Comparison between chlorhexidine and povidone-iodine solutions for flushing the preputial cavity on penile mucosa and semen bacterial counts in beef bulls.

The objectives of the present study were to estimate the number of colonies forming units (CFU) from penile mucosa and semen, the effect of two antiseptic solutions used to flush the preputial cavity to reduce the bacterial counts from those sites, and compare them. Six clinically healthy bulls between 15 and 16 mo old declared satisfactory potential breeders were used. A prospective, randomized, and controlled cross-over design was performed, in which each bull was first sampled from the penile mucosa and semen without treatment (control group) and 24 h later, after antiseptic preputial flushing (treated group). In the treated group, the preputial area was cleaned, the preputial hair was cut, urination was stimulated, prepuce area was scrubbed twice, and the preputial cavity was flushed with either 1% of povidone-iodine solution (POI; 500 mL) or 0.05% chlorhexidine digluconate (CHG; 500 mL), maintained for 10 min. Then, the preputial cavity was emptied and flushed with 500 mL of sterile saline solution. Next, the accessory sexual glands were massaged per rectum. Finally, protrusion, erection, and ejaculation were obtained by electroejaculation, and samples from penile mucosa and semen were collected for microbiological culture. The number of CFU was determined for each sample by enumerate total aerobic bacteria using Standard Plate Surface Count cultured for 48 h. In the first replicate, half of the bulls were treated with CHG, and the other half were treated with POI. After 58.8 ± 5.3 days (x ± SD) of wash-out period, the treatments were reverted, and the same protocols were applied again. In the control group, the median number of CFUs from the penile mucosa was 750,000 (range from 60,000 to 1,800,000) and the median number of CFUs in semen was 8,000,000 (700,000-45,000,000). The CFU in semen was higher than the penile mucosa (P = 0.005). Both antiseptic solutions reduced the median number of CFUs on the penile mucosa to 915 (P = 0.002) and in semen to 1,680 (P = 0.002). The antiseptic effect on the penile mucosa was higher for CHG solution (490) than for POI solution (6,650; P = 0.05). The antiseptic effect on semen of CHG was also greater (200) than for the POI solution (31,000; P = 0.05). It can be concluded that the median number of CFU was higher in semen compared with penile mucosa, and flushing the preputial cavity either with 0.05% CHG or 1% POI maintained for 10 min reduced the number of CFUs from penile mucosa and semen. The level of antiseptic activity was higher for CHG than for POI.

Campylobacter vulpis sp. nov. isolated from wild red foxes.

During a sampling of wild red foxes (Vulpes vulpes) for the detection of Epsilonproteobacteria, 14 strains were isolated from the caecal contents of 14 epidemiologically-unrelated animals. A genus-specific PCR indicated that the isolates belonged to the genus Campylobacter. Based on the results of a species-specific PCR, the isolates were initially identified as C. upsaliensis. However, multi-locus sequence typing (MLST) revealed that the isolates were significantly different from the C. upsaliensis present in the MLST database. A polyphasic study, including conventional biochemical and tolerance characteristics, morphology by transmission electron microscopy (TEM), MALDI-TOF analysis, and genetic comparisons based on partial 16S rDNA and atpA gene sequences, was undertaken. Finally, the complete genome sequence of the type strain 251/13T and the draft genome sequences of the other isolates were determined. Average nucleotide identity, average amino acid identity and in silico DNA-DNA hybridization analyses confirmed that the isolates represent a novel taxon for which the name Campylobacter vulpis sp. nov. is proposed, with isolate 251/13T (=CCUG 70587T = LMG 30110T) as the type strain. In order to allow a rapid discrimination of C. vulpis from the closely-related C. upsaliensis, a specific PCR test was designed, based on atpA gene sequences.

Incidence of Bacteremia Consequent to Different Endoscopic Procedures in Dogs: A Preliminary Study.

Endoscopic procedures are widely used in veterinary medicine, and their role in producing transient bacteremia is debatable. The growing issue of antibiotic resistance requires the correct use of antibiotics, avoiding their administration when not strictly necessary. Studies highlighting post-endoscopy bacteremia in veterinary medicine are extremely rare and often involve very few animals. This study describes the results from 74 owned dogs, brought to the Veterinary Teaching Hospital of the Department of Veterinary Medical Science of the University of Bologna, for the purpose of undergoing an endoscopic procedure. Two blood samples were taken from each dog, one before and one after the procedure, in order to assess the incidence of bacteremia linked to endoscopic procedures. Eight dogs were tested positive at the second blood culture with an Incidence Risk (IR) of 10.8%. No statistical differences were found by comparing positive and negative blood cultures with respect to sex, age, weight and anesthesia duration. In addition, no difference was found between airway and digestive tract procedures. The present findings showed that the probability of developing bacteremia after an endoscopic procedure was quite low, and additional studies confirming this are certainly recommended as well as the evaluation of categories of patients potentially considered at risk.

