Changes in the prevalence of resistant Escherichia coli in cattle receiving subcutaneously injectable oxytetracycline in addition to in-feed chlortetracycline compared with cattle receiving only in-feed chlortetracycline

Information about the prevalence of antibiotic resistance in commensal enteric bacteria is of interest because these bacteria are potential indicators of selection pressure on enteric bacteria and represent a reservoir of resistance genes in potentially pathogenic bacteria. This study reports changes in the prevalence of resistance to antibiotics in commensal Escherichia coli from cattle receiving either subcutaneously injectable oxytetracycline in addition to in-feed chlortetracycline or only in-feed chlortetracycline. Resistance to 19 antibiotics was examined. The use of injectable oxytetracycline in addition to in-feed chlortetracycline was significantly associated (P < 0.05) with an increase in the prevalence of resistance only to chloramphenicol and sulfisoxazole.     

Biofilm bacteria: formation and comparative susceptibility to antibiotics

The Calgary Biofilm Device (CBD) was used to form bacterial biofilms of selected veterinary gram-negative and gram-positive pathogenic bacteria from cattle, sheep, pigs, chicken, and turkeys. The minimum inhibitory concentration (MIC) and minimum biofilm eradication concentration (MBEC) of ampicillin, ceftiofur, cloxacillin, oxytetracycline, penicillin G, streptomycin, tetracycline, enrofloxacin, erythromycin, gentamicin, tilmicosin, and trimethoprim-sulfadoxine for gram-positive and -negative bacteria were determined. Bacterial biofilms were readily formed on the CBD under selected conditions. The biofilms consisted of microcolonies encased in extracellular polysaccharide material. Biofilms composed of Arcanobacterium (Actinomyces) pyogenes, Staphylococcus aureus, Staphylococcus hyicus, Streptococcus agalactiae, Corynebacterium renale, or Corynebacterium pseudotuberculosis were not killed by the antibiotics tested but as planktonic bacteria they were sensitive at low concentrations. Biofilm and planktonic Streptococcus dysgalactiae and Streptococcus suis were sensitive to penicillin, ceftiofur, cloxacillin, ampicillin, and oxytetracycline. Planktonic Escherichia coli were sensitive to enrofloxacin, gentamicin, oxytetracycline and trimethoprim/ sulfadoxine. Enrofloxacin and gentamicin were the most effective antibiotics against E. coli growing as a biofilm. Salmonella spp. and Pseudomonas aeruginosa isolates growing as planktonic populations were sensitive to enrofloxacin, gentamicin, ampicillin, oxytetracycline, and trimethoprim/sulfadoxine, but as a biofilm, these bacteria were only sensitive to enrofloxacin. Planktonic and biofilm Pasteurella multocida and Mannheimia haemolytica had similar antibiotic sensitivity profiles and were sensitive to most of the antibiotics tested. The CBD provides a valuable new technology that can be used to select antibiotics that are able to kill bacteria growing as biofilms.     

An investigation of exudative epidermitis (greasy pig disease) and antimicrobial resistance patterns of Staphylococcus hyicus and Staphylococcus aureus isolated from clinical cases

Exudative epidermitis (EE) is a common skin disease of young pigs, caused mainly by Staphylococcus hyicus. Increased prevalence of EE and poor response to treatment are reported. Common strategies used by Ontario pork producers to treat pigs with EE were determined using a survey. Injection of penicillin G was reported as the most common parenteral antibiotic choice. Antimicrobial resistance patterns of S. hyicus and Staphylococcus aureus isolated from clinical cases (30 herds with samples from approximately 6 pigs per farm) showed that 97% of S. hyicus isolates were resistant to penicillin G and ampicillin; 71% of these isolates were resistant to ceftiofur. Similar resistance was noted among S. aureus isolates. Antimicrobial resistance has become a problem in the treatment of EE in Ontario.     

Comparison of conventional and long-acting oxytetracyclines in prevention of induced Actinobacillus (Haemophilus) pleuropneumoniae infection of growing swine.

