Nutrient digestibility and fecal characteristics,microbiota, and metabolites in dogs fed human-grade foods

Human-grade (HG) pet foods are commercially available, but they have not been well studied. Our objective was to determine the apparent total tract digestibility (ATTD) of HG pet foods and evaluate their effects on fecal characteristics, microbiota, and metabolites, serum metabolites, and hematology of dogs. Twelve dogs (mean age = 5.5 ± 1.0; BW = 11.6 ± 1.6 kg) were used in a replicated 4 × 4 Latin square design (n = 12/treatment). The diets included 1) Chicken and Brown Rice Recipe (extruded; Blue Buffalo); 2) Roasted Meals Tender Chicken Recipe (fresh; Freshpet); 3) Beef and Russet Potato Recipe (HG beef; JustFoodForDogs); and 4) Chicken and White Rice Recipe (HG chicken; JustFoodForDogs). Each period consisted of 28 d, with a 6-d diet transition phase, 16 d of consuming 100% of the diet, a 5-d phase for fecal collection, and 1 d for blood collection. All data were analyzed using the Mixed Models procedure of SAS 9.4. Dogs fed the extruded diet required a higher (P < 0.05) daily food intake (dry matter basis, DMB) to maintain BW. The ATTD of dry matter (DM), organic matter (OM), energy, and acid-hydrolyzed fat (AHF) were greater (P < 0.05) in dogs fed the HG diets than those fed the fresh diet, and greater (P < 0.05) in dogs fed the fresh diet than those fed the extruded diet. Crude protein ATTD was lower (P < 0.05) for dogs fed the extruded diet than those fed all other diets. Dogs fed the extruded diet had greater (P < 0.05) fecal output (as-is; DMB) than dogs fed fresh (1.5–1.7 times greater) or HG foods (2.0–2.9 times greater). There were no differences in fecal pH, scores, and metabolites, but microbiota were affected by diet. Dogs fed HG beef had higher (P < 0.05) relative abundance of Bacteroidetes and lower (P < 0.05) relative abundance of Firmicutes than dogs fed the fresh or HG chicken diets. The Actinobacteria, Fusobacteria, Proteobacteria, and Spirochaetes phyla were unchanged (P > 0.05), but diet modified the relative abundance of nearly 20 bacterial genera. Similar to previous reports, these data demonstrate that the fecal microbiota of dogs fed HG or fresh diets is markedly different than those consuming extruded diets, likely due to ingredient, nutrient, and processing differences. Serum metabolites and hematology were not greatly affected by diet. In conclusion, the HG pet foods tested resulted in significantly reduced fecal output, were highly digestible, maintained fecal characteristics, serum chemistry, and hematology, and modified the fecal microbiota of dogs.     

Tooth brushing inhibits oral bacteria in dogs

In this study, scaling, polishing and daily tooth brushing were performed in 20 beagle dogs, and the number of oral bacteria was determined using a bacterial counter. The dogs were randomized into the scaling (S), scaling + polishing (SP), scaling + tooth daily brushing (SB) and scaling + polishing + tooth daily brushing (SPB) groups. Samples were collected from the buccal surface of the maxillary fourth premolars of the dogs immediately after scaling and every week thereafter from weeks 1 to 8. Throughout the study, the number of bacteria was significantly lower in the SB and SPB groups compared with the S group. The findings suggest that daily tooth brushing inhibited oral bacterial growth in the dogs.     

