Molecular Diagnosis of Footrot and Contagious Ovine Digital Dermatitis in Small Ruminants in the Iberian Peninsula.

Contagious ovine digital dermatitis (CODD) and footrot (FR), a sub-acute or acute necrotic (decaying) infectious disease involving the hoof and underlying tissues, pose economic challenges to herds in Spain and worldwide. The aetiological agent for FR is Dichelobacter nodosus, while CODD is caused by pathogenic Treponema phylogroups. We detail the findings derived from the analysis by qPCR of 105 pooled samples from 100 ovine and five caprine herds in Spain and Portugal, alongside 15 samples from healthy flocks in order to identify Dichelobacter nodosus, Fusobacterium necrophorum, Treponema spp., and three pathogenic Treponema phylogroups (T. phagedenis, T. medium, and T. pedis). Treponema spp. were detected in all 120 pools, including samples from the 15 healthy flocks where only one positive result for F. necrophorum was recorded. Mixed infections by agents different from Treponema spp. were identified in 68.57% of samples. Positive results for F. necrophorum and/or D. nodosus, were obtained for 91.4% of the pools, whereas the presence of the three pathogenic Treponema phylogroups was rare: each of them appeared in isolation in a single pool, while they were found in 18 pools in combination with other agents. While F. necrophorum was the sole finding in 16.2% of samples from affected herds, D. nodosus (the footrot causative agent) was only detected in 61% of affected farms. An improved qPCR protocol was implemented to determine the serogroups of D. nodosus in the samples and found all of them (except the G serogroup), often in combined infections (35.1%). This report concludes with comprehensive proposals for diagnosing, preventing, and treating hoof ailments, remarking the interest of the information about D. nodosus serogroups in order to improve the efficiency of immunization by choosing appropriate vaccine protocols.

Feline osteochondromatosis in a 12-year-old feline leukaemia virus-negative cat.

Feline osteochondromatosis is a spontaneous osteocartilaginous exostosis associated with feline leukaemia virus (FeLV) infection or due to a frameshift variant in the exostosin glycosyltransferase 1 (EXT1) gene. Osteochondromatosis was diagnosed in an indoor-only, 12-year-old, neutered female, Russian Blue cat. Radiographs revealed bilateral calcified proliferations in the elbow, costochondral and sternochondral joints, which distorted the normal skeletal structure. Grossly, the proliferated joints presented with consistent, rounded masses, causing complete ankylosis. The main histopathological finding was an osteocartilaginous proliferation composed of multiple irregular islands of well-differentiated hyaline cartilage surrounded and delimited by osteoid tissue. Immunohistochemistry of the osteochondromas, bone marrow and mediastinal lymph nodes, using a primary anti-FeLV gp70 antibody, and FeLV proviral DNA real-time polymerase chain reaction on bone marrow were negative. Sequencing of exon 6 of the EXT1 gene was performed and nucleotide BLAST analysis demonstrated the absence of a frameshift variant. This study reports the only case of spontaneous feline osteochondromatosis in an animal more than 10 years old.

Phylogeographic and population genetic structure of hound-like native dogs of the Mediterranean Basin.

The dog was probably the first domesticated animal. Despite extensive archaeological and genetic investigations, the origin and the evolution of the extant dogs are still being debated. Dog breeds that have over time been selected for hunting share common ancestral traits. This study represents the first comprehensive attempt to survey at the genomic and mitochondrial level eight hound-like dogs breeds indigenous to the Mediterranean Basin to determine if they share common ancient origins. Results from the microsatellite analysis indicate that all the dog populations have a low inbreeding value.The Kelb tal-Fenek has a high divergence from the current Egyptian street population, however there is not enough evidence from this study to exclude completely the potential of an ancient common relationship. Overall, the mitochondrial results indicate high frequencies of haplogroups A and B and a low representation of haplogroup C, while only one Egyptian dog could be assigned to haplogroup D. Results reveal identities and shared clades, suggesting the conservation of ancient European mitotypes in the Mediterranean hound-like breeds, especially in the Egyptian population. Although none of the dog populations/breeds participating in this study indicate to be direct descendants of the Egyptian dogs, they still have a very close morphologically resemblance to those iconic Egyptian dogs often depicted in ancient art forms and share some genetic links with the current Egyptian population. Further research is required with other markers such us complete mitogenomes and SNP panels to confirm the complex history of the Mediterranean dogs involved in this study.

