Negative seroprevalence for Toxoplasma gondii in free-living primates from Central Amazonia.

This study presents 35 negative serologies for antibodies anti-T. gondii in free-living primates from Central Amazonia. Our results suggest that these populations have not had contact with the parasite and, therefore, do not have antibodies. This was the first study surveying T. gondii in Cacajao, Callicebus, Pithecia, and Saguinus monkeys.

Apparent prevalence and risk factors for bovine tuberculosis in the state of Paraná, Brazil: an assessment after 18 years since the beginning of the Brazilian program.

Bovine tuberculosis (bTB) impacts considerably animal production and one health worldwide. To describe the prevalence, risk factors, and spatial pattern of the disease in the state of Paraná, Brazil, a cross-sectional study was conducted from September 2018 to February 2019. The area was divided into seven regions. Within each region, farms were randomly selected, and a predetermined number of cows was selected and tested by a comparative cervical tuberculin test. 17,210 animals were tested across 1757 farms. Herd prevalence of bTB-infected herds in Paraná was 2.5% [1.87-3.00%]. It has varied from 0.8 to 3.98% among seven regions, with clustering being detected in the west, central, and northeast areas. Animal prevalence was 0.35% [0.21-0.59%] and has varied from 0.08 to 0.6% among the pre-set regions. No major shifts in the prevalence of bTB were detected since 2007. Large-sized herds, dairy production, and feeding with whey were detected to be correlated with the presence of bTB. Exclusively among dairy herds, veterinary assistance from cooperatives, possession of self-owned equipment to cool milk, and feeding with whey were correlated with the disease. Considering these results, it is recommended that the state of Paraná seek to implement a surveillance system for the detection of bTB-infected herds transforming them into free ones, if possible, incorporating elements of risk-based surveillance. Health education is also recommended to inform farmers about the risks of introducing animals without testing and of feeding raw whey to calves.

Serological screening for Brucella spp. and Leptospira spp. antibodies in southern elephant seals Mirounga leonina from Elephant Island, Antarctica, in 2003 and 2004.

Brucella spp. and Leptospira spp. antibodies were surveyed in 35 southern elephant seals (SESs) Mirounga leonina at Elephant Island (South Shetland Islands), western Antarctic peninsula, in the Austral summer of 2003 and 2004. The rose Bengal test and a commercial competitive ELISA (c-ELISA) were used to detect Brucella spp. exposure, and the microscopic agglutination test (MAT) with 22 live serovars was used to determine anti-Leptospira spp. antibodies. We found evidence of Brucella spp. exposure in 3 of 35 (8.6%) SESs tested via the c-ELISA displaying high percentage inhibition (PI), similar to other studies in pinnipeds in which Brucella spp. antibodies have been determined. Two of the 3 positives were pups (PI = 70.4 and 86.6%), while the third was an adult female (PI = 48.8%). The 3 c-ELISA positive SESs were additionally tested via the serum agglutination test but were found to be negative. All individuals were negative for antibodies against 22 Leptospira spp. serovars by MAT. These results contribute to the knowledge and monitoring of zoonotic pathogens with epizootic potential in Southern Ocean pinnipeds. Given the potential impact that pathogens may have on the abundance of wild (sometimes threatened and endangered) populations, constant monitoring and surveillance are required to prevent pathogen spread, particularly under forecast climate change scenarios.

Tuberculosis caused by Mycobacterium bovis infection in a captive-bred American bullfrog (Lithobates catesbeiana).

BACKGROUND: Tuberculosis is widely known as a progressive disease that affects endothermic animals, leading to death and/or economical losses, while mycobacterial infections in amphibians are commonly due to nontuberculous mycobacteria. To the authors' knowledge, this report describes the first case of bovine tuberculosis in a poikilothermic animal. CASE PRESENTATION: An adult female captive American bullfrog (Lithobates catesbeianus Shaw, 1802) died in a Brazilian aquarium. Multiple granulomas with acid-fast bacilli were observed in several organs. Identification of Mycobacterium bovis was accomplished by culture and PCR methods. The other animals from the same enclosure were euthanized, but no evidence of mycobacterial infection was observed. CONCLUSIONS: The American bullfrog was introduced in several countries around the world as an alternative husbandry, and its production is purposed for zoological and aquarium collections, biomedical research, education, human consumption and pet market. The present report warns about an episode of bovine tuberculosis in an amphibian, therefore further studies are necessary to define this frog species' role in the epidemiology of M. bovis.

