Erratum to: Evaluation of adenosine triphosphate testing for on-farm cleanliness monitoring compared to microbiological testing in an empty pig farrowing unit.

[This corrects the article DOI: 10.5187/jast.2020.62.5.682.].

Age-dependent immune response in pigs against foot-and-mouth disease virus in vitro.

Foot-and-mouth disease, one of the most contagious diseases in cloven-hoofed animals, causes significant economic losses. The pathogenesis of foot-and-mouth disease virus (FMDV) infection is known to differ with age of the animals. In this study, we aimed to reveal the difference in immunological response in the initial stage of FMDV infection between piglets and adult pigs. Peripheral blood mononuclear cells (PBMCs) were isolated from 3 piglets (8 weeks old) and 3 pigs (35 weeks old) that were not vaccinated against FMDV. O-type FMDV (2 × 102 median tissue culture infectious dose) was inoculated into porcine PBMCs and the cells were incubated at 37.0°C under 5% CO2 for various time periods (0, 1, 3, 6, 12, 24, and 48 h). The total RNA was obtained from the FMDV-inoculated PBMCs after each time point, and the virus titer was investigated in these RNA samples. Furthermore, dynamics of mRNA expression of the six tested cytokines (interferon [IFN]-α, IFN-γ, interleukin [IL]-6, IL-8, IL-10, and tumor necrosis factor [TNF]-α) in FMDV-inoculated porcine PBMCs were evaluated by time-series analysis to determine the differences, if any, based on the age of the pigs. The PBMCs of piglets contained the highest quantity of FMDV mRNA at 6 hours post-inoculation (hpi), and the PBMCs of pigs had the highest quantity of FMDV mRNA at 3 hpi. The mean cycle threshold-value in the PBMCs steadily decreased after the peak time point in the piglets and pigs (6 and 3 hpi, respectively). The dynamics of mRNA expression of all cytokines except TNF-α showed age-dependent differences in FMDV-inoculated PBMCs. The mRNA expression of most cytokines was more pronounced in the piglets than in the pigs, implying that the immune response against FMDV showed an age-dependent difference in pigs. In conclusion, within 48 hpi, the 8-week-old piglets responded more rapidly and were more sensitive to FMDV infection than the 35-week-old pigs, which could be associated with the difference in the pathogenesis of FMDV infection among the pigs. These results provide valuable insights into the mechanisms underlying the age-dependent differences in immune response in pigs against FMDV infection.

Pathogenicity of an African swine fever virus strain isolated in Vietnam and alternative diagnostic specimens for early detection of viral infection.

BACKGROUND: African swine fever (ASF), caused by the ASF virus (ASFV), was first reported in Vietnam in 2019 and spread rapidly thereafter. Better insights into ASFV characteristics and early detection by surveillance could help control its spread. However, the pathogenicity and methods for early detection of ASFV isolates from Vietnam have not been established. Therefore, we investigated the pathogenicity of ASFV and explored alternative sampling methods for early detection. RESULTS: Ten pigs were intramuscularly inoculated with an ASFV strain from Vietnam (titer, 103.5 HAD50/mL), and their temperature, clinical signs, and virus excretion patterns were recorded. In addition, herd and environmental samples were collected daily. The pigs died 5-8 days-post-inoculation (dpi), and the incubation period was 3.7 ± 0.5 dpi. ASFV genome was first detected in the blood (2.2 ± 0.8) and then in rectal (3.1 ± 0.7), nasal (3.2 ± 0.4), and oral (3.6 ± 0.7 dpi) swab samples. ASFV was detected in oral fluid samples collected using a chewed rope from 3 dpi. The liver showed the highest viral loads, and ear tissue also exhibited high viral loads among 11 tissues obtained from dead pigs. Overall, ASFV from Vietnam was classified as peracute to acute form. The rope-based oral fluid collection method could be useful for early ASFV detection and allows successful ASF surveillance in large pig farms. Furthermore, ear tissue samples might be a simple alternative specimen for diagnosing ASF infection in dead pigs. CONCLUSIONS: Our data provide valuable insights into the characteristics of a typical ASFV strain isolated in Vietnam and suggest an alternative, non-invasive specimen collection strategy for early detection.

Dynamic Changes in Antimicrobial Resistance in Fecal Escherichia coli from Neonatal Dairy Calves: An Individual Follow-Up Study.

