Enzootiology of Trypanosoma evansi in Pantanal, Brazil

In order to better understand the enzootiology of trypanosomiasis caused by Trypanosoma evansi in the Brazilian Pantanal we examined domestic and wild mammals by microhematocrit centrifuge technique (MHCT), immunofluorescence antibody test (IFAT) and polymerase chain reaction (PCR). T. evansi infection was detected in all species sampled with exception of the sheep and the feral pig. High parasitemias were observed in capybaras (5/24), coatis (18/115), horses (31/321) and dogs (3/112). Among these species, only the capybaras did not develop anemia. Low parasitemias, only detected by PCR, were found in buffaloes (18/43), bovines (29/331), marsupials (1/4), small rodents (14/67), bats (7/18), and one armadillo (1/8). The highest prevalence of T. evansi infection was recorded in horses (73%), although no neurological signs in infected horses were observed. Diagnosis through standard parasitological tests and IFAT should be used with caution since they may overlook comprovedly infected horses. The relationship between ranch management and T. evansi infection in horse was investigated. The importance of other transmission mechanisms apart from the tabanids and reservoir hosts are discussed.     

Using PCR for unraveling the cryptic epizootiology of livestock trypanosomosis in the Pantanal, Brazil

Trypanosoma vivax and Trypanosoma evansi are livestock parasites of economic importance in Africa, Asia and South America. In the Pantanal, Brazil, they cause economic losses in both cattle and equines. Little is known of their maintenance and spread in nature, particularly in terms of reservoirs and means of mechanical transmission. Here we report for the first time the use of PCR for the detection of T. vivax and T. evansi in bovines, buffaloes and sheep. Whereas parasitological diagnosis detected only two T. vivax infections, one in buffalo and another in a cow, PCR detected infections in 34.8% buffaloes, 44.7% bovines and 37.3% sheep. Trypanozoon primers detected 41.8% infections in buffaloes and 8.1% in cattle. PCR revealed 6.9% mixed infections in buffaloes and 5.3% in cattle. The potential role of cattle and buffaloes as hosts and reservoirs of T. vivax is discussed, as well as the implications of possible extravascular foci in the maintenance of livestock trypanosomosis.     

Control of equine piroplasmosis in Brazil.

The importance of equine piroplasmosis control in endemic countries has increased in recent years and plays an important role to maintain the international market open to the horse industry. The purpose of this study was to demonstrate the occurrence of equine piroplasmosis (Babesia equi or Theileria equi and Babesia caballi) in Brazil--a country where the disease occurs endemically--in different climatic conditions, and to evaluate the results of a strategy for tick control in order to decrease infection rates. Blood samples were taken from 720 horses on 28 farms from different regions and subjected for complement fixation testing. The strategy was based on the control of the tick population by spraying the horses with acaricides, treating positive horses and preventing cattle and horses from grazing together. A significant association was found in the prevalence of antibody titres in tropical and subtropical areas. A significantly lower prevalence rate occurred on those farms where measures to control tick population were established. Farms in endemic countries may significantly reduce the prevalence of equine piroplasmosis by establishing measures to control the tick population and treating chronic carrier horses. Additional measures for controlling ticks in tropical areas are also discussed.     

Occurrence and genetic diversity of hemoplasmas in beef cattle from the Brazilian Pantanal,an endemic area for bovine trypanosomiasis in South America

Hemotropic mycoplasmas (hemoplasmas) are Gram-negative bacteria that parasitize the erythrocyte surface of a wide variety of mammals. The present study aimed at investigating the occurrence of hemoplasmas in beef cattle in the Brazilian Pantanal, an area endemic for bovine trypanosomiasis in South America. Additionally, the objective of this study was to characterize molecularly the genotypes of the found hemoplasmas. For this purpose, blood and serum samples of 400 beef cattle were collected from five properties in Corumbá, Nhecolândia sub-region, Mato Grosso do Sul, in Midwest Brazil. Blood samples underwent DNA extraction and standard 16S rRNA gene-based PCR assays for hemoplasmas. The sequences obtained were submitted to phylogenetic inferences, distance analysis, and genotype diversity. The Indirect Enzyme-Linked Immunoabsorbent Assay (iELISA) indicated the presence of anti-Trypanosoma vivax IgG antibodies in 89.75% of the animals sampled, confirming the endemicity of said agent in the studied region. Among the 400 bovine blood samples tested, 2.25% (9/400) were positive for hemoplasmas in cPCR. The phylogenetic analysis of the obtained sequences confirmed the presence of 'Candidatus Mycoplasma haemobos' and Mycoplasma wenyonii DNA in 0.5% (2/400) and 1.75% (7/400) animals, respectively. Five genotypes of M. wenyonii and one of 'Candidatus M. haemobos' were detected among the sequenced amplicons. The present study showed low molecular occurrence of haemoplasmas in beef cattle sampled in the Brazilian Pantanal, an area endemic for bovine trypanosomiasis. Despite of the conservation of the 16S rRNA gene, there was considerable diversity of hemoplasma genotypes infecting the sampled beef cattle.     

