcare professionals were 1.83 times more likely to be seropositive. The multivariate analysis showed that being male or being adult were also risk factors, while having a septic tank as a sanitary facility represented a protective factor for S. stercoralis exposure in indigenous persons. None of the variables evaluated were associated with S. stercoralis exposure in the professional group. The study herein has reported a high seroprevalence to Strongyloides stercoralis in indigenous communities of Brazil and healthcare professionals, warning for potential public health concerns of strongyloidiasis in such populations.

Bartonella spp. and Typhus Group Rickettsiae among Persons Experiencing Homelessness, São Paulo, Brazil.

Persons experiencing homelessness in São Paulo, Brazil, were seropositive for Bartonella spp. (79/109, 72.5%) and typhus group rickettsiae (40/109, 36.7%). Bartonella quintana DNA was detected in 17.1% (14/82) body louse pools and 0.9% (1/114) blood samples. Clinicians should consider vectorborne agents as potential causes of febrile syndromes in this population.

Comparative proteomic changes in rabbit vocal folds undergoing systemic dehydration and systemic rehydration.

BACKGROUND: A considerable body of clinical evidence suggests that systemic dehydration can negatively affect voice production, leading to the common recommendation to rehydrate. Evidence for the corrective benefits of rehydration, however, is limited with mixed conclusions, and biological data on the underlying tissue changes with rehydration is lacking. In this study, we used a rabbit model (n = 24) of acute (5 days) water restriction-induced systemic dehydration with subsequent rehydration (3 days) to explore the protein-level changes underlying the molecular transition from euhydration to dehydration and following rehydration using LC-MS/MS protein quantification in the vocal folds. We show that 5-day water restriction led to an average 4.3% decrease in body weight with relative increases in anion gap, Cl-, creatinine, Na+, and relative decreases in BUN, iCa2+, K+, and tCO2 compared to control (euhydrated) animals. A total of 309 differentially regulated (p < 0.05) proteins were identified between the Control and Dehydration groups. We observed a noteworthy similarity between the Dehydration and Rehydration groups, both well differentiated from the Control group, highlighting the distinct timelines of resolution of the clinical symptoms of systemic dehydration and the underlying molecular changes. SIGNIFICANCE: Voice disorders are a ubiquitous problem with considerable economic and psychological impact. Maintenance of proper hydration is commonly prescribed as a general vocal hygiene practice. There is evidence that dehydration negatively impacts phonation, but our understanding of the state of vocal folds in the context of systemic dehydration are limited, particular from a molecular perspective. Further, ours is a novel molecular study of the short-term impact of rehydration on the tissue. Given the relatively minimal difference in vocal fold proteomic profiles between the Dehydration and Rehydration groups, our data demonstrate a complex physiological response to acute systemic dehydration, and highlight the importance of considering persistent underlying molecular pathology despite the rapid resolution of clinical measures. This study sets a foundation for future research to confirm the nature of potential beneficial outcomes of clinical recommendations related to hydration.

Infection of SARS-CoV-2 in domestic dogs associated with owner viral load.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-positive pet owners are reported to be a risk factor for infection of their pets; however, the influence of the viral load and associated risks has not been fully established. This study aimed to assess potential association of viral load in owners with the presence of SARS-CoV-2 infection in their dogs. Of 20 SARS-CoV-2-positive pet owners from 13 families in Curitiba, Brazil, 5 of 22 (22.7%) dogs were positive for SARS-CoV-2. Viral presence was detected in oropharyngeal samples for 2 of 5 (40.0%) dogs at 8 and 9 days after the first positive sample. Detection of SARS-CoV-2 in these dogs was associated with higher viral loads in the owners and close owner contact. All 5 RT-qPCR-positive dogs had antibodies to at least one viral protein tested in the serological assay. Molecular detection of SARS-CoV-2 in dogs was statistically associated with clinical signs in owners such as cold, cough, or diarrhea (P = 0.039), number of positive persons in the household (P = 0.002), and higher viral load (P = 0.039). Such findings serve as a warning for risks of human to dog infection, mainly due to sharing beds and other close interactions without protection. In conclusion, people with coronavirus disease 2019 (COVID-19), particularly in households with multiple residents and high viral load, should take the same preventive measures when interacting with their dogs during self-isolation as they do with people.

Epidemiology of Rickettsia spp. in Atlantic rainforest areas of island and seashore mainland, southern Brazil.

