Evaluation of 14 PFAS for permeability and organic anion transporter interactions: Implications for renal clearance in humans.

Per- and polyfluoroalkyl substances (PFAS) encompass a diverse group of synthetic fluorinated chemicals known to elicit adverse health effects in animals and humans. However, only a few studies investigated the mechanisms underlying clearance of PFAS. Herein, the relevance of human renal transporters and permeability to clearance and bioaccumulation for 14 PFAS containing three to eleven perfluorinated carbon atoms (ηpfc = 3-11) and several functional head-groups was investigated. Apparent permeabilities and interactions with human transporters were measured using in vitro cell-based assays, including the MDCK-LE cell line, and HEK293 stable transfected cell lines expressing organic anion transporter (OAT) 1-4 and organic cation transporter (OCT) 2. The results generated align with the Extended Clearance Classification System (ECCS), affirming that permeability, molecular weight, and ionization serve as robust predictors of clearance and renal transporter engagement. Notably, PFAS with low permeability (ECCS 3A and 3B) exhibited substantial substrate activity for OAT1 and OAT3, indicative of active renal secretion. Furthermore, we highlight the potential contribution of OAT4-mediated reabsorption to the renal clearance of PFAS with short ηpfc, such as perfluorohexane sulfonate (PFHxS). Our data advance our mechanistic understanding of renal clearance of PFAS in humans, provide useful input parameters for toxicokinetic models, and have broad implications for toxicological evaluation and regulatory considerations.

Low molecular weight acids and OATP1B mediated hepatic clearance: In vitro and in vivo evaluation using novel hypoxia-inducible factor prolyl hydroxylase inhibitors (Dustats).

Organic anion transporting polypeptide (OATP1B) plays a key role in the hepatic clearance of a majority of high molecular weight (MW) acids and zwitterions. Here, we evaluated the role of OATP1B-mediated uptake in the clearance of novel hypoxia-inducible factor prolyl hydroxylase inhibitors ("Dustats"), which are typically low MW (300-400 daltons) aliphatic carboxylic acids. Five acid dustats, namely daprodustat, desidustat, enarodustat, roxadustat and vadadustat, showed specific transport by OATP1B1/1B3 in transporter-transfected HEK293 cells. Neutral compound, molidustat, was not a substrate to OATP1B1/1B3. None of the dustats showed transport by other hepatic uptake transporters, including NTCP, OAT2 and OAT7. In the primary human hepatocytes, uptake of all acids was significantly reduced by rifampin (OATP1B inhibitor); with an estimated fraction transported by OATP1B (ft ,OATP1B) of up to >80% (daprodustat). Molidustat uptake was minimally inhibited by rifampin; and low permeability acids (desidustat and enarodustat) also showed biliary efflux in sandwich culture human hepatocytes. In vivo, intravenous pharmacokinetics of all 5 acids was significantly altered by a single-dose rifampin (30 mg/kg) in Cynomolgus monkey. Hepatic clearance (non-renal) was about 4-fold (vadadustat) to >11-fod (daprodustat and roxadustat) higher in control group compared to rifampin-treated subjects. In vivo ft ,OATP1B was estimated to be ~70-90%. In the case of molidustat, rifampin had a minimal effect on overall clearance. Rifampin also considerably reduced volume of distribution of daprodustat and roxadustat. Overall, OATP1B significantly contribute to the hepatic clearance and pharmacokinetics of several dustats, which are low MW carboxylic acids. OATP1B activity should therefore by evaluated in this property space. Significance Statement Our in vitro and in vivo results suggest that OATP1B-mediated hepatic uptake play a significant role in the pharmacokinetics of low MW acidic dustats, which are being developed or approved for the treatment of anemia in chronic kidney disease. Significant active uptake mechanisms are not apparent for the neutral compound, molidustat. Characterization of uptake mechanisms is therefore important in predicting human pharmacokinetics and evaluating drug-drug interactions for low MW acids.

A case-control study of the infection risk of H5N8 highly pathogenic avian influenza in Japan during the winter of 2020-2021.

