Whole Genome Sequencing Reveals Multiple Linked Genetic Variants on Canine Chromosome 12 Associated with Risk for Symmetrical Lupoid Onychodystrophy (SLO) in the Bearded Collie.

In dogs, symmetrical lupoid onychodystrophy (SLO) results in nail loss and an abnormal regrowth of the claws. In Bearded Collies, an autoimmune nature has been suggested because certain dog leukocyte antigen (DLA) class II haplotypes are associated with the condition. A genome-wide association study of the Bearded Collie revealed two regions of association that conferred risk for disease: one on canine chromosome (CFA) 12 that encompasses the DLA genes, and one on CFA17. Case-control association was employed on whole genome sequencing data to uncover putative causative variants in SLO within the CFA12 and CFA17 associated regions. Genotype imputation was then employed to refine variants of interest. Although no SLO-associated protein-coding variants were identified on CFA17, multiple variants, many with predicted damaging effects, were identified within potential candidate genes on CFA12. Furthermore, many potentially damaging alleles were fully correlated with the presence of DLA class II risk haplotypes for SLO, suggesting that the variants may reflect DLA class II haplotype association with disease or vice versa. Strong linkage disequilibrium in the region precluded the ability to isolate and assess the individual or combined effect of variants on disease development. Nonetheless, all were predictive of risk for SLO and, with judicious assessment, their application in selective breeding may prove useful to reduce the incidence of SLO in the breed.

Genetic characterization of Addison's disease in Bearded Collies.

BACKGROUND: Primary hypoadrenocorticism (or Addison's disease, AD) is an autoimmune disease that results in destruction of the adrenal cortex and consequent adrenal insufficiency. The disease has been described in purebred and mixed breed dogs, although some breeds, including the Bearded Collie, are at increased risk for AD. Candidate gene approaches have yielded few associations that appear to be breed-specific. A single other genome-wide association study reported no significant regions of association for AD in Standard Poodles. The present study aimed to identify genomic regions of association for canine AD in Bearded Collies. RESULTS: Our study consists of the first genome-wide association analysis to identify a genome-wide significant region of association with canine AD (CFA18). Peaks of suggestive association were also noted on chromosomes 11, 16 and 29. Logistic regression analysis supported an additive effect of risk genotypes at these smaller effect loci on the probability of disease associated with carrying a risk genotype on CFA18. Potential candidate genes involved in adrenal steroidogenesis, regulation of immune responses and/or inflammation were identified within the associated regions of chromosomes 11 and 16. The gene-poor regions of chromosomes 18 and 29 may, however, harbor regulatory sequences that can modulate gene expression and contribute to disease susceptibility. CONCLUSION: Our findings support the polygenic and complex nature of canine AD and identified a strongly associated locus on CFA18 that, when combined with three other smaller effect loci, was predictive of disease. The results offer progress in the identification of susceptibility loci for canine AD in the Bearded Collie. Further studies are needed to confirm association with the suggested candidate genes and identify actual causative mutations involved with AD susceptibility in this breed.

Novel Locus Associated with Symmetrical Lupoid Onychodystrophy in the Bearded Collie.

Symmetrical lupoid onychodystrophy (SLO) is characterized by inflammation of the nail bed and nail sloughing that causes affected dogs considerable pain. Disease etiology remains unclear, although an autoimmune component is suspected. A genome-wide association study on Bearded Collies revealed regions of association on canine chromosomes (CFA) 12 and 17. The large region of association on CFA12 likely consists of two smaller linked regions, both of which are also linked to the dog leukocyte antigen (DLA) class II genes. Dogs homozygous for the alternate allele at the top CFA12 SNP also carried two DLA class II risk haplotypes for SLO, and this locus explained most of the increased risk for disease seen throughout the CFA12 region of association. A stronger peak was seen on CFA17 when analysis was done solely on dogs that carried DLA class II risk haplotypes for SLO. The majority of SLO dogs carried a homozygous alternate genotype on CFA12 and at least one CFA17 risk haplotype. Our findings offer progress toward uncovering the genetic basis of SLO. While the contribution of the CFA17 region remains unclear, both CFA12 and CFA17 regions are significantly associated with SLO disease expression in the Bearded Collie and contain potential candidate genes for this disease.

