Four intrauterine treatment strategies were evaluated for effectiveness in mares that were confirmed to be susceptible to chronic uterine infection. Pretreatment samples were obtained at detection of estrus, and a genital strain of Streptococcus zooepidemicus was infused into the uterus when a preovulatory (> 35 mm) follicle was detected. At 12 hours after inoculation, mares were assigned to 1 of 4 selected treatment groups: autologous plasma, 100 ml (n = 5); potassium penicillin, 5 million U in 100 ml of phosphate-buffered saline solution (PBSS; n = 5); 10 mg of prostaglandin F2alpha in 100 ml of PBSS (n = 5); and large-volume lavage with normal saline solution (1,000 ml increments). A fifth group, treated with vehicle alone (100 ml of PBSS), served as a negative control (n = 7). All treatments were administered into the uterus. To assess the effectiveness of the treatment, samples for culture and cytologic examination were collected at 96 hours after bacterial inoculation. An effect of treatment was observed on the number of uterine neutrophils (P = 0.02) and growth of S zooepidemicus (P < 0.01). Intrauterine treatment with potassium penicillin, prostaglandin F2alpha, and large-volume uterine lavage significantly reduced the growth of S zooepidemicus (P < 0.01) as well as the number of neutrophils (P < 0.02). Autologous plasma reduced the number of neutrophils (P < 0.05), but not growth of S zooepidemicus. There was significant correlation between the number of uterine neutrophils and growth of S zooepidemicus for each treatment group (r = 0.57; P < 0.05).

Efficacy of sulbactam, a beta-lactamase inhibitor, in combination with ampicillin, was evaluated for treatment of experimentally induced pneumonia caused by beta-lactam-resistant Klebsiella pneumoniae. Infection was experimentally induced in 18 healthy weanling foals that were randomly allocated to 3 treatment groups: sulbactam plus ampicillin (S/A, 3.3 and 6.6 mg/kg of body weight, respectively), ampicillin (6.6 mg/kg), or vehicle only. Foals were treated daily for 7 days; the observer was unaware of treatment status. Compared with ampicillin and vehicle, treatment with S/A resulted in a statistically significant (P < 0.05) decrease in severity of pneumonia, with regard to bronchoalveolar lavage cytologic findings (decreased total cell and neutrophil numbers, and increased lymphocyte numbers) and extent of macroscopic lesions in lung tissue of the noninoculated regions. Marked trends toward improvement of S/A-treated foals were observed for quantitative results of bacteriologic culture of bronchoalveolar lavage fluid samples (P < 0.07), macroscopic pathologic features of the whole lung (P < 0.1), and histopathologic variables (P < 0.07), compared with ampicillin- and vehicle-treated foals. Treatment effects were not observed for radiographic, hematologic, and blood gas abnormalities that resulted from infection. In conclusion, the combination of sulbactam plus ampicillin was found to have synergistic effects in vivo, to reduce the extent and severity of experimentally induced grain-negative lung infection in foals.

The effect of urine pH on plasma disposition of ampicillin sodium was evaluated. A single dose of 10 mg/kg of body weight was administered IV to Thoroughbreds with alkaline (pH > 8.0) or acidic (pH < 4.5) urine. Urine alkalinity was achieved and maintained by oral administration of up to 400 mg of sodium bicarbonate/kg/d, and acidity was achieved and maintained by oral administration of up to 400 mg of ammonium chloride/kg/d. Ampicillin sodium was measured in the plasma of horses by use of an agar diffusion microbiological assay with Bacillus subtilis as the test organism. The plasma disposition kinetics of ampicillin sodium best fitted a 2-exponential decay pattern, and statistically significant differences were not evident in elimination half-life, area under the plasma concentration time curve, volume of distribution, or body clearance rate between horses with alkaline or acidic urine. Results indicate that changes in urine pH over a range encountered in clinically normal horses are unlikely to affect plasma pharmacokinetic variables of ampicillin sodium after IV administration of the drug.

Vaginal aerobic bacterial flora was studied in 5 healthy bitches before, during, and after a 10-day period of treatment with ampicillin and an equally long period of treatment with trimethoprim-sulfamethoxazole. Blood variables and antimicrobial drug susceptibility also were studied. Bacteria were isolated from all bitches before the first treatment period. Bitches from which only a sparse number of bacteria were isolated had flora that varied from day to day. In most instances when bitches were given an antibiotic to which their vaginal bacterial flora was susceptible, these bacteria were eradicated after only 1 day of treatment. This was true for pasteurellae, streptococci, and, in all but one case, Escherichia coli. Staphylococcus intermedius was more difficult to eradicate, and, although susceptible in vitro, it was unaffected by antibiotic treatment in 1 bitch and it took 7 days to eradicate in another. Eradication of aerobic bacteria in the vagina was total only in the bitch that had sparse flora from the beginning. Bacteria colonized within 0 (in 4/5 bitches) to 4 days after termination of treatment with ampicillin and within 0 (in 4/5 bitches) to 3 days for trimethoprim-sulfamethoxazole. Mycoplasmas emerged during and after both treatment periods, and E coli became apparent during treatment with trimethoprim-sulfamethoxazole. Because mycoplasmas may be genital pathogens in bitches and E coli is a common uropathogen, their appearance should be an argument against widespread use of antibiotics in healthy breeding bitches. Two bitches developed a vaginal discharge during treatment or shortly after. Blood variables did not change during the study, nor did antimicrobial drug resistance of the isolated bacteria.

