Isolamento viral e diagnóstico molecular de herpevírus canino

Pós-graduação em Medicina Veterinária - FMVZ

Transtornos reprodutivos causados por agentes infecciosos em animais de canis comerciais da microrregião de Ribeirão Preto, estado de São Paulo

Pós-graduação em Medicina Veterinária - FCAV

Detecção de co-infecções por Leishmania (l.) chagasi, Trypanosoma evansi, Toxoplasma gondii e Neospora caninum em cães

Co-infections by Leishmania (L.) chagasi, Trypanosoma evansi, Toxoplasma gondii and Neospora caninum in dogs were investigated. Amastigotes forms of Leishmania spp. were detected by cytopathological analysis of lymph nodes in 46,42% (39/84) of dogs. In a male dog, adult, without defined breed, from rural area and positive for Leishmania, were observed flagellated forms of T. evansi in blood smear. By immunofluorescence antibody test, 5,95% (5/84) of dogs were considered reactive to T. gondii, with titer equal to or higher than 1:64, while 3,57% (3/84) were reactive to N. caninum, with titer ≥1:50. Among the animals with visceral leishmaniasis, one showed positive serological response to T. gondii and two for N. caninum. All dogs reactive to N. caninum were from rural area and the predominance of infection by T. gondii was in dogs from urban area. A young male dog from the rural area and seropositive for T. gondii showed Ehrlichia spp. morulae in the cytology and positive reaction for canine distemper virus. Thus, further studies are needed to assess the epidemiology of these infections in canine population, especially with respect to the reservoirs of Trypanosoma spp. in rural areas.

Participação de gambás e cães domiciliados como reservatórios de Leishmania infantum e Trypanosoma cruzi georreferenciados nos municípios da Regional de Saúde de Três Lagoas - MS

Pós-graduação em Doenças Tropicais - FMB

Retrospective evaluation of the effects of administration of tetrastarch (hydroxyethyl starch 130/0.4) on plasma creatinine concentration in dogs (2010-2013): 201 dogs

OBJECTIVE To determine changes in creatinine concentrations following the administration of 6% tetrastarch (hydroxyethyl starch [HES] 130/0.4) compared to crystalloids (CRYSs) in critically ill dogs. DESIGN Retrospective case series (2010-2013). SETTING University teaching hospital. ANIMALS Two hundred and one dogs admitted to the intensive care unit with initial plasma creatinine concentrations not exceeding laboratory reference intervals (52-117 μmol/L [0.6-1.3 mg/dL]) and receiving either CRYSs alone (CRYS group, n = 115) or HES with or without CRYSs (HES group, n = 86) for at least 24 hours. INTERVENTIONS None. MEASUREMENTS AND MAIN RESULTS Creatinine concentrations at admission to the intensive care unit (T0), and 2-13 days (T1) and 2-12 weeks (T2) after initiation of fluid therapy were analyzed. Creatinine concentrations were analyzed as absolute values and as the maximum percentage change from T0 to T1 (T1max%) and from T0 to T2 (T2max%), respectively. Creatinine concentrations were available for 192 dogs during T1 and 37 dogs during T2. The median cumulative dose of HES was 86 mL/kg (range, 12-336 mL/kg). No difference was detected between the groups for age, gender, body weight, and length of hospitalization. Outcome was significantly different between the HES (66% survived) and the CRYS (87% survived) groups (P = 0.014). No significant difference was detected between groups for creatinine concentrations at T0, T1, T2, T1max%, or T2max%. No significant difference was detected between the groups for T1max% creatinine in dogs subclassified as having systemic inflammatory response syndrome or sepsis. CONCLUSIONS HES administration in this canine population did not result in increased creatinine concentrations compared to administration of CRYSs. Further studies are needed to establish the safety of HES in critically ill dogs.

