Salmonella is one of the most important bacterial pathogens responsible for many zoonotic food-related infectious diseases. Quantitative detection of the foodborne Salmonella contamination in various food sources is therefore critical for preventing the related disease outbreaks. In this study, we developed and evaluated a reliable real-time polymerase chain reaction (RT-PCR) assay to detect the Salmonella contamination quantitatively. The experimental results showed that our invA gene-specific quantitative RT-PCR (qRT-PCR) assay provides a strong correlation between the Cq values and the direct plate counts of Salmonella species in the artificially formulated samples. Further study may be necessary to identify more accurate correlation and equation that can apply to Salmonella spp.
To evaluate alterations in fecal bacterial compositions in weaning piglets after supplementation with Phellodendron Cortex extract (PCE), piglets were allowed freely to consume feed containing 10 ppm of PCE for 7 weeks. During the feeding period, fecal samples were collected from individual piglets. A total of 578 valid sequence reads were generated from 40 fecal samples of normal piglets before supplementation with PCE. Fifteen classes were identified. Approximately 90% of classifiable sequences belonged to Clostridia and Bacilli classes. The abundance of Clostridium spp. and Lactobacillus spp. was determined based on the results of the sequencing. The supplementation with 10 ppm PCE for 3 weeks increased the numbers of Clostridium spp. and Lactobacillus spp. whereas seven weeks of supplementation with PCE reduced their abundance. In vitro, PCE inhibited the growth of E. coli and Salmonella enteritidis in a dose-dependent manner.
This study investigated the efficacy of four Brucella (B.) abortus recombinant proteins, namely adenylate kinase (Adk), nucleoside diphosphate kinase (Ndk), 50S ribosomal protein (L7/L12) and preprotein translocase subunit (SecB), as a combined subunit vaccine (CSV) against B. abortus infection in BALB/c mice. Immunoblotting assay showed that these four recombinant proteins as well as pcold-TF vector reacted individually with Brucella-positive serum, but not with Brucella-negative serum. The peripheral blood CD4+ T cell population was increased in CSV-immunized mice compared to PBS and pcold-TF vector groups. In addition, CSV and pcold-TF groups displayed induced IgG1 and IgG2a antibodies production compared to PBS and RB51 group, whereas IgG2a titer was higher than IgG1 titer in CSV group. The secretion profiles of IgG1 and IgG2a production together with an enhancement of CD4+ T cell population suggested that CSV did not only induce T helper 1 (Th1) T cell immunity but also humoral immunity. Therein, Th1 T cell immunity is more predominant in eliminating intracellular bacteria B. abortus. Furthermore, CSV immunization significantly reduced the bacterial burden in the spleen as well as the spleen weight in comparison to PBS and pcold-TF groups. Altogether, combination of these antigens could be potential to induce protective immunity against B. abortus infection in animals.
The purpose of this study was to find the change in social awareness in livestock stamping out over time. We used the text data mining method for this study. The data used in this analysis comprised approximately 5,600 newspaper articles. The data were sourced using the keywords 'livestock stamping out' 2010-2017, and classified into two-year intervals. The analysis showed a change in perspective on livestock disease and prevention activities over time. Additionally, it was confirmed that problems related to 'ethics' were continuously raised during the entire period. Prevention activities cannot succeed without the support and participation of people. Thus, the results of this study can be used to establish and implement livestock disease prevention policies.
Porcine epidemic diarrhea virus (PEDV) is a porcine coronavirus that causes enteric diseases characterized by watery diarrhea and dehydration in suckling piglets. Concentrated and highly purified viruses are required for the preparation of vaccines, diagnostics, and virus research. Currently, most protocols for virus purification require ultracentrifugation, which can be an instrumental barrier to routine operations in a laboratory. In this study, the efficacy of low-speed centrifugation for virus concentration was examined. The SM98 strain of PEDV was propagated in Vero cells and pelleted by centrifugation for 3 h at high speed (100,000 × g) or for 18 h at low speed (10,000 × g). The efficacy of virus concentration was analyzed by virus titration and western blotting. The amounts of infectious viruses and viral proteins in the pelleted samples obtained by low-speed centrifugation were comparable to those obtained by high-speed centrifugation. Interestingly, the pelleted sample impurity level was lower in low-speed than in high-speed centrifugation. In summary, we describe an efficient, easy-to-perform protocol for the preparation of purified and concentrated PEDV.
Recently, improvement of eggshell hygiene has emerged as an important issue in food industry. Various studies have continued to examine methods for controlling egg-borne pathogen, and among such methods, for table eggs, washing (with UV irradiation) is the most commonly used method. However, this method was not sufficient to control egg microbial contamination. Therefore, the purpose of this study was to verify whether it is appropriate to use ClO2 gas, which has been proven safe in this experiment model, as an alternative to the conventional washing (with UV irradiation) method. As a result, we have identified a range of optimal effectiveness in response to exposure concentrations and time of ClO2 gas. Through experimental models that reflect differences in farm size, a microbial reduction effect of approximately >2 log CFU/eggshell was achieved at 40ppm/8h for small farms, and 160ppm/30min for large farms, indicating greater effectiveness than the conventional method. However, in large-scale experiment, when bulk eggs were stacked and exposed to ClO2 gas, eggs in the depths showed a lower effect by approximately 0.8~1.5 log CFU/eggshell, as compared to the eggs in the upper section. For further study, if technical improvements are achieved in the future studies allowing the gas to better penetrate the depths of stacked eggs, it will be a model that can be more useful to the field.
Avian pathogenic Escherichia coli (APEC) is an agent associated with colibacillosis and an important primary pathogen with responsible for significant economic losses in chickens. This study investigated the molecular characteristics including virulence and antimicrobial resistance of serotype O78 APEC isolates, the predominant serotype, in Korea. Among 16 O78 APEC isolates, 13 isolates carried the genes conferring resistance to ß-lactam (blaTEM), aminoglycoside [aac(3)-II], plasmid-mediated quinolone (qnrA), tetracycline (tetA and tetB), sulfonamide (sul1 and sul2), or chloramphenicol (catA1). Three isolates showed resistance to gentamicin and carried aminoglycoside-modifying enzyme gene, aac(3)-II, simultaneously. Ten O78 APEC isolates showed resistance to nalidixic acid, but only qnrA gene among plasmid-mediated quinolone resistance genes was detected in one isolate. The tetA and tetB genes were also detected in nine and two isolates, respectively. In distribution of phylogenetic groups, four O78 APEC isolates only belonged to group D. But all isolates carried three to five essential virulence genes regardless of phylogenetic groups. Virulence factors and antimicrobial resistance of the most predominant serotype, O78, in chickens tested in this study can be significant role in persistence of APEC in Korea.