Evaluation of a novel animal milk oligosaccharide biosimilar: macronutrient digestibility and gastrointestinal tolerance,fecal metabolites,and fecal microbiota of healthy adult dogs and in vitro genotoxicity assays

Milk oligosaccharides (MO) are bioactive compounds in mammalian milk that provide health benefits to neonates beyond essential nutrients. GNU100, a novel animal MO biosimilar, was recently tested in vitro, with results showing beneficial shifts in microbiota and increased short-chain fatty acid (SCFA) production, but other effects of GNU100 were unknown. Three studies were conducted to evaluate the safety, palatability, and gastrointestinal (GI) tolerance of GNU100. In study 1, the mutagenic potential of GNU100 was tested using a bacterial reverse mutation assay and a mammalian cell micronucleus test. In study 2, palatability was assessed by comparing diets containing 0% vs. 1% GNU100 in 20 adult dogs. In study 3, 32 adult dogs were used in a completely randomized design to assess the safety and GI tolerance of GNU100 and explore utility. Following a 2-wk baseline, dogs were assigned to one of four treatments and fed for 26 wk: 0%, 0.5%, 1%, and 1.5% GNU100. On weeks 2, 4, and 26, fresh fecal samples were collected to measure stool quality, immunoglobulin A, and calprotectin, and blood samples were collected to measure serum chemistry, inflammatory markers, and hematology. On weeks 2 and 4, fresh fecal samples were collected to measure metabolites and microbiota. On week 4, total feces were collected to assess apparent total tract macronutrient digestibility. Although revertant numbers were greater compared with the solvent control in tester strain WP2uvrA(pKM101) in the presence of metabolic activation (S9) in the initial experiment, they remained below the threshold for a positive mutagenic response in follow-up confirmatory tests, supporting that GNU100 is not mutagenic. Similarly, no cytotoxicity or chromosome damage was observed in the cell micronucleus test. The palatability test showed that 1% GNU100 was strongly preferred (P < 0.05; 3.6:1 consumption ratio) over the control. In study 3, all dogs were healthy and had no signs of GI intolerance or illness. All diets were well accepted, and food intake, fecal characteristics, metabolite concentrations, and macronutrient digestibilities were not altered. GNU100 modulated fecal microbiota, increasing evenness and Catenibacterium, Megamonas, and Prevotella (SCFA producers) and reducing Collinsella. Overall, the results suggest that GNU100 is palatable and well-tolerated, causes no genotoxicity or adverse effects on health, and beneficially shifts the fecal microbiota, supporting the safety of GNU100 for the inclusion in canine diets.     

Effects of dietary cellobiose on the intestinal microbiota and excretion of nitrogen metabolites in healthy adult dogs

In order to evaluate the potential prebiotic effects of cellobiose, 10 healthy adult research beagle dogs received a complete diet containing 0, 0.5 and 1 g cellobiose/kg bodyweight (BW)/day. At the end of each feeding period, faeces, urine and blood of the dogs were collected. The results demonstrated a significant increase of faecal lactate concentrations, indicating a bacterial fermentation of cellobiose in the canine intestine. Along with this, a dose-dependent linear increase of the relative abundance of Lactobacillaceae in the faeces of the dogs was observed (p = 0.014). In addition, a dose-dependent increase (p < 0.05) of Alloprevotella, Bacteroides and Prevotella, and a linear decrease for unidentified Lachnospiraceae (p = 0.011) was observed when cellobiose was added to the diet, although the relative abundance of these genera was low (<1%) among all groups. The faecal pH was not affected by dietary cellobiose. Cellobiose seemed to modulate the excretion of nitrogen metabolites, as lower concentrations of phenol (p = 0.034) and 4-ethylphenol (p = 0.002) in the plasma of the dogs were measured during the supplementation periods. Urinary phenols and indoles, however, were not affected by the dietary supplementation of cellobiose. In conclusion, cellobiose seems to be fermented by the intestinal microbiota of dogs. Although no effect on the faecal pH was detected, the observed increase of microbial lactate production might lower the pH in the large intestine and consecutively modulate the intestinal absorption of nitrogen metabolites. Also, the observed changes of some bacterial genera might have been mediated by increased intestinal lactate concentrations or a higher relative abundance of lactobacilli. Whether these results could be considered as a prebiotic effect and used as a dietetic strategy in diseased animals to improve gut function or hepatic and renal nitrogen metabolism should be evaluated in future studies.     

