Development of a fixative protocol using formaldehyde and gluteraldehyde for preservation of microbial art on agar plates.

AIMS: Agar art bridges the gap between science and art using microbes instead of paint. Afterwards, the art can change in response to microbial fluctuation, meaning preservation of the original art is essential. Here, formaldehyde and glutaraldehyde were investigated as preservatives, involving techniques used in healthcare settings to preserve samples. METHODS AND RESULTS: Formaldehyde was tested at 1.0%, 2.0% and 3.7%, w/v, whereas glutaraldehyde was tested at 1% and 2.5%, w/v. Both compounds and respective concentrations were tested for different time periods. Escherichia coli, Serratia marcescens, Staphlococcus aureus and Micrococcus luteus were used as bacteria for "drawing" the works of art. The effectiveness of fixation was determined using integrated densities and visual assessment. Initially, both compounds showed potential promise, albeit with a loss of bacteria. Ser. marcescens was prone to colour changes and glutaraldehyde caused discolouration of agar and bacteria. These could be caused by a pH decrease in the agar, due to residual free aldehyde groups. Reduction of this was tested using 300 mM sodium metabisulfite to neutralize excess aldehydes. This initially led to reduced bacterial loss and avoided colour changes, however measurements 24 h post-fixation showed colour loss to some bacterial clusters. CONCLUSIONS: Here, at least 2% formaldehyde for a short fixation period, typically 1 min, depending on the species, was most promising for the preservation of art. Given the success of this with different bacteria, it would make a good starting combination for anyone trying to fix agar art, although methodology refinement may be needed for optimisation depending on the bacterial species used. SIGNIFICANCE AND IMPACT OF STUDY: This study shows, for the first time, successful fixation and preservation of different bacterial species on agar. The impact of this is to preserve agar art while making it safe and non-infective to those in contact with the microbial art.

MKLN1 splicing defect in dogs with lethal acrodermatitis.

Lethal acrodermatitis (LAD) is a genodermatosis with monogenic autosomal recessive inheritance in Bull Terriers and Miniature Bull Terriers. The LAD phenotype is characterized by poor growth, immune deficiency, and skin lesions, especially at the paws. Utilizing a combination of genome wide association study and haplotype analysis, we mapped the LAD locus to a critical interval of ~1.11 Mb on chromosome 14. Whole genome sequencing of an LAD affected dog revealed a splice region variant in the MKLN1 gene that was not present in 191 control genomes (chr14:5,731,405T>G or MKLN1:c.400+3A>C). This variant showed perfect association in a larger combined Bull Terrier/Miniature Bull Terrier cohort of 46 cases and 294 controls. The variant was absent from 462 genetically diverse control dogs of 62 other dog breeds. RT-PCR analysis of skin RNA from an affected and a control dog demonstrated skipping of exon 4 in the MKLN1 transcripts of the LAD affected dog, which leads to a shift in the MKLN1 reading frame. MKLN1 encodes the widely expressed intracellular protein muskelin 1, for which diverse functions in cell adhesion, morphology, spreading, and intracellular transport processes are discussed. While the pathogenesis of LAD remains unclear, our data facilitate genetic testing of Bull Terriers and Miniature Bull Terriers to prevent the unintentional production of LAD affected dogs. This study may provide a starting point to further clarify the elusive physiological role of muskelin 1 in vivo.

Characterisation of the effect of day length, and associated differences in dietary intake, on the gut microbiota of Soay sheep.

Differences in the rumen bacterial community have been previously reported for Soay sheep housed under different day length conditions. This study extends this previous investigation to other organs of the digestive tract, as well as the analysis of ciliated protozoa and anaerobic fungi. The detectable concentrations of ciliated protozoa and anaerobic fungi decreased with increased day length in both the rumen and large colon, unlike those of bacteria where no effect was observed. Conversely, bacterial community composition was affected by day length in both the rumen and large colon, but the community composition of the detectable ciliated protozoa and anaerobic fungi was not affected. Day length-associated differences in the bacterial community composition extended to all of the organs examined, with the exception of the duodenum and the jejunum. It is proposed that differences in rumen fill and ruminal 'by-pass' nutrients together with endocrinological changes cause the observed effects of day length on the different gut microbial communities.

