IDEXX Laboratories (Germany)

Pseudomonas (P.) aeruginosa is a highly adaptable, opportunistic nosocomial pathogen that poses significant risks to public health and veterinary medicine. This bacterium carries a wide range of antimicrobial resistance (AMR) determinants and produces various virulence factors that enable it to invade hosts and increase disease severity. Recognised as a One Health pathogen, P. aeruginosa can be isolated from multiple sources, including humans, animals, food, and the environment. Despite its importance in clinical settings, there are still limited genomic and epidemiological data on P. aeruginosa isolates from companion animals in Germany. To address this knowledge gap, we conducted whole-genome sequencing (WGS) of 72 P. aeruginosa isolates collected in 2023 from various companion animals, including dogs, cats, horses, and rabbits, across Germany. Phenotypic antibiotic susceptibility testing (AST) showed that 97.2% of the isolates exhibited a multidrug-resistant (MDR) phenotype. The highest resistance rates were observed for cefotaxime (98.6%), followed by chloramphenicol (93.1%) and trimethoprim/sulfamethoxazole (87.5%). Fosfomycin resistance was observed in 26.4% of the isolates, despite all isolates carrying the fosA gene. All isolates were found to be susceptible to colistin at increased exposure levels. Resistance to imipenem was detected in four dog isolates. Genome analysis revealed various AMR genes associated with resistance to β-lactams, aminoglycosides, sulfonamides, fluoroquinolones, and phenicols. Multiple virulence-associated genes were also identified, including those involved in biofilm formation, adherence, motility, immune modulation, and exotoxin A production. Moreover, 27.8% of the isolates carried plasmid contigs, and one dog isolate harboured a class 1 integron. In silico multilocus sequence typing (MLST) assigned the 72 isolates to 59 distinct sequence types (STs), including five novel STs (ST5165–ST5169). The most frequently identified STs were ST253, ST258, ST395, and ST244, each represented by three isolates. Six high-risk clones (HRCs) of P. aeruginosa, including ST308, ST277, ST244, ST395, ST253, and ST274, were identified in dog (n = 8) and cat (n = 4) isolates. This study underscores the significant genomic diversity of P. aeruginosa circulating among companion animals and emphasises the need to manage this bacterium within a One Health framework.

The authors declare no conflicts of interest.

ABSTRACTBackgroundDiarrhea in cats is common in multicat households, and fecal panels are frequently used to diagnose potential enteropathogenic bacteria.ObjectivesTo evaluate the presence of potential fecal enteropathogenic bacteria and their association with diarrhea in multicat households. The null hypothesis was that potential enteropathogenic bacteria were not related to diarrhea in the cohort.AnimalsIn total, 234 cats from 41 privately owned catteries were included.MethodsIn this prospective study, feces were evaluated for consistency according to a visual scoring system (Purina Fecal Score). Scores from 4 to 7 were defined as diarrhea. Quantitative PCR for Escherichia coli, Clostridium perfringens encoding the α toxin gene (cpa), Clostridium perfringens encoding the enterotoxin gene (cpe), Clostridioides difficile, Campylobacter jejuni/coli, and Salmonella enterica was used. Logistic regression was used to evaluate the impact of selected bacteria on diarrheic feces (fecal score ≥ 4). p values were corrected for multiple comparisons (q values in results). A q value < 0.05 was considered to indicate statistical significance.Results23/234 cats (9.8%) had diarrhea. None of the selected bacteria were significantly associated with diarrhea (cpa: q = 0.10, cpe: q = 0.20, C. jejuni: q = 0.70). E. coli was detected in all tested fecal samples (100%). The associations of Cl. difficile (detected in 2.3% of cats), C. coli (0%), and S. enterica (0.9%) with diarrhea could not be evaluated due to the small sample sizes.Conclusions and Clinical ImportanceFecal bacteria detected via PCR were not associated with clinical signs of diarrhea in multicat households. These data do not support the use of PCR for fecal bacteria for baseline routine diagnostic work‐up of diarrhea in cats.

Klebsiella (K.) pneumoniae is a One Health pathogen that has been isolated from humans, animals, and environmental sources and is responsible for a diverse range of potentially life-threatening infections. In the present study, we analyzed the genomes of 64 presumptive K. pneumoniae strains isolated in 2023 from different companion and farm animals in Germany. Using whole-genome sequencing (WGS) data, 59 isolates (92.2%) were identified as K. pneumoniae and five (7.8%) as K. quasipneumoniae. Multilocus sequence typing (MLST) assigned 53 isolates to 46 distinct sequence types (STs). Eleven isolates could not be assigned to existing STs of the Pasteur classification scheme because they contained novel alleles not previously documented. Thus, these were considered novel and designated as ST7681-ST7689 and ST7697-ST7698. Almost all isolates in this study were assigned unique STs, and only five STs were shared among multiple isolates. This research highlights the genetic diversity among K. pneumoniae strains isolated from different companion and farm animals in Germany, provides information to help in surveillance strategies to mitigate zoonotic transmission risks, and demonstrates the value of WGS and MLST in identifying novel STs of K. pneumoniae.

