Concentrations of polycyclic aromatic hydrocarbons (PAHs) in liver samples of green turtles Chelonia mydas stranded in the Potiguar Basin, northeastern Brazil.

Sea turtles are affected by pollutants worldwide, and the polycyclic aromatic hydrocarbons (PAHs) have been detected in different types of samples and at high levels in some cases. The present study brings concentrations of 37 PAHs in liver samples of 17 green turtles Chelonia mydas stranded in northeastern Brazil [four with cutaneous tumors of fibropapillomatosis (FP), being classified as FP+]. Six PAHs were detected in 100% of the liver samples, and all alkylated PAHs were frequently quantified. High levels of phenanthrene (771.20 and 794.43 ng g-1 d.w.) and fluorene (1882.36 ng g-1 d.w.) were found in three females FP- (without FP cutaneous tumors). On the other hand, one green turtle FP+ had the higher level of naphthalene (531.70 ng g-1 d.w.), compound detected in 82.35 % of the samples. Our study brings additional baseline of organic pollutants in green turtles, improving knowledge on bioaccumulation of these compounds in sea turtles.

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.

High Occurrence of Chelonid Alphaherpesvirus 5 (ChHV5) in Green Sea Turtles Chelonia mydas with and without Fibropapillomatosis in Feeding Areas of the São Paulo Coast, Brazil.

Chelonid alphaherpesvirus 5 (ChHV5) has been consistently associated with fibropapillomatosis (FP), a neoplastic disease that affects sea turtles globally. The DNA of ChHV5 has been detected in cutaneous and noncutaneous tissues (e.g., lung) of green sea turtles Chelonia mydas with (FP+) and without (FP-) clinical signs of FP, indicating a persistent ChHV5 infection. Previously published and custom primer pairs were used to amplify the fragments of ChHV5 unique long (UL) partial genes (UL30 and UL18) through end-point PCR from cutaneous tumors (n = 31), nontumored skin (n = 49), and lungs (n = 26) from FP+ (n = 31) and FP- (n = 18) green sea turtles. The DNA of ChHV5 was detected in cutaneous tumors (80.6%, 25/31), nontumored skin (74.2%, 23/31 FP+; 27.8%, 5/18 FP-), and lung samples (91.7%, 11/12 FP+; 100%, 14/14 FP-). The high occurrence of ChHV5 observed in lung samples from FP- individuals was unexpected (14/14), providing the first evidence of ChHV5 DNA presence in lungs of individuals without FP. Our results also revealed high ChHV5 occurrence among the tested cohort (93.9%, 46/49) and suggested that a large proportion (83.4%, 15/18) of FP- green sea turtles had subclinical ChHV5 infections. Hence, our findings support the hypothesis that ChHV5 requires one or more possibly environmental or immune-related co-factors to induce FP.

Analysis of the cattle trade network in the state ofMato Grosso, Brazil / Análise da rede de trânsito de bovinos no estado de Mato Grosso, Brasil

We characterized the network of cattle movements in Mato Grosso, Brazil­a state responsible for a significant amount of Brazilian beef exports. In this analysis, we were particularly interested in the importance of slaughterhouses in the network of cattle movements, in the characteristics of the trade for different purposes (especially for reproduction, fattening, and slaughter), and about the importance of the main traders. We found that the median distance of all movements was 79.0 km, with an interquartile range between 37.2 and 163.2 km, which suggests a relatively short-range trade pattern; roughly 20% of the premises purchasing more animals were responsible for 95% of the animals purchased and 20% of the premises selling more animals were responsible for 86% of the animals sold, indicating the importance of the cattle trade hubs. Fitting the degree distribution using a power-law distribution, we obtained a scaling parameter of 2.59, indicating that the cattle movement network of Mato Grosso is less dense than other observed cattle movement networks. The networks for fattening, reproduction, and slaughtering only comprised 73%, 56%, and 25% of all nodes and 52%, 30%, and 17% of all edges, respectively; and slaughterhouses had the highest in-degrees and total degrees, followed by markets and farms, playing the role of main cattle trade hubs. Therefore, slaughterhouses are potentially useful as surveillance points to detect cases, and controlling important trader premises may optimize the use of resources for controlling diseases.(AU)

