Comparison of Hounsfield Units within the Humeral Trochlea and Medial Coronoid Process in a Population of Labrador X Golden Retriever Guide Dogs and Border Collies.

OBJECTIVE: The aim of this study was to determine if variations in Hounsfield units (HU) are present within the elbow between asymptomatic dogs of two breeds of dogs susceptible to elbow dysplasia. STUDY DESIGN: Guide Dogs and Border Collies that presented for routine computed tomography (CT) screening for elbow dysplasia prior to breeding were evaluated. All dogs had no documented history of lameness. Dogs diagnosed with CT as being free of elbow dysplasia were included. The CT images were randomized and assessed by three blinded observers. A standardised approach to CT image reconstruction to create consistent image planes was used. Hounsfield units were measured within a standardised region of interest (ROI) at the humeral trochlea and medial coronoid process. The minimum, mean and maximum HU within each ROI was recorded. RESULTS: Eighty-six elbows were included in the study with 32 Guide Dogs, and 11 Border Collies. Guide Dogs had significantly higher minimum (99.75 HU, 95% confidence interval [CI]: 15.02-184.48, p = 0.022), mean (115.09 HU, 95% CI: 80.53-149.64, p < 0.01) and maximum (74.00 HU, 95% CI: 44.58-103.42, p < 0.01) difference in HU within the medial coronoid process ROI, and significantly higher mean (146.49 HU, 95% CI: 100.12-192.87, p < 0.01) and maximum (147.77 HU, 95% CI: 102.57-192.97, p < 0.01) difference in HU within the humeral trochlea ROI. CONCLUSION: In this dataset breed variations in elbow HU were present between asymptomatic Guide Dogs and Border Collies. This needs to be considered in breeding screening programmes to avoid over-interpretation of elbow sclerosis, in the absence of elbow pathology.

Presumed septic sacroiliitis in a puppy with unilateral hind limb lameness and sciatic nerve neuropathy.

A 5-month-old female entire Dachshund presented with an acute onset of left hind limb lameness following intense play. There were concurrent intermittent neurological deficits in the left hind limb, and pain in the lumbosacral region and on dorsal flexion of the tail. Computed tomography (CT) imaging revealed an asymmetric widening of the left sacroiliac joint with loss of cortical margins, accompanied by mild osteolytic changes of the adjacent ilium and sacrum highly suggestive of septic sacroiliitis. There was also perilesional steatitis in the region of the sciatic nerve. An 8-week course of antimicrobial therapy was prescribed. Complete resolution of all clinical signs was noted at a 7-month follow up. Sacroiliitis should be considered as a differential for unilateral hind limb lameness with or without intermittent sciatic neuropathy in a puppy in the absence of other orthopaedic conditions.

Clinical outcomes of patient-specific porous titanium endoprostheses in dogs with tumors of the mandible, radius, or tibia: 12 cases (2013-2016).

OBJECTIVE To characterize the processes involved in and outcomes achieved with custom-designed patient-specific implants to provide functional replacement of skeletal structures in dogs with tumors of the mandible, radius, or tibia. DESIGN Prospective case series. ANIMALS 6 dogs with mandibular tumors, 5 with tumors of the distal aspect of the radius, and 1 with a tumor in the distal aspect of the tibia treated from June 2013 to September 2016 at 3 referral centers. PROCEDURES After tumor staging, implants were designed from patients' CT scans by means of various computer-aided design applications and printed by means of selective laser melting in titanium-6 aluminum-4 vanadium alloy. A cutting jig was created in thermoplastic to ensure each osteotomy was performed as planned. Following ostectomy, the implant was secured into the defect with screws of appropriate size and length. RESULTS Initial return to normal clinical function was good to excellent for 11 of the 12 dogs. However, major complications resulted in revision of the implant or amputation of the limb in 5 dogs, and at least 3 of these complications were considered a consequence of faulty implant design or manufacturing. Infection developed in 2 dogs and was successfully treated in 1 dog. The longest-surviving dog maintained good limb function for 2 years. CONCLUSIONS AND CLINICAL RELEVANCE This is the largest reported series of dogs managed with customized 3-D-printed titanium implants. The 3-D printing allowed complex and patient-specific 3-D geometries to be fabricated, enabling function-sparing treatment of bone cancer affecting multiple anatomic sites.

Clinical outcome and complications of thoracic and pelvic limb stump and socket prostheses.

