Prevalence of Ehrlichia canis infection in thrombocytopenic dogs from Rio de Janeiro, Brazil

BACKGROUND: Infection with Ehrlichia canis causes a highly variable, multisystemic disease in dogs. Nevertheless, many clinicians in Rio de Janeiro, Brazil, use the presence of only thrombocytopenia to make a presumptive diagnosis of E canis infection. OBJECTIVE: The objective of this study was to determine the prevalence of E canis in thrombocytopenic dogs from Rio de Janeiro, Brazil, using polymerase chain reaction (PCR). METHODS: Following DNA extraction of whole blood samples from 226 dogs, PCR assays were done using primers for rickettsial DNA (including Ehrlichia spp, Anaplasma platys and A phagocytophilum) and using E canis-specific primers (16S rRNA gene). Dogs were grouped as thrombocytopenic and nonthrombocytopenic based on platelet counts. The null hypothesis that there was no difference in the prevalence of E canis in these groups was rejected at P<.05. RESULTS: Thirty-six (32.1%) of the thrombocytopenic dogs and 4 (3.5%) of the nonthrombocytopenic dogs were positive for rickettsial gene sequences (P<.0001). Further, 30 (26.8%) of thrombocytopenic dogs and 4 (3.5%) nonthrombocytopenic dogs were positive for E canis-specific gene sequences (P<.0001). CONCLUSIONS: Although the prevalence of E canis infection was higher in thrombocytopenic dogs, less than one third of these dogs had demonstrable E canis infection. Thus, thrombocytopenia is not specific for the detection of E canis infection and should not be used solely to establish a diagnosis of canine ehrlichiosis, even in a geographic area with relatively high disease prevalence.     

Genomic Typing of Canine Parvovirus Circulating in the State of Rio de Janeiro,Brazil from 1995 to 2001 Using Polymerase Chain Reaction Assay

In this study, the genomic types of canine parvovirus (CPV) circulating in the State of Rio de Janeiro, Brazil, from 1995 to 2001, were investigated using the polymerase chain reaction assay (PCR). A total of 78 faecal samples from gastroenteritic puppies, confirmed as positive for canine parvovirus by haemagglutination/haemagglutination inhibition tests or virus isolation in cell culture (MDCK), were examined. The viral DNA was extracted from faecal samples using a combination of phenol– chloroform and silica–guanidine thiocyanate methods. PCR was carried out with differential pairs of primers to distinguish the old (CPV-2) and new types of virus (CPv-2a or CPV-2b). Specific amplicons were observed for all samples using the primer pair P2ab, which detects CPV-2a and CPV-2b. Seventy-six from a total of 78 samples (97%) were considered as CPV-2b because of their reaction with the primer pair P2b. Thirty samples (30/78) were from previously vaccinated puppies and in 15 of them the enteritis symptoms began from 1 to 12 days after vaccination. PCR confirmed the infection by wild virus (CPV-2b) in 5 of these 15 puppies who had received old-type vaccines. Our results show that CPV-2b was the prevalent type circulating in the State of Rio de Janeiro from 1995 to 2001.     

Factors associated with gastrointestinal parasite infection in dogs in Rio de Janeiro, Brazil

Factors associated with parasitism by helminths and protozoans in 500 dogs presented to three veterinary clinics in the municipality of Rio de Janeiro from November 2003 to September 2004 were evaluated. Dogs were submitted to physical examination and owners were interviewed about the animal's management. One fecal sample from each dog was examined by centrifugal flotation and sedimentation methods followed by the safranin–methylene blue staining technique. Positive results for gastrointestinal parasites were detected in 46.4% of the examined samples. Infection with protozoans (29.6%) was more frequent than with helminths (23.2%). Cryptosporidium sp. (26.2%) and Ancylostoma sp. (15.2%) were the most frequent parasites. Logistic regression analysis showed that age (p < 0.001), access to soil (p < 0.001), hygiene of the environment (p = 0.001), illness (p = 0.007), owner's level of education (p < 0.006) and veterinary clinic (p = 0.043) were associated with gastrointestinal parasite infections in dogs. Treatment and control are especially important for puppies. Adult dogs should be submitted to fecal examination before treatment, placing special emphasis on those that present one or more factors associated with infection.     

