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Porcine circovirus 2 (PCV2) causes porcine circovirus-associated disease, and co-infection with porcine reproductive and respiratory syndrome virus (PRRSV) severely affects the pig breeding industry. Both viruses target the macrophages in lymphoid tissues. Various porcine pathogens enter via the nasal cavity, and the nasopharynx-associated lymphoid tissue (NALT) acts as the mucosal immune system. However, the pathological analysis has not progressed. This study aimed to histologically examine the NALT of pigs with suspected PCV2 and PRRSV infections. Six pigs were subjected to necropsy, and their NALT, tonsils, and mesenteric lymph nodes were collected. Macrophages, lymphocytic depletion, multinucleated giant cells, intracytoplasmic inclusion bodies, and neutrophil infiltration increased in the NALT. In situ hybridization revealed positive signals for PCV2 in the NALT of all pigs and PRRSV in the NALT of three pigs. PCV2-positive macrophages were mainly identified in the follicles, whereas PRRSV-positive tissues were found primarily around the crypt and directly below the epithelium. Quantitative PCR revealed 10

When a large-scale epidemic of porcine epidemic diarrhea (PED) occurred in 2013 in Japan, feedback feeding (feeding feces and gut tissues of infected piglets) was attempted to impart immunity to sows and immunize nursing piglets via breastfeeding. This study evaluated the effect of feedback feeding on PED control at 172 farms in Kagoshima and Miyazaki Prefectures. Univariable and multivariable generalized linear models were used to analyze the associations between conduct of feedback feeding and damage from the outbreak (outbreak period and the number of piglet deaths) at the farm level. The within-farm outbreak period shortened over time after the regional outbreak began on Kyushu Island (P=0.009) and was longer on large-scale farms (mean 66.0 days, P=0.003) than small-scale farms (29.4 days) and on farms that used feedback feeding (145.2 days, P=0.059) than those that did not (66.0 days). The number of dead piglets decreased over time since the first regional case (P

In Japan, reducing the use of antimicrobials in pig production is a significant problem. However, there are no published reports concerning porcine disease treatment, as related to the age of the pigs and the indications (e.g., organ system) in Japan. In this study, we analyzed the prescription records of 17 farrow-to-finish farms from 2014 to 2018 in southern Kyushu, Japan. The farms' antimicrobial usage was calculated as the active ingredient per population correction unit (PCU) or the number of treated pigs per PCU using the defined daily dose. All data were analyzed according to the indications and production stages (sows, suckling pigs, post-weaning pigs, and fattening pigs). In terms of active ingredients/PCU, tetracyclines were the most commonly used (43.2-59.3%), and the largest amounts of antimicrobials administered through feeds were for treating the respiratory organs of fattening pigs. In terms of the number of treated pigs/PCU, tetracyclines were most frequently used (16.3-31.1%), and a high frequency of antimicrobials administered through feeds was used for the treatment of respiratory organs in post-weaning pigs. In this study, it was confirmed that tetracyclines were used frequently as a herd treatment for respiratory diseases in post-weaning and fattening pigs in southern Kyushu, Japan. The findings suggest that it is necessary to improve the treatment and prevention of respiratory diseases in post-weaning and fattening pigs in order to reduce the frequencies of antimicrobial treatments.

In Japan, many pig producers buy antimicrobials using veterinarians' prescriptions. Even if veterinarians prescribe antimicrobials for treatments and metaphylaxis, producers may be using them for prophylaxis on farms with a high incidence of chronic diseases. To reduce the frequency of antimicrobial use, we applied the concept of medication adherence and launched an intervention in eight farrow-to-finish farms. For the intervention farms, we attempted to implement measures for preventing the repeated onset of chronic infections that interfered with adherence, confirmed the need to continue medications, and discontinued medications if possible. For each farm, these interventions began at different time points after 2017 and continued until 2020. We evaluated the effectiveness of this intervention based on the amount of active ingredients/population correction unit and the total number of defined daily doses for the weight group/slaughter pig. The ratio of antimicrobial use for both indicators in the year 2020 compared with the start of the intervention decreased on all farms. Among the eight non-intervention farms, six showed an increase in both indicators. These results suggest that interventions for chronic diseases administered by producers and veterinarians lead to high adherence to prescriptions and responsible and prudent use of antimicrobials on pig farms.

In this study, we investigated the occurrence of antimicrobial resistance in commensal Escherichia coli isolated from healthy Thoroughbred (TB) racehorses in Japan. A total of 212 fecal samples were individually collected from TB racehorses from March 2017 to August 2018 at Japan Racing Association training centers. E. coli was isolated by using selective agar media, deoxycholate-hydrogen sulfide-lactose (DHL) and eosin methylene blue (EMB). A total of 417 E. coli isolates were examined against 10 antimicrobial agents by using the broth microdilution method. The 417 E. coli isolates were phylogenetically grouped using a multiplex polymerase chain reaction. The highest proportion of resistance was observed for streptomycin (30.9%, 129/417) followed by ampicillin (19.4%, 81/417), trimethoprim (15.8%, 66/417), tetracycline (8.4%, 35/417), chloramphenicol (2.6%, 11/417), kanamycin (1.2%, 5/417), nalidixic acid (0.5%, 2/417), cefazolin (0.2%, 1/417), colistin (0.2%, 1/417), and gentamycin (0%). Multidrug-resistant (MDR) E. coli was detected in 7.9% (33/417) of isolates. The proportions of resistance against ampicillin, streptomycin, kanamycin, and chloramphenicol and of multidrug-resistant phenotypes in E. coli belonging to phylogenetic group B2 were significantly higher than those of other groups. This study clarified the distribution of antimicrobial-resistant (AMR) E. coli in Japanese racehorses. A continuous monitoring program for antimicrobial resistance is required to control the spread of AMR bacteria in racehorses.