Outcome in dogs with surgically resected oral fibrosarcoma (1997-2008)

Oral fibrosarcoma (FSA) is a common oral tumour in dogs, and historically reported survival times after surgical excision range from 7.0 to 12.2 months with local recurrence rates of 32-57%. The purpose of this retrospective study was to report outcome in a cohort of dogs with oral FSA treated with surgical excision with or without adjuvant radiation therapy. Twenty-nine dogs with a histological diagnosis of FSA arising from the oral cavity that underwent surgical resection of their oral FSA were included in this study. Twenty-one dogs were treated with surgical excision alone and eight dogs with both surgery and radiation therapy. The median progression-free interval was >653 days. The median survival time was 743 days. The 1- and 2-year survival rates were 87.7 and 57.8%, respectively. Seven (24.1%) dogs developed local recurrence. Seven dogs (24.1%) developed metastasis.     

The aetiopathogenesis of equine periodontal disease – a fresh perspective

Periodontal disease is a painful and highly prevalent disorder of horses that causes a significant welfare problem. Despite its importance, few scientific studies on its aetiopathogenesis have been performed. Equine periodontitis differs from the plaque‐induced periodontitis found in brachydont species where bacteria accumulating in dental plaque induce a destructive inflammatory response in the periodontium. In contrast, equine periodontitis is usually initiated by entrapment of feed between cheek teeth, which causes inflammation of periodontal tissue that likely allows bacterial infection of the periodontal tissues that is later exacerbated by the host's response. Equine oral microbiology is a neglected field of research and identification of the bacteria involved in this disorder by use of molecular bacteriology and examination of the interaction between these bacteria and the equine oral immune response should reveal important information about the pathogenesis of this disease.     

犬牙周病的临床研究进展

牙周病(periodontal disease, PD)是由细菌在牙齿表面聚集形成的牙菌斑和牙结石逐渐侵蚀牙周组织,引发周围组织炎症的一类疾病。根据疾病的发展进程,可分为齿龈炎与牙周炎2个阶段。在小动物临床疾病中牙周病尤为常见,世界范围内约有80%的成年犬患有该病。由于饲主对于犬的口腔保健意识薄弱且该疾病隐秘性较高,患犬就诊时往往已出现牙周支持组织不可逆的损伤,细菌及其毒力因子通过血液循环进入全身从而引发系统性疾病的风险大大提高。但目前犬牙周病的核心致病菌属、菌群失调的原因与发病机制尚不明确。因此,笔者基于高通量测序方法的病原学研究结果,分析与疾病发展相关的微生物群。同时,整合目前世界通用的最新犬牙周病临床诊断分级标准及犬牙周病的中西医防治方案,分析犬牙周病与全身系统性疾病之间的相关性,系统综述犬牙周病在国内外小动物临床领域的研究进展,以期为该类疾病的临床诊疗提供理论参考,推进兽医师在临床实践中更高效地开展该类疾病的诊疗工作,并为犬牙周病相关的临床研究提供思路。     

Canine oral primary melanoma cells exhibit shift to mesenchymal phenotype and phagocytic behaviour

Canine oral malignant melanoma (COMM) is a potentially lethal cancer disease. We established primary cell lines from mostly amelanotic primary COMM and metastases and assessed lesions and derived cells for Melan A, PNL2 and CD146 expression. Then, migration and invasion of CD146‐enriched vs ‐depleted COMM cells were analysed. Epithelial‐to‐mesenchymal transition (EMT) was addressed by Vimentin‐staining and MMP2/MMP9 zymography. Phagocytic behaviour was analysed by histopathological examination and phagocytosis assay. While Melan A‐ and PNL2‐staining yielded inconsistent data, 100% of COMM sections and primary cells showed CD146 expression, suggesting that this protein may serve as a prognostic marker. An overall correlation between CD146‐expression and migration/invasion was not observed. All primary cell lines consistently expressed Vimentin and secreted biologically active MMP2, indicating that they had undergone EMT. Importantly, COMM sections exhibited cell‐in‐cell structures, and all primary cell lines exhibited phagocytic activity, supporting the concept that cell cannibalism may have a role in COMM progression.     