In Japan, outbreaks of H5N8 highly pathogenic avian influenza (HPAI) were reported between November 2020 and March 2021 in 52 poultry farms. Understanding HPAI epidemiology would help poultry industries improve their awareness of the disease and enhance the immediate implementation of biosecurity measures. This study was a simulation-based matched case-control study to elucidate the risk factors associated with HPAI outbreaks in chicken farms in Japan. Data were collected from 42 HPAI-affected farms and 463 control farms that were within a 5-km radius of each case farm but remained uninfected. When infected farms were detected as clusters, one farm was randomly selected from each cluster, considering the possibility that the cluster was formed by farm-to-farm transmission within an epidemic area. For each case farm, up to three control farms were selected within a 5-km radius. Overall, 26 case farms (16 layer and 10 broiler farms) and 75 control farms (45 layer and 30 broiler farms) were resampled 1000 times for the conditional logistic regression model with explanatory variables comprising geographical factors and farm flock size. A larger flock size and shorter distance to water bodies from the farm were found to increase infection risk in layer farms. Similarly, in broiler farms, a shorter distance to water bodies increased infection risk. On larger farms, frequent access of farm staff and instrument carriages to premises could lead to increased infection risk. Waterfowl visiting water bodies around farms may also be associated with infection risk.

Assessing population structure and migration patterns of wild boar (Sus scrofa) in Japan.

Geographical wildlife patterns reflect historical range expansion and connectivity and provide insights into wildlife population management. In our large-scale phylogeographic population analysis of wild boars (Sus scrofa leucomystax) in Japan, we identified 15 clusters using 29 microsatellite markers, each structured within a range of approximately 200 km. This suggests that evolution was essentially driven by isolation by distance, and that the range of gene flow was limited. One cluster contained subpopulations located approximately 900 km apart, indicating the occurrence of past anthropogenic introductions. Moreover, we estimated effective migration to visualize the geographic genetic population diversity. This analysis identified six potential barriers, one of which involved large plains and mountainous areas in the Kanto region of eastern Japan. This barrier likely persisted in the two eastern clusters for an extended period, restricting migration to the neighboring areas. Overall, our study sheds light on the demographic history of wild boar in Japan, provides evidence of past anthropogenic introductions from distant areas, and highlights the importance of geographic barriers in shaping genetic diversity and population dynamics. This knowledge will be beneficial for forming informed wildlife management strategies toward the conservation of genetic integrity and ecological balance of wild boar populations in Japan.

Analysis of effective spatial range of oral vaccination against classical swine fever for wild boar.

Classical swine fever (CSF) re-emerged in Gifu Prefecture, central Japan, in September 2018 and is currently widespread in wild boar populations. Due to its widespread in wild boars, an oral mass vaccination strategy was initiated in March 2019, employing a commercial bait vaccine that is a live attenuated vaccine. To enhance the effectiveness of oral vaccination, it is crucial to determine the vaccine's effective spatial range. This understanding is essential for devising a comprehensive vaccination strategy, which should also include a preliminary investigation of wild boar habitats before vaccination. This study aimed to estimate the effective range of oral vaccination for wild boars against CSF by analyzing the geographical relationship between immune wild boars and vaccination points within the vaccination areas in Gifu Prefecture. This study utilized oral vaccination data from April 2021 to March 2022. The prevalence of CSF infections in wild boars remained below 5% in this period, suggesting limited disease transmission and immune wild boars were considered to be induced by the effect of vaccination. Two vaccination campaigns were conducted during this period, with almost 2000 vaccination points each. To investigate the factors associated with the intensity (i.e., density) of immune wild boar, the nearest distances to a vaccination point and to a susceptible wild boar were evaluated as explanatory variables. The Rhohat procedure and point process model were utilized to analyze the relationship between the intensity of immune wild boars and the explanatory variables. The point process model revealed a significant decrease in the intensity of immune wild boars when the distance from the nearest vaccination point exceeded 500 m, indicating that the effective spatial range of bait vaccination is within 500 m of the vaccination point. Although the distance to the nearest susceptible animal did not show significance in the model, Rhohat plots indicated that the intensity of immune wild boars decreased at distances greater than 1200 m from the nearest susceptible wild boar. This finding highlights the importance of investigating susceptible wild boar populations within a range of at least 1200 m from a vaccination point before implementation. The present study revealed the effective range of oral vaccination for wild boars against CSF and indicated the importance of investigating susceptible wild boar habitats around vaccination points before the implementation of vaccination. These findings may help improve the effectiveness of oral vaccinations.