Cataracts are not associated with retinal detachment in the Bichon Frise in the UK--a retrospective study of preoperative findings and outcomes in 40 eyes

Objective The aim of this study was to evaluate whether the Bichon Frise population in the UK is at the same risk of developing retinal detachment in association with cataract formation and following phacoemulsification as described in reports from the USA. Procedures The medical records of Bichon Frises which were presented for cataract assessment and of those which were treated with phacoemulsification at Willows Referral Service between 1997 and 2009 were reviewed. Results Forty eyes (26 dogs) with unilateral or bilateral cataracts were included in the study. There was no evidence of retinal detachment associated with the cataracts at initial presentation. Phacoemulsification was performed on 34 eyes (20 dogs). Clinically evident lens‐induced uveitis was treated preoperatively in 17/34 eyes. Artificial lens implantation was carried out in 30/34 eyes; automated anterior vitrectomy was performed in 7/34 eyes. The mean follow‐up time was 16.6 months (range 1.5–73 months). At the last re‐examination, 31/34 eyes (91.2%) were visual. Three eyes (8.8%) were blind – two (in the same dog) because of presumptive bilateral optic nerve disease and one because of uveitis and secondary glaucoma. There was no evidence of retinal detachment following phacoemulsification in any of the 34 eyes. Conclusion This study suggests that the Bichon Frise population in the UK does not appear to have a predisposition for retinal detachment in association with cataract formation or following cataract surgery. Prophylactic random transscleral laser retinopexy or transscleral cryopexy cannot therefore be routinely recommended for Bichon Frises with cataracts in the UK.     

Factors associated with bovine leptospirosis in Rio de Janeiro, Brazil

Serum samples from 379 adult cows with lowered fertility farmed in the Rio de Janeiro region of Brazil. They were examined for Leptospira antibodies. A questionnaire was completed for each herd. Antibodies were detected in 46.9% of these cows, mainly to serovar hardjo. Main risk factor associated with seropositivity was co-grazing with other species, mainly pigs (OR = 3.17; p<0.04). Absence of or infrequent veterinary assistance (OR = 1.74) has also been suggested to be associated to the overall seroprevalence to leptospirosis. We suggest that a successful control programme for bovine leptospirosis should include a complete investigation of herd management practices, which could influence in the occurrence of the infection.     

Sporotrichosis: the main differential diagnosis with tegumentary leishmaniosis in dogs from Rio de Janeiro, Brazil

Seventy-four dogs from the State of Rio de Janeiro with ulcerated cutaneous lesions were submitted to clinical, dermatological, parasitological, mycological, histopathological and cytopathological exams, a leishmanin skin test, an indirect immunofluorescence (IIF) test for leishmaniosis, and nonspecific laboratory tests such as blood count and serum biochemistry. Sporothrix schenckii was isolated from 41 dogs and Leishmania (Viannia) braziliensis was isolated from 33 animals. Most dogs with sporotrichosis were from the municipality of Rio de Janeiro (53.7%) and presented ulcerated cutaneous lesions on the head (68.3%). Laboratory alterations in these animals included anemia (58.5%), hypoalbuminemia (83%) and hyperglobulinemia (75.6%). Histopathology revealed the predominance of a chronic granulomatous inflammatory infiltrate (70.7%), and yeast-like structures were detected in 17% of the histopathological exams and in 32% of the cytological exams. Three of 41 dogs with sporotrichosis were seropositive by IIF for leishmaniosis and 2 of 20 animals tested within this group had a positive leishmanin skin test. Similarly, most of the 33 dogs with leishmaniosis were from the municipality of Rio de Janeiro (69.7%) and had ulcerated cutaneous lesions on the head (84.8%). Laboratory alterations in these animals included anemia (66.7%), hypoalbuminemia (100%) and hyperglobulinemia (91%). Histopathology showed the predominance of a chronic granulomatous inflammatory infiltrate (63.6%) and amastigote forms were detected in 30.3% of the histopathological exams and in 31.8% of the 22 cytological exams performed. About 72.7% of the dogs were seropositive by IIF and five of seven animals had a positive skin test. Due to the clinical similarities, histopathological and nonspecific laboratory results similarities, the serological and skin tests for leishmaniosis positive in dogs with sporotrichosis, and the overlapping endemic areas in Rio de Janeiro, the differential diagnosis between the two diseases requires the demonstration of their respective etiological agents.     

Leptospirosis antibodies in mammals from Rio de Janeiro Zoo,Brazil

Serum samples from 77 animals belonging to 38 species and 19 families in Rio de Janeiro Zoo, Brazil were tested for antibodies against serovars of Leptospira interrogans by microscopic agglutination test. Antibodies were detected in serum samples of 37.7% of all animals belonging to 10 families. Seropositivity was more common in the Carnivora Canidae (7/9), Procyonidae (5/9) and the Edentata Myrmecophagidae (5/9). Icterohaemorrhagiae was the most common serogroup. Seropositivity was less common in primates.     