Canine neurological diseases in a referral hospital population between 1989 and 2000 in Switzerland

OBJECTIVES: To merge clinical information from partly overlapping medical record databases of the Small Animal Teaching Hospital of the Vetsuisse Faculty, University of Berne. To describe the frequencies and localisations of neurological diseases in dogs, as well as their age, gender, breed and geographical distributions. METHODS: In this retrospective study, a new database, with specific variables and a diagnosis key list 'VITAMIN D', was created and defined. A total of 4497 dogs (average of 375 per year) with a well-documented neurological disease were included in the study. A key list for the diagnoses was developed and applied to either the presumptive or the clinical and neurohistopathological diagnosis, with a serial number, a code for localisation and a code for differential diagnoses. RESULTS: Approximately 1159 dogs (26 per cent) had a neurohistopathological diagnosis confirmed, 1431 (32 per cent) had a clinical diagnosis confirmed and 1491 (33 per cent) had a presumptive diagnosis. The most frequent breeds were mixed-breed dogs (577 of 4497, 13 per cent), followed by German shepherd dogs (466 of 4497, 10 per cent). The most common localisations were the forebrain (908 of 4497, 20 per cent) and the spinal cord at the thoracolumbar area (840 of 4497, 19 per cent). Most dogs were diagnosed with degenerative diseases (38 per cent), followed by inflammatory/infectious diseases (14 per cent). The highest number of submissions originated from geographic regions around the referral hospital and from regions with higher human population densities. CLINICAL SIGNIFICANCE: By defining closed-list fields and allocating all data to the corresponding fields, a standardised database that can be used for further studies was generated. The analysis of this study gives examples of the possible uses of a standardised database.

Lateral incisor agenesis, canine impaction and characteristics of supernumerary teeth in a South European male population

Objective: To assess the prevalence of lateral incisor agenesis impacted canines and supernumerary teeth in a young adult male population. Materials and Methods: The panoramic radiographs of 1745 military students (mean age: 18.6 ± 0.52 years) who attended the Center of Aviation Medicine of the Armed Forces of Greece during the period 1997-2011 were initially analyzed for lateral incisor agenesis by two observers. After exclusion of the known orthodontic cases, a subgroup of 1636 examinees (mean age: 18.6 ± 0.44 years) was evaluated for canine impaction and supernumerary teeth. Results: Twenty-eight missing lateral incisors were observed in 22 military students, indicating an incidence of 1.3% in the investigated population. No lateral incisor agenesis was detected in the mandibular arch. A prevalence rate of 0.8% was determined for canine impaction in the sample of young adults. The majority of impacted teeth (86.7%) were diagnosed in the maxillary arch. Thirty-five supernumerary teeth were observed in 24 examinees (prevalence rate: 1.5%). The ratio of supernumerary teeth located in the maxilla versus the mandible was 2.2:1. The most common type of supernumerary tooth was the upper distomolar. Conclusion: The prevalence of lateral incisor agenesis, canine impaction, and supernumerary teeth ranged from 0.8 to 1.5% in the sample of male Greek military students.

Allergological evaluation of a dog population in a veterinary immuno-allergology consultation: What correlates in a canine model