Cutaneous mycobacteriosis in a captive Amazonian manatee Trichechus inunguis.

An adult male Amazonian manatee Trichechus inunguis under human care presented with 3 circular cutaneous lesions on the dorsal aspect of the rostrum and between the nostrils (plenum). Initially these lesions were superficial, hypopigmented, without warmth and non-painful. Microbiological cultures of skin swabs isolated Candida sp. and Pseudomonas aeruginosa, and topical treatment with antiseptic, antifungal, anti-inflammatory and antibiotic medication was instituted. This treatment strategy did not lead to any clinical improvement, and after 6 mo, the lesions progressed to a confluent abscess (5.0 × 3.0 cm) with increased temperature and obvious discomfort on palpation. An impression smear of a cutaneous biopsy was submitted for Ziehl-Neelsen staining and after detection of acid-fast bacilli, the cutaneous biopsy and a swab from the lesion were sent for histopathology, culture and sensitivity testing. After 5 d of incubation and through PCR-restriction analysis of the isolates, Mycobacterium fortuitum and M. abscessus were identified. Sensitivity testing indicated that the isolates were susceptible to ciprofloxacin and clarithromycin, and after draining of the lesion and administration of systemic antibiotic treatment, there was rapid clinical improvement. This report describes non-healing lesions in an aquatic animal and illustrates the importance of evaluating the presence of non-tuberculous mycobacteria, opportunistic pathogens which are ubiquitous in the aquatic environment, in protracted, non-responsive cases. We also highlight the importance of a correct diagnosis and treatment approach, and we review concerns that these bacteria are zoonotic agents and are frequently resistant to conventional antibiotics.

Survey of Brucella spp. and Leptospira spp. antibodies in cetaceans and manatees of the Amazon basin and Atlantic Ocean, Brazil.

Aquatic mammals can act as sentinels of emerging and resurging pathogens in the environment. Brucella spp. and Leptospira spp. are 2 zoonotic pathogens relevant to aquatic mammals, and their detection can be used to assess pathogen exposure. In this study, serum from 84 individuals-63 cetaceans (families Iniidae, n = 37; Delphinidae, n = 22; and Kogiidae, n = 4) and 21 West Indian manatees Trichechus manatus-was tested by the Rose Bengal Test (RBT) and a commercial competitive enzyme-linked immunosorbent assay (c-ELISA) for detecting Brucella spp. antibodies, and the microscopic agglutination test (MAT) for screening Leptospira spp. exposure. Overall, 4.8% (3/63) of cetaceans were positive by RBT and 15.9% (10/63) by c-ELISA for Brucella spp. Serum from 8 c-ELISA positive cetaceans (with available serum) was further tested via serum agglutination test (SAT) and 1 individual was positive. c-ELISA was more sensitive than RBT. Exposure to Brucella spp. was found in 5 cetacean species: Clymene dolphin Stenella clymene, short-finned pilot whale Globicephala macrorhynchus, pygmy killer whale Feresa attenuata, melon-headed whale Peponocephala electra and Atlantic bottlenose dolphin Tursiops truncatus in the Atlantic Ocean, Brazil, expanding the range of known Brucella seropositive aquatic hosts. No evidence of Brucella spp. exposure was found in Iniidae and Kogiidae odontocetes and manatees. Antibodies against Leptospira spp. were not detected in cetaceans and sirenians by MAT. These results contribute to the evaluation of different Brucella spp. serological methods in cetaceans and manatees and highlight the epidemiology of zoonotic pathogens in aquatic mammals of the southwestern Atlantic Ocean and the Amazon basin.

Sero-epidemiological survey for brucellosis, leptospirosis, and toxoplasmosis in free-ranging Alouatta caraya and Callithrix penicillata from São Paulo State, Brazil.