The probability distributions of the movement of dairy and beef cattle in Japan: a data note.

OBJECTIVES: Animal movement is an important factor in the transmission of infectious diseases among livestock. A better understanding of animal movement characteristics provides a more reliable estimation of disease spread and promotes modeling studies. In Japan, all the cattle movement information is recorded in a national database called the Individual Cattle Identification Register (ICIR)." Our previous studies using this information demonstrated heterogeneity in the movement of dairy and beef cows according to location, season, and age. The present study describes the probability distributions of the movement of Japanese dairy and beef cows in the following month on a regional basis. DATA DESCRIPTION: This publication contains four probability distribution datasets for the predicted locations of dairy and beef cows in Japan in the following month, which were developed using individual cattle movement information obtained from the ICIR. These datasets provide information on cattle movement in the following month on a regional basis, given properties such as birth region, location, time, and age.

Sex-based differences in the distribution of Aujeszky's disease-seropositive Japanese wild boar.

BACKGROUND: Aujeszky's disease virus (ADV) primarily infects domestic pigs and wild boars, causing the abortion and death of young piglets due to central nervous system disorders. In Japan, the national eradication program for ADV in domestic pigs has been successful in most prefectures; however, concern has been raised regarding ADV-infected wild boars as a source of transmission to domestic pigs. RESULTS: We assessed the nationwide seroprevalence of ADV among wild boars (Sus scrofa) in Japan. Moreover, we determined the sex-based differences in the spatial clustering of seropositive animals. Serum samples were obtained from a total of 1383 wild boars acquired by hunting in 41 prefectures in three fiscal years (April-March in 2014, 2015, and 2017). Seropositivity tests for ADV using enzyme-linked immunosorbent assay, the latex agglutination and neutralization tests showed 29 boars seropositive for ADV (29/1383, 2.1% [95% confidence interval, CI: 1.4-3.0%]), with 28 of these boars originating from three prefectures in the Kii Peninsula (28/121, 23.1% [95% CI: 16.0-31.7%]). The degree of spatial clustering of these ADV-seropositive adult boars in the Kii Peninsula was evaluated using the K-function and data from sera samples of 46 (14 seropositive) male and 54 (12 seropositive) female boars. The degree of clustering among females was significantly higher in seropositive animals than in tested animals; however, such a difference was not observed for seropositive males. CONCLUSIONS: The spatial dynamics of ADV among adult wild boars may be characterized based on sex, and is likely due to sex-based differences in behavioral patterns including dispersal among wild boars.

Region-wise analysis of beef cow movements in Japan.

Animal movement is an important factor in the transmission of animal infectious diseases. A better understanding of movement patterns is therefore necessary for developing effective control measures against disease spread. In Japan, a cattle tracing system was established in 2003, following a bovine spongiform encephalopathy epidemic, and the information on all cattle movements has been stored in a national database maintained by the National Livestock Breeding Center. Using these data, we previously analyzed the movement of dairy cows, concluding that heterogeneities in cattle movement are associated with regional and seasonal factors. In the present study, we aimed to identify specific factors affecting the regional and seasonal movement patterns of beef cows in Japan. From April 2012 to March 2017, 797,553 farm-to-farm movement events were recorded. We analyzed movements by month and by cattle age and looked at the frequency of movement within and between seven regions spanning the national territory. Our results show that calf movement peaked at 9-10 months old; these movements were considered to be via the market and were frequent within and between regions. For inter-regional movements, Kyushu region was the top producer of calves for calf trading markets throughout Japan. With regard to intra-regional movements, round-trip movements for summer grazing were observed in May and October for cattle of various ages in the northern regions, especially Hokkaido and Tohoku. Moreover, the movements of Japanese Shorthorn breeds in Tohoku region exhibited consistent annual peaks in May and October/November, in accordance with their seasonal breeding practice. In the areas with high relative densities of dairy cows, such as Hokkaido, the shipping of newborn beef calves produced via embryo transfer to dairy cows was also observed. Overall, understanding the patterns of beef cow movement will help develop effective disease surveillance measures, such as pre-movement inspections focused on specific regions and types of movement.

