Molecular evidence of bovine herpesvirus 1 and 5 in cattle with suspected rabies in Rio Grande do Sul state,Brazil

Rabies and herpetic encephalitis are the main viral infections in bovines with neurological symptoms. Bovine rabies has a high prevalence in Central and South America, while bovine encephalitis associated with herpesvirus is especially important in South America. Viral isolation is the classical way to confirm herpesvirus infection, but molecular evidence of the presence of the virus in affected animals is gaining importance in the diagnosis of the disease in the laboratory. This study investigated the presence of herpesvirus type 1 and 5 (BoHV-1 and BoHV-5) in 182 encephalon of rabies-suspected cattle in Rio Grande do Sul state (RS), Brazil using multiplex real-time polymerase chain reaction (mRT-PCR). The rabies virus was investigated by direct fluorescent antibody assay and intracerebral suckling mouse inoculation. The genomes of BoHV-1 and BoHV-5 were detected in 17% of samples. BoHV-5 and BoHV-1 were detected in 100% and 19% of BoHV positive samples, respectively, indicating the circulation of the pathogens in cattle herds in RS. The high Ct values and the absence of isolation suggest viral latency. Coinfection of herpesvirus and the rabies virus was detected in 28% of samples, although no significant association between pathogens was observed. Rabies was detected in 57.7% of suspected samples, confirming the importance of the disease in the state. Concerning the method by which samples were conserved, no significant difference was observed between the number of positive results in frozen and refrigerated samples.     

Cryptosporidiosis in Horses of Urban Areas of Porto Alegre,Rio Grande do Sul,Southern Brazil

The aim of the present study was to detect the presence of Cryptosporidium spp. oocysts in the feces of horses used in military training and in horseback riding activities. A total of 90 fecal samples were collected from three sites in the city of Porto Alegre, state of Rio Grande do Sul, Brazil as follows: 71 samples were obtained from the Brazilian Army, seven from the hospital accredited with the School of Veterinary Medicine of Universidade Federal do Rio Grande do Sul, and 12 from a facility offering animal rest and training. All samples were collected between the months of January and February, 2008. They were screened microscopically for oocysts using the modified Ziehl–Neelsen staining technique. Oocysts were classified on the basis of their microscopic characteristics. The overall infection rate of horses as determined by microscopic analysis was 27.87% (25/90) and was identified only in horses raised in the Brazilian Army, with a rate of 35.21% (25/71). The morphometric analysis of oocysts revealed a mean size of 4.78 × 4.0 μm and a shape index of 1.16. None of the infected horses appeared to be clinically ill.     

Mammomonogamus laryngeus (Railliet, 1899) infection in buffaloes in Rio Grande do Sul, Brazil

Parasites of the genus Mammomonogamus affect the respiratory tract of domestic animals. The present study was carried out to determine the presence of Mammomonogamus laryngeus infection and to analyze its lesions in infected buffaloes in Rio Grande do Sul, Brazil, between April and November 1999. The infection rate was 30.5%. In 32 infected buffaloes, the worm pairs collected per animal did not exceed 20. The microscopic diagnosis showed intense polypoid to intramucosal proliferation at the entrance to the pharynx, to which the parasites had adhered, with foci of multifocal hydropic degeneration of the epithelium or individual degeneration of epithelial cells with mild intraepithelial inflammatory infiltrate. The submucosa revealed intense lymphocyte infiltrate extending into the salivary glands. The submucosa also showed formation of structures that resemble lymphoid follicles.     

Phenotypic and genetic characterization of Salmonella enterica subsp. enterica serovar Typhimurium isolated from pigs in Rio Grande do Sul,Brazil

This study aimed to investigate the relatedness of porcine Salmonella enterica subsp. enterica (S.) serovar Typhimurium strains isolated in Southern Brazil. Sixty-six isolates from pigs belonging to three commercial companies were submitted to phage typing, XbaI-macrorestriction (PFGE), IS200 hybridization, rep-PCR, antimicrobial susceptibility testing, and PCR assay targeting the spvR region. All strains presented a unique rep-PCR pattern and 63 strains had a common IS200 profile. One pulse-type (XA) was the most prevalent (39/66 strains) and included strains of phage types DT177, DT192, DT194 and RDNC. The spvR region was detected in three strains, which harboured plasmids of 90 kb. High rates of tetracycline, sulfonamide and streptomycin resistance were found. Isolates from farms located in different geographic regions but associated to the same commercial companies clustered together and presented a common resistance profile. Results suggested that clonal groups of S. Typhimurium are present in pig commercial companies in Southern Brazil.     