OBJECTIVES: To describe the use, quality of life, compliance, complications, and outcome of animals fitted with stump socket prostheses. METHODS: Medical records of dogs fitted with a stump socket prosthesis were reviewed. Functional outcome, quality of life and complications were retrospectively assessed from an owner questionnaire. RESULTS: Thirteen stump socket prostheses (12 dogs) were fitted for a variety of reasons including trauma, congenital abnormalities, and neoplasia. Eight dogs had a good outcome overall and four a poor outcome. Quality of life (QOL) remained good or excellent in 10/12 dogs. Nine complications were seen in 7/12 dogs, most were manageable; surgical wound complications (n = 2) and pressures sores (n = 4) were the most frequently encountered. One dog suffered multiple complications. Thoracic and pelvic limb stump socket prostheses had a similar complication rate, however all animals with a poor outcome had a thoracic limb stump socket prosthesis; two were small breed dogs (under 10 kg) and two had bilateral thoracic limb abnormalities. CLINICAL SIGNIFICANCE: Stump socket prostheses are feasible and versatile in animals. In correctly selected cases, good to excellent outcomes are possible. However, complications are frequent but often manageable. Further investigations are required into the risk factors for poor outcomes and prospective studies are required to assess changes in biomechanics, function, and QOL before and after fitting of a stump socket prosthesis. Until further evidence is available, careful consideration should be given before fitting bilateral thoracic limb stump socket prostheses or thoracic limb stump socket prostheses to small breed dogs.

Retrospective evaluation of recurrent secondary septic peritonitis in dogs (2000-2011): 41 cases.

OBJECTIVE: To describe the clinical characteristics of recurrent septic peritonitis in dogs. DESIGN: Multicenter retrospective observational study. ANIMALS: Client-owned dogs with recurrent septic peritonitis. SETTING: Three university emergency and referral hospitals. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Medical records from 3 veterinary university teaching hospitals were reviewed and data were collected using a standardized data collection sheet for all cases of septic peritonitis during the study period (2000-2011). Forty one dogs met the inclusion criteria for recurrent peritonitis. All dogs underwent relaparotomy. The original cause of septic peritonitis in these cases included previous surgery for gastrointestinal foreign body removal (n = 26), gastrointestinal neoplasia (n = 3), gastric or duodenal ulceration (n = 3), biliary tract leakage (n = 2), and single instance for each of the following: penetrating foreign body, hernia strangulation, intussusception, mesenteric volvulus, infection of the laparotomy incision, prostatic abscess, and trauma. Eighteen animals survived to discharge. There was no difference detected between survivors and nonsurvivors with recurrent peritonitis in terms of inciting cause, serum albumin concentration, surgical management, or provision of appropriate initial antimicrobials. The survival rate for dogs having recurrent peritonitis was 43.9% (18/41 dogs). CONCLUSIONS: This retrospective study did not identify any significant prognostic indicators for dogs with recurrent peritonitis and that the mortality rate for dogs having more than one surgery for septic peritonitis is similar to that reported for a single surgery for septic peritonitis.

Genetics of canine anal furunculosis in the German shepherd dog.

Canine anal furunculosis (AF) is characterised by ulceration and fistulation of perianal tissue and is a disease that particularly affects German shepherd dogs (GSDs). There are some similarities between AF and perianal Crohn's disease (CD) in man. An immune-mediated aetiopathogenesis for AF has been suggested due to tissue pathology, a major histocompatibility complex (MHC) association and clinical response to ciclosporin. Genome-wide association studies (GWAS) can be conducted in dogs with fewer markers and individuals than would be required in a human study. A discovery GWAS was performed on 21 affected and 25 control GSDs from the UK. No SNPs reached genome-wide significance levels at this stage. However, 127 nominally associated SNPs were genotyped in further 76 cases and 191 controls from the UK and Finland. Sequencing of these regions was undertaken to discover novel genetic variation. Association testing of these variants in the UK and Finnish cohorts revealed nine significantly associated SNPs, six of which cause non-synonymous changes in protein sequence. The ADAMTS16 and CTNND2 gene regions were most significantly associated with disease. Members of the butyrophilin protein family, important in intestinal inflammatory regulation, were also associated with disease, but their independence from the MHC region remains to be established. The CTNND2 gene region is also interesting as this locus was implicated in human ulcerative colitis and CD, albeit at a different candidate gene: DAP. We suggest that this represents a common association between inflammatory bowel disease-related conditions in both species and believe that future studies will strengthen this link.

Markers of hepatic regeneration associated with surgical attenuation of congenital portosystemic shunts in dogs.