Genotyping of Toxoplasma gondii isolates from free range chickens in the Pantanal area of Brazil

The aim of this paper was to genetically characterize Toxoplasma gondii isolates from free range chickens in regions of Brazilian territory in the state of Mato Grosso do Sul (MS) where T. gondii strains have never been studied. In total, T. gondii isolates from 22 free range chickens were included in this study. Fifty chickens from Eldorado, thirty from Rio Verde and ten from Aquidauana were sampled between January and April 2007. In relation to the genetic diversity of T. gondii isolates from chickens in MS, the magnitude of the diversity in the isolates sampled in this study was comparable to the overall diversity in a composite data set. These 22 isolates in MS revealed 11 genotypes, whereas the 321 isolates ever genotyped in Brazil have revealed 95 genotypes. The values of Simpson's Diversity Index for the whole population of T. gondii isolates in Brazil, the whole population of T. gondii isolates from chickens in Brazil and the population surveyed in this study were 0.97, 0.95 and 0.90, respectively. Seven of the 11 genotypes revealed from chicken isolates from MS are newly described genotypes and six of them each have a single isolate. In conclusion, the results obtained from isolates in MS corroborate previous studies on T. gondii isolates in Brazil, thus confirming their diversity and atypicality. Nonetheless, the applicability of PCR-RFLP markers for epidemiological inferences remains controversial.     

Bovine trypanosomosis in the Bolivian Pantanal

Trypanosomosis caused by Trypanosoma vivax has been a constraint for cattle production in the Bolivian lowlands, since it was introduced in 1996. Flooded areas like the Bolivian Pantanal have a suitable environment for the presence and transmission of Salivarian trypanosomes and farmers from that region often report trypanosomosis-like problems on their farms. The objective of the present study, therefore, was to characterize the epidemiology of bovine trypanosomosis in the Bolivian Pantanal. In order to achieve this objective, 202 cattle from the province of Angel Sandoval and 209 cattle from the province of German Busch were randomly sampled (the Pantanal is located in both provinces). Twenty-nine farms in both provinces were visited, the farmers interviewed, and biologic samples collected from their cattle. Samples were submitted for parasitological and PCR evaluation and the prevalence of bovine trypanosomosis was estimated for each province. Laboratory results were correlated with the sampled animals packed cell volume (PCV) and body condition (BC) scores and the observed T. vivax parasites measured for morphometry analysis. Results from this study show differences in morphometric measures between T. vivax parasites from each province. Differences between provinces were also observed in the T. vivax-related disease situation. While in Angel Sandoval the PCV and BC of T. vivax-affected animals were significantly lower than those of the T. vivax-negative animals, in German Busch no differences were observed in the PCV and BC of T. vivax-positive or negative animals. Animal prevalence of T. vivax in Angel Sandoval was 27.79% (95% CI: 14.52-44.28) and in German Busch was 19.03% (95% CI: 9.19-30.75). The T. evansi animal prevalence in each province was 0.99% (95% CI: 0.27-2.99) and 5.71% (95% CI: 2.43-12.19), respectively. Based on questionnaire and laboratory results, it was concluded that trypanosomosis is a primary constraint for cattle production in the Bolivian Pantanal.     

甘肃省胡枝子属植物的遗传多样性分析

采用SSR分子标记技术,研究甘肃胡枝子属10个种的遗传多样性和遗传结构,并利用10对紫花苜蓿和4对山生柳SSR引物对胡枝子进行了通用性分析,筛选出7对重复性高、条带清晰的引物,获得105个SSR多态性位点,多态性达83％.引物平均等位基因数为1.91,平均有效等位基因数为1.51.各引物的Shannon指数的多态性平均...     

Hereditary equine regional dermal asthenia in three related Quarter horses in Brazil

Hereditary equine regional dermal asthenia belongs to a group of inherited, congenital connective tissue dysplasias usually described as hyperelastosis cutis, cutaneous asthenia, dermatosparaxis, or Ehlers-Danlos-like syndrome. This report presents the clinical and histological features of three related Quarter horses affected with regional dermal asthenia. These horses had bilateral asymmetric lesions of the trunk and lumbar regions, where the skin was hyperextensible. Handling of the skin elicited a painful response and superficial trauma led to skin wounds. The skin was thinner than normal in the affected areas, with thickened borders and harder fibrotic masses (pseudotumours). The histopathological findings included thinner and smaller collagen fibrils, and a loose arrangement of collagen fibres in the middle, adventitial and deep dermis. Masson's trichrome and Calleja stains did not reveal any abnormality of collagen and elastic fibres. Electron microscopy showed no abnormalities. As in human patients, pseudotumour histopathological findings included fibroplasia and neovascularization. The pedigree chart of these animals supports an autosomal recessive type of inheritance, which has been suggested by other studies. This is the first report of this disease in Brazil. Its clinical and histological features resemble those described in horses affected with this condition in the United States.     