Phenotypic and genetic characterization of Salmonella enterica subsp. enterica serovar Typhimurium isolated from pigs in Rio Grande do Sul, Brazil

This study aimed to investigate the relatedness of porcine Salmonella enterica subsp. enterica (S.) serovar Typhimurium strains isolated in Southern Brazil. Sixty-six isolates from pigs belonging to three commercial companies were submitted to phage typing, XbaI-macrorestriction (PFGE), IS200 hybridization, rep-PCR, antimicrobial susceptibility testing, and PCR assay targeting the spvR region. All strains presented a unique rep-PCR pattern and 63 strains had a common IS200 profile. One pulse-type (XA) was the most prevalent (39/66 strains) and included strains of phage types DT177, DT192, DT194 and RDNC. The spvR region was detected in three strains, which harboured plasmids of 90 kb. High rates of tetracycline, sulfonamide and streptomycin resistance were found. Isolates from farms located in different geographic regions but associated to the same commercial companies clustered together and presented a common resistance profile. Results suggested that clonal groups of S. Typhimurium are present in pig commercial companies in Southern Brazil.     

Immunological Profile of the Yellow Clam Mesodesma mactroides (Mesodesmatidae) from the Southern Coast of Rio Grande do Sul,Brazil

The yellow clam Mesodesma mactroides (Mesodesmatidae) is a sandy beach bivalve that is distributed from Rio de Janeiro, Brazil, to the south of Buenos Aires province, Argentina. The yellow clam population has been declining in recent decades. To increase our understanding of this species, we evaluated the immunological status of yellow clams collected during different seasons from various areas in the state of Rio Grande do Sul, Brazil. We characterized the hemocytes, determined the differential hemocyte counts (DHCs), calculated the apoptotic index, and evaluated the incidence of parasites in yellow clams through histological analysis. We identified two types of hemocyte (hyaline and granular) that showed significant variation in DHCs among sampling areas during the summer and winter. The apoptotic index only exhibited significant variation during the summer. Histopathological analysis results did not significantly differ among sampling areas. This work demonstrated that environmental variation (e.g., temperature and salinity) associated with anthropogenic actions may be affecting the immune system of yellow clams. However, more studies are needed to determine the full influence of these factors on the yellow clam’s immune system and thus contribute to future management and aquaculture of the species.

Received May 10, 2015; accepted October 28, 2015.     

Neospora-like protozoan infection as a major cause of abortion in California dairy cattle

Ninety-five aborted bovine fetuses received from California dairies over a 4.5-year period had histologic lesions of focal encephalitis. Protozoa that reacted with Neospora caninum antiserum were detected in the brain of 88 of these fetuses and in the heart of 1 fetus. Sarcocystis spp schizonts were seen in the vascular endothelium of 1 fetus. It was concluded that a Neospora-like cyst-forming coccidian may be a major cause of abortion in California dairy cattle.     

Feline Sporotrichosis in the Southern Region of Rio Grande Do Sul,Brazil: Clinical,Zoonotic and Therapeutic Aspects

Clinical cases of feline sporotrichosis, originating in the Pelotas region and diagnosed at the Laboratory of Infectious Diseases (UFPel), were studied in the period from 2002 to 2006. The animals were evaluated according to the clinical forms of the mycosis, time of lesion appearance, severity of the clinical diagnosis and evolution of cutaneous lesions throughout the treatment period. Mycological analyses, carried out through direct examination, cultivation of tissue samples and exudates of feline lesions all confirmed the diagnosis of sporotrichosis in the 15 animals under study. The cutaneous dissemination form was observed in 10 animals, of which three showed prostration, anorexia and dehydration. The zoonosis occurred in 20% of case studies, and the pet owners and one attendant at a veterinary clinic were infected, developing the fixed and disseminated cutaneous forms. The treatment of mycosis was carried out with itraconazole, 10 mg kg^?1, once a day, on 12 animals. The cure of the clinical symptoms was observed on 50% of the felines. This study demonstrates a good clinical response of felines with sporotrichosis, when they were treated itraconazole and calls the attention for the incidence of human sporotrichosis on people related to the veterinary activity as well as for pet owners.     