Non-fatal cases of rickettsial infection with different clinical features than the classic BSF (Brazilian Spotted Fever) have been reported in seashore areas of Paraná state, southern Brazil. In addition, Amblyomma ovale tick infected by Rickettsia parkeri strain Atlantic rainforest has been also described in this area. Accordingly, the aim of this study was to investigate the occurrence of anti-Rickettsia spp. antibodies in human and dog populations, and Rickettsia spp. infection in ticks from oceanic islands and seashore mainland cities of southern Brazil. Serum samples were collected from 328 persons and their 282 dogs from three islands and two seashore mainland cities. A total of 211 ticks were collected from dogs, identified as A. ovale and R. sanguineus. In overall, 40 of 328 (12.2%) human samples were seropositive for Rickettsia spp., including 21 of 190 (11.1%) on islands and 19 of 138 (13.7%) on seashore mainland, and 62 of 282 (22.0%) dog samples, including 31 of 153 (20.3%) on islands and 31 of 129 (24.0%) in seashore mainland areas. In overall, nine of 82 (11.0%) ticks were positive to real-time PCR assay targeting a fragment of the rickettsial gltA gene, including two of 64 (3.1%) Rickettsia sanguineus and seven of 18 (38.9%) A. ovale, of which four were infected with the R. parkeri strain Atlantic rainforest. Despite no association between risk factors and Rickettsia spp. seropositivity was found in human beings, access to natural areas (p = .011) and tick infestation (p = .004) was significantly associated to dog seropositivity. The serological and molecular findings herein have confirmed previous tick and clinical case reports and enlarged the geographical occurrence of A. ovale infected by R. parkeri strain Atlantic rainforest in oceanic islands and seashore mainland cities of Paraná State, indicating a new likely transmission area of this new rickettsial infection in human beings and dogs of southern Brazil.

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.

Serosurvey of anti-Toxocara canis antibodies in people experiencing homelessness and shelter workers from São Paulo, Brazil.

BACKGROUND: Despite being one of the most prevalent helminth parasitic zoonoses worldwide and particularly in socioeconomically vulnerable populations, toxocariasis remains to be fully investigated in persons experiencing homelessness. Accordingly, the present study has aimed to assess the seroprevalence and associated risk factors of Toxocara spp. exposure in persons experiencing homelessness and shelter workers from a day-shelter in São Paulo city, Brazil. METHODS: Anti-Toxocara IgG antibodies were detected by enzyme-linked immunosorbent assay (ELISA). Univariable and multivariable logistic regression models were performed to assess the risks for toxocariasis. RESULTS: Overall, anti-Toxocara IgG antibodies were detected in 89/194 (45.9%, 95% CI: 39.0-52.9%) persons experiencing homelessness, twice as high (OR = 2.2; 95% CI = 1.245-3.873; P = 0.0089) than the frequency of 22/79 (27.8%, 95% CI: 19.2-38.6) in shelter workers. College education was the only protective factor for Toxocara spp. exposure (OR: 0.23; P = 0.018) revealed by logistic regression. CONCLUSIONS: Although indicating a multifactorial origin of toxocariasis, the present study has assessed a highly vulnerable population with high disease risks and premature death. Thus, the living conditions of the homeless population have influenced the high prevalence of anti-Toxocara antibodies verified here compared with domiciled shelter workers. Despite being less exposed, shelter and other outdoor workers may present an occupational risk to toxocariasis. Future studies should establish whether such environmental exposure might occur in persons experiencing homelessness in other regions worldwide.

Association of hoarding case identification and animal protection programs to socioeconomic indicators in a major metropolitan area of Brazil.

The present study assessed the identification of animal and object hoarding disorder cases by contact and mapping and the presence of animal protection programs in association with seven social-economic indicators of the metropolitan area of the ninth-biggest metropolitan area of Brazil. City Secretaries of Health and Environment provided demographic information and responded to a questionnaire. Overall, a very high level of hoarding case identification per municipality was associated with a higher Human Development Index, population, density, and income and related to distance from Curitiba, the capital of Parana State. Low and very low levels of hoarding case identification were related to greater area, higher Social Vulnerability Index (SVI), inequality, illiteracy, and rural areas. Very high identification level of animal protection programs was also associated with higher HDI, density and population, urban area, and high income, and geographical area. Similarly, low and very low levels of animal protection programs identification were major explained by low income, illiteracy, and distance related to higher population, urbanization, and higher HDI. In summary, better identification of hoarding cases and animal protection programs have shown an association with better socioeconomic indicators and higher population, density, and urban area. Whether municipalities with better human socioeconomic indicators may stimulate society's demands for identification of cases of individuals with hoarding disorder and animal programs should be further established. Regardless, animal health and welfare have been associated with improving human quality of life in a major Brazilian metropolitan area.