Using next-generation sequencing to determine diversity of horse intestinal worms: identifying the equine 'nemabiome'.

Next-generation sequencing of DNA from nematode eggs has been utilised to give the first account of the equine 'nemabiome'. In all equine faecal samples investigated, multiple species of Strongylidae were detected, ranging from 7.5 (SEM 0.79) with 99+% identity to sequences in the NCBI database to 13.3 (SEM 0.80) with 90+% identity. This range is typical of the number of species described previously in morphological studies using large quantities of digesta per animal. However, the current method is non-invasive; relies on DNA analysis, avoiding the need for specialist microscopy identification; and can be carried out with small samples, providing significant advantages over current methods.

Routine antibiotic therapy in dogs increases the detection of antimicrobial-resistant faecal Escherichia coli.

Background: Antimicrobial resistance (AMR) is a critical health problem, with systemic antimicrobial therapy driving development of AMR across the host spectrum. Objectives: This study compares longitudinal carriage, at multiple timepoints, of AMR faecal Escherichia coli in dogs undergoing routine antimicrobial treatment. Methods: Faecal samples (n = 457) from dogs (n = 127) were examined pretreatment, immediately after treatment and 1 month and 3 months post-treatment with one of five antimicrobials. Isolates were tested for susceptibility to a range of antimicrobials using disc diffusion for each treatment group at different timepoints; the presence/absence of corresponding resistance genes was investigated using PCR assays. The impact of treatment group/timepoint and other risk factors on the presence of resistance [MDR, fluoroquinolone resistance, third-generation cephalosporin resistance (3GCR) and ESBL and AmpC production] was investigated using multilevel modelling. Samples with at least one AMR E. coli from selective/non-selective agar were classed as positive. Resistance was also assessed at the isolate level, determining the abundance of AMR from non-selective culture. Results: Treatment with ß-lactams or fluoroquinolones was significantly associated with the detection of 3GCR, AmpC-producing, MDR and/or fluoroquinolone-resistant E. coli, but not ESBL-producing E. coli, immediately after treatment. However, 1 month post-treatment, only amoxicillin/clavulanate was significantly associated with the detection of 3GCR; there was no significant difference at 3 months post-treatment for any antimicrobial compared with pretreatment samples. Conclusions: Our findings demonstrated that ß-lactam and fluoroquinolone antibiotic usage is associated with increased detection of important phenotypic and genotypic AMR faecal E. coli following routine therapy in vet-visiting dogs. This has important implications for veterinary and public health in terms of antimicrobial prescribing and biosecurity protocols, and dog waste disposal.

Detecting genetic regions associated with height in the native ponies of the British Isles by using high density SNP genotyping.

Height is an important characteristic in the equine industry although little is known about its genetic control in native British breeds of ponies. This study aimed to map QTL data with the withers height in four pony breeds native to the British Isles, including two different sections within Welsh Cobs. In this study, a genome-wide analysis approach using the Illumina EquineSNP50 Infinium BeadChip was applied to 105 ponies and cobs. Analysis identified 222 highly significant height-associated SNPs (P ≤ 10-5), among which three SNPs on ECA9 have also been previously reported elsewhere. The highest number of significant SNPs associated to height in the native British horses were located on ECA1, ECA8, and ECA16.

Impact of systemic antimicrobial therapy on mucosal staphylococci in a population of dogs in Northwest England.

