Skin microbiota and allergic symptoms associate with exposure to environmental microbes.

A rural environment and farming lifestyle are known to provide protection against allergic diseases. This protective effect is expected to be mediated via exposure to environmental microbes that are needed to support a normal immune tolerance. However, the triangle of interactions between environmental microbes, host microbiota, and immune system remains poorly understood. Here, we have studied these interactions using a canine model (two breeds, n = 169), providing an intermediate approach between complex human studies and artificial mouse model studies. We show that the skin microbiota reflects both the living environment and the lifestyle of a dog. Remarkably, the prevalence of spontaneous allergies is also associated with residential environment and lifestyle, such that allergies are most common among urban dogs living in single-person families without other animal contacts, and least common among rural dogs having opposite lifestyle features. Thus, we show that living environment and lifestyle concurrently associate with skin microbiota and allergies, suggesting that these factors might be causally related. Moreover, microbes commonly found on human skin tend to dominate the urban canine skin microbiota, while environmental microbes are rich in the rural canine skin microbiota. This in turn suggests that skin microbiota is a feasible indicator of exposure to environmental microbes. As short-term exposure to environmental microbes via exercise is not associated with allergies, we conclude that prominent and sustained exposure to environmental microbiotas should be promoted by urban planning and lifestyle changes to support health of urban populations.

Changes in biomarkers in equine synovial fluid two weeks after intra-articular hyaluronan treatment: a randomised double-blind clinical trial.

BACKGROUND: Inflammatory and degenerative activity inside the joint can be studied in vivo via analysis of synovial fluid (SF) biomarkers, which are molecular markers of inflammatory processes and tissue turnover. The aim of this study was to investigate the response of selected biomarkers in the SF after an intra-articular (IA) high-molecular-weight non-animal stabilized hyaluronic acid (NASHA) treatment. Our hypothesis was that prostaglandin E2 (PGE2), substance P, aggrecan chondroitin sulfate 846 epitope (CS846), and carboxypeptide of type II collagen (CPII) concentrations in SF would decrease more in the NASHA than in the placebo group. Twenty-eight clinically lame horses with positive responses to diagnostic IA anaesthesia of the metacarpophalangeal or metatarsophalangeal joints were randomized into treatment (n = 15) and control (n = 13) groups. After collection of baseline SF samples followed by IA diagnostic anaesthesia, horses in the treatment group received 3 ml of a NASHA product IA. Those in the placebo group received an equivalent volume of sterile 0.9% saline solution. The horses were re-evaluated and a second SF sample was obtained after a 2-week period. RESULTS: CS846 concentration decreased in the NASHA group only (P = 0.010). Both PGE2 and CPII concentrations decreased within the groups (PGE2, P = 0.010 for the NASHA group; P = 0.027 for the placebo group; CPII, P < 0.001 for NASHA group; P = 0.009 for placebo group). No significant treatment effect for any biomarker was found between groups. NASHA induced an increase in white blood cell count; this was significant compared with baseline (P = 0.021) and the placebo group (P = 0.045). CONCLUSIONS: Although the SF concentration of the cartilage-derived biomarker CS846 decreased in the NASHA group, no statistically significant treatment effect of any of the biomarkers were observed between treatment groups. The significant increase in SF white blood cell count after IA NASHA may indicate a mild inflammatory response. However, as no clinical adverse effects were observed, we conclude that IA NASHA was well tolerated.

The effect of intra-articular botulinum toxin A on substance P, prostaglandin E2, and tumor necrosis factor alpha in the canine osteoarthritic joint.

