Diagnostic accuracy of a radiographic device to assess cranial tibial translation in dogs: validation protocol

This protocol describes a validation procedure to assess the diagnostic accuracy of a radiographic method using a simple device, specifically designed to quantify the cranial tibial translation in dogs

Ovine Paratuberculosis: A Seroprevalence Study in Dairy Flocks Reared in the Marche Region, Italy

In order to fulfil the seroprevalence gap on Mycobacterium avium subsp. paratuberculosis infection in ovine dairy farms of Marche region (central Italy), a stratified study was carried out on 2086 adult female sheep randomly chosen from 38 herds selected in Ancona and Macerata provinces. 73.7% flocks resulted infected by a commercial ELISA test (Pourquier, France), with a mean seroprevalence of 6.29% of sampled sheep in both provinces. A higher number of MAP seropositive ewes was recorded in the large herds' consistence than in the small and medium herds' consistence (P = 0.0269), and a greater percentage of infected sheep was obtained among female at early/late than in peak lactation stage (P = 0.0237). MAP infection was confirmed in 12.6% of infected farms by faecal culture. The true sheep-level seroprevalence was 15.1% ± 7.3%

Retrospective analysis of antibacterial resistance among uropathogen Escherichia coli in a veterinary teaching hospital (Italy, 2014-2019)

Background: Urinary tract infections (UTIs) often require long follow-up periods and repeated antimicrobial therapies that can lead to the onset of antimicrobial-resistance. Escherichia coli is the most frequent bacterium involved in uncomplicated UTIs in pet animals in which treatment is sometimes threatened by the steady increase in the number of strains bearing concurrent resistance to various antimicrobial agents. The study aimed to report the variation of antibacterial resistance of urinary pathogen E. coli (UPEC) isolated from pets in a veterinary teaching hospital, North Italy (Turin) during a 5-and-a-half-year period (2014-2019). Materials/methods: This retrospective study was carried out on E. coli strains (n= 219) collected from dogs (n=139) and cats (n=80) with UTI. Each strain was tested to 18 antibiotics belonging to 8 categories: aminoglycosides, carbapenems, folate pathway inhibitors, not-extended spectrum cephalosporins: 1st and 2nd generation, extended spectrum cephalosporins: 3rd and 4th generation, penicillins, penicillins + β-lactamase inhibitors, quinolones, following Kirby-Bauer method and interpreted according to the EUCAST guidelines. Isolates were classified as MDR (multidrug-resistant), XDR (MDR susceptible to only one or two antibiotic categories) and PDR (resistant to all agents tested). Data were analyzed using χ2 test, Pearson’s correlation among years and variance-weighted least-square regression models with STATA 15.1, choosing a significance level of α=0.05. Results: Out of 219 UPEC, 114 (52.05%) of them were MDR, of which 37 were XDR and 1 was PDR. Increasing resistance among years was seen for 4 out o 8 classes of antimicrobial agents. An overall increase in MDR proportion (coeff. = 0.074; 95CI 0.038- 0.110), and in the number of concurrent resistances (coeff. = 0.297; 95CI 0.126-0.467) were assessed. A significative difference in the baseline level of resistances and the rising of them was observed between dogs and cats. Conclusions: Approximately half of isolated strains were MDR (52.05%), but they came all from clinically ill patients, which might suggest that the prevalence in the general pet population is lower. Nevertheless the upward trend of antimicrobial resistance of UPEC to various antibiotics, the rise in the amount of concurrent resistances and the presence of XDRs and a PDR strain, poses serious public health issues

Microplastics and Associated Microorganisms in the Sea Sediment of the Sentina Regional Natural Reserve (Central Adriatic Sea, Italy)

(1) Background: The large dispersion of microplastics (MPs) in the marine environment has effects on the health of living organisms [1,2,3]. The aim of this study was to identify MPs and their associated microorganisms in Adriatic Sea sediments and to evaluate the antibiotic susceptibility patterns of the microbial communities. (2) Methods: A beach transect, parallel to the shoreline where the waves break, was identified for the samplings. A protocol to perform MP isolation from sandy sediments at different seasons, suitable for biological sample upkeeping, and based on plastic floating in high salinity water, was optimized. From floating MPs, aerobic and anaerobic cultivable microorganisms were isolated and total DNA extraction was performed for the shotgun metagenomic analysis. Susceptibility to a panel of 14 antibiotics, belonging to 12 different categories, was assessed [4,5]. Chemical characteristics of the isolated MPs were analysed using a Thermo Nicolet 6700 FT-IR Spectrometer with “Smart Orbit” diamond micro-ATR accessory and Thermo Nicolet iN10 MX FT-IR microscope. (3) Results: Via chemical analysis, polypropylene microplastics were estimated in the highest percentage, followed by polyethylene, poly-methyl acrylate, and poly-vinyl chloride. Metagenomics data revealed differences in bacterial abundances during seasons and in floating MPs with respect to total sand. The differential gene analysis showed specific metabolic pathways in MP-associated microorganisms, including antibiotic resistance. Via microbial cultivation and MALDI-TOF MS identification, bacteria that are promising for plastic degradation, such as Lysinobacillus fusiformis, Exiguobacterium sp., and Pseudomonas oleovorans, were also found, as well as potential pathogens, like Clostridium septicum, Clostridium novyi, and Shewanella putrefaciens. Only 17.2% were found to be susceptible to all the tested antibiotics. High percentages of resistance were observed for penicillins (85.7%), monobactams (80.9%), and tetracyclines (64.3%). (4) Conclusions: MPs work as a vehicle for potential pathogens and antibiotic-resistant microorganisms in the Central Adriatic Sea