Caracterizamos a rede de trânsito de bovinos em Mato Grosso, Brasil ­ um estado responsável por uma quantidade significativa das exportações brasileiras de carne bovina. Nesta análise, o interesse foi na importância de abatedouros na rede de trânsito de bovinos, nas características do comércio para diferentes finalidades (especialmente para reprodução, engorda e abate), e sobre a importância dos principais comerciantes. Encontramos que a distância mediana de todos os movimentos foi de 79,0 km, com um intervalo interquartil entre 37,2 e 163,2 km, o que sugere um padrão de comércio de alcance relativamente curto; cerca de 20% dos estabelecimentos que compram mais animais foram responsáveis por 95% dos animais comprados e 20% dos estabelecimentos que vendem mais animais foram responsáveis por 86% dos animais vendidos, o que mostra a importância dos principais comerciantes de bovinos. Ajustando a distribuição de graus usando uma distribuição de lei de potência, obtivemos um parâmetro de escala de 2,59, indicando que a rede de movimentação de bovinos de Mato Grosso é menos densa do que outras redes de trânsito bovino observadas. As redes para engorda, reprodução e abate compreenderam 73%, 56% e 25% de todos os nós e 52%, 30% e 17% de todas as arestas, respectivamente; e os frigoríficos tiveram os mais altos graus de entrada e totais, seguidos por mercados e fazendas, desempenhando o papel de principais pontos de comércio de gado. Por conseguinte, abatedouros são potencialmente úteis como pontos de vigilância para detectar casos, e o controle de estabelecimentos importantes para o comércio pode otimizar o uso de recursos para o controle de doenças.(AU)

Congenital Transmission of Toxoplasma gondii After Experimental Reinfection With Brazilian Typical Strains in Chronically Infected Sheep.

Toxoplasma gondii is a cause of congenital diseases, miscarriages and stillbirths in production animals. In Brazil, non-archetypal genotypes of the parasite may be related to severe disease. Experimental infection with T. gondii was studied in sheep to analyse congenital transmission-related parameters in reinfections with different Brazilian parasite strains. Thirteen T. gondii-seronegative sheep were orally infected with 2 × 103oocysts for the primary infection: G1 (4 animals) were inoculated with TgCatBr71 strain (Type BrI genotype) and G2 andG3 (5 and 4 animals, respectively) withTgCatBr60 strain (Type BrIII genotype). After chronification of infection, the animals were impregnated. A second infection was performed after 60 days of gestation. TheG1 andG3 animals were inoculated withTgCatBr60BrIII and the G2 animals withTgCatBr71 BrI oocysts. The effects of reinfection were compared with a control group (5 animals) through physical examination, ultrasound imaging and serology. Ovine experimental infections were evaluated using mouse bioassays, molecular analysis, serological tests, histopathology, and immunohistochemistry. No abortions occurred; a seropositive lamb and a mummified fetus from G2-BrIIIxBrI were produced. The vertical transmission rate detected in lambs from chronically infected sheep was 31.6% (6/19). It is demonstrated that reinfection and subsequent congenital transmission occured in one sheep with a primary Brl infection challenged with BrIII genotype of T. gondii. In a twin pregnancy from G2-BrIIIxBrI, congenital transmission from a latent infection was detected in both lambs. Congenital transmission could not be tracked in three lambs. Overall, previous T. gondii infection may fail to protect against congenital transmission from a reinfection and primary infection induced insufficient protection against vertical transmission which must be taken into account in decision-making for the use of seropositive animals as breeders. Similar trials with larger groups and contemplating host cellular immune response studies should be conducted to evaluate the actual impact of T. gondii reinfection involving different strains in sheep.

