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.

Detection of Toxoplasma gondii DNA in milk of dairy cows from southern Brazil.

Consumption of unpasteurized cow's milk may be a transmission route for some pathogenic microorganisms, but there is little information about the risk of Toxoplasma gondii infection. Blood and milk samples were collected in a paired and random fashion from 106 dairy cows and bulk-tank milk samples were also collected from each of the six farms, in southern Brazil. Serum anti-T.gondii antibodies (IgG) were detected by an indirect fluorescent antibody test (IFAT) with a cutoff point of 1:64. Nested PCR targeting the ITS1 was performed on milk samples to detect the Sarcocystidae family, confirmed to be T.gondii by Sanger sequencing. The occurrence of anti-T.gondii antibodies in the herds was 14.1%, (15/106) with seropositive cows in all herds. Antibody titers in positive samples ranged from 64 to 128. T.gondii DNA was detected in 2.8% (03/106) of the milk samples. The ITS1 sequences generated in this study were ON809793 - ON809794 and the sequencing revealed 98-100% identity with T. gondii DNA sequences deposited in GenBank. All cows PCR positive for T.gondii in milk were negative for IgG antibodies in serum, suggesting that naturally infected cows may shed T. gondii in milk in the acute phase of infection. The results of this study demonstrate that T. gondii DNA may be detected in raw cow's milk, so the potential risks of lactogenic infection should be considered. The presence of T. gondii DNA in milk does not confirm that the protozoa are viable and infective, and further investigations into the role of cow's milk in the epidemiology of toxoplasmosis are needed.

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.

Clinical trial and detection of SARS-CoV-2 by a commercial breath analysis test based on Terahertz technology.

Public health threats such as the current COVID-19 pandemics have required prompt action by the local, national, and international authorities. Rapid and noninvasive diagnostic methods may provide on-site detection and immediate social isolation, used as tools to rapidly control virus spreading. Accordingly, the aim of the present study was to evaluate a commercial breath analysis test (TERA.Bio®) and deterministic algorithm for detecting the SARS-CoV-2 spectral signature of Volatile Organic Compounds present in exhaled air samples of suspicious persons from southern Brazil. A casuistic total of 70 infected and 500 non-infected patients were sampled, tested, and results later compared to RT-qPCR as gold standard. Overall, the test showed 92.6% sensitivity and 96.0% specificity. No statistical correlation was observed between SARS-CoV-2 positivity and infection by other respiratory diseases. Further studies should focus on infection monitoring among asymptomatic persons. In conclusion, the breath analysis test herein may be used as a fast, on-site, and easy-to-apply screening method for diagnosing COVID-19.

Hemotropic mycoplasmas (hemoplasmas) in wild boars, hunting dogs, and hunters from two Brazilian regions.

Haemotropic mycoplasmas (haemoplasmas) are small pleomorphic bacteria infecting erythrocytes of several mammalian species, including human beings. No study to date has focused on the risk of bacteria exposure in hunting activities, particularly in natural environments of highly tick-infested areas. Accordingly, the present study aimed to assess haemoplasma occurrence in the complex encompassing wild boars, hunting dogs and hunters of Brazil. A total of 38/65 (58.5%) wild boars and 94/159 (59.1%) dogs were positive by qPCR for at least one haemoplasma. All 25 hunters were negative. Dogs with high hunting frequency were 2.4 more likely to be infected. Sequencing revealed a probable novel haemoplasma species in wild boars. Although exposure to haemoplasma species was present, the study herein found no evidence of cross-species transmission.

One Health approach on human seroprevalence of anti-Toxocara antibodies, Toxocara spp. eggs in dogs and sand samples between seashore mainland and island areas of southern Brazil.