Identification of mutations in HSF4 in dogs of three different breeds with hereditary cataracts

Cataracts are a leading cause of blindness in both dogs and humans. Mutations in several genes have been associated with inherited forms of human cataract, but no mutations have been identified as the cause of any form of canine inherited cataract. We have used a candidate gene approach to investigate 20 genes, known to be associated with cataract in humans, for their potential association with the development of hereditary cataract (HC) in dogs. We have identified mutations in the HSF4 gene in Staffordshire Bull Terriers, Boston Terriers and Australian Shepherds affected by HC. Interestingly, different mutations in this single gene may be causing a recessive form of cataract in Staffordshire Bull Terriers and Boston Terriers and a dominant cataract in Australian Shepherds. Identification of the mutations that cause HC in these three breeds provides a method of controlling the disease within populations at risk using a simple diagnostic test, and also establishes cataract in these breeds as models for their human counterparts.     

Frequencies of feline blood types in the Rio de Janeiro area of Brazil

Background: The distribution and frequency of blood types in cat populations vary according to geographic region and breed. Frequencies of feline blood types in Rio de Janeiro city, as well as in other Brazilian areas, are unknown, and the risk of unmatched transfusions and neonatal isoerythrolysis has not been estimated. Objectives: The purpose of this study was to determine the frequency of feline blood types in the area of Rio de Janeiro, Brazil. Methods: EDTA blood samples were obtained from 172 nonpedigreed domestic shorthair (DSH) cats (92 female, 80 male, 3 months-20 years old) in different sites of Rio de Janeiro city. Blood typing was performed by agglutination assays using Triticum vulgaris lectin and feline anti-A serum. The hemagglutination results for type B and AB cats were confirmed by high-performance thin-layer chromatography (HPTLC) of erythrocyte membrane gangliosides. Results: The majority (163/172, 94.8%) of cats were type A, 2.9% were type B, and 2.3% were type AB. High-titer anti-A serum agglutinated RBCs from all cats in type A and type AB blood groups, with 3+ to 4+ agglutination. The probability that a type A cat would receive type B or AB blood in a first random transfusion was calculated as 2.25% and 2.20%, respectively. HPTLC analysis of glycolipids yielded a chromatographic profile characteristic of feline gangliosides for all blood groups. Conclusions: These results indicate a high prevalence of type A cats in Rio de Janeiro, Brazil, and a low frequency of type B and AB cats, consistent with what has been observed for DSH cats in other regions of the world.     

Occurrence and risk factors associated to Toxoplasma gondii infection in sheep from Rio de Janeiro,Brazil

Toxoplasmosis is an important cause of abortion in sheep and a zoonotic risk to humans, leading to significant hazards to health and to economic losses. This study examined the seroprevalence and associated risk factors for infection with Toxoplasma gondii in 379 sheep from 12 flocks in Rio de Janeiro state, Brazil. Using the modified agglutination test (MAT), 202 (53.3 %) of 379 were seropositive with titers of 1:25 in 65, 1:50 in 40, 1:100 in 23, 1:200 in 11, 1:400 in 36, 1:800 in 7, 1:1,600 in 1, and 1:3,200 or higher in 19 sheep. The most significant factors associated with T. gondii seropositivity in sheep were age, gender, and veterinary care. Finding of T. gondii antibodies in 97 of the 202 sheep in titers of 1:100 or higher is indicative of persistently infected animals. This high level of seropositivity requires urgent control measures to reduce impact on animal productivity and public health.     

Detection of anti-lens crystallin antibody in dogs with and without cataracts

Objective To determine if antilens crystallin (ALC) serum and aqueous humor antibodies were present in normal dogs and dogs with cataracts, whether antibody incidence varied with stage of cataract, and whether antibody titer had a relationship to the presence of lens‐induced uveitis. Methods Serum and aqueous humor samples were obtained from normal dogs and dogs with cataracts. Lens crystallin was separated by SDS‐polyacrylamide gel electrophoresis (SDS‐PAGE), and antilens crystallin antibodies were detected by Western immunoblot analysis. An indirect ELISA using crystallin protein as antigen was also used to detect antilens crystallin antibodies in serum and aqueous humor. Test groups included normal, incipient, immature, mature, hypermature and diabetic cataract. Results SDS‐PAGE identified bands with molecular weights of lens crystallin subunits. Western immunoblotting demonstrated reaction between canine serum and these protein bands. The five canine serum samples that reacted with crystallin subunits on Western blots had corresponding reactivity on the ELISA. All aqueous humor samples (30) were negative. Serum ALC antibodies were detected in 59.3% (16/27) of controls, 66.7% (16/24) of incipients, 50.0% (10/20) of immatures, 37.9% (11/29) of matures, 28.6% (6/21) of hypermatures, and 26.7% (4/15) of diabetics. Serum ALC antibodies were detected in 43.1% (47/109) of all cataract samples. There was a statistically significant negative association between the presence (P = 0.004) and maturity (P = 0.004) of cataract and presence of ALC serum antibodies. In the immature and hypermature cataract groups, there was a statistically significant negative association between ALC serum antibody titer and severity of uveitis (95% confidence interval). Conclusions There is a negative association between the presence (P = 0.004) and maturity (P = 0.004) of cataract and presence of ALC serum antibodies.     

