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Medical records of 16 Japanese Black calves with respiratory signs associated with perinatal rib fracture were analyzed, retrospectively, to speculate criteria of surgical intervention for the disease. For this analysis, the severity of respiratory condition was classified into three grades: Grade 1, with no wheezing, Grade 2, with wheezing after excitement but not at rest and Grade 3, with wheezing at rest. Grade 1 (n=3) received only conservative management. Seven of Grade 2 (n=8) and all of Grade 3 (n=5) had surgical management. Clinical outcome was good in all of Grade 1, and the surgical success rate was 83% in other groups. Our result suggests that surgical intervention should be considered for cases with more the severity of Grade 2.

Two-month-old (Case 1) and one-year-old (Case 2) Japanese Black cattle presented with bilateral rostral mandibular open fractures. At presentation, the proximal edges of each fracture were discolored and had a severe stench, indicating necrosis caused by infection. In both cases, a bilateral rostral mandibulectomy over the symphysis was performed. Although the tongues of both animals prolapsed post-surgery, they had no significant problems with eating or drinking. Case 1 showed excellent growth during fattening, and Case 2 successfully became pregnant and gave birth. Rostral mandibulectomy can be an effective surgical option for the treatment of cattle with difficulty in internal or external fixation due to unfavorable necrotic cranial mandibular open fractures.

Two veterinary personnel in Japan were infected with severe fever with thrombocytopenia syndrome virus (SFTSV) while handling a sick cat. Whole-genome sequences of SFTSV isolated from the personnel and the cat were 100% identical. These results identified a nosocomial outbreak of SFTSV infection in an animal hospital without a tick as a vector.

Severe fever with thrombocytopenia syndrome (SFTS) is an emerging tick-born disease and its animal-to-human transmission has come to attention recently. During our sero-survey of SFTS virus (SFTSV) among veterinary professionals in 2018, a veterinarian and his assistant working in an animal hospital were tested positive by enzyme-linked immunosorbent assay (ELISA). An additional survey implied a cluster of SFTS cases in which four more people, a family who brought two sick dogs to the animal hospital in 2003, were involved. This study aimed at assessing the possibility of animal-to-human transmission of SFTSV in this cluster.Retrospective interviews were performed with the owner family of the dogs and their clinical records were obtained from each hospital. SFTSV-IgG were tested by ELISA and virus neutralization test using the sera collected from them in 2018.The interviews revealed that a total of six people, the two veterinary professionals and the owner family who took care of the sick dogs, suffered from SFTS-like symptoms in the same period of time in 2003. All patients did not have tick bite before the onset and all suspected causative agents were excluded by laboratory tests. The serological tests in this study revealed the four owner family members were all positive for SFTSV antibodies.Considering the extremely low seroprevalence of SFTSV antibodies among inhabitants of the region, the existence of SFTSV antibodies in all these six people presents a possibility that they were involved in an SFTS outbreak originated in the sick dogs in 2003.

We isolated two pseudorabies virus (PRV) isolates (designated OT-1 and OT-2) from two hunting dogs exhibiting neurological manifestations after eating the flesh of wild boar hunted in Oita prefecture, Kyushu Island, Japan. The isolates corresponded to a previously reported PRV (MY-1 strain) isolated from a hunting dog in neighboring Miyazaki prefecture, and it clustered into genotype II based on the glycoprotein C sequence. Our results suggest that this common PRV strain may have been maintained in wild boars on Kyushu Island even though domestic pigs in this area have attained an Aujeszky's disease-free status.