Toxocariasis, caused by Toxocara spp. nematodes, is among the top 5 neglected parasitic diseases worldwide; however, no comprehensive study to date has serologically compared infections in people and their dogs and environmentally contaminated soil or sand of mainland and island locations. Accordingly, this study aimed to assess the seroprevalence of anti-Toxocara antibodies in traditional human seashore populations, the presence of eggs in dogs' feces and hair, and the presence of eggs in environmental samples from islands compared to the adjacent mainland of southern Brazil. Overall, 212/328 (64.6%) people were positive for Toxocara spp. antibodies, including 125/190 (65.8%) island and 87/138 (63.0%) mainland residents. For dog samples, 12/115 (10.43%) were positive for the presence of Toxocara spp. eggs, all from dogs living in islands, and 22/104 (21.15%) dog hair samples contained eggs of Toxocara spp. Environmental contamination with Toxocara spp. eggs was observed in 50/130 (38.46%) samples from all sampled sites. No significant association was found between risk factors (age, sex, educational level, monthly income, owning dogs or cats, ingestion of treated water, and consumption of raw or uncooked meat) and Toxocara spp. seropositivity. The present study is the first concurrent report on people, their dogs, and environmental contamination of Toxocara spp. The high prevalence we observed in the seashore populations of both in island and mainland areas may be caused by exposure to contaminated sand and climatic factors favoring frequent exposure to Toxocara spp. In conclusion, seashore lifestyle and living conditions of both island and mainland areas may have predisposed higher contact with infected pets and contaminated soil, favoring the high prevalence of toxocariasis.

Serosurvey of anti-Leishmania (Leishmania) infantum antibodies in hunting dogs and hunters in Brazil.

BACKGROUND AND AIM: Although wild boar hunting activities and the hunting dog trade in the Atlantic Forest and Cerrado biomes of Brazil overlap both with endemic and with non-endemic areas for visceral leishmaniasis, no study to date has focused on Leishmania spp. exposure among hunting dogs and hunters. The aim of the present study was to assess the presence of Leishmania spp. antibodies in hunting dogs and hunters in different anthropized areas of two Brazilian biomes. MATERIALS AND METHODS: Blood samples were collected from 170 hunting dogs and 46 hunters between October 2016 and May 2018. The presence of antibodies against Leishmania spp. in hunting dogs was screened through a dual-path platform immunochromatographic test (DPP rapid test; Bio-Manguinhos/Fundação Oswaldo Cruz, Rio de Janeiro, Brazil) and in hunters through an rK39-based rapid immunochromatographic test. Both tests were used in accordance with Brazilian Ministry of Health recommendations. RESULTS: Overall, although antibodies were detected through the immunochromatographic test in 3/170 (0.02%) of these female asymptomatic hunting dogs, all living in anthropized areas of the Atlantic Forest biome in South Brazil, no sample was confirmed through the enzyme-linked immunosorbent assay. All the hunters were non-reactive in the rapid immunochromatographic test. CONCLUSION: Our study on three suspicious hunting dogs has suggested that Leishmania (Leishmania) infantum may circulate both in endemic and non-endemic areas in Brazil. In addition, a high rate of hunting dog replacement due to death and trade may have led to less chance of infection and transmission between animals and between animals and humans, which would corroborate the outcomes reported here. Further studies should be conducted to fully establish whether hunting dogs and hunters may be used as sentinels in other areas endemic for Leishmania spp.

Risk factors associated with ticks and Rickettsia spp. exposure in wild boars (Sus scrofa), hunting dogs, and hunters of Brazil.

BACKGROUND AND AIM: Wild boars have recently been implicated as the maintainers and carriers of Amblyomma spp. ticks, which are essential for Rickettsia spp. transmission. Consequently, wild boar hunting may increase the risk of tick exposure and subsequent human tick-borne infection and disease. Therefore, this study was conducted to evaluate the risk factors for ticks and Rickettsia spp. exposure in wild boars, hunting dogs, and hunters in Brazilian biomes. MATERIALS AND METHODS: The statistical relationship of Rickettsia spp. antibodies were evaluated using the Chi-square test in 80 wild boars, 170 hunting dogs, and 49 hunters. RESULTS: The only statistically significant difference in seropositivity found in this study was between male and female wild boars (p=0.034), probably associated with in-park exposure to Amblyomma brasiliense infected with Rickettsia spp. CONCLUSION: The absence of statistical differences in the associated risk factors for hunting dogs and hunters may indicate a random exposure to Rickettsia spp.

Seroprevalence of Anti-Brucella spp. Antibodies in Wild Boars (Sus scrofa), Hunting Dogs, and Hunters of Brazil.

All 86 wild boars (Sus scrofa), 170 hunting dogs, and 49 hunters sampled from three Brazilian regions were seronegative to Brucella spp. by the standard tube agglutination and 2-mercaptoethanol tests, suggesting a low circulation of Brucella spp. in wild boars, hunting dogs, and hunters in such areas.