BACKGROUND: Recently, intra-articular botulinum toxin A (IA BoNT A) has been shown to reduce joint pain in osteoarthritic dogs. Similar results have been reported in human patients with arthritis. However, the mechanism of the antinociceptive action of IA BoNT A is currently not known. The aim of this study was to explore this mechanism of action by investigating the effect of IA BoNT A on synovial fluid (SF) and serum substance P (SP), prostaglandin E2 (PGE2), and tumor necrosis factor alpha (TNF-α) in osteoarthritic dogs. Additionally, the aim was to compare SF SP and PGE2 between osteoarthritic and non-osteoarthritic joints, and investigate associations between SP, PGE2, osteoarthritic pain, and the signalment of dogs. Thirty-five dogs with chronic naturally occurring osteoarthritis and 13 non-osteoarthritic control dogs were included in the study. Osteoarthritic dogs received either IA BoNT A (n = 19) or IA placebo (n = 16). Serum and SF samples were collected and osteoarthritic pain was evaluated before (baseline) and 2 and 8 weeks after treatment. Osteoarthritic pain was assessed with force platform, Helsinki Chronic Pain Index, and joint palpation. Synovial fluid samples were obtained from control dogs after euthanasia. The change from baseline in SP and PGE2 concentration was compared between the IA BoNT A and placebo groups. The synovial fluid SP and PGE2 concentration was compared between osteoarthritic and control joints. Associations between SP, PGE2, osteoarthritic pain, and the signalment of dogs were evaluated. RESULTS: There was no significant change from baseline in SP or PGE2 after IA BoNT A. Synovial fluid PGE2 was significantly higher in osteoarthritic compared to control joints. Synovial fluid PGE2 correlated with osteoarthritic pain. No associations were found between SP or PGE2 and the signalment of dogs. The concentration of TNF-α remained under the detection limit of the assay in all samples. CONCLUSIONS: The results suggest that the antinociceptive effect of IA BoNT A in the joint might not be related to the inhibition of SP nor PGE2. Synovial fluid PGE2, but not SP, could be a marker for chronic osteoarthritis and pain in dogs.

Prevalence of house dust mites in the homes of atopic dogs in Finland.

BACKGROUND: The house dust mites (HDM) Dermatophagoides farinae and D. pteronyssinus are important environmental allergens implicated in the pathogenesis of human and canine atopic dermatitis. Sensitization to HDM measured by allergen-specific IgE is common in Finnish atopic dogs. Studies on HDM prevalence in Finland are few but suggest that HDM are scarce. OBJECTIVE: The aim of the present study was to evaluate the prevalence of HDM in the home environments of atopic dogs in Finland. METHODS: Dust samples were obtained from the homes of 50 atopic dogs. Samples were collected by vacuuming the owners' mattresses and each dog's bed. In each case, an area of 21 × 30 cm was vacuumed for 2 min. Samples weighing 100 mg or more were considered sufficient for determination of HDM allergen concentrations (Der f 1 and Der p 1) using standardized ELISA. Samples sufficient for further analysis were also examined by direct microscopy for the presence of mites and by multiplex PCR for HDM DNA. RESULTS: Eighty one samples were sufficient for analysis by ELISA, 59 by PCR and 29 by direct microscopy. A single sample was analysed from four homes in which the dog shared the owner's bed. Der f 1 was detected in three samples (3.7%). Der p 1 was not detected in any sample. No mites were identified on microscopy. Five samples were positive for HDM on multiplex PCR (8.4%). CONCLUSION: House dust mites seem to be uncommon in the home environment of atopic dogs in Finland despite reported frequent allergen-specific IgE antibodies.

A randomised, double-blinded, placebo-controlled clinical study on intra-articular hyaluronan treatment in equine lameness originating from the metacarpophalangeal joint.

BACKGROUND: Intra-articular inflammation resulting in lameness is a common health problem in horses. Exogenous intra-articular hyaluronic acid has been shown to provide an analgesic effect and reduce pain in equine and human osteoarthritis. High molecular weight non-animal stabilized hyaluronic acid (NASHA) has gained popularity in the treatment of human arthritic conditions due to its long-acting pain-relieving effects. The aim of this study was to compare the response to treatment of lameness localized in the equine metacarpophalangeal joint injected with non-animal stabilized hyaluronic acid (NASHA) and placebo (saline). Twenty-seven clinically lame horses with a positive response to diagnostic intra-articular anaesthesia of the metacarpophalangeal joint and with no, or at most mild, radiographic changes in this joint were included in the study. Horses in the treatment group (n = 14) received 3 mL of a NASHA product intra-articularly, and those in the placebo group (n = 13) received an equivalent volume of sterile 0.9% saline solution. RESULTS: The change in the lameness score did not significantly differ between NASHA and placebo groups (P = 0.94). Scores in the flexion test improved more in the NASHA group compared with placebo (P = 0.01). The changes in effusion and pain in flexion were similar (P = 0.94 and P = 0.27, respectively) when NASHA and placebo groups were compared. A telephone interview follow-up of the owners three months post-treatment revealed that 14 of the 21 horses (67%) were able to perform at their previous level of exercise. CONCLUSIONS: In the present study, a single IA NASHA injection was not better than a single saline injection for reducing lameness in horses with synovitis or mild osteoarthritis. However, the results of this study indicate that IA NASHA may have some beneficial effects in modifying mild clinical signs but more research is needed to evaluate whether the positive effect documented ie. reduced response in the flexion test is a true treatment effect.