Coxiella burnetii and Co-Infections with Other Major Pathogens Causing Abortion in Small Ruminant Flocks in the Iberian Peninsula.

Coxiella burnetii is an intracellular bacterium causing human Q fever and reproductive disorders in domestic ruminants. We analyzed the occurrence of C. burnetii and co-infections with six other major pathogens causing abortion in sheep (1242 cases) and goat (371 cases) flocks from Spain and Portugal. After real-time PCR detection, co-infections were established by principal component and cluster analysis that grouped cases based on the joint presence/absence of several microorganisms. C. burnetii and Chlamydia abortus were the most common abortifacient agents with approximately 75% of cases from both hosts testing positive, followed by Toxoplasma gondii, Campylobacter sp., Salmonella enterica, border disease virus and Neospora caninum. C. burnetii was significantly more common than C. abortus in goat abortions (p < 0.001). Co-infections with at least two pathogens were found in more than 66% cases of ovine abortions and 36% cases of caprine abortions testing positive for C. burnetii, mostly including mixed infections with only C. abortus. These findings indicate that both pathogens are the most significant ones to be readily prevented by vaccination in this geographical area. Biosecurity and biocontainment measures are also steadfastly recommended to prevent both the economic losses and public health risks associated with most of these abortifacient agents.

Prevalence and Associated Factors of Blastocystis sp. Infection in Patients with Gastrointestinal Symptoms in Spain: A Case-Control Study.

Blastocystis sp. is known to be the most prevalent parasite in fecal samples of humans worldwide. In the present report, a case-control study (1:9.89 (≈10)) was performed, by analyzing data from 3682 patients who attended a public hospital in the northern area of Spain showing gastrointestinal symptoms. Diagnosis was performed in human fecal samples by means of optical microscopy. The prevalence of Blastocystis sp. in patients with gastrointestinal symptoms was 9.18% (338/3682). Most of the Blastocystis sp.-infected patients tested negative for protozoa and helminths, and were underweight and foreign-born (26.4%), mainly from Africa and Central/South America. Gastrointestinal symptoms, such as abdominal pain, anorexia, halitosis, plus relative eosinophilia, as well as co-infections with pathogenic bacteria were associated with Blastocystis sp. infection. Both type 2 diabetes and treatment with immunosuppressive medicines at the time of Blastocystis sp. detection were associated with a higher proportion of infected patients. This is the first case-control study of Blastocystis sp. in humans in northern Spain and may contribute to surveillance and intervention strategies by public health authorities.

Occurrence of Rotavirus A Genotypes and Other Enteric Pathogens in Diarrheic Suckling Piglets from Spanish Swine Farms.

Species A rotavirus (RVA) is a major viral pathogen causing diarrhea in suckling piglets. Studies on its genetic heterogeneity have implications for vaccine efficacy in the field. In this study, fecal samples (n = 866) from diarrheic piglets younger than 28 days were analyzed over a two-year period (2018-2019). Samples were submitted from 426 farms located in 36 provinces throughout Spain and were tested using real-time PCR (qPCR) and reverse transcription real-time PCR (RT-qPCR) for five enteric pathogens. The individual prevalence was 89.4%, 64.4%, 44.9%, 33.7% and 4.4% for Clostridiumperfringens, Clostridioides (formerly Clostridium) difficile, species A rotavirus, species C rotavirus and porcine epidemic diarrhea virus, respectively. Most specimens (96.9%) were positive for at least one of the target pathogens, and more than 80% of samples harbored mixed infections. Nucleotide sequencing of 70 specimens positive for RVA revealed the presence of the VP7 genotypes G4, G9, G3, G5, G11 and the VP4 genotypes P7, P23, P6 and P13, with the combinations G4P7 and G9P23 being the most prevalent, and especially in the areas with the highest pig population. The study shows the extensive genetic diversity of RVA strains as well as discrepancies with the genotypes contained in the vaccine available in Spain, and multiple amino acid differences in antigenic epitopes of different G- and P- genotypes with the vaccine strains. Further investigations are needed to determine the efficacy of the vaccine to confer clinical protection against heterologous strains.