BACKGROUND: Antimicrobial-resistant bacteria are increasingly isolated from veterinary patients. OBJECTIVES: To determine risk factors for antimicrobial resistance (AMR) among canine mucosal staphylococci following routine antimicrobial treatment with cefalexin (CFX), clavulanate-amoxicillin (AC), cefovecin (CVN), clindamycin (CD) or a fluoroquinolone (FQ). ANIMALS: Mucosal swab samples (n = 463) were collected from 127 dogs pre-treatment, immediately, and at one- and three-months post-treatment. METHODS: Staphylococci were identified phenotypically and biochemically as coagulase negative (CoNS) or coagulase positive (CoPS); CoPS were speciated by nuc gene PCR. Antimicrobial susceptibility was determined using disc diffusion and mecA gene carriage by PCR. Multilevel, multivariable models examined associations between risk factors and presence/absence of CoPS, meticillin resistance (MR), multidrug-resistance (MDR) and fluoroquinolone resistance (FQR). RESULTS: The percentage of samples with CoNS increased and with CoPS (including S. pseudintermedius) decreased immediately post-treatment with CFX, CVN and CD (P ≤ 0.001) and one month post-treatment with CD (P = 0.003). By three months post-treatment, there was no significant difference compared to pre-treatment samples. Immediately post-treatment with FQs there was significantly increased risk of isolating MRS (P = 0.002), MDR (P = 0.002) or FQR (P = 0.013) staphylococci and of MDR following CFX treatment (P = 0.019). The percentage of samples with AMR staphylococci declined from immediately to three months post-treatment and there was no significant difference between resistance prevalence at one or three months post-treatment for most AMR traits and treatment groups. Exceptions include increased MDR following FQ (P = 0.048) or CFX (P = 0.021), at one and three months post-treatment, respectively. CONCLUSIONS AND CLINICAL IMPORTANCE: Systemic antimicrobials impact on mucosal staphylococci. Immediately after therapy, the mucosa may be a reservoir for AMR staphylococci that are a source of mobile genetic elements carrying AMR genes.

The effect of administration of fenbendazole on the microbial hindgut population of the horse.

Anthelmintics are used as anti-worming agents. Although known to affect their target organisms, nothing has been published regarding their effect on other digestive tract organisms or on metabolites produced by them. The current work investigated effects of fenbendazole, a benzimidazole anthelmintic, on bacteria and ciliates in the equine digestive tract and on and their major metabolites. Animals receiving anthelmintic treatment had high faecal egg counts relative to controls. Analysis was performed over two weeks, with temporal differences detected in bacterial populations but with no other significant differences detected. This suggests fenbendazole has no detectable effect on organisms other than its targets. Moreover it does not appear to make a contribution to changing the resulting metabolome.

A pilot study comparing in vitro efficacy of topical preparations against veterinary pathogens.

BACKGROUND: Topical antimicrobial agents are important for the management of cutaneous infections. For topical antimicrobial agents, in vitro efficacy data are limited. OBJECTIVES: To determine and compare the minimum bactericidal/fungicidal concentrations (MBCs/MFCs) of several topical antimicrobial agents against veterinary pathogens. MATERIALS AND METHODS: Two chlorhexidine, two oxychlorine based products (NaOCl & HOCl) lime sulfur (LS), manuka honey (MH) and hydrocortisone aceponate (HCA) were tested against American Type Culture Collection (ATCC) and clinical isolates: meticillin susceptible and resistant Staphylococcus pseudintermedius (MSSP), qac A/B carrying MSSP, antimicrobial susceptible and extended spectrum beta-lactamase producing Escherichia coli, multidrug-resistant Pseudomonas aeruginosa and Malassezia pachydermatis. The MBCs/MFCs were measured, where available, using a broth microdilution method; isolates were incubated for 3 and 10 min. RESULTS: Chlorhexidine and isopropyl alcohol (Chl(1) ) showed significantly lower MBCs (0.46 mg/L -937.50 mg/L, P = 0.027) compared to chlorhexidine and climbazole (Chl², 58.59 mg/L-1875 mg/L). NaOCl and HOCl showed excellent antimicrobial activity with HOCl having significantly lower MBCs compared to NaOCl (0.03 mg/L-1.72 mg/L and 0.03 mg/L-1.95 mg/L, respectively, P = 0.042). The detectable MBCs for LS and HCA were high, being close to the starting concentration (5,000 mg/L and 146 mg/L, respectively). The MBC/MFC for MH was not detectable. Amongst all test products there was no significant effect of contact time or isolate resistance status. CONCLUSIONS AND CLINICAL IMPORTANCE: Chlorhexidine, NaOCl and HOCl demonstrated low MBCs against tested organisms, suggesting potential in vivo efficacy. The selection of an appropriate antimicrobial agent, however, cannot be based exclusively upon MBC/MFC data; other factors should be considered.

