Survival and prevalence of Clostridium difficile in manure compost derived from pigs.

Pigs, particularly piglets, have been identified as reservoir hosts of Clostridium difficile. To examine the survival ability of this pathogen in pig feces-based manure compost, C. difficile spores, which were prepared to contain as few vegetative cells as possible, were artificially inoculated into pig feces and incubated at different temperatures. While C. difficile survived in the feces incubated at temperatures below 37 °C for over 30 days, cell numbers gradually decreased at thermophilic temperatures (over 55 °C; p < 0.05). Next, to clarify the prevalence of C. difficile in field manure compost, we isolated and characterized C. difficile from the final products of manure compost products of 14 pig farms. A total of 11 C. difficile strains were isolated from 5 of 14 (36% positive rate) samples tested. Of these 11 strains, 82% were toxigenic, with ribotype 078 being the most prevalent. Thus, the application of composted manure to land therefore poses a possible risk of C. difficile transfer to the food chain.

Serological survey of Peaton virus among wild boars (Sus scrofa) in Ehime Prefecture, Japan.

Peaton virus (PEAV) is a type of arthropod-borne virus similar to Akabane virus (AKAV), belonging to the genus Orthobunyavirus. AKAV infection is common in cattle, but previous reports have suggested that pigs may play a role in transmission cycle. In addition, antibodies against AKAV were detected in wild boars (Sus scrofa). By contrast, PEAV could infect cattle and pigs, but it remains unknown whether PEAV infects wild boars. This study aimed to reveal the possibility of PEAV infection in wild boars by conducting a serological survey. Consequently, the seropositive rate of PEAV was 26.5% in 264 free-living wild boars in Ehime Prefecture, Japan. This is the first study to report the detection of antibodies against PEAV in wild boars.

mcr-1 remains detectable in various Escherichia coli lineages isolated from healthy swine after withdrawal of colistin use on the farm.

We monitored swine-derived Escherichia coli on a Japanese farm where colistin had been used for the treatment of diseases caused by bacteria and investigated colistin resistance and the presence of mcr-1 in 36 E. coli strains isolated before and after the withdrawal of colistin use. Through the withdrawal of colistin use on the farm, the prevalence of colistin-resistant and mcr-1-positive E. coli was markedly reduced but not eradicated because mcr-1 had been maintained in multiple plasmids and various sequence types of nonpathogenic E. coli carried in healthy swine. The monitoring of sequence types of mcr-1-positive E. coli is expected to be important for controlling colistin resistance in swine or other animals.

An abnormal birth in bovine suspected of being caused by Peaton virus first occurred in Shikoku region, Japan.

Peaton virus (PEAV) is a type of arthropod-borne virus (arbovirus) belonging to the genus Orthobunyavirus, much like Akabane virus and Aino virus. These arboviruses cause stillbirth and congenital malformations of fetuses in ruminants. In Japan, abnormal birth in bovine caused by PEAV were reported in Okinawa, Kyushu, and Chugoku regions, but it has never been reported in Shikoku region. The abnormal birth occurred in 2020 in Ehime Prefecture (Shikoku region) and suspected of being caused by PEAV from results of clinical signs, pathological findings, and virus neutralization test using PEAV. However, PEAV was not detected and isolated. This report describes the case of abnormal birth in bovine suspected of being caused by PEAV first occurred in Shikoku region, Japan.

Application of a Photocatalyst as an Inactivator of Bovine Coronavirus.

Bovine coronavirus (BCoV), a major causative pathogen of bovine enteric and respiratory diseases and a zoonotic pathogen transmissible between animals and humans, has led to severe economic losses in numerous countries. BCoV belongs to the genus Betacoronavirus, which is a model of a pathogen that is threatening human health and includes severe acute respiratory syndrome coronavirus (SARS-CoV), SARS-CoV-2, and Middle East respiratory syndrome coronavirus. This study aimed to determine whether photocatalytic material effectively reduces CoVs in the environment. Using the film adhesion method of photocatalytic materials, we assessed its antiviral activity and the effect of visible light irradiation according to methods defined by the International Organization for Standardization. Consequently, photocatalytic material was found to have antiviral activity, reducing the viral loads by 2.7 log TCID50 (tissue culture infective dose 50)/0.1 mL (500 lux), 2.8 log TCID50/0.1 mL (1000 lux), and 2.4 log TCID50/0.1 mL (3000 lux). Hence, this photocatalytic material might be applicable not only to reducing CoVs in the cattle breeding environment but also perhaps in other indoor spaces, such as offices and hospital rooms. To our knowledge, this study is the first to evaluate the antiviral activity of a photocatalytic material against CoV.

Development of Genotype-Specific Anti-Bovine Rotavirus A Immunoglobulin Yolk Based on a Current Molecular Epidemiological Analysis of Bovine Rotaviruses A Collected in Japan during 2017-2020.

Bovine rotavirus A (RVA), a major causative pathogen of diarrhea in dairy and Japanese beef calves, has led to severe economic losses in numerous countries. A dual genotyping system based on genomic segments encoding VP7 (G genotype) and VP4 (P genotype), comprising the outer layer of the virion, has been used to understand the epidemiological dynamics of RVAs at the national and global levels. This study aimed to investigate occurrence frequency of G and P genotypes for multiple bovine RVAs from calf diarrheic samples collected in Japan from 2017 to 2020. After we produced anti-bovine RVA immunoglobulin yolks (IgYs) from hens immunized with the two RVAs with different genotypes (G6P[5] and G10P[11]) selected on the basis of the current epidemiological survey, we investigated cross-reactivity against bovine RVAs with different G and P combinations owing to establish a useful strategy to protect calves from RVA infections using the two IgYs. Consequently, the two produced anti-bovine IgYs showed strong cross-reactivity against bovine RVAs with the same G and/or P genotypes in neutralization assay, respectively. Therefore, our data suggest the possibility of a passive immunization to protect calves from a bovine RVA infections epidemic in Japan via oral administration of the two IgYs into calves. The findings presented herein will provide important information that IgY is one of the effective tools to prevent infections of various pathogens.

Manure Compost Is a Potential Source of Tetracycline-Resistant Escherichia coli and Tetracycline Resistance Genes in Japanese Farms.

Manure compost has been thought of as a potential important route of transmission of antimicrobial-resistant bacteria (ARB) and antimicrobial resistance genes (ARGs) from livestock to humans. To clarify the abundance of ARB and ARGs, ARB and ARGs were quantitatively determined in tetracycline-resistant Escherichia coli (harboring the tetA gene)-spiked feces in simulated composts. In the simulated composts, the concentration of spiked E. coli decreased below the detection limit at day 7. The tetA gene remained in manure compost for 20 days, although the levels of the gene decreased. Next, to clarify the field conditions of manure compost in Japan, the quantities of tetracycline-resistant bacteria, tetracycline resistance genes, and residual tetracyclines were determined using field-manure-matured composts in livestock farms. Tetracycline-resistant bacteria were detected in 54.5% of tested matured compost (6/11 farms). The copy number of the tetA gene and the concentrations of residual tetracyclines in field manure compost were significantly correlated. These results suggest that the use of antimicrobials in livestock constitutes a selective pressure, not only in livestock feces but also in manure compost. The appropriate use of antimicrobials in livestock and treatment of manure compost are important for avoiding the spread of ARB and ARGs.