In this study the tonB2 gene was cloned from Actinobacillus pleuropneumoniae JL01 (serovar 1) and expressed as a glutathione-S-transferase (GST) fusion protein in Escherichia coli BL21(DE3). The GST fusion protein was recognized by antibodies in serum positive for A. pleuropneumoniae by Western blot analysis. Purified soluble GST-TonB2 was assessed for its ability to protect BALB/c mice against A. pleuropneumoniae infection. Mice were vaccinated with GST-TonB2 subcutaneously and challenged intraperitoneally with either approximately 4.0 × 10(5) colony-forming units (CFU) or approximately 1.0 × 10(6) CFU of A. pleuropneumoniae 4074. They were examined daily for 7 d after challenge. The survival rate of the TonB2-vaccinated mice was significant higher than that of the mice given recombinant GST or adjuvant alone. These results demonstrate that A. pleuropneumoniae TonB2 is immunogenic in mice and should be further assessed as a potential candidate for a vaccine against A. pleuropneumoniae infection. In addition, an indirect enzyme-linked immunosorbent assay (ELISA) based on the GST-TonB2 recombinant protein was developed. Compared with the ApxIVA ELISA, the TonB2 ELISA provided earlier detection of antibodies in pigs at various times after vaccination with A. pleuropneumoniae live attenuated vaccine. When compared with an indirect hemagglutination test, the sensitivity and specificity of the TonB2 ELISA were 95% and 88%, respectively. The TonB2 ELISA provides an alternative method for rapid serologic diagnosis of A. pleuropneumoniae infection through antibody screening, which would be especially useful when the infection status or serovar is unknown.

The objective of this study was to determine the potential of Bdellovibrio bacteriovorus 109J as an alternative non-chemotherapeutic treatment of infectious bovine keratoconjunctivitis (IBK). To accomplish this, various parameters of B. bacteriovorus predation of Moraxella bovis were determined in vitro. Initial passage of B. bacteriovorus using M. bovis as prey required 10 d for active cultures to develop compared with 2 d for culture on normal Escherichia coli prey; however by the 5th passage, time to active predatory morphology was reduced to 2 d. This high passage B. bacteriovorus culture [1 × 10(10) plaque forming units (PFU)/mL] killed 76% of M. bovis [1 × 10(7) colony forming units (CFU)/mL] present in suspension broth in a 4 h assay. The minimal level of M. bovis supporting B. bacteriovorus predation was 1 × 10(4) CFU/mL. To assess the ability of B. bacteriovorus to kill M. bovis on an epithelial surface mimicking IBK, an in vitro assay with Madin-Darby bovine kidney (MDBK) cells inoculated with 4 × 10(7) CFU/mL M. bovis was used. Treatment with a B. bacteriovorus suspension (1.6 × 10(11) PFU/mL) decreased adherence of M. bovis to MDBK cells by 6-fold at 12 h of treatment, as well as decreased the number of unattached M. bovis cells by 1.4-fold. This study demonstrates that B. bacteriovorus has potential as an effective biological control of M. bovis at levels likely present in IBK-infected corneal epithelia and ocular secretions.

The study evaluated whether a transgenic carrot vaccine could induce a K88-specific immune response in sows and whether the resultant maternal antibody could protect piglets against enterotoxigenic Escherichia coli (ETEC) K88ac infection. Sows (n = 15) selected randomly from a farm in Korea were assigned to three groups (n = 5 per group: control [untreated]), group A (orally inoculated with a non-transgenic and transgenic carrot vaccines at 2 and 4 weeks ante partum, respectively), and group B (conventionally vaccinated according to the manufacturer’s instructions). After 7 days of lactation, 5 piglets selected randomly from each group were challenged with 1 × 10∨10 colony forming units/mL ETEC K88ac. Group C had the lowest mean fecal consistency score on post-challenge days 1 and 7. Histiologically, On post-challenge day 7, group C showed an increased duodenum and ileum villus:crypt ratio, compared to group A in the duodenum, with group B displaying the highest ratio. Groups B and C had more increased villus width than group A in the jejunum. Group C displayed the greatest increase in villus width in the ileum. The colostrums and serum from groups B and C displayed higher concentrations of IgA and IgG against ETEC K88, compared to group A. Based on the results, it was concluded that the transgenic carrot vaccine in sow per oral may have an effect on preventing piglet diarrhea as good as commercial recombinant vaccine.

Three pairs of specific primers were designed to amplify F2-1, F2-2, and XF2-2 truncated capsid protein genes of porcine circovirus type 2 (PCV-2). Amplified sequences were subcloned to pET-32a(+) vectors and expressed in Rosetta (DE3) Escherichia coli by induction of isopropy-β-D-thiogalactoside (IPTG). All of the fusion proteins had positive reactions to PCV-2 antiserum and His-XF2-2 showed the best reactivity. Proteins were used to immunize BALB/c mice to produce monoclonal antibodies (mAbs), and 7 mAbs were selected. Capsid protein N-terminal parts 55 to 96 amino acid (aa), 97 to 141 aa, and 143 to 211 aa were confirmed as binding regions of the 7 mAbs. Reactivity between His-XF2-2 and the 7 mAbs was detected, FmAb-8 showed the best reactivity. The dominant B-cell epitope was located at 97 to 141 aa. The PEPSCAN indicated that the P122–136 peptide contained the dominant B-cell epitope.

