Obesity-induced changes in gene expression in feline adipose and skeletal muscle tissue.

Indoor-confined cats are prone to developing obesity due to a sedentary life and an energy intake exceeding energy requirements. As in humans, feline obesity decreases insulin sensitivity and increases the risk of developing feline diabetes mellitus, but the pathophysiological mechanisms are currently poorly understood. Human obesity-related metabolic alterations seem to relate to changes in the expression of genes involved in glucose metabolism, insulin action and inflammation. The objective of the current study was to investigate changes in the expression of genes relating to obesity, glucose metabolism and inflammation in cats with non-experimentally induced obesity. Biopsies from the sartorius muscle and subcutaneous adipose tissue were obtained from 73 healthy, neutered, indoor-confined domestic shorthaired cats ranging from lean to obese. Quantification of obesity-related gene expression levels relative to glyceraldehyde-3-phosphate dehydrogenase was performed by quantitative real-time polymerase chain reaction. A negative association between obesity and adiponectin expression was observed in the adipose tissue (mean ± SD; normal weight, 27.30 × 10-3 ± 77.14 × 10-3 ; overweight, 2.89 × 10-3 ± 0.38 × 10-3 and obese, 2.93 × 10-3 ± 4.20 × 10-3 , p < 0.05). In muscle, the expression of peroxisome proliferative activated receptor-γ2 and plasminogen activator inhibitor-1 was increased in the obese compared to the normal-weight cats, and resistin was increased in the normal-weight compared to the overweight cats. There were no detectable obesity-related changes in the messenger RNA levels of inflammatory cytokines. In conclusion, a possible obesity-related low-grade inflammation caused by increased expression of key proinflammatory regulators was not observed. This could imply that the development of feline obesity and ensuing insulin resistance may not be based on tissue-derived inflammation, but caused by several determining factors, many of which still need further investigation.

Impact of oral amoxicillin and amoxicillin/clavulanic acid treatment on bacterial diversity and ß-lactam resistance in the canine faecal microbiota.

BACKGROUND: Aminopenicillins with or without a ß-lactamase inhibitor are widely used in both human and veterinary medicine. However, little is known about their differential impact on the gut microbiota and development of antimicrobial resistance. OBJECTIVES: To investigate changes in the faecal microbiota of dogs treated with amoxicillin or amoxicillin/clavulanic acid. METHODS: Faeces collected from 42 dogs (21 per treatment group) immediately before, during and 1 week after termination of oral treatment with amoxicillin or amoxicillin/clavulanic acid were analysed by culture and 16S rRNA gene sequence analysis. RESULTS: In both groups, bacterial counts on ampicillin selective agar revealed an increase in the proportion of ampicillin-resistant Escherichia coli during treatment, and an increased occurrence and proportion of ampicillin-resistant enterococci during and after treatment. 16S rRNA gene analysis showed reductions in microbial richness and diversity during treatment followed by a return to pre-treatment conditions approximately 1 week after cessation of amoxicillin or amoxicillin/clavulanic acid treatment. While no significant differences were observed between the effects of amoxicillin and amoxicillin/clavulanic acid on microbial richness and diversity, treatment with amoxicillin/clavulanic acid reduced the abundance of taxa that are considered part of the beneficial microbiota (such as Roseburia, Dialister and Lachnospiraceae) and enriched Escherichia, although the latter result was not corroborated by phenotypic counts. CONCLUSIONS: Our results suggest a limited effect of clavulanic acid on selection of antimicrobial resistance and microbial richness when administered orally in combination with amoxicillin. However, combination with this ß-lactamase inhibitor appears to broaden the spectrum of amoxicillin, with potential negative consequences on gut health.

Heritability and genetic trend of body weight in dogs of different breeds in Sweden.

