Characteristics of antimicrobial residues in manure composts from swine farms: Residual patterns, removal efficiencies, and relation to purchased quantities and composting methods in Japan.

Present study provides first comprehensive results on the residual levels of 19 antimicrobial (AM) residues in 12 Japanese swine manure composting facilities that use open or enclosed types of treatment methods. Tilmicosin (14000 µg/kg d.w.) and tiamulin (15000 µg/kg d.w.) were present in the highest concentrations in manure composts. Morantel (MRT) had the highest detection frequency (100%) in compost, suggesting its ubiquitous usage and resistance to degradation during composting. Sulfamethoxazole had low detection frequencies and concentrations, likely due to limited partitioning to the solid phase. A positive correlation (p < 0.05) between purchasing quantities and residue levels in manure composts was detected for fluoroquinolones (FQs). The removal efficiencies of AMs in enclosed-type facilities were lower and more inconsistent than those in open-type facilities. Tetracyclines (TCs), lincomycin, and trimethoprim were easily removed from open-type facilities, whereas FQs and MRT persisted in both facilities. After discontinuing the usage of oxytetracycline (OTC), TCs concentrations reduced drastically in input materials, remained pseudo-persistent in composts for up to 4 months, suggesting a time lag for composting and were not detected (<10 µg/kg) after 4 months of OTC withdrawal. This study emphasizes on the effectiveness of manure composting methods in reducing AM residues in swine waste.

The association between farm-level antimicrobial usage and resistance of Staphylococcus spp., as the major genus isolated from aerosol samples, in Japanese piggeries.

Bacteria are the dominant particulate matter in livestock houses and can threaten animal and public health. Antimicrobial resistance (AMR) is a crucial concern worldwide, and nationwide measures established based on the One Health approach are being implemented in many countries. This requires multidisciplinary perspectives and collaboration among the human, animal, and environmental sectors. However, information on the AMR risk in livestock house aerosol is limited, especially its association with antimicrobial usage (AMU). Therefore, this study was conducted to reveal the AMR profile of Staphylococcus, the major bacterial genus in the aerosol of the piggeries of Japanese farms, and the association between farm-level AMU and AMR. The investigation at 10 farrow-to-finish pig farms revealed that regardless of the sampling season and the piggery group, the resistance rate of isolated staphylococci for oxacillin, erythromycin, and lincomycin was more than 40% of the median and tended to be higher than that for other antimicrobials. The AMU adjusted by the defined daily dose (DDD-adjusted AMU) in the fattening piggery group was significantly higher than that in the sow piggery group (p < 0.05). Finally, for the fattening piggery group, the generalized linear mixed model revealed that the AMR rate for oxacillin, erythromycin, tetracycline, and chloramphenicol was positively associated with the corresponding class-based DDD-adjusted AMU of penicillins (odds ratio (OR) = 2.63, p = 0.03), macrolides (OR = 6.89, p = 0.0001), tetracyclines (OR = 2.48, p = 0.04), and amphenicols (OR = 3.22, p = 0.03), respectively. These significant positive associations observed in this study imply that the resistance rate for these antimicrobials may decrease by reducing the corresponding antimicrobials' use. In addition, the resistance rates for erythromycin and chloramphenicol also displayed a positive association with the AMU of antimicrobial classes other than macrolides and amphenicols, respectively. The mechanism underlying these phenomena is unclear; therefore, further evaluation will be needed. As limited studies have reported staphylococci in piggery aerosol and its AMR with quantitative AMU, these results based on on-farm investigations are expected to aid in establishing countermeasures for AMR of aerosol bacteria in pig farms.

Occurrence and the potential ecological risk of veterinary antimicrobials in swine farm wastewaters in Japan: Seasonal changes, relation to purchased quantity and after termination of oxytetracycline usage.