Serum concentration of ampicillin, a semisynthetic penicillin, was measured in mares at various time intervals up to 24 hours after intrauterine infusion of 3 g of ampicillin. Blood samples were drawn immediately before infusion and at l-, 4-, 10- and 24-hour intervals after infusion. At postinfusion hour 24, two endometrial biopsy specimens were obtained to measure endometrial concentrations of ampicillin. Blood was drawn twice as part of the 24-hour postinfusion sample collection, once before removal of the biopsy specimens and again 5 minutes after removal of the biopsy specimens. After drug infusion, more diestrous mares had detectable serum ampicillin concentration than did estrous mares for all samples, except the 24-hour prebiopsy sample. None of the 24-hour prebiopsy serum samples had detectable ampicillin concentration, but ampicillin was detected in the serum of 4 of 5 diestrous mares after endometrial biopsy. Endometrial concentrations of ampicillin were detectable at postinfusion hour 24 in estrous and diestrous mares, but were not different. All 24-hour biopsy specimens had ampicillin concentrations greater than the ampicillin minimal inhibitory concentration.

Enterococci are widespread, being part of the bacterial flora of humans and animals. The food chain can be therefore considered as the main route of transmission of antibiotic resistant bacteria between the animal and human populations. Milk in particular represents a source from which resistant bacteria can enter the human food chain. The aim of the study was to determine the occurrence and resistance to antimicrobial agents of Enterococcus spp. strains isolated from raw goat’s milk samples. A total of 207 goat’s milk samples were collected. Samples were cultivated on selective media and confirmed as E. faecium or E. faecalis and screened for selected resistance genes by PCR. Drug susceptibility determination was performed by microdilution on Sensititre EU Surveillance Enterococcus EUVENC Antimicrobial Susceptibility Testing (AST) Plates and Sensititre US National Antimicrobial Resistance Monitoring System Gram Positive CMV3AGPF AST Plates. Enterococcal strains totalling 196 were isolated, of which 40.8% were E. faecalis and 15.3% were E. faecium. All tested isolates were susceptible to linezolid, penicillin and tigecycline. For most other antimicrobials the prevalence of resistance was 0.5–6.6% while high prevalence of quinupristin/dalfopristin (51.5%), tetracycline (30%) and lincomycin (52%) resistance was observed. This study affords better knowledge concerning the safety of raw goat’s milk in terms of the enterococci possible to isolate from this foodstuff. It seems that enterococci in milk are still mostly susceptible to antimicrobials of major concern as multiply resisted drugs, such as gentamycin and vancomycin. However, the presence of multi-resistant strains in goat milk is cause for apprehension.

In vitro transferability of penicillin, streptomycin, tetracycline, and erythromycin resistance from coagulase-negative staphylococci to Staphylococcus aureus and among the former species of bovine mammary gland origin was examined by bacterial mating on filters and by mixed-culture matings in broth and in skim milk. One hundred twenty-six (42 each on filter, in broth, and in skim milk) matings were performed among 37 isolates of different Staphylococcus species. Transfer of resistance to penicillin, tetracycline, or erythromycin was not detected. Of 51 matings performed to determine streptomycin-resistance transfer, 9 (3 each on filters, in broth, and skim milk) were successful. Nine strains representing 3 species of coagulase-negative staphylococci were tested as prospective donors of streptomycin resistance. Of these, 2 strains could transfer streptomycin resistance. A double-resistant donor, S hominis, not only transferred its streptomycin resistance to an S chromogenes strain lacking resistance, but also to an S aureus strain already carrying penicillin and tetracycline resistance. The transfer of streptomycin resistance from the donor S hominis, harboring 2 plasmids, to a plasmidless S chromogenes recipient strain was associated with apparent acquisition of the smaller plasmid of the donor by the recipient. The single-resistant donor, S epidermidis 681A, transferred streptomycin resistance to a tetracycline-resistant S aureus recipient. This strain however, failed to transfer its streptomycin resistance to another S aureus, 2 S hyicus, and 1 S xylosus recipient. Frequency of transfer of streptomycin resistance ranged from 1.1 X 10(-5) to 1 X 10(-4). When transfer of resistance was successful, attempts were made to characterize the transfer process. Conjugation appeared to be the mode of streptomycin-resistance transfer. Transfer of resistance between staphylococci of bovine mammary gland origin appears to be fairly uncommon. However, in view of the limitations of the procedures used, additional in vitro and in vivo work is needed to further assess the role of coagulase-negative staphylococci in dissemination of antibiotic resistance.