Validating and reliability testing the descriptive data and three different disease diagnoses of the internet-based DOGRISK questionnaire.

BACKGROUND: The DOGRISK questionnaire is an internet-based ongoing study of canine nutrition, living environment, and disease. Here we aim to assess the performance of the questionnaire using data from the first three years in relation to some descriptive and disease variables. We used associated questions, official register records, test-retest repeatability, and email/mail contact with questionnaire respondents. RESULTS: Reliability against an official register of gender, season of birth, breed, and results of hip radiography was tested and Cohen's Kappa was between 0.95-0.99. Internal consistencies of hypothyroidism status and dog's age were calculated using Cronbach's Alpha (α = 0.95 and α = 0.99, respectively). Test-retest repeatability of ten variables among 224 participants was analyzed. Gender, season of birth, and born in owner family had Cohen's Kappa > 0.86, color of coat, vaccination status as an adult, other dogs in household, and vaccination status as a puppy had Cohen's Kappa between 0.67-0.80, and body condition score under two months of age and tidiness of household, had Cohen's kappa of 0.45 and 0.42, respectively. In addition, time spent outside had Cohen's kappa of 0.37. Of the owners contacted by email/mail to confirm their dog's atopy/allergy (skin symptoms), 8.9% reported that they had given an incorrect answer (positive predicted value 91%), but only 69% of all reaffirmed positive answers had a diagnosis set by a veterinarian. CONCLUSIONS: Our study showed that owners were diligent with basic information and with the status of three diseases. Cohen's Kappa in the reliability of the test-retest was in most variables at least 0.67. We propose that the descriptive variables and the disease variables be used as such when we generate hypotheses from the DOGRISK data.

Evaluating oxidative stress, serological- and haematological status of dogs suffering from osteoarthritis, after supplementing their diet with fish or corn oil.

BACKGROUND: Oxidative stress plays an important role in the pathogenesis of disease, and the antioxidant physiological effect of omega-3 from fish oil may lead to improvement of canine spontaneous osteoarthritis (OA). METHODS: In this prospective randomized, controlled, double-blinded study, we assessed haematological and biochemical parameters in dogs with OA following supplementation with either a concentrated omega-3 deep sea fish oil product or corn oil. Blood samples from 77 client-owned dogs diagnosed as having OA were taken before (baseline) and 16 weeks after having orally ingested 0.2 ml/Kg bodyweight/day of deep sea fish oil or corn oil. Circulating malondialdehyde (MDA), glutathione (GSH), non-transferrin bound iron (NTBI), free carnitine (Free-Car), 8-hydroxy-2-deoxyguanosine (8-OH-dG), and serum fatty acids, haemograms and serum biochemistry were evaluated. Differences within and between groups from baseline to end, were analysed using repeated samples T-test or Wilcoxon rank test and independent samples T-test or a Mann-Whitney test. RESULTS: Supplementation with fish oil resulted in a significant reduction from day 0 to day 112 in MDA (from 3.41 ± 1.34 to 2.43 ± 0.92 µmol/L; P < 0.001) and an elevation in Free-Car (from 18.18 ± 9.78 to 21.19 ± 9.58 µmol/L; P = 0.004) concentrations, whereas dogs receiving corn oil presented a reduction in MDA (from 3.41 ± 1.34 to 2.41 ± 1.01 µmol/L; P = 0.001) and NTBI (from -1.25 ± 2.17 to -2.31 ± 1.64 µmol/L; P = 0.002). Both groups showed increased (albeit not significantly) GSH and 8-OH-dG blood values. Dogs supplemented with fish oil had a significant reduction in the proportions of monocytes (from 3.84 ± 2.50 to 1.77 ± 1.92 %; P = 0.030) and basophils (from 1.47 ± 1.22 to 0.62 ± 0.62 %; P = 0.012), whereas a significant reduction in platelets counts (from 316.13 ± 93.83 to 288.41 ± 101.68 × 10(9)/L; P = 0.029), and an elevation in glucose (from 5.18 ± 0.37 to 5.32 ± 0.47 mmol/L; P = 0.041) and cholesterol (from 7.13 ± 1.62 to 7.73 ± 2.03 mmol/L; P = 0.011) measurements were observed in dogs receiving corn oil. CONCLUSIONS: In canine OA, supplementation with deep sea fish oil improved diverse markers of oxidative status in the dogs studied. As corn oil also contributed to the reduction in certain oxidative markers, albeit to a lesser degree, there was no clear difference between the two oil groups. No clinical, haematological or biochemical evidence of side effects emerged related to supplementation of either oil. Although a shift in blood fatty acid values was apparent due to the type of nutraceutical product given to the dogs, corn oil seems not to be a good placebo.