Blood Parameters and Feline Tooth Resorption: A Retrospective Case Control Study from a Spanish University Hospital.

Tooth resorption (TR; progressive destruction of hard dental tissues) varies in prevalence according to population, age, and country (29-66.1%). Our objective was twofold: describing the TR clinical presentation in Northeastern Spain, and studying 34 blood parameters to ascertain potential systemic effects associated with TR. Cases (29; presented from September 2018 to May 2019) and controls (58) were considered. Non-parametric tests were carried out to compare cases and controls for each blood parameter; those showing significant differences were chosen for multiple regression analysis (binomial logistic and hierarchical multiple regressions). In case TR was detected in 130/870 teeth (14.9%), TR stage and type were correlated (p < 0.001). Increasing CREA values (p = 0.034) and decreasing BUN/CREA and ALB/GLOB values were associated with TR presence (p = 0.029 and p = 0.03, respectively). Increasing GLOB was associated with increasing severity of TR (p < 0.01). Type 1 TR (highly related to inflammation and periodontal disease PD) was the most frequently observed type; the association of TR and inflammation biomarkers (ALB/GLOB, GLOB) are explained by this fact. The concomitant presence of PD and TR in old cats would cause TR association with kidney damage biomarkers (CREA, BUN/CREA). When affected by TR, special care in these aspects must be provided to cats.

Periodontal and Dental Status in Packs of Spanish Dogs.

While periodontal disease (PD) is the most common canine oral pathology, its prevalence varies according to diagnosis methodology, breed, and age. We intended to increase understanding of canine PD by studying dogs that are managed in a specific way: pack dogs in Spain. They received a mixed diet (home-prepared food, commercial dry food, stale bread and bones). Thirty-two conscious individuals from two packs of dogs in Northeastern Spain (30/32 crossbred hunting dogs and 2/32 Siberian Husky; 26 males and 6 females; 27.75 ± 5.807 kgs; 5.48 ± 2.818 years) received visual dental examination for assessment of absent teeth (AT), dental calculus (DC) grade, gingival recession (GR), periodontal disease (PD), tooth fracture (TF), and dental attrition (DA). DC was the most prevalent oral problem (75%), followed by TF/DA (68.75%), AT (34.37%), GR (31.25%), and, finally, PD (15.62%). Low individual affectation values were found for AT, GR, and PD (<1 tooth/individual); mean DC grade per individual was 0.06 ± 0.063; and TF and DA were found in 1.63 and 4.72 teeth/individual, respectively. Low prevalence and extent of PD was attributed to diagnosis methodology, bodyweight effect, breed, and, ultimately, diet. Individuals affected by DC remained under veterinary surveillance due to PD development.

Occurrence and genetic diversity of rotavirus A in faeces of diarrheic calves submitted to a veterinary laboratory in Spain.