Validity of a two-stage cluster sampling design to estimate the total number of owned dogs.

Estimates of owned dog population size are necessary to calculate measures of disease frequency and to plan and evaluate population management programs. We calculated the error and bias of estimates of the total number of owned dogs using a two-stage cluster sampling design. The estimates were conditioned on sample composition as well as on size and heterogeneity of the spatial distribution of owned dog populations. For this, we simulated nine cities that differed systematically in size (number of census tracts) and heterogeneity (variance of the number of dogs per census tract). Then, we defined 16 scenarios to calculate the sample composition using an algorithm that incorporated data from a pilot sample, estimates of cost, and prior specifications of the expected error and confidence level. In three additional scenarios of predefined sample composition, the numbers of primary and secondary sampling units were: 30 × 30, 50 × 20 and 65 × 15. Finally, for each city and sample composition, we selected primary sampling units (census tracts) with probability proportional to its size and with replacement, and secondary sampling units (households) by simple random sampling. For each city and composition we selected 500 samples, totaling 85500 samples. The distribution of errors conditioned on the sample composition and city showed that estimates were accurate (average mean bias = 0.006%, maximum mean bias = 0.3%). All sample compositions resulted in errors between 4% and 7% in cities with low heterogeneity. In cities with high heterogeneity, the errors for the various compositions ranged as follows: 8-11% (calculated), 11-13% (65 × 15), 12-14% (50 × 20) and 15-17% (30 × 30). The sample size of predefined compositions was between 33% and 87% lower than the sample size of calculated compositions. Therefore, the predefined compositions have an operational advantage (reduced sampling effort) and simplify the sampling design (calculation of sample composition is not needed). Furthermore, the expected error of estimates under different scenarios is known for each predefined composition. In the absence of information about the heterogeneity of the cities, the 65 × 15 is the more conservative composition.

Seroprevalence of Leptospira spp infection and its risk factors among domestic dogs in Bogotá, Colombia.

Different analytical tools were used to determine the seroprevalence of and risk factors associated with Leptospira spp infection in 192 domestic dogs (Canis familiaris) in Bogotá, Colombia. Using the microscopic agglutination test (MAT), a battery of 16 Leptospira serovars were tested. The seroprevalence of Leptospira spp was calculated as 36.46% (95% CI 0.30-0.43). A questionnaire was applied to the dogs' owners at the time of sampling and the variables "Water sources near home" and "Dog hunting rodents" were identified as risk factors for leptospirosis occurrence in the urban area of Bogotá. Geographical coordinates relating to the dogs' households were obtained in order to map out the spatial distribution of reactive and unreactive dogs. Additionally, we found that the mean annual precipitation was higher at geographical locations with reactive animals than at those with unreactive dogs (p < 0.05). Preventing exposure of dogs to rodents and waste-water bodies that could be contaminated with Leptospira might effectively reduce occurrences of leptospirosis. Moreover, promoting preventive programs and vaccination of dogs against leptospirosis in areas of higher precipitation and prior to rainy months could be an effective strategy for leptospirosis prevention.

Livestock markets play an important role in the cattle movement network in Pernambuco, Brazil / Feiras de gado desempenham papel importante na rede de trânsito de bovinos em Pernambuco, Brasil

The animal trade is an important risk factor that affects the spread of diseases among animals and herds. The goal of the present study was to characterize the cattle movement network in Pernambuco, Brazil, based on the animal movement permits (Guias de Trânsito Animal; GTAs) from 2012 to 2013, and identify the intensity of the commercial relationship between farm premises. A total of 737,950 GTAs were issued, and the movement of 3,481,185 cattle (1,688,585 in 2012 and 1,792,600 in 2013) was analyzed. Of the moved animals analyzed, 52.57% (1,829,907/3,481,185) were involved in the movement of cattle in or out of livestock markets, indicating that livestock markets played a major role in the network. Approximately 20% of the more-connected premises were responsible for approximately 87% of the movement related to sales and 95% of the movement related to purchases. Considering the important role of livestock markets and the intense cattle trade between farm premises, surveillance, and control measures could be more efficient if targeted to livestock markets and highly connected premises to prevent the spread of infectious diseases.(AU)