EXPOSURE OF WILD BOAR (SUS SCROFA) TO THE COMMON VAMPIRE BAT AND LACK OF IMMUNE PROTECTION TO RABIES VIRUS IN BRAZILIAN HUNTERS.

Rabies virus is recognized as one of the most fatal zoonotic agents affecting all mammals. Wild boars (Sus scrofa), classified as a large-size exotic invasive species in Brazil with nationwide hunting permitted, may serve as an extra blood source for the common vampire bat (Desmodus rotundus). Our aim was to document wild boar exposure to vampire bats to determine the seroprevalence of rabies virus antibodies in wild boars and to determine the immune status of hunters in southern and central-western Brazilian regions. Serum samples were collected from 80 wild boars and 49 hunters from natural and degraded areas of the Atlantic Forest biome of southern Brazil and in degraded areas of the Cerrado biome of central-western Brazil. The rabies-modified rapid fluorescent focus inhibition test was performed to detect the presence of rabies virus neutralizing antibodies in wild boars and considered seropositive when ≥0.10 IU/mL. The simplified fluorescence inhibition microtest was used for samples from hunters with a titer of ≥0.50 IU/mL and considered indicative of seroconversion. While 11% (9/80) of wild boars had serum titers for rabies exposure (≥0.10 IU/mL), 88% (43/49) of corresponding hunters lacked immune protective titers (<0.50 IU/mL). Wild boars showed serum titers for rabies likely due to contact with contaminated saliva of vampire bats or from infected carcass consumption. Additionally, Brazilian wild boars can be exposed to rabies and may play an important role in the sylvatic rabies cycle by providing a blood supply for vampire bats, highlighting the possibility of direct transmission of rabies virus to hunting dogs and hunters. These findings suggested hunters are a potential risk group for contracting rabies, and the World Health Organization may consider adding this occupation to their recommendations of who should receive the pre-exposure rabies vaccination.

Serosurvey of anti-Neospora caninum antibodies in wild boars (Sus scrofa), hunting dogs and hunters of Brazil.

The protozoan Neospora caninum is an important cause of abortion in cattle worldwide, with domestic dogs serving as the definitive hosts. Although hunting activities have been indicated as a potential risk factor for N. caninum infection in dogs, no serological evidence has so far been reported in hunting dogs, wild boars (Sus scrofa), and hunters. A total of 98 wild boars, 168 hunting dogs, and 15 hunters from three Brazilian regions were sampled and tested for anti-N. caninum antibodies by indirect immunofluorescence antibody test, resulting in 15/168 (9%) seropositive hunting dogs, and 0/98 wild boars, and 0/15 hunters seropositive. The absence of N. caninum antibodies in wild boars may suggest low exposure to oocysts shed by hunting dogs and wild canids in three different Brazilian regions. Finally, the absence of hunter seropositivity supports the current lack of evidence that N. caninum is a zoonotic parasite.

Serosurvey of Eastern, Western, and Venezuelan Equine Encephalitis Viruses in Wild Boars (Sus scrofa), Hunting Dogs, and Hunters of Brazil.

A total of 102 free-range wild boars, 170 hunting dogs, and 49 hunters from 3 Brazilian regions were sampled and tested for antibodies to eastern equine encephalitis virus (EEEV), western equine encephalitis virus, and Venezuelan equine encephalitis virus. Three of the 102 (2.9%) wild boars were positive for antibodies against EEEV by microplate serum neutralization test. Based on our data, free-range wild boars from central-western Brazil may be exposed to EEEV, and further studies are needed to evaluate the potential of incorporating serosurveys in routine arbovirus activity surveillance specifically to identify arbovirus activity foci and to help establish thresholds for epidemic transmission.

Serosurvey for Pseudorabies (Aujeszky's Disease) in Free-Range Wild Boars (Sus scrofa) of Brazil.

Free-ranging wild boars (Sus scrofa) from two different Brazilian biomes were sampled. Only one of 36 (2.8%) animals tested from central-western Brazil were positive for antibodies against pseudorabies virus (PRV) by enzyme-linked immunosorbent assay. Peccaries and other wildlife species can be exposed to PRV in these areas. In addition, wild boars could have impact on livestock and PRV eradication.