Introduction: Allergic dermatitis (AD) is the most common canine pruritic condition in veterinary dermatology. Allergic dermatitis to flea bites presents the highest prevalence, followed by atopic dermatitis and food AD. This study aimed to identify possible correlation between data from clinical signs, intradermal tests (IDT) and specific IgE levels, which are used in dog AD assessment. Methods: Fifty five dogs from the Veterinary Hospital of the University of Évora (Portugal) and Rof Codina University Hospital (Lugo, Spain) outpatient consultations were studied by means of clinical inquiry, IDT and specific IgE determination. Thirty five of the patients belonged to predisposed breeds, 30 were females and 25 males. Forty one (74%) were indoor. Results: In 82% of cases first clinical signs appeared before the age of 3 years and 24% even before 1 year old. In 70% of the individuals clinical signs included itching, which was generalized in 66%, with 78% of paw licking and chewing. Clinical profile showed seasonal worsening in 64% of cases. From the 69.1% of dogs already presenting with dermatitis, 50% also presented external otitis and 28.9% self-inflicted alopecia. "Intense itching" was found in 10.5%, "medium itching" in 81.6% and “mild itching” in 5.26% of the patients. Prevalence of positive IDT was 37.3 % to Lep d, 29.41% to Der f, 27.5% to Der p, 25.5% to Dac g and 21.6% to Malassezia sp. From the 37 dogs submitted to food IDT, 40.5% revealed positive to beef, 27% to chicken, 27% to porc and 5.4% to lamb. Specific IgE > 150 EAU was found in 84% of dogs to indoor allergen sources and in 68% to pollens. A negative correlation was found between an outdoor life and the intensity (p = 0.033) and precocity (p = 0.026) of clinical signs. Sensitization to pollens was found positively correlated with the seasonality of clinical signs (p = 0.001) and the positivity for Dac g (p = 0.007). The prevalence of chronic otitis correlated positively with alopecia and reactivity to Lep d (p = 0.008), Plantago lanceolata (p = 0.026) and Platanus acerifolia (p = 0.017). There was no correlation between the results of ITD and specific IgE. Conclusion: We can conclude that correlation between different clinical signs and positive testing for some allergenic sources may occur, as well as between sensitization to pollens and the beginning, the intensity and the seasonality of dog patient clinical signs.

Identifying Genetic Risk Factors in Canine Autoimmune Disorders

Autoimmune diseases are more common in dogs than in humans and are already threatening the future of some highly predisposed dog breeds. Susceptibility to autoimmune diseases is controlled by environmental and genetic factors, especially the major histocompatibility complex (MHC) gene region. Dogs show a similar physiology, disease presentation and clinical response as humans, making them an excellent disease model for autoimmune diseases common to both species. The genetic background of canine autoimmune disorders is largely unknown, but recent annotation of the dog genome and subsequent development of new genomic tools offer a unique opportunity to map novel autoimmune genes in various breeds. Many autoimmune disorders show breed-specific enrichment, supporting a strong genetic background. Furthermore, the presence of hundreds of breeds as genetic isolates facilitates gene mapping in complex autoimmune disorders. Identification of novel predisposing genes establishes breeds as models and may reveal novel candidate genes for the corresponding human disorders. Genetic studies will eventually shed light on common biological functions and interactions between genes and the environment. This study aimed to identify genetic risk factors in various autoimmune disorders, including systemic lupus erythematosus (SLE)-related diseases, comprising immune-mediated rheumatic disease (IMRD) and steroid-responsive meningitis arteritis (SMRA) as well as Addison s disease (AD) in Nova Scotia Duck Tolling Retrievers (NSDTRs) and chronic superficial keratitis (CSK) in German Shepherd dogs (GSDs). We used two different approaches to identify genetic risk factors. Firstly, a candidate gene approach was applied to test the potential association of MHC class II, also known as a dog leukocyte antigen (DLA) in canine species. Secondly, a genome-wide association study (GWAS) was performed to identify novel risk loci for SLE-related disease and AD in NSDTRs. We identified DLA risk haplotypes for an IMRD subphenotype of SLE-related disease, AD and CSK, but not in SMRA, and show that the MHC class II gene region is a major genetic risk factor in canine autoimmune diseases. An elevated risk was found for IMRD in dogs that carried the DLA-DRB1*00601/DQA1*005011/DQB1*02001 haplotype (OR = 2.0, 99% CI = 1.03-3.95, p = 0.01) and for ANA-positive IMRD dogs (OR = 2.3, 99% CI = 1.07-5.04, p-value 0.007). We also found that DLA-DRB1*01502/DQA*00601/DQB1*02301 haplotype was significantly associated with AD in NSDTRs (OR = 2.1, CI = 1.0-4.4, P = 0.044) and the DLA-DRB1*01501/DQA1*00601/DQB1*00301 haplotype with the CSK in GSDs (OR=2.67, CI=1.17-6.44, p= 0.02). In addition, we found that homozygosity for the risk haplotype increases the risk for each disease phenotype and that an overall homozygosity for the DLA region predisposes to CSK and AD. Our results have enabled the development of genetic tests to improve breeding practices by avoiding the production of puppies homozygous for risk haplotypes. We also performed the first successful GWAS for a complex disease in dogs. With less than 100 cases and 100 controls, we identified five risk loci for SLE-related disease and AD and found strong candidate genes involved in a novel T-cell activation pathway. We show that an inbred dog population has fewer risk factors, but each of them has a stronger genetic risk. Ongoing studies aim to identify the causative mutations and bring new knowledge to help diagnostics, treatment and understanding of the aetiology of SLE-related diseases.

