Introduction: The development of Jembrana disease vaccine is an important effort to prevent losses in the Bali cattle industry in Indonesia. This study aims to prepare a Jembrana DNA vaccine encoding the transmembrane portion of the envelope protein in pEGFP-C1 and test the success of its delivery in culture cells using a chitosan-DNA complex. Material and Methods: Cloning of the DNA vaccine was successfully performed on E. coli DH5α and confirmed by colony PCR, restriction analysis and sequencing. The plasmids were prepared as a chitosan complex using the complex coacervation method and physicochemically characterised using a particle size analyser. A transfection assay was performed in HeLa cells with 4 h exposure, and mRNA expression was assessed at 24 h post transfection. Results: With a 1:2 (wt./wt.) ratio of DNA and chitosan, the complexes have a mean diameter of 236 nm, zeta potential value of + 17.9 mV, and showed no high toxicity potential in the HeLa cells. This complex successfully delivered the DNA into cells, as shown by the presence of a specific RT-PCR product (336 bp). However, the real-time PCR analysis showed that the delivery with chitosan complex resulted in lower target mRNA expression when compared with a commercial transfecting agent. Conclusion: pEGFP-env-tm JDV as a candidate vaccine can be delivered as the chitosan-DNA complex and be expressed at the transcription level in vitro. This initial study will be used for further improvement and evaluation in vivo.

Avian pathogenicEscherichia coli (APEC) causes serious colibacillosis and significant economic losses. Data on profiles of virulence factors and antibiotic resistances among APEC strains are crucial to the control of infection. In this study, strains were isolated from eastern China, and the prevalence of virulence factors and distribution of antibiotic resistance were determined. APEC strains were isolated and characterised by PCR for O serogroups, virulence factor genes, antibiotic resistance, and phylogenetic groups. O78 was the most prevalent serogroup and type A was the most frequent phylogenetic group. ThefimH,feoB, andiron genes were the most prevalent among the isolates. All isolates were multiresistant, and all strains were resistant to ampicillin and tetracycline, which are widely used in the poultry industry in China. This study provided important data on the presence of virulence genes and antibiotic resistance profiles of APEC from poultry farms in eastern China.

Septic arthritis is an important disease in horses, necessitating aggressive and prolonged therapy. In order to guide therapy, reliable methods of detecting the eradication of infection are needed. Therefore, the objective of this study was to investigate detection of eradication of infection in an experimental model of equine septic arthritis using standard diagnostic techniques. For this purpose, 17 adult horses were assigned to 3 experimental groups. The middle carpal joint of each horse was injected with Escherichia coli (Septic group, n = 8), lipopolysaccharide (LPS) (LPS group, n = 6), or sterile saline (Control group, n = 3) at day 0. Contralateral joints were not injected. Standard therapy was applied to all joints except non-injected joints in the Control group at day 1. Sequential samples of synovial fluid (SF) were collected for bacterial culture using 3 culture media [Columbia blood agar (CBA), brain heart infusion broth (BHI), and Signal blood culture medium] and for cytological evaluation [percentage neutrophils (PN), total nucleated cell count (TNCC), and total protein (TP)]. Escherichia coli-specific polymerase chain reaction (PCR) was carried out to detect E. coli DNA in synovial fluid. Culture and PCR were positive for E. coli in all joints injected with E. coli at day 1 and 1 joint was positive on BHI at day 4. Based on the results of bacterial culture, PCR, and TNCC, the elimination of infection in our experimental model occurred by day 4 post-infection in 6 out of 7 cases. Total protein (TP) and PN remained elevated at clinical threshold used for diagnosis of septic arthritis until day 14. In our experimental model of E. coli-induced arthritis, we conclude that TP and PN may not be good indicators for detecting the eradication of bacterial infection caused by E. coli from infected and subsequently treated joints.

The objectives of this study were to describe the frequency of antimicrobial resistance (AMR) in Escherichia coli and Campylobacter spp. isolates in fecal samples from beef cow-calf herds and to examine the associations between herd management practices, reported antimicrobial use, and AMR. Baseline prevalence data are needed to evaluate the effectiveness of antimicrobial stewardship programs. A pooled fecal sample, representing 20 cows, was collected from each of 107 herds during pregnancy testing. In the 305 recovered E. coli isolates (maximum 3 per herd), resistance to ≥ 1 antimicrobial was identified in 12 isolates [4%, 95% confidence interval (CI): 2% to 7%] from 105 herds (11%, 95% CI: 7% to 19%). The most common resistances identified in E. coli isolates were to tetracycline (3%) and to both streptomycin and sulfisoxazole (3%). Only 1 E. coli isolate was resistant to an antimicrobial of very high importance to human health - amoxicillin/clavulanic acid. However, 2 E. coli isolates had intermediate susceptibility to ciprofloxacin. Resistance to 1 antimicrobial was identified in 16 of 87 Campylobacter spp. isolates (18%, 95% CI: 11% to 28%) from 87 herds. Resistance to tetracycline was reported in 15% of Campylobacter spp. isolates and to nalidixic acid in 3.4%. Herds in which cows were treated with florfenicol were more likely to have E. coli resistance to ≥ 2 antimicrobials (OR 7.1, 95% CI: 1.1 to 57, P = 0.03). Herds with calf mortality of > 5% were more likely to have E. coli with resistance to streptomycin and sulfisoxazole [odds ratio (OR): 7.8, P = 0.03]. The results of this study are consistent with previous reports from western Canada and provide a starting point for designing an ongoing antimicrobial surveillance program.