Hemotropic mycoplasmas (hemoplasmas) in free-ranging bats from Southern Brazil.

Hemotropic mycoplasmas (hemoplasmas) are bacteria distributed worldwide and affect domestic and wildlife animals and human beings. Hemoplasmas have been described infecting hematophagous and non-hematophagous bats; however, transmission risk and zoonotic potential in vampire bats remain to be fully established. This study aimed to evaluate the presence of hemotropic mycoplasma species in free-ranging bats from this area using a universal PCR protocol for hemoplasmas. Accordingly, ten blood samples were collected from six male common vampire bats (Desmodus rotundus), two male hairy-legged vampire bats (Diphylla ecaudata), and two female non-hematophagous Pallas's mastiff bats (Molossus sp.) from the Curitiba's region, Paraná State, Southern Brazil. A total of eight (8/10) blood samples were positive byconventional PCR; five (5/6) Desmodus rotundus, two (2/2) Diphylla ecaudata, and one (1/2) Molossus sp. bats. The analyses of the partial sequence of the 16S rDNA gene suggest that the hemoplasma detected in Desmodus rotundus in South Brazil has a high identity compared to the hemoplasma circulating in vampire bats from Central and South America.

Seroprevalence of anti-Toxoplasma gondii antibodies in wild boars (Sus scrofa), hunting dogs, and hunters of Brazil.

Seroprevalence of Toxoplasma gondii has been extensively studied in wild boars worldwide due to the emerging risk for human infection through meat consumption. However, this is the first study that reports toxoplasmosis seroprevalence in wild boars, wild boar hunters and their hunting dogs. The aim of the present study was to evaluate the seroprevalence of anti-T. gondii antibodies in the complex wild boars, hunting dogs and hunters, and to determine the risk factors associated with seropositivity in southern and central-western Brazil. Overall, anti-T. gondii seropositivity was observed in 15/71 (21.1%) wild boars by modified agglutination test (MAT); and 49/157 (31.2%) hunting dogs and 15/49 (32.7%) hunters by indirect immunofluorescent antibody test (IFAT). Seroprevalence of toxoplasmosis in Brazilian wild boars was within the national and international range, posting wild boars as potential environmental sentinels for T. gondii presence. In addition, the findings have comparatively shown that wild boars have been less exposed to infection than hunting dogs or hunters in both Brazilian regions. Seropositivity for T. gondii was statistically higher in 12/14 (85.7%) captured wild boars when compared to 5/57 (7.0%) free-range wild boars (p = 0.000001). Similarly, captured wild boars from anthropized areas were more likely to be seropositive than of natural regions (p = 0.000255). When in multiple regression model, dogs with the habit of wild boar hunting had significant more chance to be positive (adjusted-OR 4.62 CI 95% 1.16-18.42). Despite potential as sentinels of environmental toxoplasmosis, seroprevalence in wild boars alone may provide a biased basis for public health concerns; thus, hunters and hunting dogs should be always be included in such studies. Although hunters should be aware of potential T. gondii infection, wild boars from natural and agricultural areas may present lower protozoa load when compared to wild boars from anthropized areas, likely by the higher presence of domestic cats as definitive hosts.

Occurrence of antibodies anti -Toxoplasma gondii, Neospora caninum and Leptospira interrogans in a captive deer herd in Southern Brazil.

A large number of Brazilian zoos keep many endangered species of deer, however, very few disease surveillance studies have been conducted among captive cervids. Blood samples from 32 Brazilian deer (Blastocerus dichotomus, Mazama nana and Mazama americana) kept in captivity at Bela Vista Biological Sanctuary (Foz do Iguaçu, Brazil) were investigated for 10 ruminant pathogens, with the aims of monitoring deer health status and evaluating any potential zoonotic risk. Deer serum samples were tested for Brucella abortus, Leptospira (23 serovars), Toxoplasma gondii, Neospora caninum, bovine viral diarrhea virus, infectious bovine rhinotracheitis virus, foot-and-mouth disease virus, western equine encephalitis virus, eastern equine encephalitis virus and Venezuelan equine encephalitis virus. Antibodies against T. gondii (15.6%), N. caninum (6.2%) and L. interrogans serogroup Serjoe (3.1%) were detected. The serological results for all other infectious agents were negative. The deer were considered to be clinically healthy and asymptomatic regarding any disease. Compared with studies on free-ranging deer, the prevalences of the same agents tested among the captive deer kept at the Sanctuary were lower, thus indicating good sanitary conditions and high-quality management practices at the zoo.