The prevalence of antimicrobial-resistant (AMR) Escherichia coli is typically higher in the feces of young dairy calves than in the feces of older cattle; however, the underlying factors contributing to this difference are poorly understood. In this study, AMR fecal E. coli from neonatal calves were characterized both at phenotypic and genotypic levels by individual follow-up sampling. Antimicrobial resistance profiles of E. coli isolates from the maternal colostrum were also determined. Most of the fecal AMR E. coli emerged in the calves at 2-3 days of age. The tetB was the most prevalent resistance gene detected among AMR fecal E. coli from <7-day-old calves, and was also detected in two isolates from the maternal colostrum. Weekly sampling revealed changes in the phenotype of AMR fecal E. coli as the calves aged. More than half of the fecal E. coli isolates acquired additional resistance to beta-lactams by 21-28 days of age, and minimum inhibitory concentrations were higher in ceftiofur-exposed calves than in unexposed calves. Our findings reveal the dynamic changes in AMR fecal E. coli from neonatal calves, and suggest that the feeding of colostrum and ceftiofur administration contribute to the higher prevalence of AMR E. coli in young dairy calves.

Evaluation of adenosine triphosphate testing for on-farm cleanliness monitoring compared to microbiological testing in an empty pig farrowing unit.

Careful cleaning and disinfection of pigpens is essential to prevent disease spread and avoid the resultant economic losses. Hygiene in pigpens is generally evaluated by visual monitoring supplemented with bacteriological monitoring, which includes counting the total aerobic bacteria (TAB) and/or fecal indicator bacteria (FIB). However, these methods present drawbacks such as time and labor requirements. As adenosine triphosphate (ATP) is ubiquitous in all living organisms including microorganisms, this study aimed to directly compare the results of microbial assessment and ATP quantification, and to suggest possible detailed application methods of the ATP test for hygiene evaluation in pigpens of a farrowing unit. Before and after standard cleaning procedures, samples were collected from the floor corner, floor center, and feeding trough of four pigpens at different time points. No FIB were detected and both the TAB and ATP levels were significantly decreased in the floor center area after cleaning. FIB were continuously detected after cleaning and disinfection of the floor corners, and there was no significant ATP level reduction. The feeding trough did not show any significant difference in these values before and after cleaning, indicating insufficient cleaning of this area. The levels of TAB and ATP after cleaning were significantly correlated and the average ATP value was significantly lower in the absence of FIB than in their presence. In the absence of standard references, a more thorough hygiene management could be achieved evenly by supplementing cleaning or disinfection based on the lowest ATP results obtained at the cleanest test site, which in the present study was the floor center. Overall, these results indicate that the on-farm ATP test can be used to determine the cleanliness status, in addition to visual inspection, as an alternative to laboratory culture-based testing for the presence of microorganisms.

Live attenuated duck hepatitis virus vaccine in breeder ducks: Protective efficacy and kinetics of maternally derived antibodies.

Duck viral hepatitis type I is a rapidly spreading infection lethal in young ducklings, caused by the duck hepatitis A virus (DHAV). Vaccination of breeder ducks is a common practice to control DHAV. However, maintaining proper maternal antibody levels in large flocks is difficult. Therefore, a simple vaccination strategy that can induces stable high antibody levels through mass vaccination is desirable. We evaluated a DHAV vaccination strategy for breeder ducks involving oral administration under field conditions, and examined the kinetics of antibody response in the ducks and their progeny. The strategy included a primary intramuscular vaccination, followed by secondary and tertiary oral vaccinations. Five weeks after the primary vaccination, virus-neutralizing antibody titers increased by 8.4 ±â€¯1.3 log2. The titers remained stable at around 9.0 ±â€¯1.1 log2 for up to 36 weeks. None of the progeny died when challenged with virulent DHAV at 1, 7 or 14 days of age. The transfer percentage of antibodies from the breeder ducks to their progeny was 12.8 ±â€¯3.0%. When antibody levels of the progeny were measured from the day of hatching to 20 days of age, the levels steadily declined, reaching a mean titer of 0 log2 at 20 days. The half-life of the maternally derived antibodies against DHAV was 3.4 ±â€¯1.1 days. Our vaccination strategy might be effective in breeder ducks because it can be easily applied and induced strong immunity. Moreover, our results might provide a foundation for the mechanistic study of maternally derived antibodies in passive protection.

Protective efficacy of a bivalent live attenuated vaccine against duck hepatitis A virus types 1 and 3 in ducklings.