O comércio de animais é um importante fator de risco para a disseminação de doenças entre animais e rebanhos. O objetivo do presente estudo foi caracterizar a rede de movimentação de bovinos em Pernambuco, com base nas guias de trânsito animal (GTAs) de 2012 a 2013, e identificar a intensidade da relação comercial entre os estabelecimentos. Foram analisadas 737.950 GTAs emitidas, e as movimentações de bovinos totalizaram 3.481.185 animais (1.688.585 em 2012 e 1.792.600 em 2013). Um percentual de 52,57% (1.829.907/3.481.185) das movimentações de bovinos envolveu entrada ou saída de feiras de gado, indicando que as feiras desempenharam um papel importante na rede. Aproximadamente 20% dos estabelecimentos mais conectados foram responsáveis por aproximadamente 87% das movimentações relacionadas às vendas, e 95% das movimentações, relacionadas às compras. Considerando o importante papel das feiras e o intenso comércio de bovinos entre as propriedades, medidas de vigilância e controle podem ser mais eficientes se aplicadas em feiras e estabelecimentos altamente conectados, para prevenir a propagação de doenças infecciosas.(AU)

Challenges in Evaluating the Severity of Fibropapillomatosis: A Proposal for Objective Index and Score System for Green Sea Turtles (Chelonia mydas) in Brazil.

Fibropapillomatosis (FP) is a neoplastic disease that affects marine turtles worldwide, especially green sea turtles (Chelonia mydas). FP tumors can develop on the body surface of marine turtles and also internally in the oral cavity and viscera. Depending on their quantity, size and anatomical distribution, these tumors can interfere with hydrodynamics and the ability to feed, hence scoring systems have been proposed in an attempt to quantify the clinical manifestation of FP. In order to establish a new scoring system adapted to geographic regions, we examined 214 juvenile green sea turtles with FP caught or rescued at Brazilian feeding areas, counted their 7466 tumors and classified them in relation to their size and anatomical distribution. The patterns in quantity, size and distribution of tumors revealed interesting aspects in the clinical manifestation of FP in specimens studied in Brazil, and that FP scoring systems developed for other areas might not perform adequately when applied to sea turtles on the Southwest Atlantic Ocean. We therefore propose a novel method to evaluate the clinical manifestation of FP: fibropapillomatosis index (FPI) that provides the Southwest Atlantic fibropapillomatosis score (FPSSWA). In combination, these indexing and scoring systems allow for a more objective, rapid and detailed evaluation of the severity of FP in green sea turtles. While primarily designed for the clinical manifestation of FP currently witnessed in our dataset, this index and the score system can be adapted for other areas and compare the characteristics of the disease across regions. In conclusion, scoring systems to classify the severity of FP can assist our understanding on the environmental factors that modulate its development and its impacts on the individual and population health of green sea turtles.

Infection of mice with oocysts of Toxoplasma gondii by oral route showed differences of virulence from Brazilian RFLP genotypes BrI and BrIII.

South American strains of Toxoplasma gondii present higher genetic diversity than classical European strains. We compared the virulence of two non-archetypal Brazilian genotypes of T. gondii to mice. Oocysts of four isolates, two genotype BrI (TgCatBr71 and TgShBr11) and two BrIII (TgCatBr74 and TgCatBr60) were obtained from cats fed experimentally infected mice. After sporulation, 5.0×10(1) and 1.0×10(2) oocysts were orally administrated to Swiss albine mice in Experiments #1 and #2, respectively (4-10 mice/group). Humoral response from dead and surviving mice was analyzed on days 9 to 35 post-infection. Microscopic observations of lungs and brains were performed for tachyzoites and cysts visualization in fresh preparations. Negative results were tested by PCR. Virulence after infection with oocysts is dose dependent for genotype BrIII isolates, but not for BrI. Differences in mortality were observed among isolates from genotype BrIII on Experiment #1. Intra-genotype phenotypic variation related to the parasite stage of infection was demonstrated and this characteristic should be further studied and may influence future work regarding the role of virulence amid hosts.