A serological study on Brucella abortus, caprine arthritis-encephalitis virus and Leptospira in dairy goats in Rio de Janeiro, Brazil

In spite of the large number of goats found in several developing tropical countries, milk production remains unsatisfactory. The occurrence of infectious diseases, such as leptospirosis, brucellosis and caprine arthritis-encephalitis (CAE) may in part be responsible for sub-optimal production. In this study, 1000 serum samples were tested for leptospirosis, 953 for brucellosis and 562 for CAE. All tested flocks presented at least one seroreactive animal for leptospirosis and for CAE. Reactivity to leptospirosis was 11.1%, and serovar hardjo was the most frequently found. Anti-B. abortus agglutinins were found in 0.5% of the samples presented and 14.1% were seroreactive to CAE. Leptospirosis was considered to represent the major infectious problem in the studied goat flocks. The occurrence of infectious diseases in the tested flocks may represent an important factor contributing to the decreased productivity of the animals. These findings may be similar to those observed in other developing countries and require further study to define the relationship between seropositivity and reduced production.     

Effect of three treatment protocols on acute ocular hypertension after phacoemulsification and aspiration of cataracts in dogs

Objective  To compare the effect of topical latanoprost, intracameral carbachol, or no adjunctive medical therapy on the development of acute postoperative hypertension (POH) and inflammation after routine phacoemulsification and aspiration (PA) of cataracts in dogs.
Design  Retrospective study.
Procedures  Dogs received either one drop of topical 0.005% latanoprost (21 dogs, 39 eyes), an intracameral injection of 0.3 mL of 0.01% carbachol (15 dogs, 30 eyes), or no adjunctive therapy (46 dogs, 90 eyes) immediately following PA of cataract(s). Intraocular pressure (IOP) was measured in all dogs 2 and 4 h after surgery. IOP was measured and aqueous flare assessed at 8 am the day after surgery.
Results  Carbachol-treated dogs had significantly higher mean IOP (33.2 ± SD 20.8 mmHg) 2 h after surgery than dogs receiving no adjunctive therapy (22.0 ± SD 14.1 mmHg) ( P  =  0 .049). There were no significant differences in IOP among groups at any other time point. There were no significant differences in number of POH episodes between dogs treated with carbachol (47%), latanoprost (29%), or dogs that received no adjunctive therapy (33%). There were no significant differences in mean aqueous flare grade between eyes treated with latanoprost (1.7 ± SD 0.4) or carbachol (1.4 ± SD 0.6), and eyes that received no adjunctive therapy (1.7 ± SD 0.4).
Conclusions  Topical 0.005% latanoprost or intracameral injection of 0.3 mL of 0.01% carbachol after PA in dogs did not reduce POH or increase intraocular inflammation compared to dogs not receiving adjunctive therapy after PA of cataracts.     

High occurrence of Shiga toxin-producing Escherichia coli (STEC) in healthy cattle in Rio de Janeiro State, Brazil