Extended-spectrum β-lactamase-producing Escherichia coli (ESBLEC) have become a major health concern in both human and veterinary medicine. These bacteria could become a critical problem in equine medicine due to the limited number of antimicrobial drugs available. However, there are no previous reports of ESBLEC isolated from horses in Japan. The objectives of this study were to investigate the occurrence of ESBLEC isolated from feces in healthy Thoroughbred racehorses in Japan. Feces samples were collected from 147 healthy Thoroughbred racehorses by equine veterinarians at the Japan Racing Association (103 from Miho Training Center and 44 from Ritto Training Center) between March 2017 and April 2018. Samples were screened for ESBLECs using MacConkey agar supplemented with 1 µg/ml cefotaxime. Detection of ESBL genes was performed by PCR and confirmed by DNA sequencing. Horizontal transmission was demonstrated by conjugation assay. In this study, 24 ESBLECs were isolated from twelve horse feces samples (8.2%). All ESBLECs harbored blaCTX-M-2, and both blaTEM-1 and blaCTX-M-2 were detected in nine isolates (37.5%). ESBLECs showed resistance to all β-lactam antibiotics (100%) tested, followed by trimethoprim (66.7%), streptomycin (62.5%), tetracycline (25.0%), and oxytetracycline (25.0%). Horizontal transmission was successfully demonstrated by conjugation assay in eight of 13 isolates, and blaCTX-M-2 was detected by PCR in all transconjugants. This study showed that racehorses in Japan are potential reservoirs of ESBLECs.

Equine proliferative enteropathy (EPE) caused by Lawsonia intracellularis is characterized by hypoproteinemia. There are currently no reliable reports that provide a reference value for the total serum protein (TP) concentration to clinically diagnose EPE. The objective of this study was to statistically determine the reference value. Feces and sera of 99 foals with EPE-like clinical signs and of 35 healthy foals were obtained. The samples were used for specific-gene detection of L. intracellularis, TP measurement, and specific-antibody detection against L. intracellularis. Based on these results, the optimal reference value for the TP concentration as a clinical diagnostic index of EPE was found to be ≤ 4.8 g/dl. This clinical diagnostic index will provide an effective approach for diagnosing EPE.

We examined the effects of Eimeria pragensis infection on intestinal peristalsis, goblet cell proliferation and intestinal flora in C57BL/6 mice. Intestinal peristalsis was evaluated by radiography using barium at 7 days post-infection (p.i.). The intestinal peristalsis of E. pragensis-infected mice was significantly suppressed compared with uninfected control mice. Twenty-three mice were divided into 5 groups of 4 or 5 mice each; 2 groups of mice were infected with E. pragensis and the others were kept uninfected. At 7 days p.i., E. pragensis-infected and -uninfected mice were sacrificed to examine goblet cell numbers in the intestines, and significant decreases were observed only in the infected mice. Shiga toxin-producing Escherichia coli (STEC) O157:H7 was inoculated orally in mice both infected and uninfected with E. pragensis at 7 days p.i., with the remaining mice used as uninoculated controls. When mice were sacrificed at 2 days after STEC inoculation, STEC was only detected in the intestines of E. pragensis-infected mice. Colonization of STEC was also confirmed by immunohistochemistry on the surface of epithelial cells in concurrently infected/inoculated mice. Also, an overgrowth of residential E. coli was observed only in E. pragensis-infected mice. These results suggest that E. pragensis induces the suppression of intestinal peristalsis and modifies the intestinal environment to facilitate artificially introduced STEC colonization and multiplication, in addition to residential E. coli overgrowth.

Porcine epidemic diarrhea (PED) occurred in Japan in 2013 after an interval of 7 years. The present study assessed individual productivity of sows exposed to PED virus at different periods of the production stage. The present study was performed at three commercial farms that had PED outbreak during the month of December 2013. Herd immunization was conducted for all gilts and sows. The production records were obtained for sows that were alive during the PED outbreak at each farm. The sows were categorized into six groups based on the period in which they were exposed to PED virus between days 0–30 (G1), 31–60 (G2), 61–90 (G3), or after 91 days of pregnancy (G4), during lactation (L), and after weaning (W). The control group was assigned based on the records before the period of PED outbreak (uninfected group). The number of sow’s records obtained from farms A, B, and C were 1056, 1137, and 1035, respectively. Compared with the uninfected group, there was no reduction in the number of pigs born alive in the G1–G4 groups. Sows of the G4 and L groups, however, had 4–9 pigs fewer pigs weaned, and a 36%–77% greater pre-weaning mortality than the uninfected group (P