Monitoring techniques in the capture and adoption of dogs and cats.

The continuous improvement of the information systems of organizations that work toward the control of stray dog and cat populations facilitates the implementation of programs aimed at reducing the number of animals that roam free in public streets. This study aimed to present techniques to improve the understanding of the spatial distribution of stray dogs and cats and of people who adopt these animals and the fate of these animals in zoonosis control centers (ZCC). Ripley's K function was used with a Euclidean distance graph to detect the distribution pattern of dogs and cats captured and of the people who adopted these animals. An estimate of the kernel density was used to allow a better assessment of the spatial distribution of the phenomenon studied. The results showed that the distribution of captured animals and of the people who adopted these animals form a spatial cluster (p = 0.01). Most of the animals were captured near the premises of the ZCC and near the downtown area. Factors such as the abandonment of animals near animal control agencies and the availability of food sources are the main hypotheses associated to the distribution of the captures. The awareness of the people who live in places where there is a greater number of stray animals and the distribution of the urban population are the main hypotheses to explain the distribution of the adoptions. The results will help to implement control measures over these populations in the most problematic areas.

Dog and cat management through sterilization: Implications for population dynamics and veterinary public policies.

The present study aimed to compare different sterilization scenarios allowing the adoption of the most adequate strategy to control owned dog and cat population sizes as the official veterinary public policy for animal control in an urban area of Campinas municipality, Brazil. To achieve this goal, the vital parameters of the owned pet population were measured in a neighborhood of Campinas called Jardim Vila Olimpia through questionnaires used in two census studies performed in February 2012 and June 2013. Different hypothetical sterilization scenarios were compared with the scenario of a single sterilization campaign performed in the study area between the census studies. Using a deterministic mathematical model, population dynamics were simulated for these different scenarios. We have observed that for both owned dogs and cats, the impact on the population size achieved by a single sterilization campaign would be diluted over the years, equating to the impact achieved by the usual sterilization rate practiced before the sterilization campaign yearly. Moreover, using local and global sensitivity analyses, we assessed the relative influence on animal population evolution of each vital parameter used in the mathematical models. The more influential parameters for both species were the carrying capacity of the environment and sterilization rates of males and females (for both species). We observed that even with sterilizing 100% of the intact animals annually, it would not be possible to obtain proportions greater than 86% and 88% of sterilized dogs and cats, respectively, after 20 years due to the high introduction of new intact animals. There is no public dog and cat sterilization service in place in the city, and sporadic and local sterilization campaigns are performed with a prior communication to the owners to bring their animals to be sterilized in a selected veterinary facility. If a sterilization campaign was performed annually in the study area, it would have the most favorable cost effectiveness ratio after 20 years compared to the scenarios of 50% and 100% sterilization of intact animals annually. These results allowed the veterinary public policy stakeholders to make decisions based on scientific evidence to implement adequate control of dog and cat populations in urban areas, aiming to reduce zoonosis transmission to humans and other problems associated with uncontrolled animal populations.