These experiments tested the hypothesis that long-acting oxytetracycline (oxytetracycline-LA) was more effective than regular oxytetracycline in preventing porcine pleuropneumonia when administered either 24 or 48 h prior to experimental challenge with virulent strains of Actinobacillus pleuropneumoniae. Two experiments (1 and 2) were conducted using growing pigs (average weight 12-15 kg). Antibiotic treatments were administered once intramuscularly at 20 mg/kg body weight; controls received an equivalent volume of saline. Clinical signs were recorded over seven days, and mortality rates and pathological lesions were analyzed using analysis of variance. Serum oxytetracycline levels were compared 48 and 72 h postinjection. All pigs developed clinical disease following experimental infection. Actinobacillus pleuropneumoniae was recovered from 42% of experiment 1 pigs and all of experiment 2 pigs. The data showed that both oxytetracycline and oxytetracycline-LA given at the same dose protected pigs against experimental infection when given 24 h prior to challenge, and there was no difference between the efficacy of the two drugs in this experiment. When administered 48 h prior to challenge, only oxytetracycline-LA reduced the clinical signs and pathological changes following A. pleuropneumoniae challenge. Between 48 and 72 h postinjection, oxytetracycline-LA blood levels were significantly greater compared to oxytetracycline-treated pigs.     

Antibiotic residues in goats milk following intramammary treatment

Two lactational intramammary antibiotic preparations (cloxacillin and oxytetracycline) were each used to treat eight goatsby intramammary infusion in one half. The rate of release of the antibiotics was monitored by a diffusion assay based on the sensitive organism Bacillus stearothermophilus var. calidolactis. Cloxacillin was still detectable 156 hours afterthe final treatment while oxytetracycline was undetectable 108 hours following treatment. A small but significant amount of antibiotic was translocated between treated and untreated halves of the udder in both cases.     

Small filter membrane bags for the study of antibiotic action in the digestive tract: bioavailability and in situ activity of oxytetracycline, chloramphenicol, neomycin and gentamicin in the pig caecum

Observations were made for two consecutive days on the disposition of oxytetracycline, chloramphenicol, neomycin and gentamicin through the digestive tract of pigs given an oral drench. The effect of these antibiotics upon the colibacilli flora and upon an Escherichia coli 127 strain in small filter membrane bags placed into the caecum was also investigated. For chloramphenicol, inactivation by antibiotic resistant colibacilli and, for neomycin, losses of activity by digestive content may explain the lower concentrations obtained in the gut and the correlative lack of curative or preventive action against E coli 127 in small bags. Tetracycline and gentamicin prevented growth of E coli 127 from the second day.     

National surveillance of Salmonella enterica in food-producing animals in Japan

A total of 518 fecal samples collected from 183 apparently healthy cattle, 180 pigs and 155 broilers throughout Japan in 1999 were examined to determine the prevalence and antimicrobial susceptibility of Salmonella. The isolation rates were 36.1% in broilers, 2.8% in pigs and 0.5% in cattle. S. enterica Infantis was the most frequent isolate, found in 22.6% of broiler fecal samples. Higher resistance rates were observed against oxytetracycline (82.0%), dihydrostreptomycin (77.9%), kanamycin (41.0%) and trimethoprim (35.2%). Resistance rates to ampicillin, ceftiofur, bicozamycin, chloramphenicol and nalidixic acid were <10%. CTX-M-2 β-lactamase producing S. enterica Senftenberg was found in the isolates obtained from one broiler fecal sample. This is the first report of cephalosporin-resistant Salmonella directly isolated from food animal in Japan.     

PCR analysis of Pasteurella multocida isolates from an outbreak of pasteurellosis in Indian pigs

An outbreak of pasteurellosis with high mortality was recorded in indigenous pigs in India. The presence of Pasturella multocida in samples collected from dead pigs was detected by smear examination and isolation, and later by P. multocida specific polymerase chain reaction (PM-PCR). P. multocida was detected in all the samples collected from dead pigs, with nine strains ultimately isolated. All the isolates were positive by PM-PCR. Six isolates showed CAPA and three were of CAPD capsular types. All the isolates were negative for toxigenic gene (toxA). The isolates were sensitive to oxytetracycline, doxycycline, gentamycin, erythromycin, ampicillin, amoxycillin, chloramphenicol and enrofloxacin and resistant to sulphadiazine and cloxacillin. The PCR assays used in this study have been shown to be useful diagnostic tools for P. multocida detection and characterization.     