Seasonal changes of the prostate gland in the raccoon (Procyon lotor) inhabiting Hokkaido, Japan.

In the prostate gland of the raccoon (Procyon lotor), the morphological appearance of the epithelial cells, such as basal and luminal cells, and the expressions of p63, androgen receptor (AR), and proliferating cell nuclear antigen (PCNA) were examined histologically and immunohistochemically to clarify their seasonal dynamics throughout the year. In this study, the regression with luminal cell defluxion and the regeneration process of the prostatic glandular epithelium was revealed in the seasons with declined spermatogenesis (June to August). The expression of p63 was observed only in the basal cells. AR immunoreactivity in the luminal cells was shown in the developed and regenerating (close to developed) prostates, whereas the basal cells exhibited AR immunoreactivity all year round. PCNA expression was rare in epithelial cells of the developed prostate gland. In the regressed gland, the basal cells demonstrated proliferative ability, whereas PCNA of the luminal cells appeared for the first time in the regenerating phase. This study is the first to clarify the regression with luminal cell defluxion and restoration and the seasonal dynamics of AR expression and proliferative activity in the prostate gland of seasonal breeders.

Estimation of introduction time window of highly pathogenic avian influenza virus into broiler chicken farms during the 2020 - 2021 winter season outbreak in Japan.

When an infectious disease occurs in an area, early detection of infected farms is important to respond quickly and contain the outbreak on a small scale. Estimating the time window for the introduction of the infection is important for its prevention and control. The aim of this study was to estimate the farm-specific time window from the introduction of the highly pathogenic avian influenza (HPAI) virus into poultry farms using field data from the HPAI H5N8 outbreak in the 2020-2021 winter season in Japan. Daily mortality data from 12 broiler chicken farms during the outbreak were used for the analysis. A mathematical model (Susceptible-Exposed-Infectious-Removed, SEIR model) was applied to generate the within-flock transmission of HPAI. The model-predicted mortality was fitted to the observed excess mortality data induced by HPAI to estimate the farm-specific transmission rate and the time of virus introduction. The estimated value of the transmission rate in each farm was 1.449 day-1 in median (min: 0.661 day-1, max: 3.387 day-1). The time window from the introduction of the virus to notification in each farm was estimated at 14.0 days in median (min: 8.6 days, max: 24.1 days) in the deterministic model. In addition, in the stochastic model considering the randomness of transmission in the early phase of the outbreak, the upper value of 95 % credible interval of the time window ranged from 12 to 34 days, with a median of 21 days. The results suggest that although one to three weeks had elapsed on most farms until notification after the virus introduction, the time window could exceed three weeks considering the stochasticity of disease transmission. As for the potential farm characteristics affecting within-flock transmission, the transmission rate was smaller (p-value=0.02) and the estimated time window from introduction to notification was longer (p-value=0.02) when birds were older. This study provides reliable information for setting up a tracing period for a potential source farm and enhancing the efforts for early detection.

Pig farm vaccination against classical swine fever reduces the risk of transmission from wild boar.

In 2018, classical swine fever (CSF) re-emerged in the Gifu Prefecture, central Japan, causing an on-going outbreak among wild boars and domestic pigs in the country. Consequently, oral vaccination for wild boar and compulsory vaccination for pig farms started in 2019. We have previously shown that, before vaccination in the Gifu Prefecture, the presence of CSF-infected wild boar near pig farms increased the risk of CSF transmission. This study aimed to re-evaluate the transmission risk from wild boars to pig farms under a vaccination program. The effectiveness of vaccination was evaluated by comparing the transmission risk estimated before and after the implementation of vaccinations. In this study, we focused on two affected areas, the Kanto (eastern Japan) and Kinki (west-central Japan) regions, in which eight of 11 infected farms were detected between the start of pig farm vaccinations and April 2021. Wild boar surveillance data from an area within a 50-km radius from the infected farms were used for analysis, consisting of 18,870 1-km grid cells (207 infected cells) in the Kanto region, and 15,677 cells (417 infected cells) in the Kinki region. The transmission rates in the post-vaccination period in the Kanto and Kinki regions were much lower than that in the pre-vaccination period in the Gifu Prefecture. The values of transmission kernels (h0, transmission rate at 0 km) in the Kanto and Kinki regions decreased to 1% of the transmission kernel in the pre-vaccination period. In the pre-vaccination period, the risk of infection within 300 days was almost 95 % when one infected grid cell was detected within 1 km of a pig farm. Meanwhile, in the post-vaccination period, the risk of infection within 300 days was approximately 5% when several infected cells were detected within 1 km of a pig farm. Considering the limited effect of oral vaccination for wild boar due to distribution limitations in the Kanto and Kinki regions, vaccination on pig farms may seems to have mainly reduced the transmission risk from wild boar. However, despite the implementation of vaccination, the risk of infection on pig farms remains present due to the immunity gap of weaning pigs. Therefore, strict biosecurity measures on pig farms and an appropriate vaccination program are required to prevent and control CSF spread.