This study provides the first comprehensive investigation of the residual concentrations of eight classes of antimicrobial agents (AMs, 20 compounds) in 13 swine wastewater treatment facilities in Japan. These facilities implemented the aerobic activated sludge (AS) or its alternative methods. The maximum concentrations before treatment were found at the level of 7100, 6900, 6000, 3600, 3400, and 1400 µg/L for tilmicosin, oxytetracycline (OTC), chlortetracycline, lincomycin , sulfamethoxazole, and trimethoprim, respectively. The highest detection rate (96.3%) in influents was noted for the morantel, which was a feed additive. The seasonal difference in residual concentration was much greater for tetracyclines (TCs) and macrolides (MLs) when their residual concentrations were high, especially in the cold season. There was a positive correlation between the purchased quantity of TCs and fluoroquinolones (FQs) and their residue levels detected in the effluents (p < 0.01). The estimated removal rate of AMs was greater than 80%. In contrast, on a few occasions, it was diminished due to failing operating conditions, such as water temperature and AS rate in the aeration tank. The estimated ecological risks of AMs in effluents based on risk quotients (RQs) considered to enhance the selection pressure for drug resistance (RQs-AMR) were high for TCs and FQs, whereas ecotoxicological effects (RQs-ENV) to aquatic organisms were higher for sulfonamides and MLs. When OTC usage ceased, its concentration in wastewater decreased rapidly; however, it remained longer period in the effluents, probably due to OTC desorption from the AS. The concentrations (and respective RQs) of TCs were decreased by >99.8% and >92% in the influents and effluents, respectively. This data suggested that it is essential to reduce the amount used and introduce more efficient methods and operating conditions to constantly remove AMs during the treatment to reduce the risk of AM discharge from swine farms.

Productivity analysis of 70 farrow-to-finish swine farms in Japan from 2013 to 2018.

Improving productivity is an urgent issue in the swine industry if it is to compete internationally. However, lack of data about recent productivity transition obstructs stakeholder planning. This study investigated the yearly productivity trends among farrow-to-finish swine farms in Japan using annual productivity data from 2013-2018 obtained for 70 farms in Japan. The productivity parameters analyzed were pigs born alive per litter (PBA), preweaning mortality (PRWM), pigs weaned per litter (PWL), litters per mated female per year (LMFY), pigs weaned per mated female per year (PWMFY), post-weaning mortality (POWM) and marketed pigs per mated female per year (MP). Data were classified into three groups based on the size of the average female inventory and compared among groups. Results presented the mean PBA increased continuously over the 6-year period (P<0.001), and the PWL, PWMFY, and MP means began increasing after 2015 (P<0.001). These upward trends were particularly remarkable on large farms. The mean PRWM increased sharply in 2014, thus inhibiting the increases in PWL, PWMFY, and MP for the same year. The LMFY and POWM means did not change during the study period. Altogether, productivity in Japan improved markedly during the study period, indicating highly prolific sows were well utilized with suitable breeding techniques among farmers these days. Continued genetic improvement and sow management would aid further development in Japan.

Antimicrobial use on 74 Japanese pig farms in 2019: A comparison of Japanese and European defined daily doses in the field.

Defined daily doses (DDD) have been established in human medicine to standardize the measurement of treatment in a population. In veterinary medicine, the European Medicine Agency published defined daily dose (DDDvet) values for antimicrobial agents used in food-producing animals in 2016. National defined doses (DDDjp) for antimicrobials used for pigs in Japan have recently been determined. The aim of this study was to compare the results of calculated antimicrobial use in the field using the DDDjp and DDDvet values. Data from 74 pig farms in Japan relative to antimicrobial use in 2019 was collected. The numbers of DDDs (the weight of biomass treated in kg-days) using DDDjp and DDDvet values for each farm and for different antimicrobial classes were compared. Associations between calculated numbers of DDDjp and DDDvet on farm level were investigated. In addition, differences in antimicrobial use were investigated between different production types of farms (farrowing, finishing and farrow-to-finish farms). Using DDDjp and DDDvet values, the aggregated number of DDDs for 74 farms were 4,099,188 and 2,217,085 respectively, with the former being larger by 1.85 times than the latter. The most frequently used antimicrobial class was penicillin regardless of whether DDDjp or DDDvet was used. The absence of DDDvet values for certain antimicrobial agents used in Japan and the differences in the number of DDDjps/PCU and DDDvets/PCU indicated the need for Japanese DDDs. The number of DDDs per kg population correction unit (PCU) per farm tended to be higher in farrowing farms than in farrow-to-finish farms and finishing farms, with no significant difference (P = 0.19).