High body weight (BW) in dogs has been associated with developmental as well as degenerative diseases, but the heritability of BW in dog breeds is largely unknown. The aim of the current study was to estimate heritability and genetic change (genetic trend) for BW in a range of dog breeds in Sweden. Body weight registrations from 19 dog breeds (with n ranging from 412 to 4,710) of varying body size, type and usage were collected from 2007 to 2016. The average BW of the breeds was 8 to 56 kg. The BW registrations were performed when the dogs were 12 to 24 mo of age (18 to 30 mo for one large-sized breed) in connection with an official radiographic screening program for hip dysplasia. Collected weight records were used to estimate heritability and genetic trends for BW. Several statistical models were used. The preliminary model included the fixed effects of breed (P < 0.001), sex (P < 0.001), year of screening (P < 0.001), litter size (P = 0.06), parity of the dam (P = 0.03) and linear regression on age at screening (P < 0.001), the latter five effects all nested within breed, and the random effects of litter and dam. Season of birth and the quadratic effect of age were also tested, but were not significant (P > 0.10). For the genetic analysis, various mixed linear models were tested within breed with different combinations of random effects; the most complex model included random effects of litter, direct additive, and maternal genetic effects, and maternal permanent environmental effects. The average heritability for BW over all 19 breeds was 51%, with a range of 35% to 70%, and the additive genetic coefficient of variance was around 9%. Maternal heritability was 5% to 9% and litter variance was below 10% with one exception (15% in Shetland Sheepdogs). For nine breeds, there was a genetic trend of increasing BW, whereas seven breeds had a genetic trend of decreasing BW. The largest absolute genetic change over a 10-yr period was around 0.6 kg or about 2% of the mean. In conclusion, given the small genetic changes in spite of the high heritability, it seems that there is generally a very weak selection, if any, for BW in the included dog breeds.

High body weight in dogs is often considered to cause problems, for instance, resulting in hip and elbow diseases. Furthermore, there is a huge variation in body conformation and size between different dog breeds, which is related to breeding for specific appearances and genetic traits. The aim of this study was to investigate the genetic variation of body weight within different dog breeds. To study this, we examined 19 dog breeds with an average body weight of 8 to 56 kg. We found that on average about 50% of the total variation in body weight between dogs, within a breed, depends on genetic differences, but with a range from 35% to 70% depending on breed. There were rather small changes over time in the genetic predisposition for high or low body weight; the largest changes were 0.6 kg over a 10-yr period.

Analytical performance of a canine ELISA monocyte chemoattractant protein-1 assay for use in cats and evaluation of circulating levels in normal weight and obese cats.

BACKGROUND: In human and murine obesity, adipose tissue dwelling macrophages and adipocytes produce monocyte chemoattractant protein-1 (MCP-1) leading to systemic low-grade inflammation. The aim of the study was to validate a canine MCP-1 ELISA assay for use in cats and to investigate whether a difference in MCP-1 concentrations could be detected between: a) cats having normal or elevated circulating serum amyloid A (SAA) levels and b) normal weight and obese cats. Serum obtained from 36 client-owned cats of various breed, age and sex with normal (n = 20) to elevated SAA (n = 16) was used for the validation of the canine MCP-1 ELISA assay. As no golden standard exists for measurement of inflammation, circulating MCP-1 concentrations were compared to SAA measurements, as an indicator of systemic inflammation. Analytical precision, dilution recovery and detection limit were calculated. A possible correlation between MCP-1 concentrations and obesity related measures (body fat percentage (BF%), insulin sensitivity and cytokine expression) were investigated in another population of 73 healthy, lean to obese, neutered domestic short-haired cats. RESULTS: Intra- (2.7-4.1%) and inter-assay (2.2-3.6%) coefficient of variation and dilution recovery were acceptable, and the detection limit was 27.1 pg/mL. MCP-1 did not correlate with SAA, and there was no difference between the inflammatory (SAA > 20 mg/L) and non-inflammatory group, due to a marked overlap in MCP-1 concentrations. Circulating MCP-1 concentrations were unaffected by BF% (r2 = 2.7 × 10-6, P = 0.21) and other obesity-related markers. CONCLUSIONS: The present canine ELISA assay seems to be able to measure circulating feline MCP-1. However, further studies are needed to determine its possible use for detecting inflammation in relation to disease processes or obesity-related low-grade inflammation in cats.

Corticosteroid treatment for acute/acute-on-chronic experimental and naturally occurring pancreatitis in several species: a scoping review to inform possible use in dogs.