Objective: To determine the prevalence of selected virulence genes and the antimicrobial susceptibility of multidrug-resistant (MDR) Escherichia coli isolated from diarrheic neonatal calves. Sample: 97 E coli isolates from diarrheic neonatal calves. Procedures: E coli isolates were tested via PCR assay for 6 virulence genes and susceptibility to 17 drugs belonging to 9 classes. A 2-sample test of proportions was used to make comparisons between proportions of virulent and avirulent MDR isolates. Results: 23 of 97 (23.7%) isolates were virulent, and 74 (76.3%) were avirulent. Of the 23 virulent isolates, 15 (65.2%) were positive for K99, 14 (60.9%) for F41, 12 (52.2%) for STa, 9 (39.1%) for Stx1, 6 (26.1%) for intimin, and 0 (0%) for Stx2. Twenty of 23 (87.0%) virulent isolates expressed ≥ 2 virulence genes, and 3 of 23 (13.0%) were positive for 1 virulence factor. Eight of 23 (34.8%) virulent isolates expressed STa, K99, and F41, whereas 1 of 23 (4.4%) was positive for STa, F41, intimin, and Stx1. The second most frequent gene pattern was Stx1 and intimin. Twenty of 23 (87.0%) virulent isolates were MDR; the highest prevalence of resistance was recorded for the macrolide-lincosides, followed by the tetracyclines and penicillins. Also, 17 of 23 (74.0%) virulent isolates were resistant to sulfadimethoxine, and 10 of 23 (43.5%) were resistant to trimethoprim-sulfamethoxazole. Additionally, 60 of 74 (81.0%) avirulent isolates were MDR. Conclusions and Clinical Relevance: The prevalence of multidrug resistance was comparable for virulent and avirulent E coli isolated from diarrheic neonatal calves. Cephalosporins and aminoglycosides had reasonable susceptibility.

Few antimicrobials are licensed for use in sheep in Canada, and the range of indications is narrow. Treatment in an “extra-label” manner may be ineffective. In addition, potentially harmful drug residues in food-animal products and antimicrobial resistance in bacteria may be associated with extra-label drug use (ELDU). No data had been documented on drug use, specifically antimicrobial use (AMU), in Ontario sheep, although it was thought that much use was extra-label. This study investigated AMU and ELDU on 49 lamb-producing Ontario sheep farms. Data were prospectively collected over 12 months from the participating farms, and farm-level practices were ascertained with a questionnaire. Treatment-level and farm-level variables were investigated for associations with rates of AMU by means of Poisson rate regression models fit with a generalized estimating equation to control for clustering at the farm level. Antimicrobials with high mean exposure rates included chlortetracycline (in feed), penicillins, and oxytetracycline. The exposure rate in lambs was significantly lower (P < 0.01) with antimicrobial treatment of systemic signs, respiratory disease, or wound or injury than with treatment of other reported diseases or conditions; it was also significantly lower with 3 or more lambing periods per year (α = 0.05). The exposure rate in adult sheep was significantly lower with treatment of 5 of the 6 most prevalent diseases or conditions (α = 0.05) and significantly higher with producer decision to treat and producer experience of 20 y or greater. Rates of using antimicrobials not licensed for use in sheep were high, as was extra-label use of licensed antimicrobials. Diseases reportedly treated most often with antimicrobials (e.g., systemic signs, mastitis) were significantly associated with lower rates of ELDU (α = 0.05). Compared with the rates in adult sheep, the mean rate of use of nonlicensed antimicrobials was similar in the lambs, whereas the mean rate of extra-label use of licensed antimicrobials was lower among the lambs. The results are useful in determining if public health concerns about antimicrobial use in Ontario sheep are warranted and in creating drug use and licensure strategies for the Canadian sheep industry.

Peritoneal lavage was performed on ponies to determine the effect on peritoneal surfaces. Lavage solution (20 L) was introduced into each pony's peritoneal cavity through catheters placed in the paralumbar fossa, and the solution was removed by drainage from the ventral portion of the abdomen. Six ponies each were lavaged with sterile saline (0.9% NaCl) solution, sterile saline solution containing 5 X 10(6) U of potassium penicillin and 3 g of neomycin or povidone-iodine diluted to 3% by volume with sterile saline solution, and 3 ponies were lavaged with povidone-iodine diluted to 10% with sterile saline solution. Peritoneal lavage catheters were inserted in 3 control ponies, but lavage fluids were not administered. Peritoneal fluid specimens were collected at 6, 24, 48, and 96 hours after lavage. Nucleated cell counts, RBC counts, total protein determinations, and cytologic analysis were performed. The ponies were euthanatized at 96 hours, and representative sections of the peritoneum were examined. Lavage with saline solution and saline solution with antibiotics induced a mild, transient inflammatory response in the peritoneal fluid, with minimal or no changes observed at necropsy. Solutions containing povidone-iodine induced chemical peritonitis, which was severe in ponies lavaged with 10% povidone-iodine solutions. Peritoneal lavage with povidone-iodine solutions as dilute as 3% cannot be accomplished without causing inflammation of peritoneal surfaces.