BACKGROUND AND METHODS: Sera were tested for Brucella spp., Leptospira spp. and Toxoplasma gondii antibodies in 68 free-ranging New World monkeys from a forest fragment of the Brazilian Cerrado. RESULTS AND CONCLUSION: All animals were negative for Brucella spp. and Leptospira spp. However, 75% of Alouatta caraya and 16.6% of Callithrix penicillata were positive for T. gondii. The implications for conservation and health management are discussed.

Epidemiological Situation of Glanders in the State of Pará, Brazil.

Glanders is an anthropozoonosis caused by the bacteria Burkholderia mallei, affecting mainly equids. It has been eradicated in North America, Australia, and Western Europe, but continues to occur sporadically in countries in Asia, Africa, the Middle East, and South America. Its notification is mandatory by the World Organization for Animal Health. After 30 years, the disease reappeared in Brazil in 1999 and, thereafter, 1,413 outbreaks have been reported. However, the epidemiological situation of the disease in the country is not adequately known. Thus, 2718 animals from 654 properties in the state of Pará were randomly selected by sampling and examined using a serial protocol with Complement Fixation and Western Blot serological tests. The prevalence of properties infected with glanders in the state was estimated at 1.68% [0.84; 3.33] and of seropositive animals at 0.50% [0.27; 0.94]. The introduction of animals was individualized as a risk factor for disease introduction in the properties (OR = 5.9 [1.4; 25.5]). Despite the low prevalence of infected properties and seropositive animals, the state must review actions to fight the disease, considering that the strategies implemented have not affected the endemic balance of the disease. This process must involve all public and private agents interested in the topic.

Retrospective benefit-cost analysis of bovine brucellosis control in the state of Mato Grosso, Brazil.

From 2001, due to the launching of the National Program for the Control and Eradication of Brucellosis and Tuberculosis, Brazil began a series of studies aimed at the epidemiological characterization of bovine brucellosis in its federative units. In the state of Mato Grosso, studies conducted in 2003 and 2014 showed a reduction in the prevalence of infected herds from 41.2 % to 24 % and in the prevalence of seropositive animals from 10.2 % to 5.1 %. These results stemmed from the efficient heifer vaccination program with S19 implemented by the state. From this scenario, the present study quantified and compared the benefits and costs of bovine brucellosis control in the state through a financial analysis. When considering private and public costs, the Net Present Value (NPV) was estimated at US$ 37.5 million, the Internal Rate of Return (IRR) was calculated at 31 %, and the Benefit-Cost Ratio (BCR) was 2.3. When considering only the private costs, the NPV was US$ 51.3 million, the IRR was 46 % and the BCR was 3.43, indicating that the cattle producer in Mato Grosso had a return of US$ 3.43 for each US$ 1.00 invested. Therefore, the bovine brucellosis control actions carried out in the state of Mato Grosso between 2003 and 2014 were highly advantageous from the economic point of view. The most rational way to continue decreasing bovine brucellosis prevalences at low cost in the state is to maintain the heifer vaccination program, using the RB51 vaccine in addition to S19.

Characterization of Erysipelothrix rhusiopathiae isolated from Brazilian Tayassu pecari.

Erysipelothrix rhusiopathiae causes a variety of diseases in many animal species, including human beings. Most human infections caused by this pathogen are related to occupational exposure, and swine are considered to be the most important reservoir of E. rhusiopathiae. The white-lipped peccary (Tayassu pecari) is an ungulate that has some genetic relationship to swine, and since the demand for T. pecari meat has recently increased in Brazil and nothing is known about the relationship of this peccary with the occupational zoonotic agent, E. rhusiopathiae, an investigation on the matter was conducted. Tonsils from 21 T. pecari slaughtered in southern Brazil were examined, and one animal was positive for E. rhusiopathiae isolation. Five colonies of this positive specimen had their species identity confirmed by PCR, and were characterized by serotyping, broth microdilution susceptibility test, and pulsed field gel electrophoresis (PFGE). All colonies belonged to serotype 2b, and presented identical susceptibility profiles. Nevertheless, the five colonies showed three different PFGE profiles, demonstrating the occurrence of infection by different E. rhusiopathiae genotypes. This is the first report of E. rhusiopathiae infection in T. pecari as well as the first description of animals carrying different E. rhusiopathiae genotypes.

Hematology and Serum Chemistry of Free-Ranging Jaguars (Panthera onca) from a Floodplain Forest Site in Amazonia, Brazil.