Serological and molecular survey of Leptospira spp. among cart horses from an endemic area of human leptospirosis in Curitiba, southern Brazil.

INTRODUCTION: Cart horses are a re-emerging population employed to carry recyclable material in cities. METHODS: Sixty-two horses were sampled in an endemic area of human leptospirosis. The microscopic agglutination test (MAT) and real-time polymerase chain reaction (qPCR) were performed. RESULTS: A seropositivity of 75.8% with serovar Icterohaemorrhagiae in 80.8% of the horses was observed. Blood and urine were qPCR negative. MAT showed positive correlations with rainfall (p = 0.02) and flooding (p = 0.03). CONCLUSIONS: Although horses may be constantly exposed to Leptospira spp. in the environment mostly because of rainfall and flooding, no leptospiremia or leptospiruria were observed in this study.

An outbreak of tuberculosis by Mycobacterium bovis in coatis (Nasua nasua).

Mycobacterium tuberculosis complex, which includes Mycobacterium bovis, infrequently causes severe or lethal disease in captive wildlife populations. A dead coati from a wildlife triage center showing pulmonary lesions compatible with tuberculosis had raised suspicion of a potential disease caused by mycobacteria species and was further investigated. Four native coatis (Nasua nasua) with suspected mycobacterial infection were sedated, and bronchoalveolar lavages and tuberculin skin tests (TSTs) were performed. All animals tested positive upon TST. Mycobacterial culturing, Ziehl-Neelsen staining, and genetic testing were performed on postmortem samples and the etiologic agent was identified as M. bovis. Molecular genetic identification using a polymerase chain reaction panel was crucial to achieving a definitive diagnosis.

Detection of Neospora sp. antibodies in cart horses from urban areas of Curitiba, Southern Brazil.

Neospora caninum is a protozoan parasite which affects dogs as definitive hosts and several mammalian species as intermediate hosts mainly causing abortions and central nervous system disorders. The reemerging population of cart horses for carrying recycling material in urban areas of major cities in Brazil may have an impact on disease spreading, and these animals may be used as sentinels for environmental surveillance. Thus, the present study investigated the frequency of Neospora sp. antibodies in cart horses from Curitiba and surrounding areas, Paraná State, Southern Brazil. IgG antibodies against Neospora sp. were detected using indirect fluorescence antibody test (IFAT), and titers equal to or higher than 1:50 were considered reactive. Of all samples, 14/97 (14.4%) were positive: 2/29 (6.9%) were younger than 5; 5/26 (19.2%) between 6 and 9; and 6/31 (19.4%) older than 10 years of age. One of the 11 animals with unknown age was positive (9.1%). Cart horses are likely to be more exposed to dog feces and to Neospora sp. oocyst contamination in urban settings and a lower frequency of disease in dogs may have a negative impact on horse infection risk in these areas.

Detection of RD(Rio) strain of Mycobacterium tuberculosis in tapirs (Tapirus terrestris) from a zoo in Brazil.

Tuberculosis is a chronic infection caused by strains of the Mycobacterium tuberculosis complex and occurs in both animal and human populations. The death of a tapir showing purulent material and a hard mass in the lungs at necropsy raised suspicion of a potential disease caused by mycobacteria species in a Brazilian zoo. Later, two other tapirs with similar signs died and were further investigated. Polymerase chain reaction (PCR) from bronco-alveolar lavages was performed, and both animals tested positive for the RD(Rio) strain of M. tuberculosis, which is a recently discovered Latin American-Mediterranean sublineage and the main cause of human tuberculosis in Rio de Janeiro, Brazil. To investigate the possibility of human infection and the source of transmission, all 50 zoo employees underwent tuberculin skin testing; four were reactive, but radiographic exams and direct sample staining did not suggest tuberculosis. Thus, direct human to animal transmission was not proven. However, the presence of RD(Rio) M. tuberculosis in tapirs highlights the lack of attention to diseases that human beings may transmit to wildlife.