Sexual dimorphism in canine length of woolly spider monkeys (Brachyteles arachnoides, E. Geoffroy 1806)

We measured canine teeth from 28 woolly spider monkeys (Brachyteles arachnoides) to assess sexual dimorphism and population differences. The specimens are from the Brazilian states of Bahia, Minas Gerais, Espírito Santo, Rio de Janeiro, and São Paulo. We found strong sexual dimorphism in canine length for individuals belonging to populations south of 22°00′ latitude but no sexual dimorphism in canine length from individuals of populations north of 21°00′ latitude. Canine length did not vary among females of northern and southern populations. However, southern males had significantly longer canines than northern males. This geographical difference in canine morphology, together with the presence or absence of thumbs and published accounts of differences in genetics and social structure between northern and southern populations, suggests that Brachyteles arachnoides may be composed of at least two subspecies, which appear to be separated by the rivers Grande and Paraiba do Sul and the Serra da Mantiqueira. © 1993 Plenum Publishing Corporation.

Modulation of anti-pathogenic activity in canine-derived Lactobacillus species by carbohydrate growth substrate

Aims: To investigate the effect of various carbon sources on the production of extracellular antagonistic compounds against two Escherichia coli strains and Salmonella enterica serotype Typhimurium by three canine-derived lactobacilli strains. Methods and Materials: Cell-free preparations, pH neutralized, were used in antibiotic disc experiments as an initial screening. The bacteria/carbohydrate combinations that showed inhibition of the growth of those pathogens, were further investigated in batch co-culture experiments. The cell-free supernatants of the cultures, that decreased the population number of the pathogens in the co-culture experiments to log CFU ml(-1) less than or equal to 4, were tested for inhibition of the pathogens in pure cultures at neutral and acidic pH. Conclusions: The results showed that the substrate seems to affect the production of antimicrobial compounds and this effect could not just be ascribed to the ability of the bacteria to grow in the various carbon sources. L. mucosae, L. acidophilus and L. reuteri, when grown in sugar mixtures consisting of alpha-glucosides (Degree of Polymerization (DP) 1-4) could produce antimicrobial compounds active against all three pathogens in vitro. This effect could not be attributed to a single ingredient of those sugar mixtures and was synergistic. This inhibition had a dose-response characteristic and was more active at acidic pH. Significance and Impact of the Study: Knowledge of the effect that the carbon source has on the production of antimicrobial compounds by gut-associated lactobacilli allows the rational design of prebiotic/probiotic combinations to combat gastrointestinal pathogens.

Isolation, Characterization, and Differentiation Potential of Canine Adipose-Derived Stem Cells

Adipose tissue may represent a potential source of adult stem cells for tissue engineering applications in veterinary medicine. It can be obtained in large quantities, under local anesthesia, and with minimal discomfort. In this study, canine adipose tissue was obtained by biopsy from subcutaneous adipose tissue or by suction-assisted lipectomy (i.e., liposuction). Adipose tissue was processed to obtain a fibroblast-like population of cells similar to human adipose-derived stem cells (hASCs). These canine adipose-derived stem cells (cASCs) can be maintained in vitro for extended periods with stable population doubling and low levels of senescence. Immunofluorescence and flow cytometry show that the majority of cASCs are of mesodermal or mesenchymal origin. cASCs are able to differentiate in vitro into adipogenic, chondrogenic, myogenic, and osteogenic cells in the presence of lineage-specific induction factors. In conclusion, like human lipoaspirate, canine adipose tissue may also contain multipotent cells and represent an important stem cell source both for veterinary cell therapy as well as preclinical studies.