Comparison of Productivity and Fecal Microbiotas of Sows in Commercial Farms.

Sow productivity, that is, the number of weaned piglets per sow per year, depends on their health status. The gut microbiota is considered a crucial factor in the health of pigs and may affect sow productivity. In the present study, we aimed to investigate the relationship between productivity and the fecal microbiotas of sows in different farms. Feces of sows were collected from 18 farms (10 samples/farm). A total of 90 fecal samples of high-reproductive performance farms were labeled as group H, and 90 fecal samples from low-reproductive performance farms were labeled as group L. Fecal microbiotas were analyzed by 16S rRNA metagenomics, and the organic acids and putrefactive metabolites of the microbiotas were measured. ß-diversity was significantly different between groups H and L (P < 0.01), and the relative abundances of 43 bacterial genera, including short-chain fatty acid-producing and fiber-degrading bacteria such as Ruminococcus, Fibrobacter and Butyricicoccus, significantly differed between groups (P < 0.05). In addition, the concentrations of acetate, propionate and n-butyrate were significantly higher in group H than in group L (P < 0.05). In conclusion, sow productivity in farms was likely associated with the compositions of the fecal microbiotas.

Impact of a porcine epidemic diarrhea outbreak on swine productivity in Japan: a retrospective cohort study.

The objective was to investigate porcine epidemic diarrhea (PED) outbreak that occurred in 2014 in Japan and its effects on herd-level productivity using a data recording system (PigINFO). The study herds were selected from farrow-to-finish herds (n=99) that entered in the PigINFO system between July 2013 and March 2015. From 1 April to 30 June 2014 (PED epidemic), any herds with clinical signs of PED and feces positive for porcine epidemic diarrhea virus (PEDV) on polymerase chain reaction analysis and/or immunohistochemical staining were defined as PED-positive (n=38). They were further classified into those with long PED periods (L-PED-positive; n=28) and those with short PED periods (S-PED-positive; n=10). Herds with no clinical signs of PED were classified as PED-negative (n=61). Herd-level production data, including preweaning mortality (%; PRWM), postweaning mortality (%; POWM), pigs weaned per litter (PWL), pigs born alive per litter, litters per mated female per year and pigs marketed per sow (MP), were calculated every 3 months during study period. During the PED epidemic, L-PED-positive herds had significantly higher PRWM and POWM than PED-negative herds, and L-PED-positive and S-PED-positive herds had significantly lower PWL. During October-December 2014, L-PED-positive herds had significantly fewer MP than PED-negative herds. The PED outbreak increased mortality and consequently reduced the numbers of marketed pigs. The rapid control of an outbreak is important for reducing the financial losses arising from PED infections.

Aujeszky's disease and the effects of infection on Japanese swine herd productivity: a cross-sectional study.

Pseudorabies virus (PRV) is endemic in some regions of Japan. We investigated the effects of PRV infection status on herd productivity. Serum samples were obtained from 48 swine herds in Japan. Within each herd, three serum samples were obtained from growing pigs at four different ages, as well as from sows in low and high parity groups. Sera were tested for antibodies against wild-type PRV via competitive ELISA. Herds were classified into PRV positive and negative groups based on serological results. Herds infected with PRV exhibited postweaning mortalities (6.84%) that were significantly (P=0.0018) higher than those in unaffected herds (4.73%). Because of the reduced productivity in PRV positive herds, the current PRV eradication program must be strengthened.

Epidemics of emerging animal diseases and food-borne infection problems over the last 5 years in Japan.