Summary Porcine epidemic diarrhoea virus (PEDV) is the aetiologic agent of porcine epidemic diarrhoea (PED), a highly contagious enteric disease that is threatening the swine industry globally. Since PED was first reported in Southern Vietnam in 2009, the disease has spread throughout the country and caused substantial economic losses. To identify PEDVs responsible for the recent outbreaks, the full‐length spike (S) gene of 25 field PEDV strains collected from seven northern provinces of Vietnam was sequenced and analysed. The sequence analysis revealed that the S genes of Vietnamese PEDVs were heterogeneous and classified into four genotypes, namely North America and Asian non‐S INDEL, Asian non‐S INDEL, new S INDEL and classical S INDEL. This study reported the pre‐existence of US‐like PEDV strains in Vietnam. Thirteen Vietnamese variants had a truncated S protein that was 261 amino acids shorter than the normal protein. We also detected one novel variant with an 8‐amino acid insertion located in the receptor‐binding region for porcine aminopeptidase N. Compared to the commercial vaccine strains, the emerging Vietnamese strains were genetically distant and had various amino acid differences in epitope regions and N‐glycosylation sites in the S protein. The development of novel vaccines based on the emerging Vietnamese strains may be contributive to the control of the current PED outbreaks.

Extended-spectrum &beta, lactamase (ESBL)- and AmpC &beta, lactamase (AmpC)-producing Klebsiella spp. have become a major health problem, leading to treatment failure in humans and animals. This study aimed to evaluate the presence of ESBL/AmpC-producing Klebsiella spp. isolated from racehorses in Japan. Feces samples from 212 healthy Thoroughbred racehorses were collected from the Japan Racing Association Training Centers between March 2017 and August 2018. ESBL/AmpC-producing Klebsiella spp. were isolated using selective medium containing 1 &micro, g/mL cefotaxime. All isolates were subjected to bacterial species identification (MALDI-TOF MS), antimicrobial susceptibility test (disk diffusion test), characterization of resistance genes (PCR), conjugation assay, and genetic relatedness (multilocus sequence typing/MLST). Twelve ESBL/AmpC-producing Klebsiella pneumoniae (ESBL/AmpC-KP) were isolated from 3.3% of horse samples. Antimicrobial resistance profiling for 17 antimicrobials showed all ESBL/AmpC-KP were multidrug-resistant (MDR). Only 1 isolate was confirmed as an ESBL producer (blaCTX-M-2-positive), whereas the other 11 isolates were plasmid-mediated AmpC (pAmpC) producers (blaCMY positive). On the basis of MLST analysis, the ESBL-KP isolate was identified as sequence type (ST)-133 and four different STs among AmpC-KP isolates, ST-145, ST-4830, ST-4831, and ST-4832, were found to share six of the seven loci constituting a single-locus variant. This is the first study to show K. pneumoniae carrying MDR pAmpC isolated from a racehorse.

The objective of the present study was to assess the productivity of pigs to investigate the time-dependent change in productivity and compare productivity according to breeding company group and porcine reproductive and respiratory syndrome (PRRS) status on commercial swine farms in southern Kyushu, Japan. Data used in the present study were 245 annual productivity records from 2014 to 2018 obtained from 72 farms. Average sow inventory was 481.2 sows. Mean numbers of pigs weaned per sow per year and market pigs sold per sow per year were 23.7 and 21.6, respectively. Pigs born alive per litter increased from 11.1 to 11.9 pigs from 2014 to 2018 (p < .05). Farms using domestic breeding companies had similar numbers in 2016 to those using international breeding companies, but fewer pigs in 2014, 2015, 2017, and 2018 (p < .05). Farms with an unknown or unstable PRRS status had fewer pigs born alive per litter and pigs weaned per sow per year than those with stable or negative PRRS statuses (11.2 ± 0.06 vs. 11.8 ± 0.08 pigs and 22.6 ± 0.38 vs. 25.0 ± 0.22 pigs, respectively; p < .05). These results can be used to establish feasible targets and standards of performance to identify problem areas and improve production.

The objectives of this study were to assess abortion risk (AR) and the number of piglets that died during suckling periods per litter (DP) in farms infected with porcine epidemic diarrhea (PED) in relation to herd immunization procedures. Data were obtained from 91 farms in Japan that had PED infection during 2013 to 2014. The 91 PED-positive farms were asked the number of abortions that occurred and DP for 3 months (1 month before PED outbreak (previous month), 1 month after PED outbreak (the month of PED), and from 1 month after PED outbreak to 2 months after PED outbreak (following month)). AR in each month was calculated as the number of abortions divided by sow inventory. Both AR and DP in the month of PED were higher than those in the previous and following months (p