Is Salmonella enterica shared between wildlife and cattle in cattle farming areas? An 11-year retrospective study in Tokachi district, Hokkaido, Japan.

BACKGROUND: Salmonella enterica in cattle has long been problematic and suspected to be transmitted by wildlife in Tokachi, Hokkaido, a major cattle farming area in Japan. Understanding the role of wildlife in S. enterica transmission would be helpful for developing control strategies of bovine salmonellosis. OBJECTIVES: We aimed to elucidate the possibility of S. enterica transmission between sympatric wildlife, including raccoons and crows and cattle, in Tokachi from 2008 to 2018 by analysing S. enterica detection records, and the genetic relatedness of serotypes shared between wildlife and cattle. METHODS: S. enterica detection records were based on the results of a field survey and existing cattle records at relevant organisations, including clinical reports, a monitoring survey and quarantine for introduced calves at growing farms and public calving farms. S. enterica was identified by polymerase chain reaction assay and serotyped by agglutination assay. The detection records were organised chronologically to investigate whether common serotypes in wildlife and cattle were detected in the same year. The isolates corresponding to detection records were assessed for their genetic patterns by pulsed-field gel electrophoresis. RESULTS: The prevalence of S. enterica in raccoons and crows was 10.7% (17/159) and 5.7% (55/967), respectively. The following serotypes were detected from both wildlife and cattle: Braenderup, Dublin, Infantis, Mbandaka, Montevideo, 4,[5],12:i:- and Typhimurium. Genetically similar isolates for S. Braenderup, S. Dublin, S. Montevideo and S. 4,[5],12:i:- were detected from both species in the same year. CONCLUSIONS: Our long-term retrospective observations supported that S. enterica was shared between wildlife and cattle. Wildlife invasions should be controlled at farms to prevent inter-species transmission of S. enterica from livestock farms.

Subgrouping and analysis of relationships between classical swine fever virus identified during the 2018-2020 epidemic in Japan by a novel approach using shared genomic variants.

Classical swine fever (CSF) is a worldwide devastating disease of the pig industry caused by classical swine fever virus (CSFV). In September 2018, an outbreak of CSF occurred in Japan where the disease had been eradicated and was officially designated a CSF-free country since 2015. Following the detection of the first 2018 case on a farm in Gifu Prefecture, the disease spread among both farm pigs and wild boars and still continues. Epigenome analysis using whole-genome information is helpful in identifying the infection route, but the current approaches provide an insufficient resolution. In this study, a novel method of using single-nucleotide variants (SNVs) was employed to identify the associations among 158 isolates (65 from farms and 93 from wild boars). The identified groups of CSFV strains were plotted in different colours on a map, identifying the location where each strain was collected. The lack of an SNV set shared between the index case and the other strains suggested the first infection in Japan during the outbreak occurred in wild boars, not at the index farm. For the Atsumi Peninsula outbreaks, where nine farms were found infected within a 10-km radius area, the farm strains were assembled into three groups, suggesting these outbreaks resulted from at least three different infection events in this area. For the infections in the area around Saitama Prefecture, an area remote from the epicentre, strains from both the farms and wild boars were identified as being in the same group, suggesting they resulted from one viral introduction. Likewise, seven infected farms in Okinawa Prefecture, almost 1,500 km from Gifu Prefecture, were identified as being in a common, but separate group. By demonstrating the variety of transmission routes and possibility of long-distance infection, these results will help improve disease control measures.