A total of 237 faecal specimens from diarrheic calves younger than two months were collected and submitted for diagnosis of enteropathogens over a two-year period (2017-2018) to a veterinary laboratory. Samples originated from 193 dairy and beef farms in 29 provinces distributed throughout Spain, and were tested for the occurrence of three target enteric pathogens by reverse transcription real-time PCR (RT-qPCR): bovine rotavirus A (RVA), Cryptosporidium parvum and bovine coronavirus (BCoV). RT-PCR and nucleotide sequencing analysis were used to determine the G (VP7 gene) and P (VP4 gene) genotypes of 26 specimens positive for RVA. A total of 188 specimens (79.3 %) were positive for at least one of the three target enteric pathogens, and 101 samples (42.6 %) harbored mixed infections. The individual prevalence was 57.8 %, 50.6 % and 23.6 % for C. parvum, RVA and BCoV, respectively. Molecular analysis of selected RVA strains revealed the presence of the G6, G10, G3, P[5] and P[11] genotypes, with the combinations G6P[5] and G6P[11] being the most prevalent. Alignments of nucleotide sequences of the VP7 and VP4 markers showed a high frequency of single nucleotide polymorphisms (SNPs), with up to 294 SNPs found in 869bp of sequence at the G6 genotype (0.338 SNPs/nt), which reveals the extensive genetic diversity of RVA strains. Phylogenetic analysis of the VP7 gene of the G6 strains revealed four distinct lineages, with most strains clustering in the G6-IV lineage. The discrepancies between the RVA genotypes circulating in the sampled cattle farms and the genotypes contained in commercial vaccines currently available in Spain are discussed. We believe that this is the first study on the molecular characterization of rotavirus infecting cattle in Spain.

Tooth Resorption in Spanish Domestic Cats: Preliminary Data.

The objective of this study was to collect preliminary data about tooth resorption (TR) from cats treated at the Odontology Service (September 2016-June 2018), part of a University Veterinary Hospital in Spain, with specific emphasis on TR distribution per tooth. Diagnosis was based on visual/tactile inspection and intraoral dental radiographs. This cross-sectional study was carried out on 59 adult cats (27 females, 32 males). TR occurred in 39/59 cats (66.1%; 95% CI: 54.0%-78.2%). The median number of lesioned teeth per TR-affected animal was 3. A highly significant but weak correlation was found for age and number of TR-affected teeth per individual (Spearman´s correlation ρ = 0.381, P = .003, power = 0.853; N = 59). No TR cases were detected in incisors (0/708, 0%) but TR occurred in canines (21/236, 8.9%; 95% CI: 5.4%-12.4%), premolars (78/590, 13.2%; 95% CI: 10.5%-15.9%), and molars (33/236, 14%; 95% CI: 9.7%-18.3%). A significant age influence on TR was found. The greatest TR occurrence corresponded to 307 (21/59; 35.6%; 95% CI: 23.4%-47.8%) followed by 409 (17/59; 28.8%; 95% CI: 17.2%-40.3%), 407 (16/59; 27.1%; 95% CI: 15.7%-38.4%), and 309 (16/59; 27.1%; 95% CI: 15.7%-38.4%). These teeth would be considered as TR-sentinels in the studied population. These findings are relevant for veterinarians working in dental clinics, where the TR prevalence may be high especially in older cats.

Interaction effect of tegmen tympani and superior semicircular canal statuses on the thickness of the roof of the glenoid fossa: a cross-sectional descriptive study.

PURPOSE: Homogeneous development of temporal bone structures is explained by their ontogenic origin; tegmen tympani (TT) and superior semicircular canal (SSC) are related with the glenoid fossa at the temporomandibular joint (TMJ). Therefore, our objective was to determine a possible relationship between TT status (dehiscence or integrity) and the roof of the glenoid fossa (RGF) thickness; SSC status has also been considered. METHODS: This cross-sectional descriptive study was conducted in two tertiary hospitals on 95 patients (109 ears) presenting hypoacusia, facial palsy, vertigo, tinnitus, and other single or combined symptoms, and submitted to a thin-section multidetector-row computed axial tomography (CT) scan. RESULTS: A significant interaction effect of TT × SSC statuses on RGF thickness was found (p = 0.049). A significant difference in RGF thickness was found only for SSC integrity status between TT integrity and TT dehiscence (p = 0.004). The TT dehiscence increased the risk for RGF dehiscence 12.047 times (p = 0.002). CONCLUSIONS: There is an interaction effect of the statuses of both TT and SSC on the thickness of the RGF, instead of an independent effect of the TT status. When RGF dehiscence is found, TT and SSC statuses should be assessed, to discard associated dehiscences.