Acute pancreatitis in dogs is a prevalent disease characterised by mild to severe inflammation. Treatment with anti-inflammatory corticosteroids has been widely debated but is not generally recommended in veterinary medicine. The objective of the present study was to present current evidence on the effect of corticosteroid treatment for acute/acute-on-chronic pancreatitis across species. These findings were then used to evaluate if and how corticosteroid treatment could influence disease outcome in canine acute/acute-on-chronic pancreatitis. A scoping review was performed by searching the Agricola, CAB Abstracts, MEDLINE and Embase databases to identify relevant articles published before June 24, 2021. The inclusion criteria were English language, original research published in a peer-reviewed journal, and investigation of corticosteroid treatment effects on acute/acute-on-chronic pancreatitis by the outcome parameters clinical score, circulating CRP level, hospitalisation duration, mortality and pancreas histopathology. Research on any species was considered. Studies were rated based on the level of evidence, and methodological quality was evaluated based on similarity between groups at baseline, risk of bias and study group size. The reporting method was based on the PRISMA extension for scoping reviews. One thousand nine hundred fifty-four studies were identified, and 31 met the inclusion criteria. Five were canine studies, with 4 investigating experimentally induced pancreatitis; 5 were human clinical studies; and 21 were rodent studies of experimentally induced pancreatitis. The level of evidence ranged between randomised controlled trials and case series, the estimated risk of bias ranged from low to high, and the sample sizes ranged from very small to moderate. Evidence indicates that adding corticosteroid to symptomatic treatment of acute/acute-on-chronic canine pancreatitis could have a positive influence on disease outcome. However, the analysed evidence was based on several species, including both naturally occurring and experimentally induced pancreatitis; thus, the authors suggest that large randomised controlled studies should be performed in dogs with spontaneously occurring acute/acute-on-chronic pancreatitis to further elucidate a potential benefit of corticosteroid treatment.

A retrospective study of 237 dogs hospitalized with suspected acute hemorrhagic diarrhea syndrome: Disease severity, treatment, and outcome.

BACKGROUND: Few studies have investigated management and outcome in dogs with acute hemorrhagic diarrhea syndrome (AHDS), and there is a paucity of data on dogs with concurrent signs of sepsis. OBJECTIVES: To report outcome in dogs with suspected AHDS according to disease severity and antimicrobial treatment, and to evaluate effect of fluid resuscitation on clinical criteria. ANIMALS: Two hundred thirty-seven dogs hospitalized with suspected AHDS. METHODS: Retrospective study based on medical records. Disease severity was evaluated using AHDS index, systemic inflammatory response syndrome (SIRS) criteria, and serum C-reactive protein (CRP) according to 3 treatment groups: No, 1, or 2 antimicrobials. RESULTS: Sixty-two percent received no antimicrobials, 31% received 1 antimicrobial, predominantly aminopenicillins, and 7% received 2 antimicrobials. At admission, median AHDS index was 13 (interquartile range, 11-15), which decreased significantly after the first day's hospitalization (P < .001) for all groups. Compared with no antimicrobials (7%), more dogs had ≥2 SIRS criteria in the antimicrobial groups (15% and 36%, respectively). C-reactive protein (CRP) correlated positively with AHDS index at hospitalization (P < .001). Across treatment groups, rehydration markedly reduced number of clinical SIRS criteria. Survival to discharge was 96%, lower for dogs receiving 2 antimicrobials (77%, P < .05). CONCLUSIONS AND CLINICAL IMPORTANCE: The majority of dogs hospitalized with suspected AHDS improve rapidly with symptomatic treatment only, despite signs of systemic disease on initial presentation. The often-used SIRS criteria might be a poor proxy for identifying dogs with AHDS in need of antimicrobial treatment, in particular when hypovolemic. The role of CRP in clinical decision-making or prognostication warrants further investigation.

Diabetic cats have decreased gut microbial diversity and a lack of butyrate producing bacteria.