Occurrence of antibodies anti-Toxoplasma gondii and anti-Neospora caninum in dogs from Natal, RN, Brazil / Ocorrência de anticorpos contra Toxoplasma gondii e Neospora caninum em cães de Natal, RN, Brasil

The occurrence of anti-Toxoplasma gondii and Neospora caninum in dogs from the municipality of Natal, RN, Brazil, was determined. Information about the presence of these coccidia in this species was not known. Blood samples were collected from 29 domiciled dogs which inhabit areas that surround two important protected areas of Atlantic Forest (Parque da Cidade and Parque das Dunas) and another 73 dogs that were sacrificed due to Leishmania spp. infection, in Center for Control of Zoonosis (CCZ) for the municipality of Natal. It was only possible to obtain information about the gender of dogs that live in the parks area. The presence of antibodies against each parasite was determined by indirect Fluorescence Antibody Test (IFAT), with a cutoff of 16 for T. gondii and 50 for N. caninum. Of the 102 dogs examined, 13 (12.7%, 95% CI 7.0-20.8%) were T. gondii positive and three (2.9%, CI 0.6-8.4%) for N. caninum. Association between: localities of obtaining samples (parks x CCZ) and sex of animals, with the occurrence of antibodies against each of the parasites, was determined by the Fisher exact test. For T. gondii association was found with males (p = 0.027) and dogs living close to parks (p = 0.008) had higher rates of infection. Associations were not observed in relation to N. caninum.

A ocorrência de anticorpos anti-Toxoplasma gondii e anti-Neospora caninum foi determinada em cães do município de Natal, RN, onde informações sobre a presença destes coccídios nessa espécie não era conhecida. Para tanto, foram utilizados 29 cães domiciliados, que habitam áreas que circundam duas importantes Unidades de Conservação de Mata Atlântica presentes no município (Parque da Cidade e Parque das Dunas), e outros 73 cães que foram sacrificados no Centro de Controle de Zoonoses de Natal por serem positivos a Leishmania spp. Somente em relação aos cães que vivem próximos aos parques foi possível a obtenção de informações sobre o sexo dos animais. A presença de anticorpos contra cada um dos coccídios foi determinada com a Reação de Imunofluorescência Indireta com ponto de corte de 16 para T. gondii e 50 para N. caninum. Dos 102 cães examinados, 13 (12,7%, IC 95% 7,0-20,8) foram positivos para T. gondii e 3 (2,9%, IC 0,6-8,4%) para N. caninum. Associação entre: localidades de obtenção das amostras (parques x CCZ) e sexo dos animais com a ocorrência de anticorpos contra cada um dos parasitos, determinada através do teste exato de Fisher, foram positivas para T. gondii, com os machos (p = 0,027) e cães que habitam próximos aos parques (p = 0,008) apresentando maiores taxas de infecção. Associações não foram observadas em relação a N. caninum.

Predictive qualitative risk model of bovine rabies occurrence in Brazil.

Bovine rabies remains endemic in Brazil and despite control efforts, the disease still spreads insidiously. The main vector is the hematophagous bat, Desmodus rotundus. The present work aimed to create a predictive qualitative model of the occurrence of bovine rabies in each municipality in 25 of the 27 Brazilian States. The risk of rabies transmission from bats to bovine was estimated using decision-tree models of receptivity and vulnerability. Questionnaires, which covered a number of questions related to the surveillance of possible risk factors, such as bovine rabies outbreaks in the previous year, the presence of bat roosts, bat rabies positivity and environmental changes, were sent to the local veterinary units of each State. The bovine density and geomorphologic features were obtained from national databases and geographic information systems. Of the 433 municipalities presenting bovine rabies outbreaks in 2010, 178 (41.1%) were classified by the model as high risk, 212 (49.0%) were classified as moderate risk, 25 (5.8%) were classified as low risk, whereas the risk was undetermined in 18 municipalities (4.1%). An ROC curve was built to determine if the risk evaluated by the model could adequately discriminate between municipalities with and without rabies occurrence in future years. The risk estimator for the year 2011 was classified as moderately accurate. In the future, these models could allow the targeting of rabies control efforts, with the adoption of control measures directed to the higher risk locations and the optimization of the field veterinary staff deployment throughout the country. Additionally, efforts must be made to encourage continuous surveillance of risk factors.