Long-term functional outcome after surgical repair of cranial cruciate ligament disease in dogs.

BACKGROUND: Cranial cruciate ligament (CCL) rupture is a very common cause of pelvic limb lameness in dogs. Few studies, using objective and validated outcome evaluation methods, have been published to evaluate long-term (>1 year) outcome after CCL repair. A group of 47 dogs with CCL rupture treated with intracapsular, extracapsular, and osteotomy techniques, and 21 healthy control dogs were enrolled in this study. To evaluate long-term surgical outcome, at a minimum of 1.5 years after unilateral CCL surgery, force plate, orthopedic, radiographic, and physiotherapeutic examinations, including evaluation of active range of motion (AROM), symmetry of thrust from the ground, symmetry of muscle mass, and static weight bearing (SWB) of pelvic limbs, and goniometry of the stifle and tarsal joints, were done. RESULTS: At a mean of 2.8 ± 0.9 years after surgery, no significant differences were found in average ground reaction forces or SWB between the surgically treated and control dog limbs, when dogs with no other orthopedic findings were included (n = 21). However, in surgically treated limbs, approximately 30% of the dogs had decreased static or dynamic weight bearing when symmetry of weight bearing was evaluated, 40-50% of dogs showed limitations of AROM in sitting position, and two-thirds of dogs had weakness in thrust from the ground. The stifle joint extension angles were lower (P <0.001) and flexion angles higher (P <0.001) in surgically treated than in contralateral joints, when dogs with no contralateral stifle problems were included (n = 33). In dogs treated using the intracapsular technique, the distribution percentage per limb of peak vertical force (DPVF) in surgically treated limbs was significantly lower than in dogs treated with osteotomy techniques (P =0.044). CONCLUSIONS: The average long-term dynamic and static weight bearing of the surgically treated limbs returned to the level of healthy limbs. However, extension and flexion angles of the surgically treated stifles remained inferior to healthy joints, and impairment of AROM and weakness in thrust from the ground in the surgically treated limbs were frequently present. Ground reaction forces may be inadequate as a sole method for assessing functional outcome after cranial cruciate ligament repair.

Scent Detection Threshold of Trained Dogs to Eucalyptus Hydrolat.

Dogs' (Canis lupus familiaris) sense of smell is based on a unique anatomy and physiology that enables them to find and differentiate low concentrations of odor molecules. This ability is exploited when dogs are trained as search, rescue, or medical detection dogs. We performed a three-part study to explore the scent detection threshold of 15 dogs to an in-house-made Eucalyptus hydrolat. Here, decreasing concentrations of the hydrolat were tested using a three-alternative forced-choice method until the first incorrect response, which defined the limit of scent detection for each tested dog. Quantitative proton nuclear magnetic resonance spectroscopy was used to identify and measure the contents of ten commercial Eucalyptus hydrolats, which are used in a dog scent training sport called "nose work". In this study, the dogs' limit of detection initially ranged from 1:104 to 1:1023 but narrowed down to 1:1017-1:1021 after a training period. The results show that, with training, dogs learn to discriminate decreasing concentrations of a target scent, and that dogs can discriminate Eucalyptus hydrolat at very low concentrations. We also detected different concentrations of eucalyptol and lower alcohols in the hydrolat products and highlight the importance of using an identical source of a scent in training a dog for participation in canine scent sport competitions and in olfactory research.

Distinct healthy and atopic canine gut microbiota is influenced by diet and antibiotics.