Duck hepatitis A virus (DHAV) infection is characterized by an acute, rapidly spreading that affects young ducklings. DHAV-1 or DHAV-3 infection is prevalent, and simultaneous co-infection with both viruses has recently become increasingly frequent in the domestic duck farms. In this study, we developed a bivalent live attenuated vaccine (DHV-HSBP100 and AP-04203P100) for DHAV-1 and DHAV-3 and reported the protective efficacy and safety of the vaccine. At 1-day-old, the ducklings received a bivalent vaccine via intramuscular injection. The immunized ducklings showed effective and rapid protection against virulent DHAV-1 and DHAV-3 at 2 or 3 days post vaccination. Moreover, the ducklings showed a potent humoral immune response that peaked at 3 weeks and were maintained at 6 weeks after vaccination. The bivalent vaccine was safe; ducklings administered 10 doses of bivalent vaccines showed no clinical signs, mortality, gross lesions, and body weight changes compared with those observed in the negative controls. Ducklings vaccinated with a bivalent vaccine were evaluated for tissue tropism and viral replication of vaccine strains. Both bivalent vaccine strains were detected in various organs, and the highest virus replication was detected in the kidneys, among the tested organs. No interference occurred during the replication of both vaccine strains. Thus, these experiments suggest that bivalent vaccines would be useful as a promising and practical strategy for control DHAV outbreaks caused by DHAV-1 and DHAV-3 in duck farms.

Effects of Various LED Light Colors on Growth and Immune Response in Broilers.

We evaluated the effects of different light-emitting diode (LED) colors between blue and green on growth performance and the immune response in broilers. A total of 1,200 1-day-old Ross broilers were divided randomly into six groups and exposed to pure blue (PB), bright blue (BB), sky blue (SB), greenish blue (GB), pure green (PG), or white (W) using LEDs for 6 weeks. Consequently, body weights were higher in chickens reared under PB and GB on day (d) 7 and SB on d 21 than the other groups. Chickens in the PB group on d 42 were the heaviest among the groups, followed by the BB group and were significantly heavier than the W group. Splenocyte proliferation was significantly enhanced in chickens reared under PB followed by BB on d 42 and proliferation of peripheral blood mononuclear cells was significantly enhanced in chickens reared under BB on d 42. In addition, chickens in the BB group showed significantly elevated nitric oxide production on d 42, indicating activation of macrophages. These results suggest that immune function and growth of broilers can be improved at the later stage by rearing under shorter wavelength LEDs such as PB and BB.

Surveillance and characterization of Riemerella anatipestifer from wild birds in South Korea.

We conducted surveillance for Riemerella anatipestifer (RA) in wild birds along the East Asian-Australasian flyway in South Korea. Detected RA were characterized by serotype, antibiotic susceptibility, and sequence analysis of the 16S rRNA gene. We collected 944 wild birds of 34 species from 19 of South Korea's major migratory wild bird habitats between 2011 and 2012. We identified RA by PCR and rRNA gene sequence in 71/102 (69.6%) pharyngeal swabs and 19/944 (2.0%) cloacal swabs of wild birds. Most RA positives (71/75 [95%] pharyngeal and 19/704 [(2.6%] cloacal) were from three duck species (family Anatidae): Mallard Duck (Anas platyrhynchos), Northern Pintail (Anas acuta), and Spot-billed Duck (Anas poecilorhyncha). Thirty-three RA isolates obtained and examined were highly resistant to aminoglycosides: kanamycin (100%), gentamicin (94%), amikacin (91%), neomycin (88%), and streptomycin (82%). Six isolates were identified as serotype 4 by agar gel precipitation. Serotypes 1 and 7, which are known virulent serotypes, were also identified in three isolates from wild duck species.

Antimicrobial susceptibility profiles and molecular typing of Campylobacter jejuni and Campylobacter coli isolates from ducks in South Korea.