There have been several emerging animal diseases and food-borne infection problems occurring in Japan over the last 5 years. We describe brief pictures of these epidemics and our control activities. As acute contagious and/or emerging animal diseases, the foot and mouth disease (FMD) outbreak caused by the Pan-Asian topotype of the type O virus occurred in March 2000 after 92 years of FMD-free status. In 2004, four cases of the highly pathogenic avian influenza (HPAI), which was the first outbreak after 79 years, and caused by the H5N1 subtype, were identified. As part of the responses against these outbreaks, all the animals in the affected farms were destroyed, and movement control areas were established around the infected premises, and a nation-wide intensive survey for FMD and HPAI was performed. As for food-borne or feed-borne infections, the first bovine spongiform encephalopathy (BSE) was identified in September 2001 and 19 more cases have been reported until June 2005. A large outbreak of food-borne infection caused by low-fat milk contaminated with enterotoxin A produced by Staphylococcus aureus, involving more than 13,000 patients, occurred in 2000. In 2003, people who consumed uncooked liver and meat from wild boar and deer developed clinical signs of hepatitis caused by the hepatitis E virus. Pork is also suspected as natural source of virus transmission. Early detection of the first cases and rapid action in preventing and controlling the spread of infections are very important combined with proper risk communication about correct information of the diseases.

A quantitative assessment of the risk of exposure to bovine spongiform encephalopathy via meat-and-bone meal in Japan.

The feeding of meat-and-bone meal (MBM) derived from cattle infected with bovine spongiform encephalopathy (BSE) is a major source of BSE infection. The risks of BSE infection via MBM in Japan were examined quantitatively to estimate infectivity to cattle via MBM derived from a single clinically infected animal being rendered. Three routes of exposure were modeled: (i) feeding cattle concentrates containing MBM as an ingredient, (ii) feeding cattle concentrates contaminated with MBM from non-ruminant feed at feed plants and (iii) directly feeding MBM in supplemental form to cattle on farms. The effectiveness of measures designed to restrict the feeding of ruminants with ruminant MBM (feed restriction) as well as differences in the risk of exposure among regions were examined using the model. The model revealed that the median total infectivity fed to dairy cattle via MBM derived from one infected animal was approximately 0.49 cattle oral ID(50) (5th percentile=0.43ID(50), 95th percentile=0.54ID(50)). This value was reduced by 55% after the addition of MBM to cattle concentrates was restricted in 1996. The risk of exposure in dairy cattle was twice that in beef cattle. Comparisons of regional differences in exposure risk indicated that the risk was highest in a region where 14 of the 20 BSE cases reported to date were born. Our model suggested that the routes of exposure via MBM were unlikely to result in increased propagation of BSE in Japan. Furthermore, despite some regional variation, the risk of exposure declined further after the feed restriction was imposed in 1996.

Cross-sectional survey of ixodid tick species on grazing cattle in Japan.

Ixodid tick species were collected from cattle in 60 grazing fields throughout Japan. Haemaphysalis longicornis was mainly recovered in the western and southern regions, while Ixodes species were collected mainly in the central to northern regions. Other tick species such as Amblyomma testudinarium, Boophilus microplus, H. flava and H. kitaokai were identified from a few fields in the central and southern regions. Haemaphysalis longicornis were recovered in the fields with higher temperatures and annual rainfall, whereas I. ovatus and I. persulcatus were collected in fields with lower temperatures and annual rainfall. Some of these tick species are capable of transmitting pathogens harmful to cattle and humans, so proper control strategies are required.

Nationwide seroprevalence of Neospora caninum among dairy cattle in Japan.

Serum samples from 2420 clinically healthy dairy cattle, randomly selected from stored sera in 18 districts of Japan, were tested for the presence of Neospora caninum antibodies using an indirect fluorescent antibody test (titer > or =1:200). Nationwide seroprevalence is estimated at 5.7% (139/2420). Seropositive cattle were detected in all surveyed districts despite the evidence of confirmed case reports of bovine neosporosis, showing that N. caninum is widely distributed throughout Japan. Age-specific seroprevalence did not increase with cattle age, suggesting that Neospora infection is likely to be transmitted vertically rather than horizontally in Japan. Considering that N. caninum seropositive cows are thought to be more likely to abort, substantial fetal losses may be induced by N. caninum infection in Japan. Devising strategies are needed to reduce the economic impact on the Japanese dairy industry. This is the first study to investigate the nationwide seroprevalence of N. caninum in cattle in Asia.