The rising trend in non-communicable chronic inflammatory diseases coincides with changes in Western lifestyle. While changes in the human microbiota may play a central role in the development of chronic diseases, estimating the contribution of associated lifestyle factors remains challenging. We studied the influence of lifestyle-diet, antibiotic use, and residential environment with housing and family-on the gut microbiota of healthy and owner-reported atopic pet dogs, searching for associations between the lifestyle factors, atopy and microbiota. The results showed that atopic and healthy dogs had contrasting gut microbial composition. The gut microbiota also differed between two breeds, Labrador Retriever and Finnish Lapphund, selected for our study. Among all lifestyle factors studied, diet was most significantly associated with gut microbiota but only weakly with atopic symptoms. Thus, diet- and atopy-associated changes in the microbiota were not interrelated. Instead, the severity of symptoms was positively associated with the usage of antibiotics, which in turn was associated with the microbiota composition. Urban lifestyle was significantly associated with the increased prevalence of allergies but not with the gut microbiota. Our results from pet dogs supported previous evidence from humans, demonstrating that antibiotics, gut microbiota and atopic manifestation are interrelated. This congruence suggests that canine atopy might be a promising model for understanding the aetiology of human allergy.

The effect of puppyhood and adolescent diet on the incidence of chronic enteropathy in dogs later in life.

Diet has a key role in the homeostasis of the gut microenvironment, influencing the microbiome, the gut barrier, host immunity and gut physiology. Yet, there is little information on the role of early diet in the onset of inflammatory gastrointestinal disorders later in life, especially in dogs. Therefore, the aim of the present cross-sectional, epidemiological study with longitudinal data, was to explore associations of companion dogs' early life diet style and food items with owner-reported chronic enteropathy (CE) incidence in later life. Food frequency questionnaire data from Finnish companion dogs was analyzed using principal component analysis and logistic regression. We found that feeding a non-processed meat-based diet and giving the dog human meal leftovers and table scraps during puppyhood (2-6 months) and adolescence (6-18 months) were protective against CE later in life. Especially raw bones and cartilage as well as leftovers and table scraps during puppyhood and adolescence, and berries during puppyhood were associated with less CE. In contrast, feeding an ultra-processed carbohydrate-based diet, namely dry dog food or "kibble" during puppyhood and adolescence, and rawhides during puppyhood were significant risk factors for CE later in life.

Psychometric Testing and Validation of the Italian Version of the Helsinki Chronic Pain Index (I-HCPI) in Dogs with Pain Related to Osteoarthritis.

Pain assessment is of paramount importance for properly managing dogs with osteoarthritis (OA) pain. The aim of the present study was to develop and psychometrically validate the Italian version of the Helsinki Chronic Pain Index (I-HCPI). Owners of OA painful (n = 87) and healthy dogs (n = 40) were administered the I-HCPI once or twice after an eight-week meloxicam treatment. Sixty-nine owners of healthy and OA dogs also completed the Italian version of the Canine Brief Pain Inventory (I-CBPI). Pain on palpation on a 0-4 scale was assessed on all recruited dogs. Construct validity was tested both with hypothesis testing and principal component analysis, confirming the I-HCPI accurately measured chronic pain. Good convergent and criterion validity were shown through correlations with I-CBPI subscores and distribution among pain on palpation scores (p < 0.0001). The significant difference between the pre- and post-treatment I-HCPI scores (p < 0.0001) and Cohen's effect size (2.27) indicated excellent responsiveness. The I-HCPI was shown to be reliable through communalities (range 0.47-0.90) and Cronbach α (≥0.95). Discriminative ability and cut-off point, as tested through Receiver Operating Characteristic analysis, showed excellent diagnostic accuracy with a threshold value of 11 (specificity 0.98 and sensitivity 0.94). The I-HCPI was confirmed to be a valid, sensitive, reliable, and accurate tool to discriminate between dogs with and without pain.

Mineral, trace element, and toxic metal concentration in hair from dogs with idiopathic epilepsy compared to healthy controls.