We provide hematologic and biochemical information for 13 jaguars (Panthera onca) from the Amazon. Most showed increased lymphocytes, monocytes, eosinophils, aspartate aminotransferase, and alanine aminotransferase; all individuals showed lower mean red blood cell (RBC) counts compared to findings from other biomes. Males had a significantly higher RBC count and alkaline phosphatase than did females.

Canine Leproid Granuloma (CLG) Caused by Mycobacterial Species Closely Related to Members of Mycobacterium Simiae Complex in a Dog in Brazil.

This report describes the clinical features and molecular diagnosis of a case of canine leproid granuloma (CLG) caused by mycobacterial strains of the Mycobacterium simiae complex in Brazil. A 12-year-old non-neutered male Labrador Retriever dog was presented with a 2-week history of progressive painless cutaneous lesions. Ulcerated nodules with hematic crusts were observed on the dorsal surface of the right and left pinna and on the metacarpal, metatarsal, and digits. Complete blood count, serum biochemistry, aspiration cytology of cutaneous lesions, biopsy for histopathological evaluation, culture for aerobic and anaerobic bacteria, polymerase chain reaction and DNA sequencing to identify mycobacterial species were performed. According to the clinical and histopathological findings, a diagnosis of CLG was established. Despite the negative result of the bacterial culture, mycobacterial identification was made by sequencing the hsp65 gene. Our findings highlight that mycobacterial species closely related to members of the M simiae clade can be causative agents of CLG.

Exposure of free-ranging wild carnivores, horses and domestic dogs to Leptospira spp in the northern Pantanal, Brazil.

Leptospirosis is a zoonotic disease affecting most mammals and is distributed throughout the world. Several species of domestic and wild animals may act as reservoirs for this disease. The purpose of this study was to assess the exposure of free-ranging wild carnivores, horses and domestic dogs on a private reserve located in the northern Pantanal (Brazil) and the surrounding areas to Leptospira spp from 2002-2006, 75 free-ranging wild carnivores were captured in the Pantanal and serum samples were collected. In addition, samples from 103 domestic dogs and 23 horses in the region were collected. Serum samples were tested for the presence of Leptospira antibodies using the microscopic agglutination test. Thirty-two wild carnivores (42.7%) were considered positive with titres ≥ 100, and 18 domestic dogs (17.5%) and 20 horses (74.1%) were also found to be positive. Our study showed that horses, dogs and several species of free-ranging wild carnivores have been exposed to Leptospira spp in the Pantanal, suggesting that the peculiar characteristics of this biome, such as high temperatures and an extended period of flooding, may favour bacterial persistence and transmission. In this region, wild carnivores and horses seem to be important hosts for the epidemiology of Leptospira species.

[Risk model to assess livestock rabies exposure in the state of São Paulo, Brazil]. / Modelo de risco para circulação do vírus da raiva em herbívoros no estado de São Paulo, Brasil.

OBJECTIVE: To propose a qualitative risk assessment model for the study of livestock exposure to rabies virus from the vampire bat Desmodus rotundus (antigenic variant 3) in the Paraíba do Sul river valley, state of São Paulo, Brazil. METHODS: Based on scenario trees generated considering rabies exposure and its spread, we estimated the probability of rabies cases in large livestock and its association with the geographic location of livestock farms. RESULTS: Assessment of the historical series of rabies focal points in the first semester of 2006, which was used to validate the risk assessment model, revealed that 81.8% of the focal points were adequately foreseen by the model and could have been prevented with strategic vaccination in high-risk areas. CONCLUSIONS: The adoption of control measures specifically targeting high-risk areas might entail a substantial decrease in the number of rabies focal points, at a low cost and with optimal movement of field teams.

Multispacer Sequence Typing for Mycobacterium bovis Genotyping.