Multilocus fragment analysis of Cryptosporidium parvum from pre-weaned calves in Colombia.

The intra-species genetic diversity of Cryptosporidium parvum in dairy cattle farms in the central area of Colombia was investigated using a multilocus fragment typing approach with nine variable-number tandem-repeat (VNTR) loci and the gp60 gene. Genomic DNA of 70 C. parvum isolates from pre-weaned calves in 32 farms was analysed. Most markers showed two (ML1, MSB, CP47, and MSC6-7) or three alleles (5B12, Cgd2_3850, and Cgd6_5400), although they exhibited a major allele accounting for more than 69% of specimens, which explains their low discriminatory index. The TP14 microsatellite was monomorphic while a total of six alleles were found at the ML2 microsatellite. The two novel allelic variants (219bp, 245bp) exhibited by more than 36% of specimens at the latter locus were a remarkable finding. The 10-markers typing tool provided a Hunter-Gaston discriminatory value of 0.940 (95% CI, 0.918 - 0.961) and differentiated 22 multilocus subtypes (MLTs). Nevertheless, the combination of the three most informative markers (ML2, gp60, and Cgd2_3850) differentiated 68% of MLTs and hardly impaired the discriminatory index. The fact that many MLTs (13/22) were distinctive for individual farms provides evidence for the endemic nature of the infection and the major role played by transmission within farms. The eBURST algorithm suggested a low degree of genetic divergence. All but three MLTs were clustered in a clonal complex with a star-like topology typical of clonal expansion, however linkage analysis did not find evidence of linkage disequilibrium. Bayesian analysis also identified a genetic structure with K = 3 being the best estimation of ancestral clusters, although a large proportion of isolates (35%) could not be allocated to a single population, which indicates their mixed origin. The results confirm the genetic distinctiveness of C. parvum in cattle farms in this geographical area. This is the first multilocus analysis on the intra-specific variability of Cryptosporidium from calves in South America.

Influence of Sex and Age on Posterior Semicircular Canal Thickness.

OBJECTIVE: The aim of this study is to determine whether sex and age influence posterior semicircular canal (PSC) thickness. METHODS: This observational study was conducted in 3 tertiary hospitals. The minimal distance between the PSC and the posterior cranial fossa (PSC thickness) was estimated by thin-section multidetector row computed axial tomography (CAT) scan of the temporal bones. Nonselected consecutive patients of all ages (607 temporal bones) were considered. RESULTS: A significant effect was only detected for sex (F = 5.418, p = 0.020); PSC thickness showed a higher mean value in women (mean difference ± SE: 0.224 ± 0.096 mm). A significant and negative r value was detected for males aged >45 years (-0.173, p = 0.026); in that group of patients, PSC thickness decreased as age increased (0.018 ± 0.008 mm/year). For females aged ≤45 years, a significant and positive r value was found (0.198, p = 0.022); in that group, PSC thickness increased as age increased (0.020 ± 0.008 mm/year). CONCLUSIONS: PSC thickness did not significantly evolve with age in young males (≤45 years) but it decreased from age 45 years onwards. On the other hand, PCS thickness increased with age in women until the age of 45 years and it did not significantly change in older females.

Oral Disease and Microbiota in Free-Roaming Cats.

Oral health status was evaluated in 34 free-roaming cats that were submitted to a Trap-Neuter-Return program. Only 38.2% of individual cats showed oral disease. Periodontal disease (PD) was present in 6 cats ( 17.6%). Four cats (11.8%) showed feline gingivostomatitis, and clinical evidence of feline odontoclastic resorptive lesions was detected in only 3 animals (8.8%) by probing, but radiological studies would be necessary for reliable detection. Signs of PD occurred in 31.6% of teeth; only the less severe stages of PD were present. No retention of deciduous teeth, malocclusion, tumors, or dental fractures were found. Calicivirus was detected in 50% of the cats. The most frequent bacteria found were α-hemolytic Streptococcus (23.5%), Staphylococcus (17.6%), Neisseria (15.7%), and Pasteurella (11.8%). A significant association between age and oral disease was found (P = .001). The mean of bacterial strains/genus was higher in healthy mouths than in diseased mouths (4.5 vs. 2.1). In diseased mouths, Staphylococcus was the most frequently isolated genus (23.1%), whereas α-hemolytic Streptococcus was most prevalent in healthy mouths (33.3%). A significant association was detected between Porphyromonas and fungal isolation, with a fear concordance coefficient (κ = .334; P = .016).