Teicoplanin and Vancomycin resistance in Methicillin-Sensitive Staphylococcus aureus isolated in a rabbit farm in Italy

Objectives Teicoplanin and vancomycin are increasingly important drugs against the human MRSA infections. In veterinary medicine, most studies have focused on glycopeptide-resistant MRSA while few studies have been conducted on MSSA strains. The aims of this study were the typing of MSSA strains circulating in a rabbit farm and the assessment of susceptibility to glycopeptides. Materials and methods Out of 595 MSSA isolated from meat rabbits of different age and farm-workers [1], 96 strains were randomly selected and characterized using spa-typing [2]. Susceptibility to 16 antibiotics, teicoplanin and vancomycin included, was tested [3]. Finally, profiles of antibiotic resistance were combined with spa-types, identifying different pathotypes. Results Twenty-three antibiotic-resistance profiles and 5 different spa-types (t2802, t491, t094, t2036, t605) were identified. The two most frequent spa-types were: t2802 (53.1%), detected in both animals and farmers, and t491 (38.5%). In total, 26 pathotypes were obtained. Resistance to glycopeptides was observed in 17/26 (65.4%) pathotypes. Among them, only one was susceptible to teicoplanin, but resistant to vancomycin (clone t491); 14 were resistant to teicoplanin (t2802, t491, t094, t2036, t605), while being susceptible to vancomycin; two were resistant to both antimicrobials (t491, t094). Conclusion Many different MSSA clones, with different patterns of antibiotic-resistance, were present in this rabbit farm. The same spa-type was observed in both animals and workers, showing that S. aureus strains can easily circulate in a community. More than half of the MSSA clones exhibited resistance to glycopeptides. This data should not be underestimated as these antibiotics are among drugs of choice for treatment of MRSA infections, making it important to extend the surveillance network also to niche zootechnical species

The Effect of Age and Sampling Site on the Outcome of Staphylococcus aureus Infection in a Rabbit (Oryctolagus cuniculus) Farm in Italy

Abstract: A study in an intensive Italian rabbit farm was carried out to assess the eect of age category and anatomical site on detection of S. aureus and to describe the diversity of spa-types within farm, including workers. On 400 rabbits of dierent age, 2066 samples from the ear, nose, axilla, groin, perineum and lesions were analyzed and 595 S. aureus were isolated. In total, 284 (71.0%) were colonized by S. aureus in at least one anatomical site and 35 animals (8.8%) had lesions. S. aureus prevalence was higher in adults than other age categories. Ear (29.4%) and nose (27.2%) were the most frequently colonized sites. The presence of lesions due to S. aureus was strongly associated with the colonization of at least one body site and the risk of lesions was proportionate to the number of sites colonized by S. aureus. In addition, a genotypic characterization was performed for 96 S. aureus strains randomly selected. All isolates resulted as low-virulence methicillin-sensitive-Staphylococcus aureus (LV-MSSA). Five dierent clonal lineages were found: spa-t2802, the most frequently detected (53.1%), also in human samples; spa-t491 (38.5%); spa-t094; t605; and spa-t2036. Strains t094, t491 and t2036 have not been isolated in Italy before

Positive Influence of a Probiotic Mixture on the Intestinal Morphology and Microbiota of Farmed Guinea Fowls (Numida meleagris)