The influence of disease on feed and water consumption and on pharmacokinetics of orally administered oxytetracycline in pigs

In the present study the feed and water consumption and pharmacokinetic parameters of orally administered oxytetracycline were compared in clinically healthy pigs and in the same pigs following a challenge with Actinobacillus (Haemophilus) pleuropneumoniae toxins. Endobronchial challenge with A. pleuropneumniae toxins was accompanied by anorexia, increased lassitude, labored breathing, fever, and increased white blood cell counts. Pleuropneumonia was evident in all pigs on autopsy. Following the challenge, both feed and water consumption were markedly reduced. In contrast to recommendations in the literature, it is concluded that drugs should not be administered to pneumonic pigs via water. In healthy pigs the oral bioavailability of oxytetracycline (50 mg/kg), given on an empty stomach, was 4.8% and the elimination half-life (t1/2 beta) was 5.92 h. After challenge, the pigs showed great variation in oxytetracycline plasma concentrations. In addition, the mean computed elimination rate constant (beta), t1/2 beta, the area under the plasma concentration-time curve (AUC), and clearance in pneumonic pigs differed significantly (P less than .05) from the values found in healthy pigs. The elimination half-life (t1/2 beta), AUC, and volume of distribution (Vd area) were increased. In diseased pigs the mean of maximum plasma concentrations (.87 micrograms/ml) was reached after 7 h, in contrast to 1.74 h (1.87 micrograms/ml) in the healthy pigs.     

Influence of antibiotics used as feed additives on the immune effect of erysipelas live vaccine in swine

To investigate the influence of antibiotics used as feed additives on the immune response to erysipelas live vaccine, the pig inoculation test was applied. Avilamycin, oxytetracycline quaternary salt, enramycin, virginiamycin and tylosin phosphate were selected as test antibiotics. Five experimental feeds containing each antibiotic at the highest concentration permitted for feed additives in Japan, and the basal diet lacking antibiotics were examined. Twenty-nine pigs were divided into six groups. At first all the groups were fed with the antibiotic-free basal diet for 7 days, and then each group received the experimental feeds. On the 14th day after feeding with test feeds all the pigs, except for one control pig in each group, were immunized with the vaccine and all the pigs were then challenge-exposed to a virulent strain of Erysipelothrix rhusiopathiae 14 days after vaccination. The clinical response was observed every day for 14 days. In all the groups, most of the vaccinated pigs did not develop any clinical signs of acute erysipelas after the challenge exposure, whereas non-vaccinated control pigs died or showed severe generalized erythema with profound depression and anorexia. No differences in the protection against the challenge exposure were observed among the groups. Therefore, the present results suggest that these selected antibiotics would not interfere with the immune effect of the vaccine if given at the usual concentrations used for feed additives.     

Development and validation of an endoscopic myringotomy technique to treat otitis media and interna in a case series of three guinea pigs (Cavia porcellus)

Background: Medical treatment of clinical otitis media and interna in guinea pigs is often unsatisfactory. Total ear canal ablation and lateral bulla osteotomy are used successfully in dogs, cats, and rabbits to treat chronic, medically nonresponsive ear disease. However, in guinea pigs this surgery can cause death. This study aimed to evaluate and describe a standardized myringotomy technique to treat guinea pigs with otitis media and interna. Methods: Ear dissections and endoscopically assisted myringotomy were evaluated on guinea pig carcasses to choose the most suitable endoscope and optics. Three client-owned guinea pigs with chronic otitis media and interna had a myringotomy with bacteriological sampling and therapeutic irrigation of the tympanic cavity performed. Anesthesia for the procedures was 25 minutes, the patients were recovered and eating within 2–6 hours, and discharged on targeted antibiotic therapy within 24 hours. Videotoscopy was performed at 7- and 14-days follow-up to assess the ear canal, healing of the tympanic membrane, and to evaluate the tympanic cavity. Repeat lavaging of the tympanic cavity was performed until the tympanic membrane incision was healed. Further follow-up occurred at 21- and 154-days postmyringotomy. Results: Two out of three guinea pigs had complete resolution of clinical signs, and the other showed clinical improvement from a 90° head tilt to a 30° head tilt. Conclusions and case series relevance: Videotoscopy assisted myringotomy under general anesthesia was a quick and straightforward endosurgical technique in three guinea pigs with chronic otitis media and interna. It allowed the collection of fluid for bacteriological culture and provided symptomatic relief.     