Longer prolapsed rectum length increases recurrence risk after Delorme's procedure.

PURPOSE: Risk factors for recurrence of rectal prolapse after surgery remain unclear. Delorme's procedure is often selected for relatively small-sized rectal prolapse, but there are few reports discussing the association between prolapsed rectum length and prolapse recurrence after Delorme's procedure. We hypothesized that patients with longer rectal prolapses are at a higher risk of recurrence after Delorme's procedure. METHODS: The study population comprised patients with rectal prolapse who underwent Delorme's procedure between January 2014 and December 2019 at Tokyo Yamate Medical Center. We extracted data on patient age, sex, body mass index, previous history of anal surgery, previous history of surgery for rectal prolapse, and length of prolapse, to identify risk factors for prolapse recurrence. RESULTS: Altogether, 96 patients were eligible for analysis. The median length of the prolapsed rectum was 3.0 cm (range, 1.0-6.6 cm). Twenty-four patients (25.0%) experienced recurrence after Delorme's procedure after a median of 7.5 months (interquartile range, 3.2-20.9 months). Multivariate analysis revealed that longer prolapsed rectum length increased the risk of recurrence after Delorme's procedure (hazard ratio, 6.28; 95% confidence interval, 1.83-21.50; P<0.001). CONCLUSION: The length of the prolapsed rectum should be measured before Delorme's procedure for rectal prolapse, because length is associated with a risk of recurrence after the surgery.

Additive Bayesian network analysis of the relationship between bovine respiratory disease and management practices in dairy heifer calves at pre-weaning stage.

BACKGROUND: Nursery farms that accept nursing and growing pre-weaned heifer calves from private dairy farms must work to prevent bovine respiratory disease (BRD). Knowledge of the BRD-associated risk factors related to calf management and calves' condition will help to develop appropriate neonatal management practices at original farms and to identify calves at higher risk for BRD at nursery farms. In this study, the relationship between BRD and calf management practices (colostrum feeding, dam parity, serum total protein concentration at introduction (TP), body weight at introduction, introduction season, and daily average growth) was investigated using observational data from pre-weaned dairy calves introduced into a nursery farm in Hokkaido, Japan between 2014 and 2018 (n = 3185). Using additive Bayesian network (ABN) analysis, which is a multivariate statistical modelling approach, the direct and indirect associations between these factors were assessed. RESULTS: Colostrum feeding contributed to an increase in TP (correlation 1.02 [95 % CI, 0.94;1.10]), which was negatively associated with BRD directly (log odds ratio - 0.38 [- 0.46;-0.31]) and indirectly through increasing daily growth (correlation 0.12 [0.09;0.16]). Calves of multiparous dams had higher body weight at introduction (correlation 0.82 [0.74;0.89]), which indirectly reduced BRD risk through the increasing daily growth (correlation 0.17 [0.14;0.21]). Calves introduced during winter had the highest risk for BRD (log odds ratio 0.29 [0.15;0.44]), while those introduced in summer had the lowest risk (log odds ratio - 0.91 [- 1.06;-0.75]). The introduction season was also associated with BRD indirectly through dam parity, body weight at introduction, and daily growth. CONCLUSIONS: The following calf management practices are recommended for preventing BRD in pre-weaned calves at nursery farms: (1) encouraging colostrum feeding to neonatal calves at their original farms; and (2) identifying calves with higher BRD risk, i.e., those without feeding colostrum, born to primiparous cattle, with low body weight at introduction, and/or introduced in winter, and paying intensive attention to the calves for rapid detection of BRD. ABN analysis applied enabled us to understand the complex inter-relationships between BRD incidence and the risk factors, which will help to reduce BRD incidence and to rear healthy calves at nursery farms.

Region-wise analysis of dairy cow movements in Japan.