Obesity and inactivity are major risk factors of feline diabetes mellitus (FDM) and human type II diabetes mellitus (T2DM). In recent years, changes in the gut microbiota have been suggested as a contributing factor to T2DM. Whether the gut microbiota (GM) composition plays a role in FDM remains unknown. The aim of the current study was firstly a cross-sectional comparison of the GM of diabetic cats, to that of lean, and of obese/overweight non-diabetic cats of a similar age. Specifically, fecal samples from 82 privately-owned cats from Denmark and Switzerland were sequenced using 16S rRNA gene amplicon metabarcoding. Secondly dietary intervention data was generated, by obtaining additional samples from a subset of cats after placing them on a high-protein diet for four weeks. The GM diversity of diabetic cats was lower than that of lean cats in the cross-sectional study, and lower compared to lean and to overweight/obese cats after diet intervention. Diabetic cats also exhibited fewer Anaerotruncus, Dialister, and unknown Ruminococcaceae than lean cats. Serum fructosamine levels correlated negatively with Prevotellaceae abundance and positively with Enterobacteriaceae abundance. In summary the intestinal microbiota of diabetic cats was characterized by decreased GM diversity and loss of butyrate producing bacterial genera.

European dog owner perceptions of obesity and factors associated with human and canine obesity.

Obesity is a common nutrition-related disorder leading to reduced life expectancy in both humans and dogs. With the aim of identifying new prevention and control options, the study objectives were (1) to investigate dog-owner perceptions about obesity in terms of themselves and their dogs, and (2) to identify factors associated with obesity and possible social, environmental and economic drivers for its development in dog owners and their pets. A cross-sectional questionnaire-based study was performed across multiple countries. The questionnaire focused on human and canine obesity, associated factors and potential drivers, and was distributed online and in the form of hard copies among dog owners in 11 European countries. In total, 3,185 responses from ten countries were included in multivariable analyses. Between 19.1% and 48.8% of the dog owners reported to be overweight/obese. Owner-reported overweight/obesity in dogs ranged from 6.0% to 31.3% based on body condition score charts, and 31.8% to 69.4% based on body fat index charts. Common factors associated with obesity in owners and their dogs were age, gender and owners' attitudes to diet and physical activity. Dog owners who did not consider obesity to be a disease were more likely to have obese dogs.

Insulin detemir treatment in diabetic cats in a practice setting.

Insulin detemir is a long-acting insulin analogue and may represent a valuable treatment option for diabetic cats. So far, only one study addressing detemir treatment of diabetic cats has been published, and this was based on an intensive blood glucose monitoring protocol. The aim of the current, retrospective study was to evaluate the effect of detemir therapy in diabetic cats in a general clinical setting. Fourteen diabetic cats with a follow-up period of at least 3 months were included. Data were collected from medical records at the University Hospital for Companion Animals, University of Copenhagen, Denmark. Thirteen of 14 cats achieved moderate or excellent control of clinical symptoms within the initial 3 months of detemir therapy, including five cats previously treated unsuccessfully with other types of insulin. Clinical improvements were noted after 1 month of therapy and continued over time. Three cats achieved remission within the initial 3 months and none experienced a diabetic relapse during the study period. One cat achieved remission after 13 months of therapy. Improvements in clinical symptoms were markedly better than indicated by blood glucose and serum fructosamine concentrations. The safety of detemir was very high, with only two reported episodes of clinical hypoglycaemia, neither of which required veterinary attention. Based on these results detemir can be recommended for the treatment of diabetic cats, including cats previously treated unsuccessfully with other types of insulin.

Exploring the feasibility and synergistic value of the One Health approach in clinical research: protocol for a prospective observational study of diagnostic pathways in human and canine patients with suspected urinary tract infection.