Sweat osmolarity shows intra-animal regional variation in the horse.

BACKGROUND: Sweating is important in regulating body temperature but can be a source of loss of both fluids and electrolytes. Although the process has been studied in horses, the variation in sweat osmolarity across the body has not. OBJECTIVES: This work describes an investigation to determine if there is regional variation in the osmolarity of sweat across different anatomical regions of the horse. ANIMALS: Ten horses were used in the study and were animals either stabled for riding lessons or had livery on-site. METHODS: Sweat samples were collected from five regions on each horse following exercise and the osmolarity measurements were made using an Osmomat 030 (Gonotec, Berlin, Germany). Values were analysed by paired t-tests and analysis of variance. RESULTS: Samples from the back and ears had statistically (P < 0.05) lower osmolarity values than those seen for the neck and forelimb, with thigh values intermediate between the other two sets of values. CONCLUSIONS AND CLINICAL IMPORTANCE: Previous studies have used osmolarity values based on the sweat collected from the horse's back. The current work demonstrates that these values are probably an underestimation of electrolyte loss, which may have implications for the composition and administration of rehydration compounds.

Antimicrobial resistance and characterisation of staphylococci isolated from healthy Labrador retrievers in the United Kingdom.

BACKGROUND: Coagulase-positive (CoPS) and coagulase-negative (CoNS) staphylococci are normal commensals of the skin and mucosa, but are also opportunist pathogens. Meticillin-resistant (MR) and multidrug-resistant (MDR) isolates are increasing in human and veterinary healthcare. Healthy humans and other animals harbour a variety of staphylococci, including MR-CoPS and MR-CoNS. The main aims of the study were to characterise the population and antimicrobial resistance profiles of staphylococci from healthy non-vet visiting and non-antimicrobial treated Labrador retrievers in the UK. RESULTS: Nasal and perineal samples were collected from 73 Labrador retrievers; staphylococci isolated and identified using phenotypic and biochemical methods. They were also confirmed by matrix-assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI-TOF-MS), PCR of the nuc gene and PCR and sequencing of the tuf gene. Disc diffusion and minimum inhibitory concentration (MIC) susceptibility tests were determined for a range of antimicrobials. In total, 102 CoPS (S. pseudintermedius n = 91, S. aureus n = 11) and 334 CoNS isolates were detected from 99% of dogs in this study. In 52% of dogs CoNS only were detected, with both CoNS and CoPS detected in 43% dogs and CoPS only detected in 4% of dogs. Antimicrobial resistance was not common among CoPS, but at least one MDR-CoNS isolate was detected in 34% of dogs. MR-CoNS were detected from 42% of dogs but no MR-CoPS were isolated. S. epidermidis (52% of dogs) was the most common CoNS found followed by S. warneri (30%) and S. equorum (27%), with another 15 CoNS species isolated from ≤ 15% of dogs. S. pseudintermedius and S. aureus were detected in 44% and 8% of dogs respectively. CONCLUSIONS: MR- and MDR-CoPS were rare. However a high prevalence of MR- and MDR-CoNS were found in these dogs, even though they had no prior antimicrobial treatment or admission to veterinary premises. These findings are of concern due to the potential for opportunistic infections, zoonotic transmission and transmission of antimicrobial resistant determinants from these bacteria to coagulase positive staphylococci.