DLA class II risk haplotypes for autoimmune diseases in the bearded collie offer insight to autoimmunity signatures across dog breeds.

BACKGROUND: Primary hypoadrenocorticism (Addison's disease, AD) and symmetrical lupoid onychodystrophy (SLO) are two clinical conditions with an autoimmune etiology that occur in multiple dog breeds. In man, autoimmunity is associated with polymorphisms in immune-related genes that result in a reduced threshold for, or defective regulation of, T cell activation. The major histocompatibility complex (MHC) class II genes encode molecules that participate in these functions, and polymorphisms within these genes have been associated with autoimmune conditions in dogs and humans. Bearded collies have a relatively high prevalence of autoimmune diseases, particularly AD and SLO. Our study assessed the relationship between particular MHC (dog leukocyte antigen, DLA) class II haplotypes and the two autoimmune diseases most common in this breed. Moreover, five unrelated breeds at increased risk for AD were studied for comparative purposes and analyzed in the context of extant literature. RESULTS: A single DLA class II three-locus haplotype, determined by sequence-based typing, was associated with increased risk for AD (DLA-DRB1*009:01/DQA1*001:01/DQB1*008:02) in bearded collies. Comparative analysis with the five additional breeds showed limited allele sharing, with DQA1*001:01 and DQB1*002:01 being the only alleles observed in all breeds. A distinct three-locus risk haplotype (DLA-DRB1*001:01/DQA1*001:01/DQB1*002:01) was associated with AD in the West Highland white terrier and Leonberger. Two different risk haplotypes were associated with increased risk for SLO in the bearded collie (DLA-DRB1*018:01/DQA1*001:01/DQB1*002:01 and DLA-DRB1*018:01/DQA1*001:01/ DQB1*008:02). CONCLUSION: Two-locus DQ haplotypes composed of DLA-DQA1*001:01 in association with DLA-DQB1*002:01 or DLA-DQB1*008:02 make up the four risk haplotypes identified in the present study and are also found in other risk haplotypes previously associated with diabetes mellitus and hypothyroidism across different dog breeds. Our findings build upon previously published data to suggest that this two-locus (DQ) model serves as a good indicator for susceptibility to multiple organ-specific autoimmune diseases in the canine population. However, it is also clear that additional loci are necessary for actual disease expression. Investigation of affected and unaffected dogs carrying these predisposing DQ haplotype signatures may allow for the identification of those additional genetic components that determine autoimmune disease expression and organ specificity.

Individual signatures and environmental factors shape skin microbiota in healthy dogs.

BACKGROUND: The individual, together with its environment, has been reported as the main force driving composition and structure of skin microbiota in healthy dogs. Therefore, one of the major concerns when analyzing canine skin microbiota is the likely influence of the environment. Despite the dense fur covering, certain skin diseases exhibit differential prevalence among skin sites, dog breeds, and individuals. RESULTS: We have characterized the normal variability of dog skin microbiota in a well-controlled cohort of a large number of Golden-Labrador Retriever crossed dogs (N = 35) with similar ages, related genetic background, and a shared environment. We found that the individual drives the skin microbiota composition and structure followed by the skin site. The main bacterial classes inhabiting dog skin in this cohort are Gammaproteobacteria and Bacilli. We also detected bacteria associated to the environment on different dog skin sites that could be reflecting the different degrees of exposure of each skin site and each dog. Network analyses elucidated bacterial interactions within and between skin sites, especially in the chin, abdomen, axilla, and perianal region, with the highly shared interactions probably representing an anatomical, behavioral, or environmental component. When analyzing each skin site independently to assess host-specific factors, we found that temporality (season of birth and time spent in the kennel) affected all the skin sites and specially the inner pinna. The most abundant taxon driving this difference was Sphingomonas. We also found taxonomic differences among male and female dogs on the abdomen, axilla, and back. CONCLUSIONS: We observed a large inter-individual variability and differences among skin sites. Host-specific variables, such as temporality or sex, were also shaping skin microbiota of healthy dogs, even in an environmental homogenous cohort.