BACKGROUND: The One Health approach is emerging in response to the development of bacterial resistance. To the best of our knowledge, the possibility to use this approach in a clinical context has not yet been explored. Thus, in this paper, we report the procedures to implement a prospective observational study of diagnostic pathways in human and canine patients with suspected urinary tract infection as a means to assess the feasibility and synergistic value of setting up One Health clinical research projects and interventions. METHODS/DESIGN: A prospective observational study will compare different diagnostic pathways (i.e., 16 possible combinations of diagnostic tools) to gold standard in human and veterinary primary care practice in Denmark. Fifty primary care practices and 100 veterinary clinics will each consecutively include 20 human patients or 8-10 dogs, respectively. Data will be collected at practice and patient level comprising (a) information about the organization of the practice and access to different diagnostic tools, (b) information about clinical history, diagnostic path and treatment during the index consultation, (c) information about severity of symptoms during the 7-10 days following inclusion, and (d) urine culture (type of microorganism and susceptibility test). The feasibility and synergistic value of conducting future research, and/or designing common interventions, will be assessed by evaluating the comparability of human primary care and veterinary primary care with respect to study implementation and study results. DISCUSSION: Results from this study will give an insight into the feasibility and synergistic value of setting-up One Health research projects in a clinical context. This is crucial if we are to embrace the One Health approach, as a legitimate strategy to implement common interventions aimed at influencing the diagnostic process in human and canine patients in order to decrease inappropriate use of antibiotics. TRIAL REGISTRATION: The study in humans has been registered in ClinicalTrials.gov NCT02249273.

Quantitative assessment of faecal shedding of ß-lactam-resistant Escherichia coli and enterococci in dogs.

Quantitative data on faecal shedding of antimicrobial resistant bacteria are crucial to assess the risk of transmission from dogs to other animals as well as humans. In this study we investigated prevalence and concentrations of ß-lactam-resistant Escherichia coli and enterococci in the faeces of 108 dogs presenting at a veterinary hospital in Denmark. The dogs had not been treated with antimicrobials for 4 weeks prior to the study. Total E. coli and enterococci were quantified by counts on MacConkey and Slanetz-Bartley, respectively. Resistant E. coli and enterococci were counted on the same media containing relevant antibiotic concentrations, followed by species identification using MALDI-TOF. Ampicillin- and cefotaxime-resistant E. coli were detected in 40% and 8% of the dogs, respectively, whereas approximately 15% carried ampicillin-resistant enterococci, mainly Enterococcus faecium. In the faeces of the carriers, the proportion of resistant strains in the total bacterial species population was on average 15% for both ampicillin-resistant E. coli (median faecal load 3.2×10(4)cfu/g) and E. faecium (5.8×10(2) cfu/g), and 4.6% for cefotaxime-resistant E. coli (8.6×10(3) cfu/g). Cefotaxime resistance was associated with the presence of blaCTX-M-1 (n=4), blaCMY-2 (n=4) or multiple mutations in the promoter and coding region of chromosomal ampC (n=1). Altogether the results indicate that the risks of zoonotic transmission of ß-lactam-resistant bacteria via human exposure to canine faeces greatly vary amongst individual dogs and are influenced by unidentified factors other than recent antimicrobial use.

Preterm birth makes the immature intestine sensitive to feeding-induced intestinal atrophy.

Preterm birth and formula feeding predispose to small intestinal dysfunction, which may lead to necrotizing enterocolitis (NEC). In piglets, we tested whether the physiological and environmental transitions occurring at birth affect the response of the immature intestine to enteral feeding. Pig fetuses (106 days gestation, term = 115 days) were prepared with esophageal feeding tubes and fed either sow's colostrum (n = 8) or infant formula (n = 7) in utero. After 24 h of oral feeding, the pig fetuses were delivered by cesarean section and their gastrointestinal morphology and function were compared with those of preterm newborn (NB) littermates that were not fed (n = 8) or fed colostrum (n = 7) or formula (n = 13) for 24 h after birth. Before birth, both colostrum and formula feeding resulted in marked increases in intestinal mass, brush-border enzyme activities, and plasma glucagon-like peptide 2 concentrations, to levels similar to those in NB colostrum-fed piglets. In contrast, NB formula-fed piglets showed reduced intestinal growth, decreased brush-border enzyme activities, and intestinal lesions, reflecting NEC. NB formula-fed pigs also showed impaired enterocyte endocytotic function and decreased antioxidative capacity, whereas brush-border enzyme mRNA levels were unaltered, relative to NB colostrum-fed pigs. Our results indicate that the feeding-induced growth and enzyme maturation of the immature intestine are not birth dependent. However, with a suboptimal diet (milk formula), factors related to preterm birth (e.g., microbial colonization and metabolic and endocrine changes) make the immature intestine sensitive to atrophy and development of NEC.