Evaluation of graded levels of Bacillus coagulans GBI-30, 6086 on apparent nutrient digestibility,stool quality,and intestinal health indicators in healthy adult dogs

Bacillus coagulans GBI-30, 6086 is a commercially available spore-forming non-toxigenic microorganism approved for use in dog foods with high resiliency to stresses associated with commercial manufacturing. The objectives of this research were to examine the effect of B. coagulans on stool quality, nutrient digestibility, and intestinal health markers in healthy adult dogs. Extruded diets containing graded levels of B. coagulans applied either to the base ration before extrusion or to the exterior of the kibble as a topical coating after extrusion were randomly assigned to 10 individually housed adult beagle dogs (7 castrated males and 3 spayed females) of similar age (5.75 ± 0.23 yr) and body weight (12.3 ± 1.5 kg). The study was designed as a 5 × 5 replicated Latin square with 16-d adaptation followed by 5-d total fecal collection for each period. Five dietary treatments were formulated to deliver a dose of 0-, 6-, 7-, 8-, and 9-log₁₀ colony-forming units (CFU) per dog per day for the control (CON), extruded B. coagulans (PEX), and low, moderate, and high B. coagulans coating levels (PCL, PCM, and PCH), respectively. Food-grade TiO₂ was added to all diets at a level of 0.4% to serve as an indigestible dietary marker for digestibility calculations. Data were analyzed using a mixed model through SAS (version 9.4, SAS Institute, Inc., Cary, NC) with treatment as a fixed effect and room (i.e., replicate), period, and dog(room) as random effects. Apparent total tract digestibility of organic matter, crude protein, crude fat, and gross energy calculated by the marker method were numerically greatest for dogs fed the 9-log₁₀ dose treatment with increases (P < 0.05) observed in gross energy and organic matter digestibility compared with the negative control. No significant differences were observed in food intake, stool quality, fecal pH, fecal ammonia, fecal short-chain fatty acids, or branched-chain fatty acids for the extruded B. coagulans treatment (PEX) or the coated B. coagulans treatments (PCL, PCM, and PCH) compared with CON. These results suggest that B. coagulans has a favorable impact on nutrient digestibility and no apparent adverse effects when added to extruded diets at a daily intake level of up to 9-log₁₀ CFU in healthy adult dogs.     

Physical activity patterns of free living dogs diagnosed with osteoarthritis

Osteoarthritis (OA) affects about 90% of dogs > 5 yr of age in the United States, resulting in reduced range of motion, difficulty climbing and jumping, reduced physical activity, and lower quality of life. Our objective was to use activity monitors to measure physical activity and identify how activity counts correlate with age, body weight (BW), body condition score (BCS), serum inflammatory markers, veterinarian pain assessment, and owner perception of pain in free-living dogs with OA. The University of Illinois Institutional Animal Care and Use Committee approved the study and owner consent was received prior to experimentation. Fifty-six client-owned dogs (mean age = 7.8 yr; mean BCS = 6.1) with clinical signs and veterinary diagnosis of OA wore HeyRex activity collars continuously over a 49-d period. Blood samples were collected on day 0 and 49, and dog owners completed canine brief pain inventory (CBPI) and Liverpool osteoarthritis in dogs (LOAD) surveys on day 0, 21, 35, and 49. All data were analyzed using SAS 9.3 using repeated measures and R Studio 1.0.136 was used to generate Pearson correlation coefficients between data outcomes. Average activity throughout the study demonstrated greater activity levels on weekends. It also showed that 24-h activity spiked twice daily, once in the morning and another in the afternoon. Serum C-reactive protein concentration was lower (P < 0.01) at day 49 compared to day 0. Survey data indicated lower (P < 0.05) overall pain intensity and severity score on day 21, 35 and 49 compared to day 0. BW was correlated with average activity counts (P = 0.02; r = −0.12) and run activity (P = 0.10; r = −0.24). Weekend average activity counts were correlated with owner pain intensity scores (P = 0.0813; r = −0.2311), but weekday average activity count was not. Age was not correlated with total activity count, sleep activity, or run activity, but it was correlated with scratch (P = 0.03; r = −0.10), alert (P = 0.03; r = −0.13), and walk (P = 0.09; r = −0.23) activities. Total activity counts and activity type (sleep, scratch, alert, walk, and run) were not correlated with pain scored by veterinarians, pain intensity or severity scored by owners, or baseline BCS. Even though the lack of controls and/or information on the individual living conditions of dogs resulted in a high level of variability in this study, our data suggest that the use of activity monitors have the potential to aid in the management of OA and other conditions affecting activity (e.g., allergy; anxiety).     