Experimental acute canine monocytic ehrlichiosis: clinicopathological and immunopathological findings

Clinical signs, humoral and cellular immune responses, and microscopic and gross tissue alterations resulting from acute experimental Ehrlichia canis infection in dogs were studied. Four dogs were inoculated with E canis and four were used as uninfected controls. After a 10-14-day incubation period, infected dogs developed pyrexia up to 41 degreesC for 6-8 days. Antibody titers to E. canis antigen were demonstrable in all inoculated dogs at 30 days post-infection. Necropsy of infected animals revealed pale mucous membranes, generalized lymphadenopathy, splenomegaly, edema and ascites. Microcopically, the main lesions were: lymphoreticular hyperplasia in cortical areas of lymph nodes and spleenic white pulp, periportal accumulation of mononuclear cells and centrolobular fatty degeneration of the liver. Kidneys presented with glomerulonephritis characterized by interstitial, mononuclear infiltration. Immunophenotyping of lymphocytes from lymph nodes and spleen sections displayed alterations in IgG, IgM, CD3+ and CD8+ cells population in infected dogs. (C) 2003 Elsevier B.V. All rights reserved.

Maxillary canine-first premolar transposition - Restoring normal tooth order with segmented mechanics

Tooth transpositions present at a relatively low incidence in the world population and primarily affect maxillary canines and premolars. Treatment of this disturbance should take into account aspects such as facial pattern, age, malocclusion, tooth-size discrepancy, stage of eruption, and magnitude of the transposition. Mechanics for correction should be entirely individualized, reducing the risks and adverse effects. Practitioners often select simpler options, indicating extraction of permanent teeth, which is an irreversible procedure that may bring about damages to the patient. This study presents a case report and treatment of unilateral transposition of maxillary canine and premolar with repositioning of affected teeth to their respective normal positions.

Canine bartonellosis: serological and molecular prevalence in Brazil and evidence of co-infection with Bartonella henselae and Bartonella vinsonii subsp berkhoffii

The purpose of this study was to determine the serological and molecular prevalence of Bartonella spp. infection in a sick dog population from Brazil. At the São Paulo State University Veterinary Teaching Hospital in Botucatu, 198 consecutive dogs with clinicopathological abnormalities consistent with tick-borne infections were sampled. Antibodies to Bartonella henselae and Bartonella vinsonii subsp. berkhoffii were detected in 2.0% ( 4/197) and 1.5% ( 3/197) of the dogs, respectively. Using 16S-23S rRNA intergenic transcribed spacer ( ITS) primers, Bartonella DNA was amplified from only 1/198 blood samples. Bartonella seroreactive and/or PCR positive blood samples ( n = 8) were inoculated into a liquid pre-enrichment growth medium ( BAPGM) and subsequently sub-inoculated onto BAPGM/blood-agar plates. PCR targeting the ITS region, pap31 and rpoB genes amplified B. henselae from the blood and/or isolates of the PCR positive dog ( ITS: DQ346666; pap31 gene: DQ351240; rpoB: EF196806). B. henselae and B. vinsonii subsp. berkhoffii ( pap31: DQ906160; rpoB: EF196805) co-infection was found in one of the B. vinsonii subsp. berkhoffii seroreactive dogs. We conclude that dogs in this study population were infrequently exposed to or infected with a Bartonella species. The B. henselae and B. vinsonii subsp. berkhoffii strains identified in this study are genetically similar to strains isolated from septicemic cats, dogs, coyotes and human beings from other parts of the world. To our knowledge, these isolates provide the first Brazilian DNA sequences from these Bartonella species and the first evidence of Bartonella co-infection in dogs.