The rate and role of pseudogenes of the Mycobacterium tuberculosis complex.

Whole-genome sequence analyses have significantly contributed to the understanding of virulence and evolution of the Mycobacterium tuberculosis complex (MTBC), the causative pathogens of tuberculosis. Most MTBC evolutionary studies are focused on single nucleotide polymorphisms and deletions, but rare studies have evaluated gene content, whereas none has comprehensively evaluated pseudogenes. Accordingly, we describe an extensive study focused on quantifying and predicting possible functions of MTBC and Mycobacterium canettii pseudogenes. Using NCBI's PGAP-detected pseudogenes, we analysed 25 837 pseudogenes from 158 MTBC and M. canetii strains and combined transcriptomics and proteomics of M. tuberculosis H37Rv to gain insights about pseudogenes' expression. Our results indicate significant variability concerning rate and conservancy of in silico predicted pseudogenes among different ecotypes and lineages of tuberculous mycobacteria and pseudogenization of important virulence factors and genes of the metabolism and antimicrobial resistance/tolerance. We show that in silico predicted pseudogenes contribute considerably to MTBC genetic diversity at the population level. Moreover, the transcription machinery of M. tuberculosis can fully transcribe most pseudogenes, indicating intact promoters and recent pseudogene evolutionary emergence. Proteomics of M. tuberculosis and close evaluation of mutational lesions driving pseudogenization suggest that few in silico predicted pseudogenes are likely capable of neofunctionalization, nonsense mutation reversal, or phase variation, contradicting the classical definition of pseudogenes. Such findings indicate that genome annotation should be accompanied by proteomics and protein function assays to improve its accuracy. While indels and insertion sequences are the main drivers of the observed mutational lesions in these species, population bottlenecks and genetic drift are likely the evolutionary processes acting on pseudogenes' emergence over time. Our findings unveil a new perspective on MTBC's evolution and genetic diversity.

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.

One Health Index (OHI) applied to Curitiba, the ninth-largest metropolitan area of Brazil, with concomitant assessment of animal, environmental, and human health indicators.

One Health has been defined as a comprehensive approach to human, animal, and environmental health, but unsuccessful in providing a specific index for city, state, or country assessment. Accordingly, the present study aimed to calculate the One Health Index (OHI) in 29 cities of the Curitiba metropolitan area, the ninth largest in Brazil. Animal and Environmental health indicators were obtained from the city secretary of environment. The social vulnerability index (SVI) was used as the overall human health indicator. The indicators were parameterized following a binary logic Principal component analysis (PCA) was applied in association with city population, per capita income, and distance from the capital Curitiba. Permutational multivariate analysis of variance (PERMANOVA) was applied using the three first coefficients of the principal components obtained from Principal Component Analysis Plot (PCA) and exploring a pairwise comparison between the scenario ranges. A value of p less than 0.05 was considered significant. Overall, a higher OHI was associated with a higher city population and income, and shorter distance from the capital, and tendency of low-income cities to present lower OHI compared to higher-income cities. In conclusion, the OHI proposed herein portrays a holistic representation of a city's overall health. Moreover, animal issues should be considered part of local to global sustainability, considering OHI to calculate sustainability indexes.

Novel duplex RT-qPCR for animal rabies surveillance.

Rabies is a lethal zoonosis affecting mammals worldwide. Diagnosis of rabies follows international standard protocols, primarily relying on direct immunofluorescence (DI) followed by mouse inoculation test (MIT). WHO recommends molecular biology techniques such as RT-qPCR for replacing MIT to diagnose rabies in animal samples. Recently, a real-time PCR protocol that detects all rabies virus variants identified worldwide was validated. This assay is a pan-Lyssavirus TaqMan quantitative RT-PCR called LN34. A modified LN34 assay protocol was tested at the Paraná State Reference Laboratory (Lacen/PR) using animal samples previously tested by DI and MIT, the gold standard (GS). This method has been changed to a RT-qPCR duplex format to better fit the diagnostic routine. The new assay was called duplex LN34 and ß-actin RT-qPCR. All the 88 samples evaluated using the GS test, modified pan-Lyssavirus TaqMan RT-qPCR and duplex LN34 and ß-actin RT-qPCR showed 100% agreement with each other. This novel duplex RT-qPCR protocol has shown adequate diagnostic performance and may be used in research and surveillance purposes, replacing the standard MIT and ending mice use for rabies diagnosis.