Dogs with congenital portosystemic shunts (CPSS) have liver hypoplasia and hepatic insufficiency. Surgical CPSS attenuation results in liver growth associated with clinical improvement. The mechanism of this hepatic response is unknown, although liver regeneration is suspected. This study investigated whether markers of liver regeneration were associated with CPSS attenuation. Dogs treated with CPSS attenuation were prospectively recruited. Residual liver tissue was collected for gene expression analysis (seven genes) from 24 CPSS dogs that tolerated complete attenuation, 25 dogs that tolerated partial attenuation and seven control dogs. Relative gene expression was measured using quantitative polymerase chain reaction (qPCR). Blood samples were collected before, 24 h and 48 h post-surgery from 36 CPSS dogs and from 10 control dogs. Serum hepatocyte growth factor (HGF) concentration was measured using a canine specific enzyme-linked immunosorbent assay (ELISA). HGF mRNA expression was significantly decreased in CPSS compared with control dogs (P = 0.046). There were significant increases in HGF (P = 0.050) and methionine adenosyltransferase 2 A (MAT2A; P = 0.002) mRNA expression following partial CPSS attenuation. Dogs with complete attenuation had significantly greater MAT2A (P = 0.024) mRNA expression compared with dogs with partial attenuation. Serum HGF concentration significantly increased 24 h following CPSS attenuation (P < 0.001). Hepatic mRNA expression of two markers of hepatocyte proliferation (HGF and MAT2A) was associated with the response to surgery in dogs with CPSS, and serum HGF significantly increased following surgery, suggesting hepatocyte proliferation. These findings support the concept that hepatic regeneration is important in the hepatic response to CPSS surgery.

Polymorphisms in the TLR4 and TLR5 gene are significantly associated with inflammatory bowel disease in German shepherd dogs.

Inflammatory bowel disease (IBD) is considered to be the most common cause of vomiting and diarrhoea in dogs, and the German shepherd dog (GSD) is particularly susceptible. The exact aetiology of IBD is unknown, however associations have been identified between specific single-nucleotide polymorphisms (SNPs) in Toll-like receptors (TLRs) and human IBD. However, to date, no genetic studies have been undertaken in canine IBD. The aim of this study was to investigate whether polymorphisms in canine TLR 2, 4 and 5 genes are associated with IBD in GSDs. Mutational analysis of TLR2, TLR4 and TLR5 was performed in 10 unrelated GSDs with IBD. Four non-synonymous SNPs (T23C, G1039A, A1571T and G1807A) were identified in the TLR4 gene, and three non-synonymous SNPs (G22A, C100T and T1844C) were identified in the TLR5 gene. The non-synonymous SNPs identified in TLR4 and TLR5 were evaluated further in a case-control study using a SNaPSHOT multiplex reaction. Sequencing information from 55 unrelated GSDs with IBD were compared to a control group consisting of 61 unrelated GSDs. The G22A SNP in TLR5 was significantly associated with IBD in GSDs, whereas the remaining two SNPs were found to be significantly protective for IBD. Furthermore, the two SNPs in TLR4 (A1571T and G1807A) were in complete linkage disequilibrium, and were also significantly associated with IBD. The TLR5 risk haplotype (ACC) without the two associated TLR4 SNP alleles was significantly associated with IBD, however the presence of the two TLR4 SNP risk alleles without the TLR5 risk haplotype was not statistically associated with IBD. Our study suggests that the three TLR5 SNPs and two TLR4 SNPs; A1571T and G1807A could play a role in the pathogenesis of IBD in GSDs. Further studies are required to confirm the functional importance of these polymorphisms in the pathogenesis of this disease.

Interleukin-2 and interferon-gamma mRNA expression in canine anal furunculosis lesions and the effect of ciclosporin therapy.