Tris-EDTA significantly enhances antibiotic efficacy against multidrug-resistant Pseudomonas aeruginosa in vitro.

BACKGROUND: Multidrug-resistant Pseudomonas aeruginosa commonly complicates chronic bacterial otitis in dogs. HYPOTHESIS/OBJECTIVES: The aim of this in vitro study was to determine the effect of ethylenediaminetetraacetic acid-tromethamine (Tris-EDTA) on the minimal bactericidal concentrations (MBCs) and minimal inhibitory concentrations (MICs) of marbofloxacin and gentamicin for multidrug-resistant P. aeruginosa isolates from cases of canine otitis. METHODS: Eleven isolates were identified as multidrug resistant on disc diffusion; 10 were resistant to marbofloxacin and two were resistant to gentamicin. Isolates were incubated for 90 min with each antibiotic alone and in combination with Tris-EDTA at concentrations of 0.075 µg/mL to 5 mg/mL for marbofloxacin, 0.001 µg/mL to 10 mg/mL for gentamicin and 17.8:4.7 to 0.14:0.04 mg/mL for Tris-EDTA. Positive and negative controls were included. Aliquots of each antibiotic and/or Tris-EDTA concentration were subsequently transferred to sheep blood agar to determine the MBCs, and tryptone soy broth was added to the remaining suspensions to determine the MICs. RESULTS: Tris-EDTA alone was bacteriostatic but not bactericidal at any concentration. The addition of Tris-EDTA significantly reduced the median MBC (from 625 to 468.8 µg/mL; P < 0.001) and MIC (from 29.3 to 2.4 µg/mL; P = 0.008) of marbofloxacin, and the median MBC (from 625 to 39.1 µg/mL) and MIC (from 19.5 to 1.2 µg/mL) of gentamicin (both P < 0.001). CONCLUSIONS AND CLINICAL IMPORTANCE: Tris-EDTA significantly reduced the MBCs and MICs of marbofloxacin and gentamicin for multidrug-resistant P. aeruginosa in vitro. This may be of use to clinicians managing these infections in dogs.

Cross-reaction and co-sensitization among related and unrelated allergens in canine intradermal tests.

BACKGROUND: Intradermal tests are used to identify allergens for avoidance and immunotherapy in atopic dogs. HYPOTHESIS/OBJECTIVES: To evaluate cross-reaction or co-sensitization among 53 intradermal test allergens. ANIMALS: Six hundred and fifty-one client-owned dogs with atopic dermatitis. METHODS: Intradermal tests were performed with 53 house dust/storage mite, epidermal, insect, tree, weed and grass pollen and mould allergens. Pairwise comparisons were used to calculate the odds ratios (ORs), 95% confidence intervals (CIs) and statistical significance for the results of each allergen pair, with significance at P < 0.0006 (Holm-Bonferroni correction to reduce the false-detection rate). RESULTS: Apart from cotton, cockroach, red clover, grain smut and Penicillium, the results for the allergens within each group were statistically associated [ORs from 4.7 (CI 2.5-8.9) to 1229.4 (CI 166.5-1795.1); P = 0.0005 to P < 0.0001]. Excluding red clover and cotton, 94% of results between tree, weed and grass pollens were also statistically associated [ORs from 8.3 (CI 3.6-24.7) to 117 (CI 29.1-341); P = 0.0005 to P < 0.0001]. In contrast, few allergens from unrelated groups were statistically associated [ORs from 0.12 (CI 0.03-1.1) to 27.7 (CI 0.2-93); P = 1.0 to P < 0.0001]. The mean (SD) of the log e transformed ORs for the related and statistically associated allergens [5.3 (1.3)] was significantly greater than those for related but nonstatistically associated [1.7 (1.6)] or unrelated allergens [1.4 (1.4); P < 0.0001]. CONCLUSIONS AND CLINICAL IMPORTANCE: This suggests that there is cross-reaction or co-sensitization between related allergens. This could have implications for allergen selection in testing and immunotherapy, but further studies are required to differentiate cross-reaction from co-sensitization.