Severe fever with thrombocytopenia syndrome (SFTS), caused by Dabie bandavirus, generally called SFTS virus (SFTSV), is an emerging zoonosis in East Asia. In Japan, 50-100 cases of SFTS have been reported each year since the first case was reported in 2013. SFTS is a tick-borne infectious disease, and SFTSV has been isolated from ticks in China and South Korea. Haemaphysalis longicornis and Amblyomma testudinarium are considered the primary vectors in Japan. However, the other tick species seldom feeding on humans might also play an important role in maintaining the virus in nature. In this study, we collected ticks on vegetation around the location where two SFTS patients were estimated to have been infected in Miyazaki Prefecture, Japan, isolated live SFTSV, and performed a phylogenetic analysis. A total of 257 ticks were collected, and SFTSV RNA was detected in 19.5% (9/46) of tick pools. A total of 10 infectious SFTSVs were successfully isolated from A. testudinarium, Haemaphysalis flava, Haemaphysalis formosensis, Haemaphysalis hystricis, and Haemaphysalis megaspinosa. Furthermore, the whole viral sequences isolated from ticks were highly homologous to sequences isolated from SFTS patients in the same sampling area in the past. These results suggest that SFTSVs are maintained in these tick species in the sampling area and sporadically transmitted to humans. Surveillance of SFTSV in ticks provides important information about the risk of incidental transmission to humans.

Severe fever with thrombocytopenia syndrome virus (SFTSV) is the causative agent of SFTS, an emerging tick-borne disease in East Asia, and is maintained in enzootic cycles involving ticks and a range of wild animal hosts. Direct transmission of SFTSV from cats and dogs to humans has been identified in Japan, suggesting that veterinarians and veterinary nurses involved in small-animal practice are at occupational risk of SFTSV infection. To characterize this risk, we performed a sero-epidemiological survey in small-animal-practice workers and healthy blood donors in Miyazaki prefecture, which is the prefecture with the highest per capita number of recorded cases of SFTS in Japan. Three small-animal-practice workers were identified as seropositive by ELISA, but one had a negative neutralization-test result and so was finally determined to be seronegative, giving a seropositive rate of 2.2% (2 of 90), which was significantly higher than that in healthy blood donors (0%, 0 of 1000, p &lt, 0.05). The seroprevalence identified here in small-animal-practice workers was slightly higher than that previously reported in other high-risk workers engaged in agriculture and forestry in Japan. Thus, enhancement of small-animal-practice workers’ awareness of biosafety at animal hospitals is necessary for control of SFTSV.

Osteochondrosis (OC) is not well recognized in cattle. Japanese Black (JB) cattle are valued for their beef and studies regarding OC are scarce. Hence, the aim of this study was to retrospectively discuss clinical and radiographical findings, diagnosis and treatment options for OC in JB cattle for successful outcomes. Medical records of 11 calves affected with OC were reviewed, retrospectively. Clinical and radiographic findings, treatment and outcomes and postmortem examination findings were analyzed. All calves had lameness. Stifle (4/14) and tarsal (4/14) were the most affected joints. Subchondral bone lucencies were observed in all joints (14/14) and accompanied with sclerosis (10/14). Radiographic grading was performed in calves. Calves were treated surgically (8/11) or conservatively (1/11). Surgically treated calves were auctioned (5/8). Postmortem examination was performed (2/11). Gross findings revealed ulcerative lesion at articular surface. Histopathologically, cartilage layer was defected at lesion. Inflammation (2/2) and granulation tissue (1/2) was also observed. In conclusion, OC in JB cattle can be diagnosed on the basis of radiography with history and clinical signs in clinical setting. Surgery should be performed in mild cases for successful outcomes. A modified radiographic grading criteria for clinical cases may hold potential in evaluating prognosis and outcomes.

In a two-year survey of the 24 Japanese broiler chicken flocks at 9 farms from 2013 to 2014, C. jejuni/C. coli prevalence was assessed in a total of 131 slaughtered broiler chicken cecal samples by conventional culture methods and loop-mediated isothermal amplification (LAMP) assay. While 93 samples were C. jejuni / C. coli -negative, 38 (29.0%) showed Campylobacter loads of between 6.4 and 9.0 log CFU/g of ceca in conventional culture methods. The performance of LAMP assay was 100% accurate in terms of diagnostic sensitivity (38/38), specificity (93/93). Furthermore, LAMP assay enabled direct screening of C. jejuni and C. coli in cecal samples from broiler chicken chickens as rapid and cost-effective detection within 90 min and less than 1 US dollar, which can help monitor release of Campylobacter -contaminated chicken into the food chain, thereby reducing the incidence and public health risk of campylobacteriosis. Seasonal changes in C. jejuni and C. coli prevalence in broiler chicken ceca were significantly correlated with the frequency of food poisoning incidents caused by these bacteria in Japan.