BACKGROUND: Altered trace element status is associated with epilepsy in humans and dogs with idiopathic epilepsy (IE). OBJECTIVES: Compare hair element concentrations in epileptic and healthy dogs. ANIMALS: Sixty-three dogs with IE (53 treated, 10 untreated) and 42 controls. METHODS: Case-control study using ICP-MS to determine hair calcium, magnesium, phosphorus, sodium, potassium, iron, copper, manganese, zinc, selenium, chromium, lead, mercury, cadmium, arsenic, aluminum, and nickel concentration. Groups were compared using nonparametric tests. Results were controlled for diet, sex, age, and hair color using generalized linear mixed models. RESULTS: Compared to healthy controls, dogs with IE had lower hair phosphorus (mean ± SD; IE: 286.19 ± 69.62 µg/g, healthy: 324.52 ± 58.69 µg/g; P = .001), higher hair copper (IE: 10.97 ± 3.51 µg/g, healthy: 8.41 ± 1.27 µg/g; P < .001), zinc (IE: 158.25 ± 19.64 µg/g, healthy: 144.76 ± 32.18 µg/g; P < .001), copper/zinc ratio (IE: 0.07 ± 0.02, healthy: 0.06 ± 0.01; P = .003), selenium (IE: 1.65 ± 0.43 µg/g, healthy: 0.94 ± 0.73 µg/g; P < .001), and arsenic (IE: 0.40 ± 0.78 µg/g, healthy: 0.05 ± 0.08 µg/g; P < .001). When comparing treated and untreated epileptic dogs with healthy dogs, the differences in phosphorus and selenium remained significant for both groups, whereas the differences in copper, zinc, and arsenic were significant only for treated dogs. Potassium bromide treatment was strongly associated with high hair arsenic (P = .000). CONCLUSIONS AND CLINICAL IMPORTANCE: Altered trace element status could be involved in the pathophysiology of IE in dogs. Antiseizure drugs might affect trace element and arsenic metabolism.

Assessing the association between supplemented puppyhood dietary fat sources and owner-reported epilepsy in adulthood, among Finnish companion dogs.

Introduction: Epilepsy is a serious and common neurological condition in dogs, despite the wide number of antiepileptic drugs available, in approximately one third of the patients, epilepsy remains unsatisfactorily controlled. We aim to analyze whether feeding dietary fat sources during puppyhood was associated with canine epilepsy in adulthood. Methods: A nested case-control study was compiled from the validated DogRisk food frequency questionnaire (DogRisk FFQ). DogRisk FFQ collected feeding, disease, and background data about the dog. The study sample consisted of 108 owner-reported epileptic cases and 397 non-epileptic controls. Each case was matched with up to four controls for the key confounding factors of sex, breed, and age. We analyzed associations between feeding as a puppy and owner-reported epilepsy as an adult dog using Cox regression. We tested 55 different food variables. Results: We found that feeding fish fat from dietary sources at least once a week during puppyhood was inversely associated with epilepsy in later life in the unadjusted analysis [OR 0.46 (95% CI 0.25-0.83), p=0.01], while when adjusting for keeping conditions and dog characteristics the association was [OR 0.45 (95% CI 0.23-0.88), p=0.02]. When adjusted for keeping conditions, dog characteristics, and other feeding factors, the association was of similar magnitude but not significance [OR 0.56 (95% CI 0.27-1.15), p=0.12]. Discussion: The study indicates possible protective associations of feeding the dog with dietary sources of fish fat against epilepsy, although the result could be confounded by other feeding factors. Findings are compatible with current knowledge regarding the role of omega-3 fatty acids and ketogenic diet, a low carbohydrate, high fat diet as supportive treatments of epilepsy. As our findings are based on observations, we suggest the possibility of causality but do not prove it. Dietary intervention studies should now be conducted to confirm our findings.

Early life programming by diet can play a role in risk reduction of otitis in dogs.