Bacteriophage Therapy to Control Bovine Mastitis: A Review.

Bovine mastitis is a polymicrobial disease characterised by inflammation of the udders of dairy and beef cattle. The infection has huge implications to health and welfare of animals, impacting milk and beef production and costing up to EUR 32 billion annually to the dairy industry, globally. Bacterial communities associated with the disease include representative species from Staphylococcus, Streptococcus, Enterococcus, Actinomyces, Aerococcus, Escherichia, Klebsiella and Proteus. Conventional treatment relies on antibiotics, but antimicrobial resistance, declining antibiotic innovations and biofilm production negatively impact therapeutic efficacy. Bacteriophages (phages) are viruses which effectively target and lyse bacteria with extreme specificity and can be a valuable supplement or replacement to antibiotics for bovine mastitis. In this review, we provide an overview of the etiology of bovine mastitis, the advantages of phage therapy over chemical antibiotics for the strains and research work conducted in the area in various model systems to support phage deployment in the dairy industry. We emphasise work on phage isolation procedures from samples obtained from mastitic and non-mastitic sources, characterisation and efficacy testing of single and multiple phages as standalone treatments or adjuncts to probiotics in various in vitro, ex vivo and in vivo bovine mastitis infection models. Furthermore, we highlight the areas where improvements can be made with focus on phage cocktail optimisation, formulation, and genetic engineering to improve delivery, stability, efficacy, and safety in cattle. Phage therapy is becoming more attractive in clinical medicine and agriculture and thus, could mitigate the impending catastrophe of antimicrobial resistance in the dairy sector.

A Systematic Review on the Role of Wildlife as Carriers and Spreaders of Campylobacter spp.

Campylobacter spp. are important zoonotic pathogens and can cause one of the main bacterial diarrheal diseases worldwide. Research in the context of infection arising from transmission from other humans and other vertebrates has been extensive. A large fraction of these investigations has focused on domestic animals; however, there are also a number of publications which either totally, or at least in part, consider the role of wild or feral animals as carriers or spreaders of Campylobacter spp. Here, we carry out a systematic review to explore the role played by wild vertebrates as sources of Campylobacter spp. with a compilation of prevalence data for more than 150 species including reptiles, mammals and birds. We found that numerous vertebrate species can act as carriers of Campylobacter species, but we also found that some host specificity may exist, reducing the risk of spread from wildlife to domestic animals or humans.

Comparative in vitro efficacy of antimicrobial shampoos: a pilot study.

This study compared the antimicrobial efficacy of shampoos against meticillin-sensitive Staphylococcus pseudintermedius (MSSP), meticillin-resistant S. pseudintermedius (MRSP), antibiotic-sensitive Pseudomonas aeruginosa (PA), multidrug-resistant P. aeruginosa (MDR-PA) and Malassezia pachydermatis. Three isolates were incubated for 10, 30 and 60 min with each shampoo diluted in phosphate-buffered saline. Aliquots were then incubated for 16-18 h on sheep blood agar (bacteria) or for 3 days on Sabouraud's dextrose agar (Malassezia). The minimal bactericidal concentrations (MBCs) for chlorhexidine products (Malaseb(®), Pyoderm(®)/Microbex(®) and Hibiscrub(®)) were 1:1,024-1:2,048 for MSSP and MRSP, 1:512-1:1,024 for PA and MDR-PA, and 1:2,048-1:5,096 for Malassezia at all time points. The MBCs for benzoyl peroxide (Paxcutol(®)) for MSSP and MRSP were 1:2-1:8 at 10 min, and 1:256 after 30 and 60 min. A 1:2 dilution was effective against Pseudomonas, and 1:512-1:1,024 dilutions were effective against Malassezia at all time points. The MBCs for ethyl lactate (Etiderm(®)) for MSSP and MRSP were 1:2 at 10 min, and 1:2-1:16 after 30 and 60 min. A 1:2 dilution was effective against Pseudomonas, and a 1:512 dilution was effective against Malassezia at all time points. Chloroxylenol (Coatex(®)) and acetic acid-boric acid (Malacetic(®)) were not effective against MSSP, MRSP or Pseudomonas. Both were effective against Malassezia at 1:8-1:16 dilution at 10 min, and at 1:8-1:32 dilution after 30 and 60 min. In conclusion, chlorhexidine appeared to be the most effective topical biocide, and MRSP and MDR-PA were no less susceptible than antibiotic-sensitive organisms. These results should, however, be confirmed with larger numbers of isolates.