The bovine MHC (BoLA) class II DRB3 alleles are associated with polyclonal expansion of lymphocytes caused by bovine leukemia virus (BLV) infection in cattle. To examine whether the DRB3*0902 allele, one of the resistance-associated alleles, is associated with the proviral load, we measured BLV proviral load of BLV-infected cattle and clarified their DRB3 alleles. Fifty-seven animals with DRB3*0902 were identified out of 835 BLV-infected cattle and had significantly lower proviral load (P

To understand how the latest dominant bovine leukemia virus (BLV) strains were introduced and spread in the Miyazaki prefecture, we collected blood samples from 3 geographic areas (north, central and south) and carried out sequence analysis of the BLV env gene. Two genotypes, genotype I, and III, were identified and the majority of the strains belonged to genotype I (71/74). To clarify a route of BLV introduction, we divided the strains into 20 subgenotypes based on their nucleotide sequences and performed phylogenetic analysis. Our study indicated that common BLV strains were comparatively evenly distributed even in the area, where the farmers have not introduced cattle from other areas and the cattle have limited exposure to BLV infection in grazing fields.

How pathogens evolve their virulence to humans in nature is a scientific issue of great medical and biological importance. Shiga toxin (Stx)–producing Escherichia coli (STEC) and enteropathogenic E. coli (EPEC) are the major foodborne pathogens that can cause hemolytic uremic syndrome and infantile diarrhea, respectively. The locus of enterocyte effacement (LEE)–encoded type 3 secretion system (T3SS) is the major virulence determinant of EPEC and is also possessed by major STEC lineages. Cattle are thought to be the primary reservoir of STEC and EPEC. However, genome sequences of bovine commensal E. coli are limited, and the emerging process of STEC and EPEC is largely unknown. Here, we performed a large-scale genomic comparison of bovine commensal E. coli with human commensal and clinical strains, including EPEC and STEC, at a global level. The analyses identified two distinct lineages, in which bovine and human commensal strains are enriched, respectively, and revealed that STEC and EPEC strains have emerged in multiple sublineages of the bovine-associated lineage. In addition to the bovine-associated lineage-specific genes, including fimbriae, capsule, and nutrition utilization genes, specific virulence gene communities have been accumulated in stx- and LEE-positive strains, respectively, with notable overlaps of community members. Functional associations of these genes probably confer benefits to these E. coli strains in inhabiting and/or adapting to the bovine intestinal environment and drive their evolution to highly virulent human pathogens under the bovine-adapted genetic background. Our data highlight the importance of large-scale genome sequencing of animal strains in the studies of zoonotic pathogens.

Aflatoxins are toxic by-products of fungi contaminating maize and other crops; they can be carried over into milk, meat and eggs when livestock eat aflatoxin-contaminated feed or fodder. People who consume such animal products are exposed to the toxins. To assess the aflatoxin contamination status in marketed raw milk and associated risk factors in peri-urban Nairobi, we conducted a census of raw milk retailers in Dagoretti Division, Nairobi, Kenya. Structured questionnaires were filled in by face-to-face interviews with all retailers who agreed to participate in this study. Small portions of milk were purchased from each respondent and tested for aflatoxin M1 (AFM1) using competitive enzyme-linked immunosorbent assay (cELISA) tests. Geographic coordinates of each eligible retailer were recorded. A total of 350 milk retailers were visited and 344 were plotted on a map. Of these, 250 retailers answered the questionnaire and a milk sample from each of 200 retailers underwent cELISA analysis for AFM1. Four types of businesses were found: kiosks (71%), dairy shops (21%), street or mobile vendors (3%) and grocery stands (1%); for 4% the business type was not identified. Milk was mainly sourced directly from dairy farms (59%) or from intermediate distributors (35%). Most retailers sold less than 20 litres per day of raw milk. The mean daily milk consumption of the milk retailers’ households was 940 ml for adults and 729 ml for children. Although 58% of retailers had heard about aflatoxins and the majority of them agreed that aflatoxins could be present in milk, only 29% believed that “milk safety cannot be solely judged by sight or taste” and only 6% that “milk is not completely safe even after boiling”. The cELISA test found the mean concentration of AFM1 was 128.7 parts per trillion (ppt) (median=49.9; 95% confidence interval=3.0–822.8) with a maximum of 1675 ppt. Overall, 55% of samples exceeded the European Union maximum level of 50 ppt and 6% exceeded the recommended maximum level of the United States Food and Drug Administration of 500 ppt. Compared to milk from street vendors, a significantly higher amount of AFM1 was detected in milk from kiosks and dairy shops, especially when the milk was sourced from farms without an intermediate distributor. Our findings indicate the need to better understand and manage aflatoxin in milk in Nairobi. Keywords: Aflatoxin M1, Kenya, milk, dairy value chain, milk retailers, Dagoretti, mycotoxin, informal milk marketing