Present status of the conservation of livestock genetic resources in Brazil

Brazil has various species of domestic animals, which developed from breeds brought by the Portuguese settlers soon after the discovery of the country. After being submitted to a long process of natural selection, these breeds present characteristics adapted to specific Brazilian environmental conditions, and are known as “criollo”, “local” or naturalized breeds. From the beginning of the 20th century, some exotic breeds, selected in temperate regions, have begun to be imported, and, although more productive, they do not have adaptive traits, such as resistance to disease and parasites found in the naturalized breeds. Even so, they gradually replaced the native breeds, to such an extent that the latter are in danger of extinction. To avoid further loss of this important genetic material, in 1983 the National Research Center for Genetic Resources and Biotechnology (Cenargen) of the Brazilian Agricultural Research Corporation (Embrapa) decided to include conservation of animal genetic resources among its priorities. The conservation activities have been conducted under the Brazilian Genetic Resources Network — Renargen, and is carried out by various research centres of Embrapa, universities and State research institutions, as well as by private farmers, with a single coordinator at national level, Cenargen. The in situ conservation of cattle, horses, buffaloes, donkeys, goats, sheep and pigs is being carried out by Conservation Nuclei, located in the animal's original habitat. Ex situ conservation is centred at the Brazilian Animal Germplasm Bank (AGB), kept at Cenargen. This is responsible for the storage of semen and embryos of various breeds of domestic animals threatened with extinction. Presently the AGB has almost 60,000 doses of semen and more than 250 embryos, as well as over 7000 DNA samples. An important challenge for this program is to increase awareness within the different segments of society in terms of the importance of the conservation of animal genetic resources. The development and evolution of Renargen and its efforts to facilitate the conservation of genetic resources of livestock in Brazil are described in this paper.     

Diagnosis of equine piroplasmosis in Brazil by serodiagnostic methods with recombinant antigens.

Serum samples from horses in the States of Sao Paulo and Mato Grosso do Sul, Brazil were examined for diagnosis of equine piroplasmosis by both the latex agglutination test (LAT) and enzyme-linked immunosorbent assay (ELISA) with recombinant antigens. Of the 47 samples analyzed, 38 (81%) and 42 (90%) samples were positive for B. equi infection and B. caballi infection, respectively. In addition, 35 (75%) samples were positive for both B. equi and B. caballi infections. These results indicate that equine piroplasmosis is widespread and therefore a cause for serious concern in the States of Sao Paulo and Mato Grosso do Sul, Brazil.     

Microsatellite analysis of genetic diversity in the Catalonian donkey breed

A total of 111 individuals (79 females and 32 males) of the endangered Catalonian donkey breed were analysed by using a commercial equine paternity polymerase chain reaction typing kit. Eleven of the 12 horse microsatellites were amplified when using donkey's DNA. One locus, ASB2, did not amplify in any sample. The allele range for HTG4 overlapped with HMS7 and their results could not be interpreted. The mean number of alleles detected per locus was 7.7 (±1.0), being the average of genetic diversity detected in the population, and measured as the unbiased average expected heterozygosity ( H _e), of 0.712 ± 0.038. All the analysed loci showed disagreement with Hardy–Weinberg proportions except HTG7 (from permutation tests). The within-population-inbreeding estimate was highly significant (p < 0.001) and equal to 15.4% (as measured by F _IS-statistic). The cumulative exclusion probability was 0.999. The deficit of heterozygotes can be partly explained by a population subdivision effect ( F _ST=3.5%), although the main factor that has provoked this lack of heterozygotes can be attributed to consanguinity.     

Prevalence of Leptospira interrogans antibodies in free-ranging Tayassu pecari of the Southern Pantanal, Brazil, an ecosystem where wildlife and cattle interact

We surveyed a wild population of white-lipped peccaries (Tayassu pecari) in the Brazilian Pantanal for evidence of Leptospira interrogans. Serum samples from 71 free-ranging T. pecari were obtained between 2003 and 2005 in the southern Pantanal of Mato Grosso do Sul state. We used microscopic microagglutination to test for antibodies against 14 L. interrogans serovars (antibody titers ≥1:100 were considered seropositive). Seventy percent of captured animals tested positive for leptospirosis antibodies. Antibodies against icterohaemorrhagiae and autumnalis serovars were the most prevalent. We used log-linear analyses to test for associations among seropositivity, age class, and sex of captured animals. Seropositivity was strongly associated with animal age class, but independent of sex. Forty-six percent of animals less than 2 years old, 63% of adults during peak reproductive years, and 100% of the oldest age class were seropositive. A nonparametric multivariate procedure (MRPP) showed that the composition of serovar antibody types changed with age, and ANOVA models demonstrated that antibody titers increased with age, suggesting long-term exposure to a greater number and variety (i.e., serovar types) of L. interrogans infections. This study presents the first quantitative survey of antibodies against L. interrogans serovars in a T. pecari population of the Pantanal. The high prevalence of leptospirosis antibodies in free-ranging white-lipped peccaries and the potential impacts on reproduction and population dynamics emphasize the need for further studies investigating the roles of Pantanal wildlife and livestock in the transmission and maintenance of L. interrogans in the environment.