Extended-spectrum β-lactamase-producing Escherichia coli (ESBLEC) have become a major health concern in both human and veterinary medicine. These bacteria could become a critical problem in equine medicine due to the limited number of antimicrobial drugs available. However, there are no previous reports of ESBLEC isolated from horses in Japan. The objectives of this study were to investigate the occurrence of ESBLEC isolated from feces in healthy Thoroughbred racehorses in Japan. Feces samples were collected from 147 healthy Thoroughbred racehorses by equine veterinarians at the Japan Racing Association (103 from Miho Training Center and 44 from Ritto Training Center) between March 2017 and April 2018. Samples were screened for ESBLECs using MacConkey agar supplemented with 1 µg/ml cefotaxime. Detection of ESBL genes was performed by PCR and confirmed by DNA sequencing. Horizontal transmission was demonstrated by conjugation assay. In this study, 24 ESBLECs were isolated from twelve horse feces samples (8.2%). All ESBLECs harbored blaCTX-M-2, and both blaTEM-1 and blaCTX-M-2 were detected in nine isolates (37.5%). ESBLECs showed resistance to all β-lactam antibiotics (100%) tested, followed by trimethoprim (66.7%), streptomycin (62.5%), tetracycline (25.0%), and oxytetracycline (25.0%). Horizontal transmission was successfully demonstrated by conjugation assay in eight of 13 isolates, and blaCTX-M-2 was detected by PCR in all transconjugants. This study showed that racehorses in Japan are potential reservoirs of ESBLECs.

In this study, the occurrence of antimicrobial-resistant (AMR) enterococci was evaluated in Thoroughbred (TB) racehorses in Japan. Fecal samples were collected from 212 healthy TB racehorses at the Miho and Ritto Training Centers of the Japan Racing Association from March 2017 to August 2018. Isolation and identification were performed by enterococcus selective medium and confirmed to the species using MALDI-TOF MS. Enterococcus faecium and E. faecalis isolates were subjected to antimicrobial susceptibility test against 11 antimicrobials by minimum inhibitory concentration based on recommendation from Clinical and Laboratory Standards Institute guidelines. Among 583 enterococcus isolates, E. faecium and E. faecalis were identified for 48.2% (281/583) and 7.4% (43/583), respectively. One isolate that was representing E. faecium (153 isolates) and E. faecalis (31 isolates) from each sample was selected for antimicrobial susceptibility test. The highest rate of resistance for E. faecium isolates was observed against enrofloxacin (57.5%; 88/153), followed by streptomycin (32.0%; 49/153), kanamycin (18.3%; 28/153), gentamycin (5.9%; 9/153), erythromycin (5.9%; 9/153), and oxytetracycline (4.6%; 7/153). For E. faecium isolates, the highest resistance was observed against streptomycin (90.3%; 28/31), followed by kanamycin (41.9%; 13/31), gentamycin (29.0%; 9/31), lincomycin (9.7%; 3/31), oxytetracycline (6.5%; 2/31), erythromycin (6.5%; 2/31), tylosin (6.5%; 2/31), enrofloxacin (6.5%; 2/31), and chloramphenicol (3.2%; 1/31). The results indicated that enrofloxacin and aminoglycosides were highly resistant among tested antimicrobials. Continuous monitoring studies are useful to increase the awareness of the potential for AMR bacteria to arise from imprudent use of antimicrobials in TB racehorses in Japan.

Porcine epidemic diarrhea (PED) is an emerging and/or re-emerging disease of pigs in several countries, with high morbidity and mortality in suckling piglets. Farms affected with PED perform various intervention practices to control and/or eliminate the PED virus. The objectives of the present study were to assess the effect of biosecurity measures and intervention practices to control PED on time to absence of clinical signs (TAC) and number of dead suckling piglets during TAC. A questionnaire was administered to 120-PED affected farms located across Japan between 2013, when the first case was reported in Japan, and 2014. Farms were asked to provide information on farm characteristics and internal or external biosecurity measures during PED outbreak, as well as on intervention practices to control PED. The TAC was defined as the number of days from the date that clinical PED signs appeared to the date that clinical PED signs disappeared. The number of dead piglets per sow (DP/S) was calculated as the number of dead suckling piglets during TAC divided by the sow inventory. Regarding the effect of biosecurity measures during PED outbreak on TAC and DP/S, longer TAC was observed in Actinobacillus pleuropneumoniae-positive farms and farms outsourcing pig transport to the slaughterhouse (p

Summary Mycoplasma bovis is a microorganism associated with pneumonia, mastitis, arthritis and otitis media of cattle; however, there are no reports of this organism causing bovine endocarditis. Five adult cattle with endocarditis characterized by caseated lesions (diameter 5–12 cm) of the endocardial surface of the left atrium, but without lesions in heart valves or affecting the right side of the heart, were identified in slaughterhouses in Japan. M. bovis was successfully isolated from the lesions and M. bovis antigen was detected immunohistochemically within the lesions. The results suggest that the lesions may have been associated with M. bovis alone. To our knowledge, this is the first demonstration of bovine endocarditis associated with M. bovis.

We improved a loop-mediated isothermal amplification (LAMP) assay permitting sensitive and rapid Mycoplasma bovis detection. A total of 55 bacterial strains were examined in this study, including 33 M. bovis strains, 14 non-M. bovis mycoplasmas and eight non-mycoplasma bacterial strains. M. bovis was successfully detected by the LAMP assay within 60 min without cross-reaction to any other bacteria. Furthermore, a total of 135 nasal swab samples were tested directly using our LAMP assays, the previously reported LAMP assay, conventional PCR assay without pre-culture and comparing standard culture methods. The improved LAMP assay showed sensitivity and specificity of 97.2% and 90.9%, respectively (with a kappa coefficient of 0.8231), and the sensitivity of our revised LAMP assay was increased compared to existing methods.