Protein expression and genetic variability of canine Can f 1 in golden and Labrador retriever service dogs.

BACKGROUND: Valued for trainability in diverse tasks, dogs are the primary service animal used to assist individuals with disabilities. Despite their utility, many people in need of service dogs are sensitive to the primary dog allergen, Can f 1, encoded by the Lipocalin 1 gene (LCN1). Several organizations specifically breed service dogs to meet special needs and would like to reduce allergenic potential if possible. In this study, we evaluated the expression of Can f 1 protein and the inherent variability of LCN1 in two breeds used extensively as service dogs. Saliva samples from equal numbers of male and female Labrador retrievers (n = 12), golden retrievers (n = 12), and Labrador-golden crosses (n = 12) were collected 1 h after the morning meal. Can f 1 protein concentrations in the saliva were measured by ELISA, and the LCN1 5' and 3' UTRs and exons sequenced. RESULTS: There was no sex effect (p > 0.2) nor time-of-day effect; however, Can f 1 protein levels varied by breed with Labrador retrievers being lower than golden retrievers (3.18 ± 0.51 and 5.35 ± 0.52 µg/ml, respectively, p < 0.0075), and the Labrador-golden crosses having intermediate levels (3.77 ± 0.48 µg/ml). Although several novel SNPs were identified in LCN1, there were no significant breed-specific sequence differences in the gene and no association of LCN1 genotypes with Can f 1 expression. CONCLUSIONS: As service dogs, Labrador retrievers likely have lower allergenic potential and, though there were no DNA sequence differences identified, classical genetic selection on the estimated breeding values associated with salivary Can f 1 expression may further reduce that potential.

Seroprevalence of Toxoplasma gondii (Nicole & Manceaux, 1909) and retroviral status of client-owned pet cats (Felis catus, Linnaeus, 1758) in Rio de Janeiro, Brazil.

Cats, as definitive host, play an important role in the transmission of Toxoplasma gondii. This study aimed to establish the seroprevalence of anti-T. gondii immunoglobulins G and M, and determine the frequency of oocysts in the feces of the domestic cat population in Rio de Janeiro, Brazil. We also aimed to study the association between T. gondii infection and age, sex, breed, lifestyle, diet and retroviral infection. A total of 108 cats were included in the study and fecal samples of 54 of those cats were obtained. Only 5.6% of the cats were seropositive for anti-T. gondii immunoglobulins using the indirect hemagglutination test. None of the 54 cats presented oocysts in their fecal samples. Although not statistically significant, males, mixed-breed, free-roaming and cats aged two years and older were found to be more exposed. Age, lifestyle and the use of litter boxes were found to play an important role as risk factors. Anemia and retroviral infections were independent of T. gondii infection. No antibodies were detected in the majority of cats (94.4%), indicating that those cats had never been exposed to the parasite and, therefore, once infected, they could present the risk of shedding large numbers of oocysts into the environment.