The European Community's (EC) Key, which is also called Bendixen's Key, is a well-established bovine leukemia virus (BLV) diagnostic method that classifies cattle according to the absolute lymphocyte count and age. The EC Key was originally designed for dairy cattle and is not necessarily suitable for Japanese Black (JB) beef cattle. This study revealed the lymphocyte counts in the BLV-free and -infected JB cattle were significantly lower than those in the Holstein cattle. Therefore, applying the EC Key to JB cattle could result in a large number of undetected BLV-infected cattle. Our proposed hematological key, which was designed for JB cattle, improves the detection of BLV-infected cattle by approximately 20%. We believe that this study could help promote BLV control.

A key virulence factor of enterohaemorrhagic Escherichia coli (EHEC) is the bacteriophage-encoded Shiga toxin (Stx). Stxs are classified into two types, Stx1 and Stx2, and Stx2-producing strains are thought to cause more severe infections than strains producing only Stx1. Although O26 : H11 is the second most prevalent EHEC following O157 : H7, the majority of O26 : H11 strains produce Stx1 alone. However, Stx2-producing O26 strains have increasingly been detected worldwide. Through a large-scale genome analysis, we present a global phylogenetic overview and evolutionary timescale for E. coli O26 : H11. The origin of O26 has been estimated to be 415 years ago. Sequence type 21C1 (ST21C1), one of the two sublineages of ST21, the most predominant O26 : H11 lineage worldwide, emerged 213 years ago from one of the three ST29 sublineages (ST29C2). The other ST21 lineage (ST21C2) emerged 95 years ago from ST21C1. Increases in population size occurred in the late 20th century for all of the O26 lineages,but most remarkably for ST21C2. Analysis of the distribution of stx2-positive strains revealed the recent and repeated acquisition of the stx2 gene in multiple lineages of O26, both in ST21 and ST29. Other major EHEC virulence genes, such as type III secretion system effector genes and plasmid-encoded virulence genes, were well conserved in ST21 compared to ST29. In addition, more antimicrobial-resistance genes have accumulated in the ST21C1 lineage. Although current attention is focused on several highly virulent ST29 clones that have acquired the stx2 gene, there is also a considerable risk that the ST21 lineage could yield highly virulent clones. KEYWORDS: Shiga toxin; antimicrobial-resistance gene; enterohaemorrhagic Escherichia coli; genome evolution; population structure PMID: 29208163 PMCID: PMC5729918 DOI: 10.1099/mgen.0.000141, A key virulence factor of enterohaemorrhagic Escherichia coli (EHEC) is the bacteriophage-encoded Shiga toxin (Stx). Stxs are classified into two types, Stx1 and Stx2, and Stx2-producing strains are thought to cause more severe infections than strains producing only Stx1. Although O26 : H11 is the second most prevalent EHEC following O157 : H7, the majority of O26 : H11 strains produce Stx1 alone. However, Stx2-producing O26 strains have increasingly been detected worldwide. Through a large-scale genome analysis, we present a global phylogenetic overview and evolutionary timescale for E. coli O26 : H11. The origin of O26 has been estimated to be 415 years ago. Sequence type 21C1 (ST21C1), one of the two sublineages of ST21, the most predominant O26 : H11 lineage worldwide, emerged 213 years ago from one of the three ST29 sublineages (ST29C2). The other ST21 lineage (ST21C2) emerged 95 years ago from ST21C1. Increases in population size occurred in the late 20th century for all of the O26 lineages, but most remarkably for ST21C2. Analysis of the distribution of stx2-positive strains revealed the recent and repeated acquisition of the stx2 gene in multiple lineages of O26, both in ST21 and ST29. Other major EHEC virulence genes, such as type III secretion system effector genes and plasmid-encoded virulence genes, were well conserved in ST21 compared to ST29. In addition, more antimicrobial-resistance genes have accumulated in the ST21C1 lineage. Although current attention is focused on several highly virulent ST29 clones that have acquired the stx2 gene, there is also a considerable risk that the ST21 lineage could yield highly virulent clones.