Resolution of persistent oral papillomatosis in a dog after treatment with a recombinant canine oral papillomavirus vaccine

This report describes a 16‐month‐old female, otherwise seemingly healthy, Siberian husky dog with severe oral papillomatosis that did not regress spontaneously and was refractory to surgical treatment over a 6‐month period. Regression of the papillomas was achieved by administering a series of experimental vaccinations starting at the time of the last surgery. The vaccine consisted of systemically administered canine oral papillomavirus major coat protein L1 that has been shown to self‐assemble into virus‐like particles. They cause a humoral response that has been shown to prevent the onset and development of papillomas. In this case, however, following unsuccessful surgical treatment, the vaccine acted therapeutically, causing the papillomas that had regrown to shrink. No side‐effects were noted.     

Clinical pharmacokinetics and outcomes of oral fluconazole therapy in dogs and cats with naturally occurring fungal disease

This multi-institutional study was designed to determine the clinical pharmacokinetics of fluconazole and outcomes in client-owned dogs (n = 37) and cats (n = 35) with fungal disease. Fluconazole serum concentrations were measured. Pharmacokinetic analysis was limited to animals at steady state (≥72 hr of treatment). The mean (range) body weight in 31 dogs was 25.6 (2.8–58.2) kg and in 31 cats was 3.9 (2.4–6.1) kg included in pharmacokinetic analyses. The dose, average steady-state serum concentrations (C_SS), and oral clearance in dogs were 14.2 (4.5–21.3) mg/kg/d, 26.8 (3.8–61.5) µg/mL, and 0.63 ml min⁻¹ kg⁻¹, respectively, and in cats were 18.6 (8.2–40.0) mg/kg/d, 32.1 (1.9–103.5) µg/mL, and 0.61 ml min⁻¹ kg⁻¹, respectively. Random inter-animal pharmacokinetic variability was high in both species. Two dogs had near twofold increases in serum fluconazole when generic formulations were changed, suggesting lack of bioequivalence. Median C_SS for dogs and cats achieving clinical remission was 19.4 and 35.8 µg/ml, respectively. Starting oral doses of 10 mg/kg q12h in dogs and 50–100 mg total daily dose in cats are recommended to achieve median C_SS associated with clinical remission. Due to the large pharmacokinetic variability, individualized dose adjustments based on C_SS (therapeutic drug monitoring) and treatment failure should be considered.     

Profiling of plasma metabolites in canine oral melanoma using gas chromatography-mass spectrometry

Malignant melanoma is one of the most common and aggressive tumors in the oral cavity of dog. The tumor has a poor prognosis, and methods for diagnosis and prediction of prognosis after treatment are required. Here, we examined metabolite profiling using gas chromatography-mass spectrometry (GC-MS) for development of a discriminant model for evaluation of prognosis. Metabolite profiles were evaluated in healthy and melanoma plasma samples using orthogonal projection to latent structure using discriminant analysis (OPLS-DA). Cases that were predicted to be healthy using the OPLS discriminant model had no advanced lesions after radiation therapy. These results indicate that metabolite profiling may be useful in diagnosis and prediction of prognosis of canine malignant melanoma.     