BackgroundEscherichia coli is an opportunistic pathogen which colonizes various host species. However, to what extent genetic lineages of E. coli are adapted or restricted to specific hosts and the genomic determinants of such adaptation or restriction is poorly understood.ResultsWe randomly sampled E. coli isolates from four countries (Germany, UK, Spain, and Vietnam), obtained from five host species (human, pig, cattle, chicken, and wild boar) over 16 years, from both healthy and diseased hosts, to construct a collection of 1198 whole-genome sequenced E. coli isolates. We identified associations between specific E. coli lineages and the host from which they were isolated. A genome-wide association study (GWAS) identified several E. coli genes that were associated with human, cattle, or chicken hosts, whereas no genes associated with the pig host could be found. In silico characterization of nine contiguous genes (collectively designated as nan-9) associated with the human host indicated that these genes are involved in the metabolism of sialic acids (Sia). In contrast, the previously described sialic acid regulon known as sialoregulon (i.e. nanRATEK-yhcH, nanXY, and nanCMS) was not associated with any host species. In vitro growth experiments with a Δnan-9 E. coli mutant strain, using the sialic acids 5-N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc) as sole carbon source, showed impaired growth behaviour compared to the wild-type.ConclusionsThis study provides an extensive analysis of genetic determinants which may contribute to host specificity in E. coli. Our findings should inform risk analysis and epidemiological monitoring of (antimicrobial resistant) E. coli.

IntroductionDiarrhea is the second most common cause of mortality in shelter kittens. Studies examining prevention strategies in this population are lacking. Probiotics are of particular interest but studies in cats are largely limited to healthy adults or those with induced disease. Only one study in domestic cats describes the use of host-derived bacteria as a probiotic. We previously identified Enterococcus hirae as a dominant species colonizing the small intestinal mucosa in healthy shelter kittens. Oral administration of a probiotic formulation of kitten-origin E. hirae (strain 1002-2) mitigated the increase in intestinal permeability and fecal water loss resulting from experimental enteropathogenic E. coli infection in purpose-bred kittens. Based on these findings, we hypothesized that administration of kitten-origin E. hirae to weaned fostered shelter kittens could provide a measurable preventative health benefit.MethodsWe conducted a randomized, placebo-controlled, blinded clinical trial to determine the impact of a freeze-dried E. hirae probiotic on body weight gain, incidence of diarrhea, carriage of potential diarrheal pathogens, and composition of the intestinal microbiota in weaned fostered shelter kittens.ResultsOne-hundred thirty kittens completed the study. Fifty-eight kittens received the probiotic and 72 received the placebo. There were no significant differences in age, weight upon initiation of the study, number of days in the study, average daily gain in body weight, or weight at completion of the study. Kittens treated with E. hirae were 3.4 times less likely to develop diarrhea compared to kittens treated with placebo (odds ratio = 0.294, 95% CI 0.109–0.792, p = 0.022). A significant impact of E. hirae was not observed on the presence or abundance of 30 different bacterial, viral, protozoal, fungal, algal, and parasitic agents in feces examined by qPCR. With exception to a decrease in Megamonas, administration of the E. hirae probiotic did not alter the predominant bacterial phyla present in feces based on 16S rRNA gene amplicon sequencing.DiscussionDecreased incidence of diarrhea associated with preventative administration of E. hirae to foster kittens supports a rationale for use of E. hirae for disease prevention in this young population at high risk for intestinal disease though additional studies are warranted.

Zusammenfassung: Normalerweise ist der Pferdeharn hell

In contrast to sheep, chronic copper oversupply or toxicity represent infrequent events in cattle. This case report describes a chronically elevated exposure to copper arising from excessive supplementation of mineral feedstuff leading to a herd health problem in a German Holstein dairy herd. For diagnosis on herd level, 10 cows of both the close-up (VB) and high-yielding (HL) groups were selected and blood as well as urine samples were collected. Clinical chemistry investigation of these samples only revealed slightly raised activities of liver enzymes in cows of the HL group. Analysis of the total mixed ration (TMR) of both groups revealed a significant oversupply with copper, zinc, cobalt, and selenium. On the basis of these findings, the copper content of urine, feces, pigmented hair as well as plasma ceruloplasmin activity were measured additionally. Liver biopsies were performed in cows of the HL group and in heifers, the latter of which had received a mineral feed for young cattle according to the manufacturer's instructions. Results indicated increased fecal copper in both HL and VB groups as well as above normal or high-normal values of hepatic copper and cobalt resp. selenium, zinc, and manganese in the HL cows. In contrast, heifers had physiological amounts of trace elements in their liver tissues. This report demonstrates the suitability of fecal analysis and TMR for a reliable assessment of copper supply, while blood, urine, and hair failed to represent appropriate sample types. In order to attain a dependable diagnosis of potential copper oversupply, a liver biopsy - which may also be performed in buiatric practice - is deemed necessary.