BACKGROUND: Animal movement is considered the most significant factor in the transmission of infectious diseases in livestock. A better understanding of its effects would help provide a more reliable estimation of the disease spread and help develop effective control measures. If the movement pattern is heterogeneous, its characteristics should be considered in epidemiological analyses, such as when using simulation models to obtain reliable outputs. In Japan, following the bovine spongiform encephalopathy epidemic, a traceability system for cattle was established in 2003, and the registration of all cattle movements in the national database began. This study is the first to analyze cattle movements in Japan. We examined regional and seasonal heterogeneity in dairy cow movements, which accounted for most Japanese breeding cattle. RESULTS: In the 14 years from April 2005 to March 2018, 4,577,709 between-farm movements of dairy cows were recorded, and the number of movements was counted by month and age for both inter- and intra-regional movements. As a result, two characteristic round-trip movements were observed: one was non-seasonal and inter-regional movements related to cattle-breeding ranches in Hokkaido (the northern region of Japan), which consists of the movement of cows around ages 6 to 8 and 21 to 23 months old. In addition, the seasonal movement of heifers for summer grazing within Hokkaido occurred in May and October at the peak ages of 13 to 14 and 19 to 20 months old, respectively. The observed heterogeneity seemed to reflect the suitability of raising the Holstein breed in Hokkaido and the shortage of supply of replacement heifers and available farming areas outside Hokkaido. CONCLUSIONS: Understanding the patterns of dairy cow movements will help develop reliable infectious disease models and be beneficial for developing effective control measures against these diseases.

Epidemiological analysis of classical swine fever in wild boars in Japan.

BACKGROUND: Classical swine fever (CSF) is a contagious disease of pigs and wild boars that is transmitted through direct/indirect contact between animals or CSF virus-contaminated fomites. When the disease re-emerged in 2018 in Japan, a CSF-infected wild boar was reported shortly after the initial pig farm outbreak; subsequently, the disease spread widely. To control the disease spread among wild boars, intensive capturing, fencing, and oral bait vaccination were implemented with concomitant virological and serological surveillance. This study aimed to describe the disease spread in the wild boar population in Japan from September 2018, when the first case was reported, to March 2020, based on the surveillance data. We conducted statistical analyses using a generalized linear mixed model to identify factors associated with CSF infection among wild boars. Moreover, we descriptively assessed the effect of oral bait vaccination, which started in March 2019 in some municipalities in the affected areas. RESULTS: We observed a faster CSF infection spread in the wild boar population in Japan compared with the CSF epidemics in European countries. The infection probability was significantly higher in dead and adult animals. The influence of the multiple rounds of oral bait vaccination was not elucidated by the statistical modeling analyses. There was a decrease and increase in the proportion of infected and immune animals, respectively; however, the immunization in piglets remained insufficient after vaccination for 1 year. CONCLUSIONS: Conditions regarding the wild boar habitat, including forest continuity, higher wild boar population density, and a larger proportion of susceptible piglets, were addressed to increase the infection risk in the wild boar population. These findings could improve the national control strategy against the CSF epidemic among wild boars.

First nationwide survey of the seroprevalence of Toxoplasma gondii in wild boars in Japan.

The increase in some wildlife species is of global concern. The utilization of wildlife meat as food and feed represents a potential source of food-borne pathogens; this is particularly a potential concern for the use of wild board as a food source in Japan. Regarding food safety and an animal infectious disease control, however, little is known about the infection level of zoonotic pathogens including Toxoplasma gondii (T. gondii) in wild boar populations in their natural habitats in Japan. A total of 1279 blood samples from 41 prefectures were collected from apparently healthy wild boars during the three hunting periods (September to February) of 2014-2015, 2015-2016, and 2017-2018. Out of these sera, 461 samples tested positive by a commercial indirect ELISA for T. gondii, and the total apparent and adjusted true seroprevalence were estimated to 36.0% (95% confidence interval [CI], 33.4-38.7) and 31.3% (95% CI, 33.1-38.9), respectively. The seroprevalence was significantly higher in yearlings and adults than in piglets (P < 0.05); however, no significant difference according to gender was noted. These results indicate the importance of adequate heating of wild boar meat before consumption to prevent transmission of T. gondii to humans. Furthermore, freezing meat for several days before cooking is recommended to reduce the risk of infection. In addition, although the incidence of toxoplasmosis in pig farms is relatively low in Japan, biosecurity measures against the felids and the varied intermediate hosts should be strengthened, especially, at the farms located in the wild boar habitats to prevent livestock infection.