Comparable efficacy of a topical 0.0584% hydrocortisone aceponate spray and oral ciclosporin in treating canine atopic dermatitis.

This study compared the efficacy of a 0.0584% hydrocortisone aceponate (HCA) spray (Cortavance(®); Virbac SA) and ciclosporin (Atopica(®); Novartis Animal Health) in canine atopic dermatitis in a single-blind randomized controlled trial. Dogs received HCA (two sprays/100 cm(2); n=24) or ciclosporin (5 mg/kg; n=21). Canine Atopic Dermatitis Extent and Severity Index (CADESI)-03, pruritus (visual analog scale with grade descriptors) and owner scores (5-point scales) were recorded every 28 days for 84 days. Intention-to-treat data were analysed. CADESI-03 and pruritus significantly decreased over time (P<0.0001), but there was no difference between the treatment groups (P=0.91 and P=0.52, respectively). Similar proportions of HCA- and ciclosporin-treated dogs achieved ≥50% reductions in CADESI-03 and pruritus scores at 28 days (CADESI-03 58.3 and 57.1%, P=0.76; pruritus 33.3 and 38.1%, P=1.0), 56 days (CADESI-03 70.8 and 81.0%, P=1.0; pruritus 62.5 and 57.1%, P=1.0) and 84 days (CADESI-03 75 and 85.7%, P=0.72; pruritus 65.2 and 57.1%, P=0.76). The CADESI-03 and pruritus scores were close to equivalence (0.47 and 0.51, respectively). By 84 days, every-other-day or twice-weekly therapy was achieved in 13 of 24 HCA- and 12 of 21 ciclosporin-treated dogs (P=0.85). There were no significant differences in scores for efficacy (P=0.82), tolerance (P=0.62) and ease of administration (P=0.25). Scores for tolerance (0.49) and administration (0.46) were close to equivalence. The score for efficacy favoured HCA (0.68). Mild adverse events were noted in six of 21 ciclosporin and none of 24 HCA dogs (P=0.008). Five HCA-treated dogs and three ciclosporin-treated dogs were prematurely withdrawn (P=0.7). In conclusion, HCA and ciclosporin proved equally effective in treating canine atopic dermatitis for up to 84 days.

Efficacy of a 0.0584% hydrocortisone aceponate spray in presumed feline allergic dermatitis: an open label pilot study.

This study evaluated the efficacy of a 0.0584% hydrocortisone aceponate (HCA) spray (Cortavance(®); Virbac SA) in 10 cats with presumed allergic dermatitis. The cats initially received two sprays/100 cm(2) of skin once daily. Clinical lesions (a Feline Dermatitis Extent and Severity Index; FeDESI), pruritus (10 cm visual analog scale with grade descriptors) and owner assessments of efficacy, tolerance and ease of use (from 1=very poor to 5=excellent) were assessed every 14 days. The frequency of treatment was reduced after day 28 in cats with a >50% reduction in FeDESI and pruritus scores. One cat was lost to follow up at day 28 and two at day 42. Intention-to-treat data were analysed. The FeDESI [mean (SD): day 0, 42.2 (15.7) and day 56, 9.9 (11.7); P<0.0001] and pruritus scores [day 0, 61.2 mm (20.1) and day 56, 14.6 mm (16.1); P<0.0001] significantly decreased throughout the trial. The owner scores for tolerance [median (range): day 14, 4 (1-5) and day 56, 4 (3-5); P=0.003] and ease of administration [day 14, 3 (2-5) and day 56, 4 (2-5); P=0.02] significantly increased during the trial, but there was no significant change in efficacy scores [day 14, 4 (3-5) and day 56, 4 (2-5); P=0.5]. There were no adverse effects attributable to the HCA spray, no significant changes in weight [mean (SD): day 0, 5.0 kg (1.4) and day 56, 5.0 kg (1.6); P=0.51] and no significant changes in haematology, biochemistry or urinalysis (n=4). Six cats required every-other-day treatment and four required daily treatment. In conclusion, HCA spray appeared to be effective and safe in these cats, although it is not licensed for use in this species.