Introduction: Otitis in dogs is often chronic while local treatment primarily consists of flushing, antibiotics, and/or antifungals. We were interested in finding early life variables that associate with otitis later in life, preferably some that could be modified. Methods: A cross-sectional hypothesis-driven study with longitudinal data was performed to search for associations between pre- and postnatal exposures, and the incidence of owner-reported otitis in dogs at over 1 year of age. The multivariate logistic regression analysis study included data from 3,064 dogs and explored 26 different early life variables at four early life stages: prenatal, neonatal, postnatal, and puppyhood. We compared two feeding patterns, a non-processed meat-based diet (NPMD, raw) and an ultra-processed carbohydrate-based diet (UPCD, dry). Results: We report that eating a NPMD diet significantly decreased the risk of otitis later in life, while eating a UPCD diet significantly increased the risk. This was seen in different life stages of mother or puppy: The maternal diet during pregnancy (p=0.011) and the puppies' diet from 2 to 6 months of age (p=0.019) were both significantly associated with otitis incidence later in life, whereas the puppies' first solid diet, was associated in the same way, but did not reach significance (p=0.072). Also, analyzing food ratios showed that when puppies were consuming >25% of their food as NPMD it significantly decreased their incidence of otitis later in life, while a ratio of >75% UPCD in their diet significantly increased their risk of otitis. Also, if the dog was born in the current family, was exposed to sunlight for more than 1 hour daily, and was raised on a dirt floor during puppyhood, there was a lower risk of otitis development later in life. Discussion: The findings only suggest causality, and further studies are required. However, we propose that veterinarians, breeders, and owners can impact otitis risk by modifying factors such as diet and environment.

GATA-3 expression and its correlation with prognostic factors and survival in canine mammary tumors.

Introduction: The transcription factor GATA-3 plays a significant role in mammary gland development and differentiation. Recent studies on human oncology have demonstrated its association with favorable pathologic factors in breast cancer. Canine mammary tumours, proposed as comparative and translational study models, have epidemiological, clinical, biological, and genetic characteristics similar to those of human breast cancers. Methods: Here, we evaluated the frequency of GATA-3 expression in mammary tumors of dogs and its relationship with prognostic factors and survival. Tumor samples were obtained from 40 female dogs and grouped according to histological type into benign tumors (n = 10), carcinoma in mixed tumors (CMTs) (n = 20), and aggressive tumors (n = 10). CMTs were further separated according to histological grade, and data on clinical staging and diagnosis, histopathological grading, and survival rate were collected. Results: GATA-3 and estrogen receptor (ER) expression were higher in benign and well-differentiated carcinomas than in aggressive tumors, which showed greater Ki-67 expression. The expression rate of ER in the studied groups was equivalent to that of GATA-3. We identified a strong positive correlation between GATA-3 and ER expression frequencies and a negative correlation between those of GATA-3 and Ki-67. There were associations between GATA-3 (p < 0.001), Ki-67 (p = 0.003), tumor size (p < 0.001), clinical stage (p = 0.002), lymph node metastasis (p < 0.001), and histological grade (p < 0.001) by univariate survival analysis. The parameters ER (p = 0.015) and GATA-3 (p = 0.005) also influenced survival in a multifactorial manner. Discussion: Kaplan-Meier analysis of survival curves validated our previous findings that dogs with GATA-3 expression in ≥79.4% of cells had significantly higher survival rates (p < 0.001). The performance analysis showed that the expression of GATA-3 in ≥79.4% of cells effectively predicted survival or death in dogs with mammary tumors. Collectively, these results suggest that GATA-3 can be a relevant marker in the study of mammary tumor progression and has potential as a prognosis marker for predicting outcomes in canine mammary tumors.

An un-commissioned randomized, placebo-controlled double-blind study to test the effect of deep sea fish oil as a pain reliever for dogs suffering from canine OA.

BACKGROUND: An un-commissioned randomized, double-blinded, placebo controlled clinical study was planned using a deep sea fish oil product for pets. Seventy-seven client-owned dogs with osteoarthritis were randomly assigned to supplement the food with either the fish oil product or corn (=placebo) oil. Our main outcome variables were force platform variables peak vertical force (PVF) and impulse, the validated Helsinki Chronic Pain Index (HCPI) and the use of rescue NSAIDs. Secondary outcome variables were a locomotion visual analog scale (VAS), a Quality of life VAS, a comparative questionnaire, a veterinary assessment, owners' final assessment of outcome and guessing the product given. RESULTS: When comparing the two test groups at the end of the trial (16 weeks) there was no significant difference in any of the main outcome variables but owners of dogs that had taken fish oil were significantly happier with the treatment at the end visit and did significantly better at guessing what group their dogs had been in, compared to the placebo group. When comparing variables within the fish oil group as change from baseline to trial end, there were significant positive changes in PVF, HCPI, NSAID use, Quality of life VAS, as well as in all three scores in the comparative questionnaire (locomotion, every-day situations, and skin & coat). There were similar positive trends in force platform impulse and in the veterinary assessment variables, although they did not reach significance. Within the placebo group there were significant positive changes only in the HCPI and a significant deterioration according to veterinary assessment. CONCLUSIONS: When compared to placebo, there was not a major statistically significant benefit in using deep sea fish oil as a pain reliever in our study population of dogs suffering from osteoarthritis. However, the fish oil treated patients improved significantly in many of the variables, when comparing baseline values to the study-end values within the group, indicating a true but small relief in symptoms. Deep sea fish oil supplementation could be considered a part of the multimodal pain relieving approach currently recommended for dogs suffering from OA, especially for individuals that do not tolerate anti-inflammatory drugs.