Prolongation of survival of dogs with oral malignant melanoma treated by en bloc surgical resection and adjuvant CSPG4‐antigen electrovaccination

Reported post‐surgery 1‐year survival rate for oral canine malignant melanoma (cMM) is around 30%; novel treatments are needed as the role of adjuvant chemotherapy is unclear. This prospective study regards adjuvant electrovaccination with human chondroitin sulfate proteoglycan‐4 (hCSPG4)‐encoded plasmid in 23 dogs with resected II/III‐staged CSPG4‐positive oral cMM compared with 19 dogs with resected only II/III‐staged CSPG4‐positive oral cMM. Vaccination resulted in 6‐, 12‐, 18‐ and 24‐month survival rate of 95.6, 73.9, 47.8 and 30.4%, respectively [median survival time (MST) 684 days, range 78–1694, 8 of 23 dogs alive] and 6‐, 12‐, 18‐ and 24‐month disease‐free interval (DFI) rate of 82.6, 47.8, 26.1 and 17.4%, respectively (DFI 477 days, range 50–1694). Non‐vaccinated dogs showed 6‐, 12‐, 18‐ and 24‐month survival rate of 63.2, 26.3, 15.8 and 5.3%, respectively (MST 200 days, range 75–1507, 1 of 19 dogs alive) and 6‐, 12‐, 18‐ and 24‐month DFI rate of 52.6, 26.3, 10.5 and 5.3%, respectively (DFI 180 days, range 38–1250). Overall survival and DFI of vaccinated dogs was longer in those <20 kg. In vaccinated and non‐vaccinated dogs local recurrence rate was 34.8 and 42%, respectively while lung metastatic rate was 39 and 79%, respectively.     

Effects of dietary macronutrient profile on apparent total tract macronutrient digestibility and fecal microbiota,fermentative metabolites,and bile acids of female dogs after spay surgery

Obesity and estrogen reduction are known to affect the gut microbiota and gut microbial-derived metabolites in some species, but limited information is available in dogs. The aim of this study was to determine the effects of dietary macronutrient profile on apparent total tract macronutrient digestibility, fecal microbiota, and fecal metabolites of adult female dogs after spay surgery. Twenty-eight adult intact female beagles (age: 3.02 ± 0.71 yr, BW: 10.28 ± 0.77 kg; BCS: 4.98 ± 0.57) were used. After a 5-wk baseline phase (week 0), 24 dogs were spayed and randomly allotted to one of three experimental diets (n = 8 per group): 1) control (CO) containing moderate protein and fiber (COSP), 2) high-protein, high-fiber (HPHF), or 3) high-protein, high-fiber plus omega-3 and medium-chain fatty acids (HPHFO). Four dogs were sham-operated and fed CO (COSH). All dogs were fed to maintain BW for 12 wk after spay and then allowed to consume twice that amount for 12 wk. Fecal samples were collected at weeks 0, 12, and 24 for digestibility, microbiota, and metabolite analysis. All data were analyzed using repeated measures and linear mixed models procedure of SAS 9.4, with results reported as a change from baseline. Apparent organic matter and energy digestibilities had greater decreases in HPHF and HPHFO than COSH and COSP. Increases in fecal acetate, total short-chain fatty acids, and secondary bile acids were greater and decreases in primary bile acids were greater in HPHF and HPHFO. Principal coordinates analysis of weighted UniFrac distances revealed that HPHF and HPHFO clustered together and separated from COSH and COSP at weeks 12 and 24, with relative abundances of Faecalibacterium, Romboutsia, and Fusobacterium increasing to a greater extent and Catenibacterium, Bifidobacterium, Prevotella 9, Eubacterium, and Megamonas decreasing to a greater extent in HPHF or HPHFO. Our results suggest that high-protein, high-fiber diets alter nutrient and energy digestibilities, fecal metabolite concentrations, and fecal gut microbiota, but spay surgery had minor effects. Future research is needed to investigate how food intake, nutrient profile, and changes in hormone production influence gut microbiota and metabolites of dogs individually and how this knowledge may be used to manage spayed pets.