The objective of this study was to describe bacterial culture and antibiotic susceptibility results in 476 dogs presenting with suspected bacterial keratitis in Iowa and surrounding Midwestern states, further detailing trends in patient characteristics, seasonality, and antimicrobial resistance. Corneal swabs yielded 465 bacterial isolates and 220 cultures (46.2%) with no apparent growth (0–5 isolates per culture). The most frequent bacterial genera were Staphylococcus (32.3%), Streptococcus (19.1%), and Pseudomonas (12.5%), while the most common bacterial species were Staphylococcus pseudintermedius (26.7%), Streptococcus canis (12%), and Pseudomonas aeruginosa (7.5%). Compared to mixed-breed dogs, canine breeds most likely to be examined for ulcerative keratitis included Boston terrier, Cavalier King Charles spaniel, miniature pinscher, pug, rat terrier, Saint Bernard, shih tzu, and silky terriers. In summer, the likelihood to yield a negative culture was reduced while the likelihood to culture Pseudomonas species was increased. Bacteria considered multidrug resistant (MDR, resistant to ≥ 3 antibiotic classes) represented 20% of all canine isolates and were most prevalent for Staphylococcus species (33%). An alarming, escalating trend of MDR prevalence was noted between 2016 (5%) and 2020 (34%). Individual ophthalmic preparations (i.e., single antibiotics or commercially available antibiotic combinations) with highest efficacy against all bacterial isolates included chloramphenicol (83%), ceftiofur (79%), amikacin (77%), neomycin-polymyxin B-bacitracin (77%), and gentamicin (74%). Efficacy of systemic antibiotics and combinations of ophthalmic preparations was also evaluated. Based on the present findings, triple antibiotic (Neo-Poly-Bac) is recommended as empirical monotherapy for prophylactic antibiotic therapy in dogs with simple corneal ulcers, while a chloramphenicol-ciprofloxacin combination is empirically recommended for therapeutic management of infected corneal ulcers. Pending culture and susceptibility results, appropriate selection of empiric antibiotic therapy is important to enhance therapeutic outcome and reduce antibacterial resistance in dogs with corneal ulceration.

The objective of this experiment was to investigate the impact of an F18 enterotoxigenic Escherichia coli (ETEC) challenge on growth performance, aspects of intestinal function, and selected immune responses of piglets, as well as to evaluate potential protective effects of direct-fed microbial (DFM) blends. Seventy-two weaned piglets (6.4 ± 0.2 kg body weight [BW]; ~21 d of age) were assigned to one of four treatments: 1) NC: Nonchallenged (n = 10), 2) positive challenged control (PC): F18 ETEC-challenged (n = 10), 3) PC + DFM1 (n = 8; three strains of Bacillus amyloliquefaciens; 7.5 × 105 colony-forming units [cfu]/g), or 4) PC + DFM2 (n=8; 2 strains of B. amyloliquefaciens and one strain of Bacillus subtilis; 1.5 × 105 cfu/g). Feed intake and BW were recorded on day 0, 7, and 17. Pigs were sham-infected either with 6 mL phosphate-buffered saline or inoculated with 6 mL F18 ETEC (~1.9 × 109 cfu/mL) on day 7 (0 d postinoculation [dpi]). All ETEC-challenged pigs were confirmed to be genetically susceptible to F18. Pigs had ad libitum access to feed and water throughout the 17-d trial. Fecal scores were visually ranked and rectal temperatures were recorded daily. To evaluate ETEC shedding, fecal swabs were collected on dpi 0, 1, 2, 3, 5, 7, and 10. Blood samples were collected on dpi 0, 1, 2, 4, 7, and 10. Ileal tissues were collected at necropsy on dpi 10. All challenged treatments had lower final BW, decreased average daily gain (ADG), and average daily feed intake (ADFI) during the 10-d postchallenge period (P < 0.01). The DFM2 treatment increased E. coli shedding on dpi 2 and decreased iton dpi 7 (P < 0.05) compared with the PC. Rectal temperature decreased across all challenged treatments (P < 0.01). Ileal mRNA abundance of occludin (OCLN) and zonula occludens-1 (ZO-1) decreased in PC and DFM1 compared with NC (P < 0.05). Pigs fed DFM2 had intermediate ileal mRNA abundance of OCLN and increased ZO-1 mRNA compared with pigs in PC (P < 0.05). Interleukin 8 (IL-8) increased in the plasma of PC and DFM2 on dpi 2 compared with NC (P < 0.05). Mucosal IL-8 increased in PC compared with NC (P < 0.05). All challenged treatments tended to have elevated tumor necrosis factor-α (TNF-α) mRNA abundance compared with NC (P < 0.10). Challenged pigs had reduced secretory immunoglobulin A and villus height compared with NC pigs (P < 0.05). The impact of an ETEC challenge on intestinal function and the immune system has been revealed, information critical to developing improved treatment regimes.