A farm trial was conducted to evaluate the effect of in-water iodine on piglet growth, the incidence of diarrhea, and the development of deleterious oral and dental conditions. A total of 208 weaned piglets were included in the study. Piglets were weighed 3 times: within 24 h of weaning, and 3 wk and 6 wk after weaning. A concentration of 1 ppm iodine was provided in their drinking water. Swabs were taken from all water nipples and water lines and pooled fecal samples were collected from all pen floors. Fecal samples were also collected from sows at weaning. The swabs and fecal samples were tested for the presence of Salmonella and Escherichia coli. Within 24 h of each weighing, a complete oral examination was performed on each piglet. No significant difference in growth (P > 0.05) or dental conditions (P > 0.05) was found among treatment groups during the period that iodine was added to the drinking water. After weaning, all deleterious oral conditions increased (oral lesions from weaning to 6 wk, staining and caries from weaning to 3 wk, gingivitis from 3 wk to 6 wk; P < 0.05). Only gingivitis was found to be negatively associated with piglet weight (P < 0.05). Salmonella was cultured only twice from fecal samples and never from water nipples. Only 1 sow tested positive for Salmonella and E. coli O139: K82 and O157:K“V17 were cultured only rarely from the water nipples. No signs of diarrhea were noted throughout the study. Adding an aqueous iodine supplement to nursery pigs, therefore, did not provide an advantage for either growth or oral condition. Deleterious oral conditions do increase after weaning, with gingivitis being associated with lower piglet weight.

This study established a disease model and protocol for bacterial challenge with Escherichia coli serogroup O2 strain EC317 in Japanese quail. Five groups of 10 birds each were injected subcutaneously in the breast with 200 μL of a brain–heart infusion (BHI) culture containing 1 × 10(8), 1 × 10(7), 1 × 10(6), 1 × 10(5), or 1 × 10(4) colony-forming units/mL of the test organism, which had been isolated from a turkey with cellulitis and septicemia. Birds in a 6th group were controls that received sterile BHI alone. Localized lesions of cellulitis developed in all of the birds that received E. coli. The morbidity and mortality rates were highest (100%) in the birds receiving the highest dose of E. coli and decreased linearly with decreasing dose (P < 0.05). Severity of disease, including lesions of pericarditis and perihepatitis, was also directly proportional to the dose of E. coli. These findings indicate that this disease challenge protocol can be used to study disease resistance and immunologic consequences of contaminant exposure or other stressors in birds.

Objective—To evaluate the impact of oxytetracycline exposure on horizontal transfer of an antimicrobial resistance plasmid. Sample—Populations of Salmonella enterica subsp enterica serovar Typhimurium and Escherichia coli. Procedures—Mixed populations of plasmid donor (Salmonella Typhimurium) and recipient (E coli) bacteria were assigned to 1 of 2 simulated oxytetracycline dosing regimens (high peak concentration-short elimination half-life [HC-SHL] or low peak concentration—long elimination half-life [LC-LHL]) or served as untreated control replicates. Donor, recipient, and transconjugant (E coli that acquired the plasmid) bacteria populations were quantified at 12, 24, and 36 hours after oxytetracycline administration by use of culture on selective bacterial growth media. Results—The ratio of transconjugant to donor bacteria was significantly reduced in the oxytetracycline-exposed replicates, compared with the ratio for the control replicates, at 12 hours. At 24 and 36 hours, results for the HC-SHL regimens were not significantly different from results for the respective control replicates, and results for the LC-LHL regimens also were not significantly different from results for the respective control replicates. The oxytetracycline concentration at these time points (12 hours in the HC-SHL regimen and all 3 time points in the LC-LHL regimen) were in excess of the minimum inhibitory concentration of the recipient bacteria. Conclusions and Clinical Relevance—Transfer of antimicrobial resistance plasmids may be suppressed in vitro by oxytetracycline exposure at concentrations greater than the minimum inhibitory concentration of the recipient bacteria.

This study investigated the occurrence and antimicrobial resistance profiles of Escherichia coli and Salmonella sp. isolated from swine samples submitted to the Minnesota Veterinary Diagnostic Laboratory (MVDL) in Saint Paul, Minnesota from 1995 to 2004. During this time period, a total of 5072 E. coli and 2793 Salmonella sp. was isolated. Most of these isolates were found to be resistant to the tetracycline and beta-lactam group of antibiotics. Resistance to spectinomycin was also frequently observed. An increasing trend in ampicillin resistance and a decreasing trend in apramycin resistance were seen in both pathogens, although ampicillin resistance was relatively higher in E. coli than in Salmonella. Aminoglycoside (amikacin) and quinolone (enrofloxacin) were the only antimicrobials to which minimum or no resistance was observed. The resistance of pig pathogens to several antibiotics indicates the need to routinely monitor the use of these antimicrobials and their associated resistance in pig populations.