The molecular typing of Mycobacterium bovis, which causes bovine tuberculosis, can be accomplished by combining different polymorphic markers, contributing to its epidemiological investigation. Multispacer sequence typing (MST) is a sequencing-based method that employs intergenic regions susceptible to higher mutation rates given the low selection pressure. It has been applied to M. tuberculosis, but not to M. bovis. The aim of this study was to evaluate a MST for M. bovis. A total of 58 strains isolated from tissues with lesions suggestive of bovine tuberculosis, coming from cattle herds in six Brazilian states and four standard samples of M. bovis were typified employing the MST technique. Fourteen intergenic regions were used, and four types of genetic events were reported: single nucleotide mutation (SNP), insertion, deletion, and tandem repeat (TR). Seven loci were chosen for typing. Twenty-eight type sequences (ST) were identified, indicating type sequences (ST) were identified, indicating a 92.9% HGDI (Hunter Gaston Discriminatory Index). The data were used to analyze the evolutionary patterns of these isolates and correlate them to phylogeographic lineages based on the formation of clonal complexes generated from eBURST software. Later, we associated the MST with spoligotyping technique, currently considered the gold standard for classification of M. bovis. The results support the MST as an alternative method for genotyping of M. bovis. The method has the advantage of sequencing and the availability of sequences analyzed in public databases, which can be used by professionals around the world as a tool for further analysis. This was the first study to identify the variability of isolates of M. bovis by the MST method.

Seroepidemiological investigation of animal leptospirosis and molecular characterization of the first Leptospira strain isolated from Fernando de Noronha archipelago, Brazil.

Leptospirosis has been widely reported in insular environments worldwide, characterizing a major public health threat. Although low-genetic biodiversity is expected in these regions, the introduction of domestic and synanthropic mammals may contribute to the wider diversity of leptospiral strains in insular settings. This study proposes a large-scale seroepidemiological investigation of Leptospira infection in animals from Fernando de Noronha archipelago and describes the characterization of the first leptospiral strain ever isolated from an insular setting in Brazil. A total of 1,265 blood samples from domestic (n = 682), synanthropic (n = 133) and wild (n = 450) animals were collected between 2007 and 2014, totalling 12 species. The presence of anti-Leptospira spp. antibodies was investigated by the microscopic agglutination test (MAT), and kidney samples from 20 synanthropic rodents were collected for the isolation of Leptospira spp. The leptospires recovered were further characterized by serogrouping with polyclonal antibodies, whole-genome sequencing and multilocus sequence typing (MLST). The MAT results revealed the presence of agglutinins in 90 samples (7.1%) and the most frequently found serogroup was Icterohaemorrhagiae (n = 57) in practically all species included. Viable leptospires were recovered from one brown rat, and characterization revealed that the isolate belongs to L. interrogans serogroup Pyrogenes. The results suggest that synanthropic rodents might play an important role in leptospiral infection among wildlife and domestic species in the archipelago.

Evolutionary analysis of Mycobacterium bovis genotypes across Africa suggests co-evolution with livestock and humans.

Mycobacterium bovis is the pathogenic agent responsible for bovine tuberculosis (bTB), a zoonotic disease affecting mostly cattle, but also transmittable to humans and wildlife. Genetic studies on M. bovis allow to detect possible routes of bTB transmission and the identification of genetic reservoirs that may provide an essential framework for public health action. We used a database with 1235 M. bovis genotypes collected from different regions in Africa with 45 new Mozambican samples. Our analyses, based on phylogeographic and population genetics' approaches, allowed to identify two clear trends. First, the genetic diversity of M. bovis is geographically clustered across the continent, with the only incidences of long-distance sharing of genotypes, between South Africa and Algeria, likely due to recent European introductions. Second, there is a broad gradient of diversity from Northern to Southern Africa with a diversity focus on the proximity to the Near East, where M. bovis likely emerged with animal domestication in the last 10,000 years. Diversity indices are higher in Eastern Africa, followed successively by Northern, Central, Southern and Western Africa, roughly correlating with the regional archaeological records of introduction of animal domesticates. Given this scenario M. bovis in Africa was probably established millennia ago following a concomitant spread with cattle, sheep and goat. Such scenario could translate into long-term locally adapted lineages across Africa. This work describes a novel scenario for the spread of M. bovis in Africa using the available genetic data, opening the field to further studies using higher resolution genomic data.

High discrimination of Mycobacterium bovis isolates in Brazilian herds by spoligotyping.