Therapeutic responses of piglets to experimentally induced colibacillosis.

The purpose of these experiments was to determine the effects of prior exposure to antimicrobials on subsequent treatment of artificially induced colibacillosis in pigs. One- to two-week-old piglets were given 10(7) multiple antibiotic resistant Escherichia coli orally. Two groups of the piglets received priming doses at different levels of the antimicrobial preparation, ASP-250 before challenge. The remaining group received no antimicrobial. Piglets ill as a result of the challenge were treated with chloramphenicol of chlortetracycline, or received no treatment. Chloramphenicol was significantly more effective than chlortetracycline in terminating colibacillosis in the primed and unprimed groups. There were fewer deaths and relapse cases in those groups which received chloramphenicol treatment. Results in piglets treated with chlortetracycline were not significantly better than those which received no treatment.     

Comparison of two pseudorabies virus vaccines,that differ in capacity to reduce virus excretion after a challenge infection,in their capacity of reducing transmission of pseudorabies virus

Pseudorabies virus (PRV) vaccines are often compared for their capacity to reduce virus excretion after a challenge infection. Vaccines, used for the eradication of PRV, however, should reduce transmission of PRV among pigs. The purpose of this study was to investigate whether the amount of virus excreted after a challenge infection is an accurate measure of the capacity of a vaccine to reduce transmission of PRV among pigs. Two experiments were carried out, each using two groups of 10 pigs. The pigs in group one were intramuscularly vaccinated once with the glycoprotein E (gE)-negative vaccine X, the pigs in group two with the gE-negative strain 783. Eight weeks later, 5 pigs in each group were inoculated with wild-type PRV. A gE-ELISA was used to detect PRV infection. The transmission of PRV was estimated from the number of contact infections and expressed as the reproduction ratio R. The inoculated pigs vaccinated with vaccine X shed significantly more virus than the inoculated pigs vaccinated with strain 783. However, despite the difference in virus excretion, the transmission of PRV between the two groups did not differ. We conclude that virus excretion is not an accurate measure for determining vaccine effectiveness. However, R of vaccine X (R = 0.98) was not significantly below one, whereas R of vaccine 783 (R = 0) was significantly below one. Consequently, we cannot exclude the possibility that major outbreaks of PRV occur among pigs vaccinated with vaccine X.     

The acute phase response in pigs experimentally infected with Escherichia coli and treated with systemic bactericidal antibiotics

The objective of this study was to monitor the acute phase response (APR) in pigs experimentally infected with a known enterotoxogenic strain of Escherichia coli (E. coli) followed by treatment with a systemic bactericidal antibiotic to evaluate a model of acute enterotoxemia. The APR was determined by measuring rectal temperature (RT) and serum concentration of cortisol (CS), C-reactive protein (CRP), haptoglobin (HG), tumor necrosis factor-alpha (TNF-α), and interferon-gamma (IFN-γ). Twenty-four single source, male, 24-day-old, crossbred pigs were used for this study. Twelve of the pigs were non-surgically cannulated for blood collection and 12 were used for RT monitoring. All pigs received an oral dose of E. coli K88 (2.4×10⁸ colony-forming units). Five hours later, six pigs in each group received an intramuscular injection of 0.5 ml of saline and the remaining six received an intramuscular injection of antibiotic (25 mg of Ceftiofur HCl in 0.5 ml). Data collection occurred hourly from −1 to 5 h post-E. coli, then every 30 min between 5 and 8 h, and at 24 h post-E. coli. Prior to antibiotic treatment, RT increased (P<0.0001) in both treatment groups. After antibiotic treatment, RT, CS, and CRP were all higher in the antibiotic treatment groups (P<0.05) compared to the saline groups. An increase in HG and IFN-γ (P<0.01) was observed in both groups, while TNF-α remained unchanged. Results indicate that systemic bactericidal antibiotic treatment post-E. coli induces an APR similar to that in other studies using different models of enterotoxemia. Therefore, we propose that this approach of eliciting an acute enterotoxemia provides a more natural scenario encountered in a swine production unit.     