Intra-Species Genetic Diversity and Clonal Structure of Cryptosporidium parvum in Sheep Farms in a Confined Geographical Area in Northeastern Spain.

A multilocus fragment typing approach including eleven variable-number tandem-repeat (VNTR) loci and the GP60 gene was used to investigate the intra-farm and intra-host genetic diversity of Cryptosporidium parvum in sheep farms in a confined area in northeastern Spain. Genomic DNA samples of 113 C. parvum isolates from diarrheic pre-weaned lambs collected in 49 meat-type sheep farms were analyzed. Loci exhibited various degrees of polymorphism, the finding of 7-9 alleles in the four most variable and discriminatory markers (ML2, Cgd6_5400, Cgd6_3940, and GP60) being remarkable. The combination of alleles at the twelve loci identified a total of 74 multilocus subtypes (MLTs) and provided a Hunter-Gaston discriminatory index of 0.988 (95% CI, 0.979-0.996). The finding that most MLTs (n = 64) were unique to individual farms evidenced that cryptosporidial infection is mainly transmitted within sheep flocks, with herd-to-herd transmission playing a secondary role. Limited intra- host variability was found, since only five isolates were genotypically mixed. In contrast, a significant intra-farm genetic diversity was seen, with the presence of multiple MLTs on more than a half of the farms (28/46), suggesting frequent mutations or genetic exchange through recombination. Comparison with a previous study in calves in northern Spain using the same 12-loci typing approach showed differences in the identity of major alleles at most loci, with a single MLT being shared between lambs and calves. Analysis of evolutionary descent by the algorithm eBURST indicated a high degree of genetic divergence, with over 41% MLTs appearing as singletons along with a high number of clonal complexes, most of them linking only two MLTs. Bayesian Structure analysis and F statistics also revealed the genetic remoteness of most C. parvum isolates and no ancestral population size was chosen. Linkage analysis evidenced a prevalent pattern of clonality within the parasite population.

Intra-Species Diversity and Panmictic Structure of Cryptosporidium parvum Populations in Cattle Farms in Northern Spain.

The intra-herd and intra-host genetic variability of 123 Cryptosporidium parvum isolates was investigated using a multilocus fragment typing approach with eleven variable-number tandem-repeat (VNTR) loci and the GP60 gene. Isolates were collected from intensively farmed diarrheic pre-weaned calves originating from 31 dairy farms in three adjoining regions in northern Spain (País Vasco, Cantabria and Asturias). The multilocus tool demonstrated an acceptable typeability, with 104/123 samples amplifying at all twelve loci. The ML2, TP14, GP60 and the previously un-described minisatellite at locus cgd2_3850 were the most discriminatory markers, while others may be dismissed as monomorphic (MSB) or less informative (CP47, ML1 and the novel minisatellites at loci Cgd1_3670 and Cgd6_3940). The 12-satellite typing tool provided a Hunter-Gaston index (HGDI) of 0.987 (95% CI, 0.982-0.992), and differentiated a total of 70 multilocus subtypes (MLTs). The inclusion of only the four most discriminatory markers dramatically reduced the number of MLTs (n: 44) but hardly reduced the HGDI value. A total of 54 MLTs were distinctive for individual farms, indicating that cryptosporidiosis is an endemic condition on most cattle farms. However, a high rate of mixed infections was detected, suggesting frequent meiotic recombination. Namely, multiple MLTs were seen in most farms where several specimens were analyzed (90.5%), with up to 9 MLTs being found on one farm, and individual specimens with mixed populations being reported on 11/29 farms. Bayesian Structure analysis showed that over 35% of isolates had mixed ancestry and analysis of evolutionary descent using the eBURST algorithm detected a high rate (21.4%) of MLTs appearing as singletons, indicating a high degree of genetic divergence. Linkage analysis found evidence of linkage equilibrium and an overall panmictic structure within the C. parvum population in this discrete geographical area.