Bovine tuberculosis is an infectious disease caused by Mycobacterium bovis (M. bovis), that leads to economic losses in infected herds and it is also considered an important zoonosis. The molecular typing methods of M. bovis isolates are fundamental for the bovine tuberculosis surveillance system, and spoligotyping is the standard genotyping technique for this species. Thus, the aim of the present study is to analyze the spatial and cluster distribution of M. bovis strains from several regions of Brazil through molecular typing. Spoligotyping technique was applied on 422 isolates identified as M. bovis, and Ripley's K function was used to perform the spatial and cluster analysis of each identified profile. Forty-three (43) different profiles were identified and spoligotype SB0121 was the most frequent and showed a uniform pattern in the spatial distribution while spoligotypes SB0295, SB1380 and SB1050 formed clusters. In addition, three novel spoligotype profiles (SB2361, SB2362, SB2364) were identified in different herds. In this perspective, it is believed that molecular identification and typing can significantly improve the performance of surveillance systems for bovine tuberculosis in Brazil.

Experimental leptospirosis in capybaras (Hydrochaeris hydrochaeris) infected with Leptospira interrogans serovar Pomona.

Capybara (Hydrochaeris hydrochaeris), the largest rodent in the world, is widely distributed in South America. These animals live in areas with abundant water, which makes them a potential reservoir for Leptospira. The objective of this study was to investigate seroconversion, leptospiremia, and leptospiruria in capybaras experimentally infected with a virulent strain of Leptospira interrogans serovar Pomona. Seven capybaras were used: one control and six infected. Agglutinins against serovar Pomona were initially detected in serum 6 or 7 day after innoculation with Leptospira (10(9)-10(11) organisms, given i.v.), peaked (titer, approximately 3,200) between 9 and 27 day, and were still present at 83 day (end of study). The earliest and latest isolation of leptospires from the blood was from 2-12 day and from urine, 9-19 day after exposure. However, polymerase chain reaction and isolation results from kidney and liver samples were negative for leptospires. The control animal tested negative on all diagnostic tests. Hence, the capybara can serve as a host for Leptospira.

Molecular, serological, pathological, immunohistochemical and microbiological investigation of Brucella spp. in marine mammals of Brazil reveals new cetacean hosts.

Brucella-exposure and infection is increasingly recognized in marine mammals worldwide. To better understand the epidemiology and health impacts of Brucella spp. in marine mammals of Brazil, molecular (conventional PCR and/or real-time PCR), serological (Rose Bengal Test [RBT], Competitive [c]ELISA, Serum Agglutination Test [SAT]), pathological, immunohistochemical (IHC) and/or microbiological investigations were conducted in samples of 129 stranded or by-caught marine mammals (orders Cetartiodactyla [n = 124], Carnivora [n = 4] and Sirenia [n = 1]). Previous serological tests performed on available sera of 27 of the 129 animals (26 cetaceans and one manatee), indicated 10 seropositive cetaceans. Conventional PCR and/or real-time PCR performed in cases with available organs (n = 119) and/or blood or swabs (n = 10) revealed 4/129 (3.1%) Brucella-infected cetaceans (one of them with positive serology; the remaining three with no available sera). Pathological, IHC and/or microbiological analyses conducted in PCR/real-time PCR and/or seropositive cases (n = 13) revealed Brucella-type lesions, including meningitis/meningoencephalitis, pneumonia, necrotizing hepatitis, pericarditis and osteoarthritis in some of those animals, and positive IHC was found in all of them (excepting two live-stranded animals without available organs). Brucella spp. culture attempts were unsuccessful. Our results demonstrated exposure, asymptomatic, acute and chronic Brucella sp. infection in several cetacean species in the Brazilian coast, highlighting the role of this pathogen in stranding and/or death, particularly in Clymene dolphin (Stenella clymene) and short-finned pilot whale (Globicephala macrorhynchus) off Ceará State. Novel hosts susceptible to Brucella included the franciscana (Pontoporia blainvillei), the Guiana dolphin (Sotalia guianensis) and the spinner dolphin (Stenella longirostris). Additionally, three coinfection cases involving Brucella spp. and cetacean morbillivirus, Edwarsiella tarda and Proteus mirabilis were detected. To the best of our knowledge, this is the first long-term and large-scale survey of Brucella spp. in marine mammals of South America, widening the spectrum of susceptible hosts and geographical distribution range of this agent with zoonotic potential.