We developed a multiplex polymerase chain reaction (PCR) method using primers specific to the capsular polysaccharide biosynthesis genes (cps) to type Actinobacillus pleuropneumoniae serovars 1, 2, 5, 7, and 15. This multiplex PCR method may be useful for the typing of serovar 15, which is recently becoming more prevalent, as well as the most prevalent serovars 1, 2, 5, and 7 in Japan. Discipline: Animal health Additional key words: pig, pleuropneumonia, serotyping * Corresponding author: e-mail itohiroy@affrc.go.jp Received 7 October 2014; accepted 24 December 2014. Introduction Actinobacillus pleuropneumoniae is an etiologic agent of porcine pleuropneumonia that causes serious economic losses in the pig-rearing industry (Gottschalk 2012). Fifteen serovars of A. pleuropneumoniae have been identified to date, mainly based on the antigenic diversity of their capsular polysaccharides (Blackall et al. 2002, Perry et al. 1990). The prevalent local serovars vary among countries (Gottschalk 2012); in Japan, serovars 2 (59%), 1 (28%), 5 (8%), and 7 (3%) account for approximately 98% of the strains isolated from pigs (Asawa et al. 1995, Fukuyasu et al. 1991, 1996, Kume & Nakai 1988, Morioka et al. 2006, 2008, Suzuki et al. 1989, Yoshimura et al. 2002). However, the incidence of isolated cases of serovar 15, which is the most recently identified serovar (Blackall et al. 2002), has been recently increasing in Japan (Ito 2013, Morioka et al. 2008). Because the virulence of strains differs depending on their serovars and differences in serovars affect the effectiveness of vaccines (Gottschalk 2012), serotyping is widely performed in veterinary diagnostic laboratories. However, cross-reactions are often observed among different serovars, for example, among serovars 1, 9, and 11 (Gottschalk 2012), serovars 3, 6, 8, and 15 (Gottschalk 2012), serovars 4 and 7 (Gottschalk 2012), and serovars 7 and 15 (Blackall et al. 2002, Koyama et al. 2007), preventing the accurate and rapid typing of field strains. In the last decade, serovar-specific polymerase chain reaction (PCR) typing methods based on the sequence variation in genes involved in capsular polysaccharide biosynthesis (cps) have been developed for serovars 1-3, 5-8, 10, 12, and 15 (Angen et al. 2008, Bosse et al. 2014, Ito 2010, Jessing et al. 2003, Lo et al. 1998, Schuchert et al. 2004, Turni et al. 2014, Zhou et al. 2008). These PCR typing methods facilitate the reliable typing of A. pleuropneumoniae serovars without cross-reactions and mistyping. In the present study, we developed a multiplex PCR method for cps typing of A. pleuropneumoniae serovars 1, 2, 5, and 7, which are the most prevalent in Japan (Asawa et al. 1995, Fukuyasu et al. 1991, 1996, Kume & Nakai 1988, Morioka et al. 2006, 2008, Suzuki et al. 1989, Yoshimura et al. 2002), as well as serovar 15, isolated cases of which have been increasing (Ito 2013, Morioka et al. 2008). Materials and methods We used the A. pleuropneumoniae serovar reference strains serovar 1, 4074; serovar 2, S1536; serovar 3, S1421; serovar 4, M62; serovar 5a, K17; serovar 6, Femo, serovar 7, WF83, serovar 8, 405; serovar 9; CVJ13261; serovar 10, D13039; serovar 11; 56153; serovar 12, 8329; serovar 13, N273; serovar 14, 4906; and serovar 15, HS143, to determine suitable conditions for multiplex PCR. Seventy

Nucleotide sequence determination and analysis of the cps gene involved in the capsular polysaccharide biosynthesis of Actinobacillus pleuropneumoniae serotype 15 revealed the presence of three open reading frames, designated as cps15ABC genes. At the protein level, Cps15A and Cps15B showed considerably high homology to CpsA (67.0 to 68.7%) and CpsB (31.7 to 36.8%), respectively, of A. pleuropneumoniae serotypes 1, 4 and 12, revealing the common genetic organization of the cps among serotypes 1, 4, 12 and 15. However, Cps15C showed no homology to any proteins of A. pleuropneumoniae serotypes, indicating that cps15C may be specific to serotype 15. This study will provide the basic molecular knowledge necessary for the development of diagnostics and a vaccine for A. pleuropneumoniae serotype 15.

Porcine epidemic diarrhea (PED) occurred in Japan in 2013 after an interval of 7 years. The present study assessed individual productivity of sows exposed to PED virus at different periods of the production stage. The present study was performed at three commercial farms that had PED outbreak during the month of December 2013. Herd immunization was conducted for all gilts and sows. The production records were obtained for sows that were alive during the PED outbreak at each farm. The sows were categorized into six groups based on the period in which they were exposed to PED virus between days 0-30 (G1), 31-60 (G2), 61-90 (G3), or after 91 days of pregnancy (G4), during lactation (L), and after weaning (W). The control group was assigned based on the records before the period of PED outbreak (uninfected group). The number of sow's records obtained from farms A, B, and C were 1056, 1137, and 1035, respectively. Compared with the uninfected group, there was no reduction in the number of pigs born alive in the G1-G4 groups. Sows of the G4 and L groups, however, had 4-9 pigs fewer pigs weaned, and a 36%-77% greater pre-weaning mortality than the uninfected group (P 0.05). There was no difference in farrowing rate and number of pigs born alive at subsequent parities among the sow groups. There were no interactions between sow groups and parity for sow productivity.