Horizontal transmission is recognized as a major infection route for bovine leukemia virus (BLV), and cattle with high viral loads are considered to be a major infectious source in a herd. However, a correlation between viral loads and the risk of infection has been insufficient to use as a foundation for BLV control strategies. In this report, we examined the epidemiology of BLV infection and the infectious source in a local area. In 2013-2014, BLV infection was investigated in 1,823 cattle from 117 farms in two adjacent districts, Miyazaki, Japan. Seropositive samples for BLV were detected with 88 cattle and in 14 farms. Phylogenetic analysis revealed that 94% of the isolates clustered into genotype I and the remaining isolate into genotype III. Among genotype I, genetically distinct strains were spread at each farm, and cattle infected with less than 3 copies/100 cells did not transmit BLV to other cattle for more than thirty months. This is the first report of concrete data of viral load in relation to viral horizontal transmission under the field condition. The data facilitate farmers and veterinarians understanding the status of BLV infected cattle. This research contributes to BLV infection control and the development of effective BLV eradication programs.

To understand how the latest dominant bovine leukemia virus (BLV) strains were introduced and spread in the Miyazaki prefecture, we collected blood samples from 3 geographic areas (north, central and south) and carried out sequence analysis of the BLV env gene. Two genotypes, genotype I, and III, were identified and the majority of the strains belonged to genotype I (71/74). To clarify a route of BLV introduction, we divided the strains into 20 subgenotypes based on their nucleotide sequences and performed phylogenetic analysis. Our study indicated that common BLV strains were comparatively evenly distributed even in the area, where the farmers have not introduced cattle from other areas and the cattle have limited exposure to BLV infection in grazing fields.

An epidemiological study and control trial were conducted to assess taeniid infection in farm dogs in Qinghai Province, China. To improve egg detection by fecal examination, a deworming step with praziquantel was incorporated into the sampling methodology. As a result, a marked increase in the number of egg-positive samples was observed in samples collected at 24 hr after deworming. Then, the fecal examination and barcoding of egg DNA were performed to assess the prevalence of taeniid species in dogs from Xinghai, Haiyan, Gangcha and Chengduo counties. Analysis of 277 dog feces revealed that taeniid cestodes, including Taenia spp. and Echinococcus granulosus, were highly prevalent in Xinghai (34.4%), but eggs were not found in Haiyan where a control trial on canine echinococcosis had been conducted 20 years previously. A control trial involving the administration of 5–10 mg/kg praziquantel to 90 farm dogs at 45-day intervals was conducted in Xinghai. The prevalence of taeniid cestodes in the dogs was reduced to 9.6% and 4.9% after one and two years, respectively, indicating that some dogs were not administered praziquantel properly. A questionnaire survey of farmers in Xinghai and Haiyan revealed that most farmers in Xinghai were not familiar with echinococcosis or the transmission route of the disease, while most farmers in Haiyan had a more thorough understanding of the disease. The findings implied that a program for educating local farmers would be important for efficiently controlling canine taeniid infection in the region.