Proportion of abortions due to neosporosis among dairy cattle in Japan.

A retrospective cohort study was conducted to clarify the association between seropositive reactions to Neospora caninum and subsequent reproductive disorders among dairy cattle in Japan. A statistically significant association between Neospora seropositive reactions and abortions was observed (P=0.016), and seropositive cattle were 6.1 times more likely to abort compared to seronegative cows. No significant differences were observed between seropositive reactions and other reproductive disorders such as conception failure, perinatal death and calf mortality. As indicated by estimation of the attributable fraction, 83.6% of abortions in Neospora seropositive animals may be attributed to N. caninum. Considering seroprevalence of N. caninum in the cattle which aborted in Japan, 21.8% of abortions were estimated to be caused by neosporosis in Japan.

Seroprevalence of Neospora caninum in dairy and beef cattle with reproductive disorders in Japan.

Serum samples from 145 dairy and 65 beef cattle with reproductive disorders and 54 normally calving dairy cattle (controls) in Japan were tested for presence of Neospora caninum antibodies by use of an indirect fluorescent antibody test (IFAT, titer 1:200). Overall, seroprevalence of N. caninum was significantly higher (P < 0.001) in dairy cattle (20.0%, 29/145) than in beef cattle (1.5%, 1/65). In cattle which aborted, seroprevalence of N. caninum was significantly higher (P = 0.041) in dairy cattle (26.1%, 23/88, compared with controls (3.7%, 2/54)) than in beef cattle (5.0%, 1/20), indicating that neosporosis might be a more common problem in dairy cattle than in beef cattle in Japan. Seropositive cattle were 9.2 times more likely to abort compared to seronegative cows. Abortions associated with N. caninum seropositivity in this study were most frequently observed in the second trimester, and the mean gestational age of the fetuses aborted from seropositive dams was 5.7 months. In conclusions, N. caninum seems to be causing serious economic losses in the dairy industry in Japan. This is the first report on an objective comparison of seroprevalence of dairy and beef cattle with reproductive disorders in Asia.

Loss of milk yield due to Akabane disease in dairy cows.

Akabane disease is an infection with clinical signs of congenital malformation and abortion in ruminants. Abnormal parturitions caused by Akabane disease result in great economic loss. The purpose of this study is to estimate the reduction in the milk yield from abnormal parturition due to Akabane disease. The data were collected from 33 Holstein cows on 11 farms. The animals had abnormal parturitions during the period from September 1998 to March 1999, and were diagnosed as having Akabane disease. The mean and standard deviation of the rate of reduction in the milk yield of 33 cows after abnormal parturition caused by Akabane disease was -11.4 +/- 14.9%. The means and standard deviations of the rate of reduction of four cows calving abnormally at 220-239 days of gestation, nine cows calving abnormally at 240-270 days of gestation, and 20 cows calving abnormally at 271-300 days of gestation were -26.6 +/- 24.7%, -14.7 +/- 11.0%, and -6.9 +/- 12.3%, respectively. In this study, we demonstrated that the rate of reduction in the milk yield in cows affected by Akabane disease was -11.4 +/- 14.9%, but values as high as -26.6 +/- 24.7% were reached in the comparison with the milk yield obtained after normal parturition.

Application of proteomics for comparison of proteome of Neospora caninum and Toxoplasma gondii tachyzoites.

Protein profiles of two isolates of Neospora caninum (KBA-2 and JPA1) and Toxoplasma gondii RH strain were investigated by proteomic approach. Approximately, 78% of protein spots on two-dimensional gel electrophoresis (2-DE) profiles and 80% of antigen spots on 2-DE immunoblotting profiles were exhibited to share the same pI and M(r) between KBA-2 and JPA1 of N. caninum. On the other hand, a total of 30 antigen spots of T. gondii were recognized on 2-DE immunoblotting profile using rabbit antiserum against N. caninum KBA-2. A number of homologue proteins, such as heat shock protein 70, tubulin alpha- and beta-chain, putative protein disulfide isomerase, actin, enolase and 14-3-3 protein homologue are believed as the conserved proteins in both N. caninum and T. gondii. On the contrary, NcSUB1, NcGRA2 and NCDG1 (NcGRA7) might be the species-specific proteins for N. caninum tachyzoites. The present study showed that the high degree of similarity between N. caninum isolates (KBA-2 and JPA1), whereas large differences between N. caninum and T. gondii were noticed by proteome comparisons.