We investigated the associations of hind limb claw lesions with claw measures and reproductive performance of sows on a commercial breeding farm. We used a five-point scale (score of 0-4) for each hind limb claw region to generate a claw lesion score (CLS). Of 110 sows, 1.8% had no hind claw lesions, and the remaining 98.2% had at least one. The proportions of sows with a CLS of 2 and 3 were 49.1% and 3.6%, respectively. The junction region had the highest frequency of a CLS of ≥ 2 (P < 0.05). The mean toe length was 45.0 ± 0.4 to 46.4 ± 0.4 mm. Sows with the highest CLS of ≥ 2 in the lateral claw had longer sole dimensions than did sows with the highest CLS of ≤ 1 (P < 0.05). There were no significant differences in claw width or height between sows with the highest CLS of ≥ 2 and ≤ 1. There was also no difference in reproductive performance between sows with the highest CLS of ≥ 2 and ≤ 1. In conclusion, most sows had hind claw lesions. The association between CLS and claw measures indicates that the sole length can be used as an indicator of claw lesions.

1. The present study determined descriptive values of the main production measurements of flocks and assessed the relationship between these measurements and related management factors in Japanese commercial broiler farms. 2. The data set included 5060 flock records from 183 farms. The production index was calculated as follows: liveability × average daily gain/feed conversion ratio × 10. Management factors included in the analysis were broiler breeder age, the time interval between successive flocks, the season of placement and stocking density. 3. The mean (±SD) production index was 283.9 ± 28.83. Management factors significantly associated with a decreased production index were low broiler breeder age, flocks placed in summer and high stocking density (P < 0.05). 4. In regard to an interaction for the production index, flocks with high stocking density had a lower production index than those with low stocking density in flocks with a low broiler breeder age (P < 0.05). In summer, flocks with a short time interval between successive flocks had a lower production index than those with an intermediate or long time interval (P < 0.05). 5. The present study identified factors related to flock performance. The knowledge obtained from this analysis will contribute to improve flock performance by optimising management.

We investigated the reproductive and lifetime performances of Kagoshima Berkshire gilts and sows. We examined 20 605 parity records of 4419 pigs for 2008 to 2012 on a farrow-to-finish commercial farm. The mean parity (± SD) of all animals was 3.0 ± 2.1. For farrowing performance, the highest numbers of total pigs born and pigs born alive were found in sows with parities 5 and 6 and with parity 3-6, respectively (P < 0.05). Regarding weaning and mating performance, sows with parity 2 had the lowest preweaning mortality (P < 0.05). The longest weaning-to-first-mating interval was found in parity 1 pigs, and the interval decreased as parity increased (P < 0.05). Parities 0 and 1 pigs had the lowest farrowing rate and those with parity 4 had the highest farrowing rate (P < 0.05). The mean parity at culling, total number of pigs born alive in a lifetime, and nonproductive days in a lifetime were 5.5 ± 2.93, 49.2 ± 24.72 pigs, and 132.1 ± 83.34 days, respectively. These animals had a lower litter size and fertility that the F1 crossbred sows mainly used in Japan, but a similar tendency for performance by parity.

Porcine epidemic diarrhea (PED) is an emerging disease of pigs in several countries. In the present study, individual sow productivity of Berkshire sows exposed to PED virus at different stages of production was compared. On a commercial farrow-to-finish farm in Kagoshima Prefecture, Japan, the clinical presence of PED was observed in the farrowing barn on January 6, 2014, and all gilts and sows were immunized on January 9, except those in the farrowing barn. The sows were categorized into six groups based on the period in which they were exposed to PED virus: between days 0–30 (G1), 31–60 (G2), 61–90 (G3), or after 91 days of pregnancy (G4), during lactation (L), and after weaning (W). The control group was not exposed to PED during the period of PED outbreak. The study was based on 574 production records. The sows of the G4 and L groups had the fewest piglets weaned (4.8 ± 0.4, and 4.0 ± 0.3 pigs, respectively; P