Organic Anion-Transporting Polypeptide 1B1/1B3-Mediated Hepatic Uptake Determines the Pharmacokinetics of Large Lipophilic Acids: In Vitro-In Vivo Evaluation in Cynomolgus Monkey.

It is generally presumed that uptake transport mechanisms are of limited significance in hepatic clearance for lipophilic or high passive-permeability drugs. In this study, we evaluated the mechanistic role of the hepato-selective organic anion-transporting polypeptides (OATPs) 1B1/1B3 in the pharmacokinetics of compounds representing large lipophilic acid space. Intravenous pharmacokinetics of 16 compounds with molecular mass ∼400-730 Da, logP ∼3.5-8, and acid pKa <6 were obtained in cynomolgus monkey after dosing without and with a single-dose rifampicin-OATP1B1/1B3 probe inhibitor. Rifampicin (30 mg/kg oral) significantly (P < 0.05) reduced monkey clearance and/or steady-state volume of distribution (VDss) for 15 of 16 acids evaluated. Additionally, clearance of danoprevir was reduced by about 35%, although statistical significance was not reached. A significant linear relationship was noted between the clearance ratio (i.e., ratio of control to treatment groups) and VDss ratio, suggesting hepatic uptake contributes to the systemic clearance and distribution simultaneously. In vitro transport studies using primary monkey and human hepatocytes showed uptake inhibition by rifampicin (100 µM) for compounds with logP ≤6.5 but not for the very lipophilic acids (logP > 6.5), which generally showed high nonspecific binding in hepatocyte incubations. In vitro uptake clearance and fraction transported by OATP1B1/1B3 (ft,OATP1B) were found to be similar in monkey and human hepatocytes. Finally, for compounds with logP ≤6.5, good agreement was noted between in vitro ft,OATP1B and clearance ratio (as well as VDss ratio) in cynomolgus monkey. In conclusion, this study provides mechanistic evidence for the pivotal role of OATP1B-mediated hepatic uptake in the pharmacokinetics across a wide, large lipophilic acid space. SIGNIFICANCE STATEMENT: This study provides mechanistic insight into the pharmacokinetics of a broad range of large lipophilic acids. Organic anion-transporting polypeptides 1B1/1B3-mediated hepatic uptake is of key importance in the pharmacokinetics and drug-drug interactions of almost all drugs and new molecular entities in this space. Diligent in vitro and in vivo transport characterization is needed to avoid the false negatives often noted because of general limitations in the in vitro assays while handling compounds with such physicochemical attributes.

Epidemiology of Classical Swine Fever in Japan-A Descriptive Analysis of the Outbreaks in 2018-2019.

This study describes the epidemiological characteristics of classical swine fever (CSF) outbreaks in Japan. The first case was confirmed in September 2018, 26 years after the last known case. Outbreaks occurred on 39 farms, 34 commercial farms, and 5 non-commercial farms, between September 2018 and August 2019. In this study, a descriptive analysis was conducted of the epidemiological data on the characteristics of the affected farms, clinical manifestations, intra-farm transmission, association with infected wild boars, and control measures implemented on the farms. Twenty-eight of the 34 affected commercial farms were farrow-to-finish farms. It was assumed that the major risk factors were frequent human-pig interactions and the movement of pigs between farms. Fever and leukopenia were commonly observed in infected pigs. In 12 out of 18 farms where clinical manifestations among fattening pigs was the reason for notification, death was the most frequent clinical manifestation, but the proportion of dead animals did not exceed 0.5% of the total number of animals at most of the affected farms. Therefore, the clinical form of CSF in Japan was considered to be sub-acute. Twenty-three of the 29 farms (79%) with pigs at multiple stages (i.e., piglets, fattening pigs, and sows), had infection across the multiple stages. Many of these farms were within 5 km of the site where the first infected wild boars had been discovered, suggesting that infected wild boars were the source of infection. Infections still occurred at farms that had implemented measures at their farm boundaries to prevent the introduction of the virus into their farms, such as disinfection of vehicles and people, changing boots of the workers, and installation of perimeter fences. It is necessary to continue to strengthen biosecurity measures for farms located in areas with infected wild boars and to continue monitoring the distribution of infected wild boars so that any abnormalities can be reported and inspected at an early stage.