Association Between Tegmen Tympani Status and Superior Semicircular Canal Pattern.

OBJECTIVE: Detecting and quantifying the possible association between tegmen tympani (TT) status and superior semicircular canal (SSC) pattern. DESIGN: Observational study. SETTING: Study conducted in three tertiary Spanish hospitals. PATIENTS: Nonselected consecutive patients of all ages (607 temporal bones). INTERVENTIONS: Thin-section multidetector row computed axial tomography (CAT scan) of the temporal bones. MAIN OUTCOME MEASURE: Thickness of SSC bone coverture adjacent to the middle fossa, and TT status as a dichotomous variable: dehiscence (TTD) or integrity (TTI). RESULTS: The observed SSC patterns were dehiscence (3.79%), papyraceous or thin (11.20%), normal (76.77%), thick (4.94%), and pneumatized (3.29%). The observed TT statuses were TTD (10.87%) and TTI (89.13%). TTD was associated with SSCD and papyraceous patterns, and TTI percentages were higher in normal and thick patterns (χ2 = 11.102; p = 0.001). The TTD probability was estimated as a function of SSC pattern and age by a multivariate binary logistics regression model (χ2 = 45.939; p < 0.001). CONCLUSION: SSC pattern was significantly associated with TT status. Age influenced this association. The risk for TTD increased by 4.1% per each year of increasing age, did not differ significantly for normal and thick patterns, and increased 12 times and 20 times for papyraceous and SSCD patterns, respectively.

Bilateral Ovotestes in a 78, XX SRY-Negative Beagle Dog.

This report describes a disorder of the sexual development in a beagle dog resulting in an intersex condition. A 6 mo old beagle was presented for evaluation of a protruding structure from the vulva consistent with an enlarged clitoris. Ultrasonographic examination revealed the presence of both gonadal and uterine structures. Retrograde cystourethrovaginogram showed the presence of an os clitoris and severe vaginal stenosis. Histological studies revealed the presence of bilateral ovotestes and uterus. The gonad had interstitial cells within seminiferous-like tubules lined only with Sertoli cells and abundant interstitial cells among primordial, primary, and secondary follicles. Hormone assays completed before and after gonadohysterectomy showed an elevation in the levels of progesterone and dihydrotestosterone that returned to baseline 3 mo after surgery. Testosterone levels that were within the male reference ranges before surgery decreased to basal levels postsurgically. 17-ß-Estradiol levels showed little variation and values were always within the reference ranges for a male. Cytogenetic analysis showed a normal female karyotype (2n = 78, XX) and polymerase chain reaction analysis revealed the absence of the sex-determining region Y gene. In summary, the dog presented bilateral ovotestes and a 2n = 78, XX chromosomal complement lacking the sex determining region Y gene, consistent with a diagnosis of true hermaphroditism.

Ovine congenital myotonia associated with a mutation in the muscle chloride channel gene.

Congenital myotonia (CM) is characterised by a delay in muscular relaxation after sudden contractions. In a recent outbreak of ovine CM affecting 1% of new-born lambs in a Spanish flock of Rasa Aragonesa sheep, a comparative pathology approach was taken: because a mutation in the muscle chloride channel gene (CLCN1) was identified as responsible for CM in goats, the same gene was sequenced in the affected lambs. A non-synonymous single nucleotide variation (SNV) in the second exon of CLCN1 was associated with this pathology. Rams carrying this SNV heterozygously were thereafter identified and replaced by wild-type homozygous young males. No additional CM cases were detected in subsequent lambing seasons.