German Shepherd Dogs have an increased incidence of anal furunculosis (AF), which is a disease characterised by inflammation and ulceration of the perianal tissues. Ciclosporin, an immunosuppressive drug, has been successfully used to treat AF, suggesting that the pathogenesis of disease is likely to have an immune-mediated component. Previous research has shown that the cytokine mRNA profile in AF lesions is consistent with T cell-mediated inflammation. The aims of the current study were to quantify IL-2 and IFNgamma mRNA expression in AF biopsies taken before and after treatment with ciclosporin and to compare cytokine expression with lesion severity. Twenty-two dogs with AF were recruited into the study and lesional biopsies were taken prior to ciclosporin therapy. Lesion severity was graded using a visual analogue scale. All dogs were evaluated after 4 weeks of ciclosporin therapy and, in 10 dogs with persistent disease, residual lesions were resected. RNA was extracted from AF-lesional tissue and control perianal tissue samples (n=10), which was used as the template for RT-PCR. Analysis of IL-2 and IFNgamma mRNA expression was performed using real-time PCR. IL-2 and IFNgamma mRNA was consistently detected in pre-treatment AF biopsies and, when quantified, this was significantly increased compared to control tissue (P<0.05). However, no correlation was seen between lesion severity and pre-treatment cytokine mRNA expression. In the ten paired pre- and post-treatment samples, IL-2 mRNA expression was significantly reduced in residual disease tissue following ciclosporin therapy (P=0.013). Treatment with ciclosporin seemed to result in decreased expression of IFNgamma mRNA in AF lesions but this was not statistically significant. In six of the 10 dogs with persistent disease, T cell cytokine mRNA could still be detected in the tissues, suggesting that there was inadequate immunosuppression. The absence of a correlation between T cell cytokine expression and the severity of disease suggests that tissue destruction observed in AF might be a consequence of other inflammatory mediators or downstream effects of T cell activation.

Pattern-recognition receptor mRNA expression and function in canine monocyte/macrophages and relevance to canine anal furunuclosis.

Pattern-recognition receptors (PRRs) are important components of the innate immune system, enabling early detection of infection. Defective PRR function has been implicated in several infectious and immune-mediated diseases of human beings, including Crohn's disease (CD). Anal furunculosis (AF) is an immune-mediated disease which primarily occurs in German shepherd dogs (GSD) and could result from a similar type of PRR dysfunction. The aim of the current study was to investigate canine PRR responses in vitro and to test the hypothesis that these were altered in AF-affected GSD. The pattern-recognition receptors TLR1, TLR2, TLR4, TLR6, TLR9, NOD1 (nucleotide-binding oligomerisation domain) and NOD2 were evaluated in the DH82 canine monocyte/macrophage cell line. These cells were found to express mRNA for all the selected PRRs with TLR2 mRNA the most and TLR5 mRNA the least abundant. A similar pattern of expression was found in canine blood-derived monocyte/macrophages. Stimulation of DH82 cells and blood-derived monocyte/macrophages using specific PRR-ligands, resulted in expression of pro-inflammatory cytokine mRNA. Quantification of TNFalpha mRNA and protein secretion from stimulated cells demonstrated variable responses with lipopolysaccharide (TLR4 ligand) and PAM(3)CSK4 (TLR1/2 ligand) proving to be the most potent and CpG DNA (TLR9 ligand) the least potent. Comparing PRR responses in blood-derived monocyte/macrophages from healthy blood-donor dogs with those from AF-affected GSD showed a deficiency in the latter in response to LD-MDP (NOD2 ligand) at the mRNA level but not at the protein level. It is possible that dysfunctional NOD2 responses by cells of the monocyte/macrophage lineage are involved in the pathogenesis of AF.

Evaluation of the effect of two dose rates of cyclosporine on the severity of perianal fistulae lesions and associated clinical signs in dogs.

OBJECTIVE: To investigate the effect of cyclosporine (2 or 5 mg/kg every 24 hours) on perianal fistulae (PAF) lesions. STUDY DESIGN: Blinded randomized, prospective trial. ANIMALS: Dogs (n = 20) with perianal fistulae. METHODS: Dogs were randomly assigned to administration of either 2 mg/kg (n = 10) or 5 mg/kg (n=10) of cyclosporine orally every 24 hours for 8 weeks. Lesion surface area was measured, lesion severity was graded using a visual analog scale, and the presence and severity of clinical signs recorded every 2 weeks. RESULTS: Lesion variables were significantly reduced in both groups after 8 weeks and owners also reported a reduction in clinical sign severity. The 5 mg/kg dose rate significantly accelerated lesion resolution compared with 2 mg/kg. In the 2 mg/kg group, 20% of dogs had complete resolution of clinical signs and 10% had resolution of lesions. In the 5 mg/kg group, 40% of dogs had complete resolution of clinical signs and 60% had resolution of lesions. CONCLUSIONS: A dose rate of 5 mg/kg every 24 hours was more effective at reducing the surface area and severity of PAF lesions than 2 mg/kg every 24 hours but less effective at resolving PAF lesions than previous studies using dose rates > or =5 mg/kg every 12 hours. CLINICAL RELEVANCE: Cyclosporine at 5 mg/kg every 24 hours may be useful for the palliation of PAF lesions.