Effect of Feeding Insoluble Fiber on the Microbiota and Metabolites of the Caecum and Feces of Rabbits Recovering from Epizootic Rabbit Enteropathy Relative to Non-Infected Rabbits.

This study aimed to investigate the effect of feeding insoluble fiber on the microbiota and metabolites of the caecum and feces of rabbits recovering from epizootic rabbit enteropathy relative to non-infected rabbits. Rabbits that had either recovered from epizootic rabbit enteropathy or ones that had never had epizootic rabbit enteropathy were fed on a diet of 32% or 36% neutral detergent fiber until they were 70 days of age. At this point, the short-chain fatty acid and ammonia levels were measured in caecotroph and fecal samples and compared using 2 × 2 ANOVA. The microbial composition of the samples was also analyzed using next-generation sequencing and compared by PERMANOVA. Caecotrophic samples from previously affected rabbits on lower fiber diets had higher short-chain fatty acid contents and higher species diversity index values for some indices (p < 0.05), although the fecal samples showed lower species diversity levels (p < 0.05). In addition, the PERMANOVA analyses demonstrated that differences were detected in the microbial composition of both fecal and caecotrophic samples, depending on the disease status at the outset of the experiment (p < 0.05). The results of this work show that, although there is some potential in the use of high-fiber diets for the treatment of rabbits that have had epizootic rabbit enteropathy, they are not able to produce the same digestive tract properties as those seen in rabbits that have never had the condition. This is true even after the rabbits have recovered from epizootic rabbit enteropathy.

House dust and storage mite contamination of dry dog food stored in open bags and sealed boxes in 10 domestic households.

Dry pet food is a potential source of exposure to house dust and storage mite allergens in canine atopic dermatitis. This study evaluated contamination of house dust and dry dog food stored in paper bags, sealable plastic bags and sealable plastic boxes in 10 households for 90 days using Acarex(®) tests for guanine, a Der p 1 ELISA and mite flotation. Acarex(®) tests were negative in all the food samples but positive in all the house dust samples. The Der p 1 levels and mite numbers significantly increased in food from paper bags (P = 0.0073 and P = 0.02, respectively), but not plastic bags or boxes. Mite numbers and Der p 1 levels were 10-1000 times higher in house dust than the corresponding food samples (P < 0.0001). There were significant correlations between Der p 1 in house dust and food from the paper (P < 0.0001) and plastic bags (P = 0.003), and mite numbers in house dust and food from the paper bags (P = 0.0007). Bedding and carpets were significantly associated with Der p 1 levels in house dust (P = 0.015 and P = 0.01, respectively), and food from the paper (both P = 0.02) and plastic bags (P = 0.03 and P = 0.04, respectively). Mites were identified in six of 10 paper bag, three of 10 plastic bag, one of 10 plastic box and nine of 10 house dust samples. These comprised Dermatophagoides (54%), Tyrophagus (10%; all from food) and unidentified mites (36%). Storage of food in sealable plastic boxes largely prevented contamination for 3 months. Exposure to mites and mite proteins in all the stored food, however, appeared to be trivial compared with house dust.