Effect of intermittent administration of human parathyroid hormone on bone mineral density and arthritis in rats with collagen-induced arthritis.

OBJECTIVE: To investigate the effect of intermittent administration of human parathyroid hormone (PTH) on bone mineral density (BMD) and arthritis in rats with collagen-induced arthritis (CIA). METHODS: Seven-month-old female Sprague-Dawley rats were divided into 4 groups: rats without CIA (control), rats with CIA treated with vehicle, rats with CIA treated with PTH for 4 weeks, and rats with CIA treated with PTH for 6 weeks. PTH (20 mug/kg) was injected subcutaneously 3 times per week. BMD in the proximal metaphysis and the diaphysis of the tibia was measured by peripheral quantitative computed tomography every 2 weeks until week 8. Eight weeks after initial sensitization, the animals were killed, and the BMD and mechanical properties of excised limbs were evaluated. Histomorphometric analysis of tibiae and histologic evaluation of arthritis were also performed. RESULTS: In the PTH-treated rats with CIA, the incidence and severity of arthritis were macroscopically and histologically similar to the findings in the vehicle-treated rats with CIA. The decrease of BMD caused by CIA was suppressed by treatment with human PTH, in a manner that was dependent on the duration of administration. In the histomorphometric analysis, bone formation parameters were higher and bone resorption parameters were lower in the PTH-treated arthritic rats compared with vehicle-treated arthritic rats. Mechanical properties were also maintained in the PTH-treated rats. CONCLUSION: Our findings indicate that, in an animal model of arthritis, intermittent PTH administration activates bone formation, resulting in increased BMD and preventing deterioration of mechanical properties. However, PTH has no effect on the arthritis itself.

Effect of minodronic acid (ONO-5920) on bone mineral density and arthritis in adult rats with collagen-induced arthritis.

OBJECTIVE: To study the effect of minodronic acid (ONO-5920) on bone loss and arthritis in rats with collagen-induced arthritis (CIA) treated according to 2 different schedules. METHODS: Four groups of female Sprague-Dawley rats (7 months old) were studied: rats without CIA treated with vehicle (controls), CIA rats treated with vehicle (CIA-V), CIA rats treated therapeutically with minodronic acid (CIA-T), and CIA rats treated prophylactically with minodronic acid (CIA-P). Minodronic acid was administered orally at 0.2 mg/kg 3 times a week, beginning 2 weeks after initial sensitization in the CIA-T rats and beginning the day after initial sensitization in the CIA-P rats. Bone mineral density (BMD) was measured by peripheral quantitative computed tomography in the proximal metaphysis and diaphysis of the tibia every 2 weeks until week 8, when the rats were killed. The BMD and bone microstructure of the excised femur were evaluated by dual x-ray absorptiometry and microfocal computed tomography, respectively. Histomorphometry of the proximal tibia was also performed. RESULTS: In CIA-P rats, the incidence of arthritis and the severity of posterior limb swelling were reduced early after sensitization, and the decrease in BMD was prevented throughout the observation period. Bone and joint destruction evaluated by radiography of the foot was reduced in CIA-P rats. The eroded surface was reduced and the microstructure was maintained in CIA-P rats compared with CIA-V rats. The mineral apposition and bone formation rates were not reduced in the CIA-P rats. In CIA-T rats, however, the inflammation was not suppressed and the inhibitory effect on bone loss was smaller than that in CIA-P rats. CONCLUSION: Minodronic acid suppressed the decrease in BMD and the deterioration of the bone microstructure caused by arthritis. Prophylactic administration of minodronic acid had a preventive effect on arthritis at the early stage, although not throughout the observation period.