Hospitalized Pets as a Source of Carbapenem-Resistance.

The massive and irrational use of antibiotics in livestock productions has fostered the occurrence and spread of resistance to "old class antimicrobials." To cope with that phenomenon, some regulations have been already enforced in the member states of the European Union. However, a role of livestock animals in the relatively recent alerts on the rapid worldwide increase of resistance to last-choice antimicrobials as carbapenems is very unlikely. Conversely, these antimicrobials are increasingly administered in veterinary hospitals whose role in spreading bacteria or mobile genetic elements has not adequately been addressed so far. A cross-sectional study was carried out on 105 hospitalized and 100 non-hospitalized pets with the aim of measuring the prevalence of carbapenem-resistant Gram-negative bacteria (GNB) colonizing dogs and cats, either hospitalized or not hospitalized and estimating the relative odds. Stool samples were inoculated on MacConkey agar plates containing 1 mg/L imipenem which were then incubated aerobically at 37°C ± 1 for 48 h. Isolated bacteria were identified first by Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and were confirmed by 16S rRNA sequencing. The genetic basis of resistance was investigated using PCR methods, gene or whole genome sequencing (WGS). The prevalence of pets harboring carbapenem-resistant bacteria was 11.4 and 1.0% in hospitalized and not-hospitalized animals, respectively, with an odds ratio of 12.8 (p < 0.01). One pet carried two diverse isolates. Overall, 14 gram-negative non-fermenting bacteria, specifically, one Acinetobacter radioresistens, five Acinetobacter baumannii, six Pseudomonas aeruginosa and two Stenotrophomonas maltophilia were isolated. The Acinetobacter species carried acquired carbapenemases genes encoded by bla NDM-1 and bla OXA-23. In contrast, Pseudomonas phenotypic resistance was associated with the presence of mutations in the oprD gene. Notably, inherent carbapenem-resistant isolates of S. maltophilia were also resistant to the first-line recommended chemotherapeutic trimethoprim/sulfamethoxazole. This study estimates the risk of colonization by carbapenem-resistant non-fermenting GNB in pets hospitalized in veterinary tertiary care centers and highlights their potential role in spreading resistance genes among the animal and human community. Public health authorities should consider extending surveillance systems and putting the release of critical antibiotics under more strict control in order to manage the infection/colonization of pets in veterinary settings.

Antimicrobial Susceptibility of Campylobacter Cuniculorum Isolated from Rabbits Reared in Intensive and Rural Farms.

The present study aimed to investigate the antimicrobial susceptibility in Campylobacter cuniculorum. To do so, 29 isolates from rabbits reared in 18 intensive and 11 rural farms not epidemiologically correlated were tested. Minimum inhibitory concentration of 8 antimicrobial agents was determined using the agar dilution method recommended by the Clinical and Laboratory Standards Institute (Wayne, PA, USA), modified - for what supplements in the base medium and incubation conditions concern - for C. cuniculorum isolates. The isolates obtained from rural farming resulted susceptible to all the antimicrobial agents tested, with the exception of one isolate resistant to nalidixic acid. All the isolates obtained from intensively farmed rabbits were sensitive to chloramphenicol and ampicillin; 16 isolates were resistant to tetracycline; 15 to nalidixic acid and erythromycin; 13 and 10 isolates to ciprofloxacin and enrofloxacin, respectively; and only 1 to gentamicin. The resistance of several isolates to macrolides and fluoroquinolones, which are the drugs of choice in treatment of human campylobacteriosis, could pose a risk to human health if a pathogenic role of C. cuniculorum was demonstrated.

European multicenter study on antimicrobial resistance in bacteria isolated from companion animal urinary tract infections.