Presentation of postweaning Escherichia coli diarrhea in southern Ontario, prevalence of hemolytic E. coli serogroups involved, and their antimicrobial resistance patterns

Post-weaning Escherichia coli diarrhea (PWECD) in Ontario was investigated using a case-control study involving 50 Ontario nurseries. The clinical signs and the impact on productive parameters were determined by means of a producer survey. The hemolytic E. coli serogroups involved in PWECD (O149:K91:K88) were examined in this study. Based on a polymerase chain reaction test, the hemolytic E. coli from 82% of the case herds were positive for 3 enterotoxins (STa, STb, and LT), those from 12% of the case herds were positive for STb and LT only, and those from one herd (6%) were positive for 3 enterotoxins, as well as for verotoxin and F18 pili. The E. coli involved in disease were resistant to multiple antibiotics. Case farms commonly used a wide variety of antibiotics either in the feed or water, or as injectable drugs. The most common antibiotic used to treat PWECD on the study farms was apramycin, but evidence of resistance to this antibiotic was noted. The PWECD problem was commonly seen within a week of weaning but onset of diarrhea was reported as late as the grower-finisher stage. Growth rate was poorer in case herds and mortality was higher than in control herds, demonstrating that PWECD is an economically important disease in Ontario.     

Evaluation of Petrifilm? Select E. coli Count Plate medium to discriminate antimicrobial resistant Escherichia coli

Background

Screening and enumeration of antimicrobial resistant Escherichia coli directly from samples is needed to identify emerging resistant clones and obtain quantitative data for risk assessment. Aim of this study was to evaluate the performance of 3M™ Petrifilm™ Select E. coli Count Plate (SEC plate) supplemented with antimicrobials to discriminate antimicrobial-resistant and non-resistant E. coli.

Method

A range of E. coli isolates were tested by agar dilution method comparing the Minimal Inhibitory Concentration (MIC) for eight antimicrobials obtained by Mueller-Hinton II agar, MacConkey agar and SEC plates. Kappa statistics was used to assess the levels of agreement when classifying strains as resistant, intermediate or susceptible.

Results

SEC plate showed that 74% of all strains agreed within ± 1 log₂ dilution when comparing MICs with Mueller-Hinton II media. High agreement levels were found for gentamicin, ampicillin, chloramphenicol and cefotaxime, resulting in a kappa value of 0.9 and 100% agreement within ± 1 log₂ dilution. Significant variances were observed for oxytetracycline and sulphamethoxazole. Further tests showed that the observed discrepancy in classification of susceptibility to oxytetracycline by the two media could be overcome when a plate-dependent breakpoint of 64 mg/L was used for SEC plates. For sulphamethoxazole, SEC plates provided unacceptably high MICs.

Conclusion

SEC plates showed good agreement with Mueller-Hinton II agar in MIC studies and can be used to screen and discriminate resistant E. coli for ampicillin, cephalothin, streptomycin, chloramphenicol, cefotaxime and gentamicin using CLSI standardized breakpoints, but not for sulphamethoxazole. SEC plates can also be used to discriminate oxytetracycline-resistant E. coli if a plate-dependent breakpoint value of 64 mg/L is used.     

High prevalence and widespread distribution of multi-resistant Escherichia coli isolates in pigs and poultry in China

Escherichia coli play an important ecological role within resistant bacteria populations, and can be used as a bio-indicator of antimicrobial resistance. The aim of the present study was to use this feature of E. coli to investigate the prevalence of antimicrobial resistance and the degree of cross-species transmission of bacteria in pigs and poultry in China. A total of 592 E. coli strains, isolated from pigs and poultry (healthy and diseased animals), were tested for resistance to 22 antimicrobials representing eight antimicrobial drug types.E. coli isolates had high rates of resistance to ampicillin (99.5%), doxycycline (95.6%), tetracycline (93.4%), trimethoprim–sulfamethoxazole (74.3%), amoxicillin (65.1%), streptomycin (54.7%), and chloramphenicol (50.2%). Resistance to cephalosporins, quinolones, and aminoglycosides was also quite prevalent. The majority (81%) of isolates demonstrated multi-antimicrobial resistance, most commonly to 5–6 different antimicrobial types. One isolate was resistant to all 22 antimicrobials. Twenty-two cultures exhibiting multi-antimicrobial resistance were analysed by pulsed-field gel electrophoresis (PFGE) to assess their distribution between farms. Three distinct PFGE types were identified, indicating inter-farm transmission of multi-antimicrobial resistant bacteria. The study confirmed the presence and transmission of multi-antimicrobial-resistant E. coli strains amongst pigs and poultry in China and highlights the urgent need for appropriate monitoring programmes.     