Seroprevalence of Neospora caninum in dairy cattle in Bahia, Brazil.

Sera collected from 447 dairy cattle on 14 dairy farms were tested for Neospora caninum antibodies by use of an immunofluorescent antibody technique. Positive reactions with titres > or =1:200 were found in 63 (14.09%) of animals. Neospora positive sera were also tested for Toxoplasma gondii antibodies by using a commercial latex agglutination test. Antibodies to T. gondii were detected in 3 (4.76%) of 63 N. caninum positive sera. These results indicate that N. caninum infection is widespread among dairy cattle in Bahia state.     

Occurrence of Neospora caninum in dogs and its correlation with visceral leishmaniasis in the urban area of Campo Grande, Mato Grosso do Sul, Brazil

Neospora caninum is an obligate intracellular protozoan that can infect domestic and wild canids, as well as ruminants and equines, and is described as causing neuromuscular alteration and death in dogs. Visceral leishmaniasis (VL) is an infectious disease that affects both humans and animals, being caused by protozoan parasites of the genus Leishmania, of which Leishmania (Leishmania) chagasi is found in Brazil -- transmitted by sand flies, such as Lutzomyia longipalpis, in most of the American continent. The immunosuppression caused by VL can promote the occurrence of co-infections with other agents. In order to determine the frequency of N. caninum and its relationship to VL in Campo Grande, MS, Brazil, 345 blood sera were collected from dogs. The sera were submitted to an indirect fluorescent antibody test (IFAT) for detection of anti-N. caninum antibodies and VL antibodies. N. caninum was found in 26.5% of VL-negative dogs and in 29% of VL-positive ones. Among males, it was found in 30.7% of VL-negative animals and in 30.4% of VL-positive ones; among females, in 21% of VL-negative animals and in 27.7% of VL-positive ones. Among juvenile dogs (under 1 year), N. caninum was detected in 10.5% of VL-negative animals and in 11.2% of VL-positive ones. For adult dogs (1 year and older) the results were 31.4% for VL-negative animals and 28.8% of VL-positive ones. The study revealed a statistically significant association with age (chi(2)=9.76, P<0.05) in the N. caninum results for VL-negative animals. No significant correlation in N. caninum seroprevalence was found when VL-positive or VL-negative dogs were compared (chi(2)=0.21, P=0.64). The findings suggest that in Campo Grande N. caninum and VL co-infection is common in dogs, though VL does not appear to enhance susceptibility to N. caninum.     

Incidence of leptospiral abortion in Brazilian dairy cattle

We examined dams' paired serum samples and foetal kidneys (histopathologically and attempting leptospiral cultures) from 120 Brazilian Holstein abortions from 10 herds near Sao Paulo, Brazil, in 1996. Leptospiras was isolated from 15 foetuses. The Leptospira interrogans serovar hardjo was obtained from four, pomona from three and wolffi from eight foetuses. Ten of these 15 foetuses showed a four-fold rise in titres between the day of abortion and the second samples taken 15 days later. Fifty-seven other foetuses had histologic evidence of leptospires; 27 of their dams (47%) had four-fold rises in titres. In total, 72 of 120 aborted foetuses had evidence of leptospiral infection.     