Porcine epidemic diarrhea (PED) was detected for the first time in seven years in Japan in October 2013 in Okinawa Prefecture. By December 2013, PED had spread into Miyazaki and Kagoshima Prefectures in the southern part of Kyushu, one of the regions with the highest farm density in the country. The objective of the study here was to assess the spatial dynamics of PED spread during the 8 months of the epidemic in the southern part of Kyushu between December 2013, the month observed first case in the studied region, and July 2014. Information on location and capacity of all farms in the prefectures (n=1269) was obtained from a government database containing demographic information for livestock producers. Additionally, data on PED detection (positive or negative) was obtained from the regional Livestock Hygiene Service Center. The Cuzick-Edwards (CE) test, the Knox test, the directional test, and the permutation model of the scan statistic were used to assess the spatio-temporal distribution of the epidemic. PED cumulative farm level incidence was 19.5% (248/1269) through the study period. The highest density of positive farms was observed in the most farm-populated areas of the prefecture. The CE test revealed an extensive degree of spatial clustering, with clustering of positive sites being significant (P0.01) up to the 35th level of neighborhood (approximately 5km in the studied data). The observed-to-expected ratio of cases was maximized at short spatio-temporal distances, with values of the observed-to-expected ratio of cases maximized when the thresholds were set at 2km and 10days, respectively. A significant (P0.01) direction of spread was detected towards the northeastern direction. The permutation model detected five significant (P0.01) clusters occurring at different stages of the epidemic wave. The strong spatio-temporal clustering of PED-infected farms during the first 6 months of the epidemic in the southern part of Kyushu is consistent with results obtained elsewhere and demonstrates the rapid spread of the virus in naïve populations.

The objective of this study was to investigate factors that caused rapid spread during the early phase of the porcine epidemic diarrhea (PED) epidemic in Japan in 2013 and 2014. Anonymized datasets from all pig farms were provided by Kagoshima (709 farms) and Miyazaki Prefectures (506 farms). Semi-parametric survival analysis was conducted using the first 180 days from the first case on December 3, 2013 in Kagoshima Prefecture. To compare the hazard between different farm management types, univariable survival analysis was conducted. As farm sizes varied among different farm types, bivariable survival analysis was conducted for farm size categories and farm density per km 2 for each management type. A case-control study using a postal questionnaire survey was conducted in September 2014, and risk factor analysis was performed using generalized linear models with binomial errors. The hazard was significantly higher in farrow-to-finish farms than fattening farms [hazard ratio (HR) = 1.6, p 0.01], but was not significantly different between reproduction and fattening farms (HR = 1.3, p = 0.16). In separate bivariable survival analyses for each farm type, large- and middle-scale farms had higher hazard than small-scale farms in fattening (HR = 5.8 and 2.6, respectively, both p 0.01) and reproduction farms (HR = 4.0 and 3.6, respectively, both p 0.01). In farrow-to-finish farms, large-scale farms had higher hazard than small-scale farms (HR = 2.8, p 0.01), and higher farm density per km 2 was also a risk factor (HR = 7.6, p 0.01). In the case-control study, questionnaires were returned from 78 PED virus-infected and 91 non-infected farms. The overall response rate was 34%. Risk factors of the final model were occurrence of porcine reproductive and respiratory syndrome in the past 5 years [odds ratio (OR) = 1.97, 95% confidence interval (CI): 0.97–4.00, p = 0.054], use of a common compost station (OR = 2.51, 95%CI: 1.08–5.83, p = 0.03), and use of a pig excrement disposal service (OR = 2.64, 95%CI: 1.05–6.63, p = 0.04). High hazard in farrow-to-finish farms suggested transmission from slaughterhouses to susceptible suckling piglets. Hazard associated with large-scale farms and high density might be due to frequent vehicle entrance and transmission by roads. Improvement of farm hygiene management and avoidance of risky practices associated with contact with pig excrement were keys in preventing invasion of PED virus to a farm.

Although fibrin sealant (FS) has an advantage of high biocompatibility, its adhesive force and sealing effect have been generally considered to be inadequate. In the present study, a high adhesive force and sealing effect were obtained by first rubbing fibrinogen solution into the target tissue, attaching polyglycolic acid (PGA) felt to the treated area, and finally spraying it with FS. This method was compared with three conventional FS application methods and a method using fibrin glue-coated collagen fleece. The adhesive force resulting from the present method was 12 times higher than that for the sequential application method, 4.5 times higher than the spray method, 2.5 times higher than the rubbing and spray method, and 2.2 times higher than the use of fibrin glue-coated collagen fleece. The high adhesive force of FS with PGA felt seemed to be due the high fibrin content of the fibrin gel (FG). Light and electron microscopic observations suggested that the formation of FG in closer contact with the muscle fibers was a factor contributing to this superior adhesive force. Comparison of the sealing effect of the present method with other methods using various biomaterials in combination with FS showed that the sealing effect of FS with PGA felt was 1.4 times higher that of polyglactin 910, 1.8 times that of polytetrafluoroethylene, and 6.7 times that of oxidized regenerated cellulose.

During the 1998 to 2003 period, cecal contents of 4,024 broiler chickens from 252 flocks raised in 63 holdings were examined for Salmonella. The aims were to establish the actual status of the infection, its temporal distribution, prevalent serotype, and common genotype among broiler flocks brought at the slaughterhouse. Collected samples were preenriched in Hajna tetrathionate broth, and after 24 h of incubation, 10 μL of the broth was streaked on selective Rambach agar plate. Suspected scarlet color colonies of Salmonella were cloned on nutrient agar, confirmed through biochemical tests and sero-typed using O and H antigens. Pulsed field gel electrophoresis technique generated DNA fragments banding patterns and established their clonal relatedness. Salmonella was isolated from 563 (14%) samples in 179 (71%) flocks. The flock situation varied from Salmonella-negative holdings (n = 9), positive-flocks from persistently infected holdings (n = 21), and holdings (n = 19) that showed fluctuations with alternating negative and positive flocks for variable time periods. Fourteen holdings (negative, n = 5 and positive, n = 9) were sampled once throughout the study period. Seasonality component was not observed, and salmonellae were found colonizing broiler ceca in warm and cold months. Predominant serovar was Salmonella Infantis (93.3%; n = 525). Macrorestriction fingerprints of Salmonella Infantis using Xba I presumed the isolates to be derived from a common parent. Enhanced discrimination by Bln I digestion produced 3 banding patterns that were closely related genetically and hence epidemiologically related. Such epidemiological information may enable producers to formulate effective control action plan tailored for individual holdings with special emphasis on biosecurity, hygiene, and pest control.