Associations of antimicrobial use with antimicrobial resistance in Campylobacter coli from grow-finish pigs in Japan

To determine associations between antimicrobial use and antimicrobial resistance in Campylobacter coli, 155 isolates were obtained from the feces of apparently healthy grow-finish pigs in Japan. In addition, data on the use of antibiotics collected through the national antimicrobial resistance monitoring system in Japan were used for the analysis. Logistic regression was used to identify risk factors to antimicrobial resistance in C. coli in pigs for the following antimicrobials: ampicillin, dihydrostreptomycin, erythromycin, oxytetracycline, chloramphenicol, and enrofloxacin. The data suggested the involvement of several different mechanisms of resistance selection. The statistical relationships were suggestive of co-selection; use of macrolides was associated with enrofloxacin resistance (OR=2.94; CI(95%): 0.997, 8.68) and use of tetracyclines was associated with chloramphenicol resistance (OR=2.37; CI(95%): 1.08, 5.19). The statistical relationships were suggestive of cross-resistance: use of macrolides was associated with erythromycin resistance (OR=9.36; CI(95%): 2.96, 29.62) and the use of phenicols was associated with chloramphenicol resistance (OR=11.83; CI(95%): 1.41, 99.44). These data showed that the use of antimicrobials in pigs selects for resistance in C. coli within and between classes of antimicrobials.     

Comparison of penicillin, oxytetracycline, and trimethoprim-sulfadoxine in the treatment of acute undifferentiated bovine respiratory disease

Penicillin, oxytetracycline, and a trimethoprimsulfadoxine combination were compared as first choice antibiotics for the treatment of acute bovine respiratory disease in weaned beef calves. There was no statistical difference in the mortality losses due to respiratory disease; however, the case fatality rate in the trimethoprim-sulfadoxine treatment group (3%) was markedly lower than in the penicillin (10%) and oxytetracycline (8%) treatment groups. The trimethoprim-sulfadoxine group also had statistically fewer treatment days compared to the penicillin and oxytetracycline groups (p < 0.05). Inclusion of mortality costs in the calculation of treatment costs demonstrated that treatment of the trimethoprimsulfadoxine group was appreciably less costly than treatment of the other groups. Temperature response abnormalities, defined as either an elevation in temperature or a failure of temperature to drop from one treatment day to the next, were associated with a relapse rate of approximately 50%. In vitro antibiotic sensitivity testing of Pasteurella haemolytica isolated from pretreatment nasal swabs was not a useful predictor of treatment success of antimicrobials.     

Study of serum drug levels in calves following intramuscular administration of three tetracycline drug preparations

Three antibiotic formulations, oxytetracycline (A) in propylene glycol and oxytetracycline (B) in polyvinyl pyrrolidine and pyrrolidino-methyltetracycline in an oil suspension were given to calves by the intramuscular route. Only oxytetracycline (A) appeared to cause much pain after injection.

The half-time elimination (t_½cl) for oxytetracycline (A) was 14.000 ± 3.580 h for oxytetracycline (B) 10.290 ±5.159 h and for pyrrolidinomethyltetracycline 8.160 ± 0.920 h. The rate of elimination `beta slope” for oxytetracycline (A) was 0.052 ± 0.012 h⁻¹ for oxytetracycline (B) 0.077 ± 0.261 h⁻¹ and for pyrrolidinomethyltetracycline 0.086 ± 0.010 h⁻¹. The Y intercept of the “beta” elimination slope C_os (μg/mL) for oxytetracycline (A) was 2.490 ± 1.040, for oxytetracycline (B) 3.463 ± 1.874 and for pyrrolidinomethyltetracycline 2.852 ± 1.360.

Pyrrolidinomethyltetracycline appeared to have a two component elimination curve, however, only the “beta slope” was used for the calculations.