Prevalence of anti-Toxoplasma gondii and anti-Neospora caninum antibodies in sheep from Mossoró, Rio Grande do Norte, Brazil

Toxoplasma gondii is an obligate intracellular parasite with a variety of hosts, responsible for reproductive problems and economic losses in sheep flocks. Neospora caninum was recently identified and its clinical presentation in sheep is similar to that of toxoplasmosis, which can cause repeated abortions, though less frequently in this species. In order to confirm the prevalence of these agents in the city of Mossoró, Rio Grande do Norte, Brazil, 409 serum samples from adult sheep (364 females and 45 males) were tested by the indirect immunofluorescence antibody test, using cut-off point at a dilution of 1:64 and 1:50 for T. gondii and N. caninum, respectively. From the 35 properties examined, 23 (65.7%) had at least one seropositive animal for T. gondii and six (17.1%) for N. caninum. The prevalence of seropositive animals for T. gondii was 20.7% and for N. caninum 1.8%. There was no association between the presence of the agent’s antibody and gender, reports of reproductive problems and presence of dogs and/or cats in the properties. T. gondii is well distributed and N. caninum has low prevalence in sheep and in the properties of the studied region.     

Ivermectin and moxidectin resistance characterization by larval migration inhibition test in field isolates of Cooperia spp. in beef cattle,Mato Grosso do Sul,Brazil

Ivermectin (IVM) resistance of Cooperia spp. in cattle has become an increasing and global problem. The early detection of anthelmintic resistance (AR) is important to propose strategies to slow down the development of resistance and requires sensitive, reliable, economic high-throughput and practical tests. The purpose of the present study was to apply a larval migration inhibition test (LMIT) for evaluating IVM and MOX efficacy against well-characterized field isolates of Cooperia spp. infecting cattle in Brazil. Eight isolates were used for IVM and seven for MOX. The following EC50 values of IVM were observed for the isolates: susceptible, 1.16 ηmol; Nova Alvorada do Sul I, 4.09 ηmol (RF = 3.52); Campo Grande BNA, 3.57 ηmol (RF = 3.07); Campo Grande TBR, 4.09 ηmol (RF = 3,52); Nova Alvorada do Sul II, 2.50 ηmol (RF = 2.15); Bandeirantes, 11.35 ηmol (RF = 9.78); Campo Grande II, 6.03 ηmol (RF = 5.20); and Porto Mortinho, 8.63 ηmol (RF = 7.44). For MOX, the following EC50 values were observed: susceptible, 0.75 ηmol; Campo Grande BNA, 0.93 ηmol (RF = 1.24); Campo Grande TBR, 0.36 ηmol (RF = 0.48); Nova Alvorada do Sul II, 2.57 ηmol (RF = 3.42); Bandeirantes, 1.43 ηmol (RF = 1.90); Campo Grande II, 1.08 ηmol (RF = 1.44); and Porto Mortinho, 0.49 ηmol (RF = 0.65). The LMIT used in the present study can be a useful tool for in vitro evaluation of IVM, but not of MOX. However, such methodology cannot be used in large-scale studies yet. The isolates of Cooperia spp. showed various degrees of resistance to IVM, though remaining susceptible to MOX.     

Shiga toxin-producing Escherichia coli (STEC) isolated from healthy dairy cattle in southern Brazil

Over a period of 1 year, the production of verotoxin was investigated in 1127 Escherichia coli isolated from 243 dairy cattle from 60 small farms in southern Brazil. Vero cell assay was used to detect toxins in culture supernatants from E. coli isolated from bovine feces. Shiga toxin-producing E. coli (STEC) detection rates were 95% (57 of 60) for farms and 49% (119 of 243) for cattle. Prevalence of STEC-positive cattle in the farms ranged from 0 to 100%. Ninety-six percent (315 of 327) of the STEC isolates did not react in the panel of sera used for typing. Twelve isolates, all non-motile, belonged to serogroups previously associated with human diseases, and 67% (8 of 12) were of only two serotypes (O91:H- and sorbitol-fermenting O157:H-). These results indicate that dairy cattle from the region surveyed may be a source of STEC potentially pathogenic for humans.     

Bluetongue virus as a cause of hydranencephaly in cattle.

Hydranencephaly was produced in a foetus and a calf by intra-uterine infection with an attenuated Type 10 bluetongue virus. Laparotomy was performed on the respective dams and the foetuses, respectively 126 days and 138 days old, were inoculated intramuscularly through the uterine wall with 1 ml of a virus suspension containing 5 x 103 tissue culture infective doese. The younger feotus was aborted on Day 262, while the other one was born alive on Day 273. Both foetuses showed marked hydranencephaly.