SEROPREVALENCE OF Toxoplasma gondii (Nicole & Manceaux, 1909) AND RETROVIRAL STATUS OF CLIENT-OWNED PET CATS (Felis catus, Linnaeus, 1758) IN RIO DE JANEIRO, BRAZIL / Soroprevalência da infecção por Toxoplasma gondii (Nicole & Manceaux, 1909) e a infecção por retrovírus em população urbana de gatos domésticos (Felis catus, Linnaeus, 1758) no Rio de Janeiro, Brazil

Cats, as definitive host, play an important role in the transmission of Toxoplasma gondii. This study aimed to establish the seroprevalence of anti-T. gondii immunoglobulins G and M, and determine the frequency of oocysts in the feces of the domestic cat population in Rio de Janeiro, Brazil. We also aimed to study the association between T. gondii infection and age, sex, breed, lifestyle, diet and retroviral infection. A total of 108 cats were included in the study and fecal samples of 54 of those cats were obtained. Only 5.6% of the cats were seropositive for anti-T. gondii immunoglobulins using the indirect hemagglutination test. None of the 54 cats presented oocysts in their fecal samples. Although not statistically significant, males, mixed-breed, free-roaming and cats aged two years and older were found to be more exposed. Age, lifestyle and the use of litter boxes were found to play an important role as risk factors. Anemia and retroviral infections were independent of T. gondii infection. No antibodies were detected in the majority of cats (94.4%), indicating that those cats had never been exposed to the parasite and, therefore, once infected, they could present the risk of shedding large numbers of oocysts into the environment.

Os gatos, como hospedeiros definitivos, apresentam um papel fundamental na transmissão do Toxoplasma gondii. Nosso estudo teve como objetivo determinar a presença de imunoglobulinas G e M anti-T. gondii, e a frequência de oocistos nas fezes de uma população de gatos domésticos do Rio de Janeiro, Brasil. Objetivou-se ainda estudar a associação da infecção por T. gondii com a idade, sexo, raça, estilo de vida, dieta e infecção por retrovírus. Um total de 108 gatos foi incluído no estudo e 54 amostras de fezes foram obtidas desses animais. Somente 5,6% dos gatos foram sororreagentes para T. gondii, utilizando-se o teste de hemaglutinação indireta. Nenhum dos 54 gatos apresentou oocistos em suas amostras fecais. Embora sem comprovação estatística, machos, sem raça definida, com acesso livre às ruas e gatos com mais de dois anos de idade tenderam a ser mais expostos ao parasito. Idade, estilo de vida e uso de caixa de areia foram considerados importantes fatores de risco. Anemia e infecção por retrovírus não apresentaram relação com infecção por T. gondii. Não foram detectados anticorpos na maioria dos gatos (94,4%), indicando que esses gatos nunca foram expostos ao parasito e que, se infectados, poderão eliminar grande número de oocistos no ambiente.

Reduction of feral cat (Felis catus Linnaeus 1758) colony size following hysterectomy of adult female cats.

The size of urban cat colonies is limited only by the availability of food and shelter; therefore, their population growth challenges all known population control programs. To test a new population control method, a free-roaming feral cat colony at the Zoological Park in the city of Rio de Janeiro was studied, beginning in 2001. The novel method consisted of performing a hysterectomy on all captured female cats over 6 months of age. To estimate the size of the colony and compare population from year to year, a method of capture-mark-release-recapture was used. The aim was to capture as many individuals as possible, including cats of all ages and gender to estimate numbers of cats in all population categories. Results indicated that the feral cat population remained constant from 2001 to 2004. From 2004 to 2008, the hysterectomy program and population estimates were performed every other year (2006 and 2008). The population was estimated to be 40 cats in 2004, 26 in 2006, and 17 cats in 2008. Although pathogens tend to infect more individuals as the population grows older and maintains natural behavior, these results show that free-roaming feral cat colonies could have their population controlled by a biannual program that focuses on hysterectomy of sexually active female cats.

Prevalence of Bartonella species DNA and antibodies in cats (Felis catus) submitted to a spay/neuter program in Rio de Janeiro, Brazil.