We examined the usefulness of PCR-based restriction fragment length polymorphism (PCR-RFLP) and species-specific PCR combined with a newly devised rapid biochemical test using microplates for identifying weakly beta-hemolytic intestinal spirochetes (WBHIS) isolated from pigs. WBHIS strains showing atypical biochemical characteristics were decisively identified at the species level by PCR-RFLP and species-specific PCR. Identification of WBHIS at the species level in routine diagnostic work will certainly contribute to clarifying the pathogenicity of WBHIS.

Aim To elucidate the distribution and circulation dynamics of Campylobacter and Salmonella in Japanese chicken broiler flocks. Methods and Results A 2-year investigation of the distribution of Campylobacter and Salmonella was conducted in 25 broiler flocks at nine farms in Japan from 2013 to 2014. Campylobacter and Salmonella tested positive in 11 (44·0%) and 24 (96·0%) broiler flocks respectively. One hundred and ninety-five Campylobacter and 184 Salmonella isolates were characterized into 12 Campylobacter (including two novel genotypes) and three Salmonella MLST genotypes. Only Salmonella isolation between caecal and environmental samples were significantly correlated. Further, one litter sample tested positive for Salmonella before new chicks were introduced. The Campylobacter strains rapidly lost culturability within 2–18 days; in contrast, the Salmonella strains survived from 64–211 days in artificially inoculated water samples. Conclusion No persistent circulation-mediated Campylobacter contamination was observed. In contrast, circulation of Salmonella in broiler houses was seen, apparently due to the litter excreted from broiler flocks, as well as Salmonella-contaminated water and feed. Significance and Impact of the Study This paper provides the distribution, genotypic data and circulation dynamics of Campylobacter and Salmonella as recently observed in Japanese chicken broiler farms.

Since late 2013, outbreaks of porcine epidemic diarrhea virus (PEDV) have reemerged in Japan. In the present study, we observed a high detection rate of PEDV, with 72.5 % (148/204) of diarrhea samples (suckling, weaned, and sows) and 88.5 % (77/87) of farms experiencing acute diarrhea found to be positive for PEDV by reverse transcription PCR. Sequencing and phylogenic analyses of the partial spike gene and ORF3 of PEDV demonstrated that all prevailing Japanese PEDV isolates belonged to novel genotypes that differed from previously reported strains and the two PEDV vaccine strains currently being used in Japan. Sequence and phylogenetic analysis revealed prevailing PEDV isolates in Japan had the greatest genetic similarity to US isolates and were not vaccine-related. Unlike vaccine strains, all prevailing field PEDV isolates in Japan were found to have a number of amino acid differences in the neutralizing epitope domain, COE, which may affect antigenicity and vaccine efficacy. The present study indicates recent PEDV isolates may have been introduced into Japan from overseas and highlights the urgent requirement of novel vaccines for controlling PEDV outbreaks in Japan.

This study compared the reproductive and growth performances of pigs before and after depopulation and restocking after a foot-and-mouth disease outbreak in Japan. Data for the time period before and after depopulation and restocking were obtained from three farrow-to-finish farms. As a result of depopulation and restocking, hygiene levels were improved, and common infectious diseases, such as porcine reproductive and respiratory syndrome and Aujeszky's disease, remained undetected on the farms. Compared with before depopulation, reproductive and growth performances improved after depopulation; the number of total pigs born was higher, the postweaning mortality rate was lower, and the age at slaughter was lower (P

Porcine epidemic diarrhea (PED) is an emerging disease of pigs that has recently led to large numbers of piglet deaths in a number of countries of Eastern Asia and The Americas. The objective of the present study was to identify and compare risk factors associated with PED infection in locally and non-locally PED-exposed farms in Japan. A questionnaire was administered to a convenience selection of pig farms located throughout Japan. Questionnaires were administered between November 2013 (when the first case was reported in Japan) and August 2014. PED-positive farms (cases, n=124) were asked to provide information on their status (positive or negative) and select herd management practices for the two weeks prior to onset of PED clinical signs. Negative farms (controls, n=128) were given the same questionnaire and asked herd management practices for the two weeks prior to a given reference date. This date was assigned based on the date of PED occurrence in the town/prefecture in which the farm was located. Case and control farms were categorized as "locally exposed" if they were located within a 5km radius from a PED-infected farm and "non-locally exposed", otherwise. Logistic regression analysis was used to identify factors associated with PED infection. Two separate regressions were done for locally exposed and non-locally exposed farms using PED status (positive/negative) as the dependent variable. PED in locally-exposed farms was associated (P