BACKGROUND: There is a growing concern regarding the increase of antimicrobial resistant bacteria in companion animals. Yet, there are no studies comparing the resistance levels of these organisms in European countries. The aim of this study was to investigate geographical and temporal trends of antimicrobial resistant bacteria causing urinary tract infection (UTI) in companion animals in Europe. The antimicrobial susceptibility of 22 256 bacteria isolated from dogs and cats with UTI was determined. Samples were collected between 2008 and 2013 from 16 laboratories of 14 European countries. The prevalence of antimicrobial resistance of the most common bacteria was determined for each country individually in the years 2012-2013 and temporal trends of bacteria resistance were established by logistic regression. RESULTS: The aetiology of uropathogenic bacteria differed between dogs and cats. For all bacterial species, Southern countries generally presented higher levels of antimicrobial resistance compared to Northern countries. Multidrug-resistant Escherichia coli were found to be more prevalent in Southern countries. During the study period, the level of fluoroquinolone-resistant E. coli isolated in Belgium, Denmark, France and the Netherlands decreased significantly. A temporal increase in resistance to amoxicillin-clavulanate and gentamicin was observed among E. coli isolates from the Netherlands and Switzerland, respectively. Other country-specific temporal increases were observed for fluoroquinolone-resistant Proteus spp. isolated from companion animals from Belgium. CONCLUSIONS: This work brings new insights into the current status of antimicrobial resistance in bacteria isolated from companion animals with UTI in Europe and reinforces the need for strategies aiming to reduce resistance.

Helicobacter apri sp. nov., isolated from wild boars.

Three isolates (A19T, C21 and F12) with spiral-shaped cells and one bipolar sheathed flagellum were obtained from gastric mucosa and caecal contents of three different wild boars (Sus scrofa) and subjected to a polyphasic taxonomic study. A genus-specific PCR showed that these isolates belonged to the genus Helicobacter. Phylogenetic analysis based on 16S rRNA, 60-kDa heat-shock protein (HSP60) and atpA genes demonstrated they formed a novel lineage within this genus. Pairwise 16S rRNA, HSP60 and atpA gene sequence comparisons of the three isolates revealed 99.7, 99.4 and 99.9 % similarity, respectively, among the three isolates; the 16S rRNA gene of isolate A19T shared 98.5 % sequence similarity with its nearest validly named neighbouring species, Helicobacter mastomyrinus (to the type strain MIT 97-5577T). The taxonomic uniqueness of the wild boar isolates was confirmed by protein analysis performed by matrix-assisted laser desorption/ionization time-of-flight MS and by a distinctive biochemical profile. These data indicated that isolates A19T, C21 and F12 represent a novel taxon, for which the name Helicobacter apri sp. nov. is proposed, with isolate A19T (=DSM 28990T=LMG 28471T) as the type strain.

Typing of Campylobacter jejuni Isolated from Turkey by Genotypic Methods, Antimicrobial Susceptibility, and Virulence Gene Patterns: A Retrospective Study.

In this retrospective study, typing ability, discriminatory power, and concordance between typing results obtained on 123 Campylobacter jejuni turkey isolates, collected in 1998, within 14 different farms, applying multilocus sequence typing (MLST), pulsed-field gel electrophoresis (PFGE), antibiotic resistance profile, and virulence gene pattern, were assessed and compared. Overall, 33 sequence types, 28 pulsotypes, 10 resistotypes, and 5 pathotypes were identified. MLST and PFGE showed the better discriminatory ability (i.e., Simpson's diversity index >0.90) as well as unidirectional (i.e., Wallace and adjusted Wallace coefficients >0.86) and bidirectional (i.e., adjusted Rand coefficient >0.60) concordance. Moreover, both methods showed a good unidirectional and bidirectional concordance with the resistotype. On the contrary, the congruence of both genotyping methods and resistotype with the pathotype seemed due to chance alone. A clonal relationship was identified among 66.7% of the isolates. Furthermore, 59.7% of the investigated isolates were resistant to two or more antimicrobials and 92% to tetracycline. All the isolates harbored cadF and pldA genes, whereas a flaA gene product and a cdtB gene product were amplified from 85.4% and 79.7% of the isolates, respectively, using the primers designed by Bang et al. (2003). The results of this study clarify the level of genetic diversity among the C. jejuni originating from turkeys. MLST level of correlation with PFGE, resistotype, and pathotype is assessed. This result supports the selection of type and number of typing methods to use in epidemiological studies. Finally, the identification of clonal complexes (i.e., groups of profiles differing by no more than one gene from at least one other profile of the group using the entire Campylobacter MLST database) shared between turkey and human isolates suggests that turkeys could be a possible source of Campylobacter infection.