Part I of Finnish Agility Dog Survey: Training and Management of Competition-Level Agility Dogs.

Knowledge regarding training, competition, and management routines of agility dogs is lacking. Through a retrospective online questionnaire, Finnish owners and handlers of 745 competition-level agility dogs provided information on training routines and management of these dogs during one year free of agility-related injuries. Competition routines were collected from the national competition results database. Most dogs trained agility 1-2 times a week, with a median active training time of 18 min a week. Dogs competed in a median of 2.1 runs per month at a speed of 4.3 m/s. Common field surfaces were different types of artificial turfs and dirt surface. Warm-up and cool-down were established routines, and 62% of dogs received regular musculoskeletal care. Moreover, 77% of dogs underwent conditioning exercises, but their frequency was often low. Additionally, dogs were walked for a median of 1.5 h daily. Pearson's chi-squared and Kruskal-Wallis tests were used to evaluate the association between a dog's competition level and training and competition variables. A dog's competition level was associated with competition (p < 0.001) and training frequency (p < 0.001); dogs at higher levels compete more but train less than dogs at lower levels. This study provides information on training, competition, and management routines of competing agility dogs.

Diet and dog characteristics affect major and trace elements in hair and blood of healthy dogs.

Obtaining correct amounts of essential elements, and avoiding toxic metals are key factors in dog health. Through analyzing major and trace elements in hair and blood of 50 healthy companion dogs using ICP-MS, we study their associations with dog characteristics and diet, hypothesizing that eating the same diet long-term results in strong correlations between hair and blood element concentrations, and that dog characteristics and diet affect element status. The correlation between hair and blood was significant for Hg (R = 0.601, p = 0.000) and Pb (R = 0.384, p = 0.010). The following associations were significant (p < 0.05): Dark hair had higher Ca and Mg compared to light hair. Females had higher hair Zn, blood Mn, and blood As compared to males. Blood Mn and Se increased, while blood Pb decreased with age. Raw diet fed dogs had higher hair Zn and Se compared to dry or mixed diet fed dogs, and lower blood Mn compared to dry diet fed dogs. Dry and mixed diet fed dogs had higher blood Cd compared to raw diet fed dogs. Mixed diet fed dogs had higher hair Ca and Mg compared to raw or dry diet fed dogs, and higher hair Pb compared to dry diet fed dogs. Wild game consumption was associated with higher blood Pb, and rice consumption with higher blood As. In conclusion, hair provides an alternative for assessing Hg and Pb exposure, and major and trace elements status is affected by hair color, sex, age, and diet.

Part II of Finnish Agility Dog Survey: Agility-Related Injuries and Risk Factors for Injury in Competition-Level Agility Dogs.

Dog agility is associated with a risk for sport-related injuries, but few risk factors for injury are known. A retrospective online questionnaire was used to collect data on 864 Finnish competition-level agility dogs-including 119 dogs (14%) with agility-related injury during 2019. Data included injury details, health background, experience in agility, and sport and management routines prior to the injury. Risk factors for injury were evaluated with multivariate logistic regression. The rate of competition-related injuries was 1.44 injuries/1000 competition runs. The front limb was injured in 61% of dogs. In 65% of dogs, the injury presented as lameness. The main risk factors for agility-related injury during 2019 were multiple previous agility-related injuries (OR 11.36; 95% CI 6.10-21.13), older age when starting course-like training (OR 2.04 per one year increase; 95% CI 1.36-3.05), high training frequency, diagnosis of lumbosacral transitional vertebra, and physiotherapy every two to three months compared with never. The most important protective factors were moderate competition frequency and A-frame performance technique. These associations do not confirm causality. We identified new risk factors for injury in agility. This information can be used to improve the welfare of agility dogs.