Campylobacter is a food-borne zoonotic pathogen that causes human gastroenteritis worldwide. Campylobacter bacteria are commensal in the intestines of many food production animals, including ducks and chickens. The objective of the study was to determine the prevalence of Campylobacter species in domestic ducks, and the agar dilution method was used to determine resistance of the isolates to eight antibiotics. In addition, multilocus sequence typing (MLST) was performed to determine the sequence types (STs) of selected Campylobacter isolates. Between May and September 2012, 58 duck farms were analyzed, and 56 (96.6%) were positive for Campylobacter. Among the isolates, 82.1% were Campylobacter jejuni, 16.1% were C. coli, and one was unidentified by PCR. Of the 46 C. jejuni isolates, 87.0%, 10.9%, and 21.7% were resistant to ciprofloxacin, erythromycin, and azithromycin, respectively. Among the C. coli isolates, all 9 strains were resistant to ampicillin, and 77.8% and 33.3% were resistant to ciprofloxacin and azithromycin, respectively. The majority of the Campylobacter isolates were classified as multidrug resistant. Twenty-eight STs were identified, including 20 STs for C. jejuni and 8 STs for C. coli. The most common clonal complexes in C. jejuni were the ST-21 complex and the ST-45 complex, while the ST-828 complex predominated in C. coli. The majority of isolates were of STs noted in ducks and humans from earlier studies, along with seven STs previously associated only with human disease. These STs overlapped between duck and human isolates, indicating that Campylobacter isolates from ducks should be considered potential sources of human infection.

Prevalence of Salmonella isolates and antimicrobial resistance in poultry meat from South Korea.

Contamination of Salmonella was assessed in duck and chicken meat collected from supermarkets, traditional markets, internet shopping malls, and wholesale markets in Jeonlado, South Korea, in 2013. Salmonella contamination was found in 51.3% of duck meat samples and 3.7% of chicken meat samples. Salmonella contamination of duck meat samples differed by meat type, i.e., 69.8% of samples of whole ducks and 33.9% of samples of duck pieces. Six serotypes were identified from 64 Salmonella isolates in duck meat: Salmonella Typhimurium (37.5%), Salmonella Enteritidis (21.8%), Salmonella Stanley (3.1%), Salmonella Regent (1.6%), Salmonella Winterthur (3.1%), and Salmonella Westhampton (1.6%). All isolates were resistant to one or more antibiotics. Resistance to sulfisoxazole was most common (93.8% of isolates), followed by resistance to nalidixic acid (59.4%), ceftazidime (26.6%), and ampicillin (26.6%). To our knowledge, this study is the first to report Salmonella contamination in duck meat from Korea. Duck meat should be considered an important source of foodborne pathogens.

Prevalence of duck circovirus infection of subclinical pekin ducks in South Korea.

An investigation was carried out to determine the prevalence and infection pattern of duck circovirus (DuCV) in subclinical Pekin ducks on South Korean duck farms. A total of 147 samples collected from 92 duck farms in five provinces were examined from 2011 to 2012. The overall prevalence of DuCV PCR-positive pooled bursa of Fabricius and liver samples was 21.8% (32/147). The prevalence of DuCV PCR-positive samples increased significantly in 3-week-old ducks compared with that in 1-week-old ducks (P<0.05). DuCV in association with Riemerella and Salmonella infections (10.9%; 16/147) occurred at the same level as infection with DuCV alone (10.9%; 16/147). In comparison of the relationship between bacterial diseases (salmonellosis, Riemerella infection) and morbidity in farms with and without DuCV, morbidity was higher in circovirus-positive farms (50%; 16/32) than in circovirus-negative farms (26.1%; 30/115). Our findings provide baseline information on the degree of DuCV infection and distribution and pattern of DuCV in ducks, and it is evident that DuCV can be associated with subclinical diseases and that subclinical infection could be economically important.

Isolation and characterization of a low pathogenic duck hepatitis A virus 3 from South Korea.

The D11-JW-018 strain of duck hepatitis A virus (DHAV) was isolated from 7-day-old ducklings in Kyeonggi province, South Korea with no clinical signs of typical hepatitis. Phylogenetic analyzed of whole genome showed that D11-JW-018 strain was belonged to DHAV-3 genotype. The pathogenicity of the D11-JW-018 strain in 1-, 7-, 14-, and 21-day-old ducklings was examined. Mortality of D11-JW-018 strain was lower than DRL-62 (DHAV-1) age-dependent but incubation period was longer in 1-day-old ducklings. Unlike DRL-62 strain infection, D11-JW-018 strain induced only liver discoloration without hemorrhagic mottling and lymphocyte infiltration and bile duct hyperplasia in histological lesion. The D11-JW-018 strain was detected only in the heart, liver, spleen, gall bladder, pancreas, and kidney among 12 organs in infected 1-day-old ducklings. Serum biochemical analyses revealed a significant difference in aspartate transaminase and alanine transaminase between the D11-JW-018 strain-infected ducklings and those infected with the DRL-62 strain (P<0.05). We identified the D11-JW-018 strain in South Korean ducklings and provide the characteristics of DHAV-3.