Bacterial keratitis is a serious and vision-threatening condition in veterinary and human patients, one that often requires culture and susceptibility testing to adjust therapy and improve clinical outcomes. The present study challenges the antimicrobial susceptibility testing (AST) paradigm in ophthalmology, enabling more accurate in vitro-to-in vivo translation by incorporating factors normally present during host-pathogen interactions in clinical patients. Thirty bacteria (10 Staphylococcus pseudintermedius, 10 Streptococcus canis, 10 Pseudomonas aeruginosa) were isolated from canine patients with infectious keratitis. For each isolate, commercial plates (Sensititre™ JOEYE2) were used to assess the minimal inhibitory concentration (MIC) of 17 different antibiotics in the absence (0% albumin, control) or presence of canine albumin (0.01–2%). For Staphylococcus pseudintermedius, the experiment was repeated with actual tear fluid collected from canine eyes with ocular surface inflammation. Kruskal-Wallis, Wilcoxon signed rank test and Spearman's correlation tests were used for statistical analysis. Clinical outcomes were unfavorable in selected canine patients with bacterial keratitis (e.g., globe perforation, graft dehiscence) despite standard AST (i.e., 0% albumin in test medium) confirming that most corneal infections (93%) were susceptible to ≥1 topical antibiotics used at the initial visit. Albumin levels ≥0.05% increased MICs in a dose-dependent, bacteria-specific, and antibiotic-specific manner. No significant differences (P = 1.000) were noted in MICs of any antibiotic whether albumin or tear fluid was added to the Mueller-Hinton broth. Percent protein binding inherent to each antibiotic was associated with clinical interpretations (Spearman's rho = −0.53, P = 0.034) but not changes in MICs. Albumin in tears impacted the efficacy of selected ophthalmic antibiotics as only the unbound portion of an antibiotic is microbiologically active. The present findings could improve decision making of clinicians managing bacterial keratitis, reduce development of antimicrobial resistance, influence current guidelines set by the Clinical and Laboratory Standards Institute, and serve as a reference for bacteriological evaluations across medical fields and across species.

The present study describes the prevalence of bacterial cross-contamination in a veterinary ophthalmology setting, a serious issue that can result in healthcare-associated (or nosocomial) infections among patients and staff. Retrospective (n = 5 patients) and prospective (n = 23 patients) studies evaluated bacterial isolates in companion animals presenting with ulcerative keratitis, sampling the patients' cornea and surrounding examination room, including the environment (exam table, countertop, floor) and ophthalmic equipment (slit lamp, transilluminator, direct ophthalmoscope, indirect headset, tonometer). Results of bacterial culture and antibiotic susceptibility testing were recorded, and degree of genetic relatedness was evaluated in six pairs of isolates (cornea + environment or equipment) using pulse-field gel electrophoresis (PFGE). Overall contamination rate of ophthalmic equipment, environment, and examination rooms (equipment + environment) was 42.9% (15/35 samples), 23.7% (9/38 samples) and 32.9% (24/73 samples), respectively. Methicillin-resistant Staphylococcus pseudintermedius (MRSP), a multi-drug resistant (MDR) pathogen with zoonotic potential, was isolated in 8.2% (6/73) of samples. The patient's cornea was likely the source of cross-contamination in 50% (3/6) of MRSP pairs as evaluated by PFGE; notably, two of the three similar bacterial strains did not have an exact match of their antibiotic susceptibility profiles, highlighting the importance of advanced diagnostics such as PFGE to assess cross-contamination in healthcare facilities. Future work could examine the contamination prevalence of specific equipment or the efficacy of cleaning protocols to mitigate cross-contamination in veterinary practice.