In order to evaluate the prevalence of Shiga toxin-producing Escherichia coli (STEC) strains, 197 fecal samples of healthy cattle from 10 dairy farms, four beef farms and one slaughterhouse at Rio de Janeiro State, Brazil, were examined for Shiga toxin (Stx) gene sequences by polymerase chain reaction (PCR). For presumptive isolation of O157:H7 E. coli, the Cefixime-potassium tellurite-sorbitol MacConkey Agar (CT-SMAC) was used. A high occurrence (71%) of Stx was detected, and was more frequently found among dairy cattle (82% vs. 53% in beef cattle), in which no differences were observed regarding the age of the animals. Dot blot hybridization with stx1 and stx2 probes revealed that the predominant STEC type was one that had the genes for both stx1 and stx2 in dairy cattle and one that had only the stx1 gene for beef cattle. Three (1.5%) O157:H7 E. coli strains were isolated from one beef and two dairy animals by the use of CT-SMAC. To our knowledge, this is the first report of O157:H7 isolation in Brazil. A PCR-based STEC detection protocol led to the isolation of STEC in 12 of 16 randomly selected PCR-positive stool samples. A total of 15 STEC strains belonging to 11 serotypes were isolated, and most of them (60%) had both stx1 and stx2 gene sequences. Cytotoxicity assays with HeLa and Vero cells revealed that all strains except two of serotype O157:H7 expressed Stx. The data point to the high prevalence of STEC in our environment and suggest the need for good control strategies for the prevention of contamination of animal products.     

Diversity of the infracommunities of strongylid nematodes in the ventral colon of Equus caballus from Rio de Janeiro state, Brazil

Nematodes from the ventral colon of 31 adult horses, 24 males and 9 females, in the metropolitan region of the state of Rio de Janeiro were analysed. There were 53,444 (86.4%) adults of the total recovered strongylid nematodes. They belonged to 21 species of Cyathostominae and seven of Strongylinae. Larval forms made up 13.6% (8407) of the total recovered, and 49% of the strongylid nematodes were observed in ventral colon. The most prevalent and abundant species were Cyathostomum tetracanthum, Cylicocyclus nassatus, Cylicostephanus minutus, Cylicostephanus longibursatus, Cylicostephanus leptostomus, Cylicostephanus calicatus and Cylicostephanus goldi, corresponding to 93.2% of the total adult population; these species were classified as central species, except C. goldi; and other species were classified as secondary (n = 8) and satellite (n = 14). The low Green's aggregation index, presented to central species, indicated high dispersion because they were found in a greater number of infracommunities and were proven by the greater prevalence. Only 12.1% of the horses were uninfected or infected by a single species; the other infections were multiple, ranging from 3 to 23 species.     

Natural infection by Cryptosporidium sp., Giardia sp. and Eimeria leuckarti in three groups of equines with different handlings in Rio de Janeiro, Brazil

To detect Cryptosporidium sp., Giardia sp. and Eimeria leuckarti in horses, fecal samples were collected from three different handling horse groups from the state of Rio de Janeiro, Brazil. Group A was composed of “Mangalarga Marchador” pure breed horses, Group B was formed by horses of a Military Corporation and Group C by stray horses captured by the Center of Zoonosis Control Paulo Dacorso Filho. A total of 396 fecal samples were collected, 212 samples from Group A, 154 samples from Group B and 30 from Group C. The material was submitted to the centrifugation - flotation technique and staining by the safranin-methylene blue technique and analyzed. Oocysts of Cryptosporidium sp. were identified in 0.75% of the samples (n = 3); cysts of Giardia sp. in 0.5% (n = 2) and oocysts of E. leuckarti in 0.5% (n = 2). One case of E. leuckarti in group A and one of Cryptosporidium sp. in group B were observed. In group C were observed two cases of Cryptosporidium, two of Giardia and one of E. leuckarti,. Horses of group C were more parasitized by the three protozoans than animals from the other groups (p < 0.01). It was possible to verify that factors related to the animals, like host individual susceptibility and sanitary factors may influence the occurrence of natural infections by gastrointestinal protozoans, although the age did not have influence. This study reports, for the first time, the occurrence of Cryptosporidium sp., Giardia sp. and E. leuckarti in equines of the State of Rio de Janeiro.     

Diversity of Campylobacter jejuni and Campylobacter coli Genotypes from Human and Animal Sources from Rio de Janeiro, Brazil

To compare the genotypes of Campylobacter jejuni and Campylobacter coli isolates of human and animal origin collected in Rio de Janeiro City, 30 C. jejuni and 35 C. coli isolates from animal sources (n = 45) and human patients with gastroenteritis (n = 20) were genotyped by PCR-based techniques, namely random amplified polymorphic DNA (RAPD-PCR) and enterobacterial repetitive intergenic consensus sequence (ERIC-PCR). RAPD-PCR identified 50 types and ERIC-PCR identified 22 genotypes, among the 65 Campylobacter isolates. Both PCR methods discriminated the C. jejuni and C. coli groups of isolates. Combining the results of both methods, no single genotype was shared between isolates from human and animal sources. Two groups of two C. coli isolates each with identical genotypes were found among poultry and pig isolates. A high level of genetic diversity observed among the Campylobacter isolates suggests lack of overlap between isolates from different sources.