To understand the effectiveness of a probiotic mixture on intestinal morphology, mucus layer composition, and cecal microbiota diversity, 40 10-day-old Guinea fowls (Numida meleagris) were assigned to two groups: the control group (C), receiving drinking water, and the treated group (P), receiving water plus a commercial multi-strain probiotic (Slab51®, 2 × 1011 CFU/L). Birds were slaughtered after 4 months, and the intestines were collected. Samples from the duodenum, ileum, and cecum were processed for morphological and morphometric studies, and conventional glycohistochemistry. Cecal samples were also used to assess the microbiota by 16S metataxonomic approach. Group P showed significant increase in the villus height (p < 0.001 in the duodenum and p < 0.05 in the ileum and cecum), villus width (p < 0.05 in all investigated tracts), depth of crypts (p < 0.001 in the duodenum and cecum; p < 0.05 in the ileum), and goblet cells per villus (p < 0.001 in all investigated tracts) compared with group C. Cecal microbiota of the birds varied considerably and comparing the relative abundance of the main observational taxonomic units (OTUs), a positive enrichment of several beneficial taxa, such as Oscillospira, Eubacterium, Prevotella, and members of the Ruminococcaceae, was observed. The enrichment of those taxa can improve microbiota stability and resilience facing environmental stresses, enhancing its resistance against invading pathogens. Ruminococcaceae, which represent the most important taxon in both groups, and Prevotella have a key role in the gut physiology due to the production of short-chain fatty acids (SCFAs), which are a vital energy source for enterocytes, improve glucose metabolism, and exert an overall anti-inflammatory effect. Probiotic administration enriches the presence of Coprococcus, Oscillospira, and Eubacterium taxa that produce butyrate, which exerts a beneficial effect on growth performance, structure of villi, and pathogen control and has anti-inflammatory properties too. This study indicates that Slab51® supplementation positively affects the morphology and microbiota diversity of the guinea fowl intestine

Health assessment of wild speckled dwarf tortoises, Chersobius signatus.

BACKGROUND In free-ranging reptile populations, bacterial, fungal, viral and parasitic pathogens may affect hosts through impairment in movements, thermoregulation, reproduction, survival, and population dynamics. The speckled dwarf tortoise (Chersobius [Homopus] signatus) is a threatened species that is mostly restricted to the Succulent Karoo biome in South Africa, and little information on pathogens of this species is available yet. We derived baseline parameters for five males and five females that were captured to genetically enhance a conservation breeding program in Europe. Upon collection of the tortoises, ticks were removed and identified. Immediately upon arrival in Europe, ocular, nasal, oral and cloacal swabs were taken for viral, bacteriological and mycological examinations. Fecal samples were collected before and 1 month after fenbendazole treatment, and analyzed for parasites. A panel of PCR, aiming to detect herpesviruses, adenoviruses and iridoviruses, was carried out. RESULTS Samples were negative for viruses, while bacteriological examination yielded detectable growth in 82.5% of the swabs with a mean load of 16 × 107 ± 61 × 108 colony forming units (CFU) per swab, representing 34 bacterial species. Cloacal and oral swabs yielded higher detectable growth loads than nasal and ocular swabs, but no differences between sexes were observed. Fungi and yeasts (mean load 5 × 103 ± 13 × 103 CFU/swab) were detected in 25% of the swabs. All pre-treatment fecal samples were positive for oxyurid eggs, ranging from 200 to 2400 eggs per gram of feces, whereas after the treatment a significantly reduced egg count (90-100% reduction) was found in seven out of 10 individuals. One remaining individual showed 29% reduction, and two others had increased egg counts. In five tortoises, Nycthocterus spp. and coccidian oocysts were also identified. Soft ticks were identified as Ornithodoros savignyi. CONCLUSIONS Our baseline data from clinically healthy individuals will help future studies to interpret prevalences of microorganisms in speckled dwarf tortoise populations. The study population did not appear immediately threatened by current parasite presence

Evidence and antibiotic resistance profiles of clinical Acinetobacter calcoaceticus-Acinetobacter baumannii (ACB) and non-ACB complex members in companion animals: A 2020-2022 retrospective study

To evaluate the frequency of Acinetobacter spp., belonging to both Acinetobacter calcoaceticus-baumannii (ACB) and non-ACB complex, and their antibiotic resistance profiles in veterinary medicine, a three-year (2020–2022) retrospective study was carried out on sick companion animals. Epidemiological data from different clinical canine, feline, and equine samples, were acquired. For each strain, MALDI-TOF MS identification and susceptibility to a panel of 11 antibiotics, by Kirby-Bauer and E-test methods, were performed. Out of 628 bacteriological examinations, 2.5% resulted positive for strains belonging to Acinetobacter genus. Frequencies of 2.3%, 1.9%, and 3% were obtained from both in-visiting and hospitalized dogs, cats, and horses, respectively. Members of ACB-complex accounted for 50% of isolates. Since all strains resulted susceptible to aminoglycosides and polymyxins, no pandrug-resistant (PDR) species were recorded. While 12.5% A. baumannii resulted extensively-drug resistant (XDR), a higher percentage of multidrug-resistant strains was recorded among non-ACB strains (35.5%) than ACB strains (25%). Susceptibility was observed in the same percentage in both groups (62.5%). All ACB strains confirmed their intrinsic resistances. Non-ACB species showed lower resistances against antipseudomonal penicillins plus beta-lactamase inhibitors (P=0.1306), III generation cephalosporins (P=0.0547), and tetracyclines (P=0.0209) than ACB species. Carbapenem-resistance was observed for XDR A. baumannii (12.5%) and, in particular for MDR non-ACB complex members (25%). To our knowledge, A. lactucae represents the first description in two sick dogs in Italy. Furthermore, our results emphasize the role of non-ACB-complex species as important zoonotic pathogens, which could be reservoirs of clinically relevant resistance profiles