The prevalence of Bartonella species DNA and antibodies for Bartonella henselae were studied in 40 clinically healthy cats (Felis catus, Linnaeus 1758) submitted to a spay/neuter program in Rio de Janeiro, Brazil. Additionally, the prevalence of Bartonella species DNA was investigated in the fleas found parasitizing the subject cats. For this purpose, blood samples were obtained from all cats, and DNA extraction was performed on the blood, and blood clotted samples, as well as on pools of fleas obtained from them. Antibodies for B henselae were detected on serum samples. Bartonella species DNA was detected in 17 cats, whereas serum reactivity for B henselae was found in 19. A total of 20 cats were flea-infested and nine of these 20 had Bartonella species DNA in their blood. In four of the 20 flea-infested cats, Bartonella species DNA was detected in the fleas obtained from those cats, but only one of these four cats had Bartonella species DNA in its blood.

Characterization of ectoparasites in an urban cat (Felis catus Linnaeus, 1758) population of Rio de Janeiro, Brazil.

Ectoparasites are capable of transmitting infectious diseases and, therefore, are of zoonotic concern. Cats submitted to a spay/neuter program in the city of Rio de Janeiro were examined to determine the distribution of ectoparasites in cats from a city with a tropical climate. Independent of gender, breed, or age, 292 cats were combed and subjected to otoscopic examination. Ectoparasites were collected, and blood samples were taken to determine packed cell volume. The majority of the 292 cats were female (71%), and most of them were categorized as domestic short hair cats (92%). Different species of ectoparasites that produced both single agent and multi-agent infestations were detected in these cats. Most cats were infested by Ctenocephalides felis (60%); however, other ectoparasites were found to a lesser degree in the sampled population (Felicola subrostratus (5.4%), Rhipicephalus sanguineus (1.4%), Lynxacarus radovskyi (1%), and Otodectes cynotis (6.2%)). Within the infested cats, 16.3% were also anemic, and there was a significant association between the occurrence of anemia and flea infestation. Of all sampled cats, 65% were infested by at least one species of ectoparasites, which suggests both a heavy infestation of the environment and owner negligence. Ectoparasites are not only nuisances to both cats and owners, but they are also significant carriers of disease. Therefore, the implementation of rigorous, safe preventive measures is of great importance.

Escore de condição corporal como indicador do prognóstico de gatos com doença renal crônica / Body condition score as an indicator of prognosis for cats with chronic renal disease

A caquexia tem sido relacionada à maior mortalidade de pacientes com doença renal crônica, tanto em humanos, como em animais. O escore de condição corporal (ECC) pode ser utilizado em conjunto com o peso para melhor avaliação da composição corporal desses animais. O objetivo deste trabalho foi de correlacionar o escore de condição corporal com o prognóstico de felinos com doença renal crônica. Avaliaram-se 110 felinos idosos, dos quais 70 apresentavam-se hígidos (Grupo I) e 40 apresentavam doença renal crônica (Grupo II). No grupo I, apenas 5,7 por cento dos gatos apresentaram ECC abaixo do ideal e destes nenhum foi classificado como caquético. No grupo II, 70 por cento dos gatos apresentaram ECC abaixo do ideal, dos quais 32,5 por cento eram caquéticos. A taxa de mortalidade no grupo II foi significantemente maior naqueles caquéticos. O ECC abaixo do considerado ideal indica um prognóstico desfavorável nos pacientes com doença renal crônica.

Cachexia has been associated with higher mortality in patients with chronic renal disease both in human and veterinary medicine. Body condition score (BCS) can be used along with body weight for a better evaluation of a patient's body composition. The objective of this study was to associate body condition score with prognosis of cats with chronic renal disease. One hundred and ten elderly cats were evaluated; of which 70 were healthy (Group I) and 40 had been diagnosed with chronic renal disease (Group II). In Group I, only 5.7 percent of the cats presented a BCS below ideal, though none were found to be cachectic. In Group II, 70 percent of the cats presented a BCS below ideal, where 32.5 percent were cachectic. Mortality was significantly higher within the cachectic patients of Group II. BCS below ideal indicates a poor prognosis for patients with chronic renal disease.