Infection with Moniezia benedeni is sometimes found in confined cattle in Japan. Between October 2011 and January 2013, we monitored the fecal egg prevalence at a confined cattle farm in Miyazaki prefecture where continuous M. benedeni infection has been recognized for years to evaluate the possible infection routes. Fecal egg prevalence changed seasonally with the highest in October 2011 (27.3%: 9/33). This was followed by a gradual decrease until July 2012 (9.4%: 3/32) and then an increase between August to December 2012 when new egg-excreting cases were observed. The pattern of seasonal changes was similar to that reported previously for cattle kept in a barn with an outside playing yard. Although M. benedeni-infected mites were not found, we constantly detected an oribatid mite, Oribatula sakamorii Aoki, 1970, in the litter of cattle bedding from May to October 2012. This species belongs to a genus which has been reported to be a suitable intermediate host for M. benedeni, suggesting that M. benedeni infection may have been autonomously maintained at the farm via oribatid mites living in the cowshed. When infected cattle were treated with praziquantel, it was found that a single oral inoculation with a dose of 5 mg/kg was effective for deworming.

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.

2.3-month-old (Case 1), one-month-old (Case 2) and 6-month-old (Case 3), Japanese Black calves presented with mild to severe wheezing. All calves had histories of dystocia at birth with breech presentation. Physical examination, thoracic radiography, endoscopy or computed tomography indicated wheezing associated with tracheal collapse and stenosis caused by perinatal rib fractures. Partial resection of the fractured first and second ribs was performed on all calves. The respiration in Cases 1 and 2 immediately improved after the surgery, while Case 3 required two weeks to improve. Cases 1 and 3 grew up healthy and were sold at auction, but Case 2 had a recurrence of wheezing at three months post-discharge and showed growth retarding. Partial costectomy may be an effective solution for control of respiration, however, further cases are required to discuss the criteria for surgical management and to obtain favorable postoperative prognosis in calves with tracheal collapse and stenosis caused by perinatal rib fractures.

Aflatoxin M1 (AFM1), a human carcinogen, is found in milk products and may have potentially severe health impacts on milk consumers. We assessed the risk of cancer and stunting as a result of AFM1 consumption in Nairobi, Kenya, using worst case assumptions of toxicity and data from previous studies. Almost all (99.5%) milk was contaminated with AFM1. Cancer risk caused by AFM1 was lower among consumers purchasing from formal markets (0.003 cases per 100,000) than for low-income consumers (0.006 cases per 100,000) purchasing from informal markets. Overall cancer risk (0.004 cases per 100,000) from AFM1 alone was low. Stunting is multifactorial, but assuming only AFM1 consumption was the determinant, consumption of milk contaminated with AFM1 levels found in this study could contribute to 2.1% of children below three years in middle-income families, and 2.4% in low-income families, being stunted. Overall, 2.7% of children could hypothetically be stunted due to AFM1 exposure from milk. Based on our results AFM1 levels found in milk could contribute to an average of &minus, 0.340 height for age z-score reduction in growth. The exposure to AFM1 from milk is 46 ng/day on average, but children bear higher exposure of 3.5 ng/kg bodyweight (bw)/day compared to adults, at 0.8 ng/kg bw/day. Our paper shows that concern over aflatoxins in milk in Nairobi is disproportionate if only risk of cancer is considered, but that the effect on stunting children might be much more significant from a public health perspective, however, there is still insufficient data on the health effects of AFM1.

Bovine abortion, diarrhea, and respiratory disease complexes, caused by infectious agents, result in high and significant economic losses for the cattle industry. These pathogens are likely transmitted by various vectors and reservoirs including insects, birds, and rodents. However, experimental data supporting this possibility are scarce. We collected 117 samples and screened them for 44 bovine abortive, diarrheal, and respiratory disease complex pathogens by using Dembo polymerase chain reaction (PCR), which is based on TaqMan real-time PCR. Fifty-seven samples were positive for at least one pathogen, including bovine viral diarrhea virus, bovine enterovirus, Salmonella enterica ser. Dublin, Salmonella enterica ser. Typhimurium, and Neospora caninum; some samples were positive for multiple pathogens. Bovine viral diarrhea virus and bovine enterovirus were the most frequently detected pathogens, especially in flies, suggesting an important role of flies in the transmission of these viruses. Additionally, we detected the N. caninum genome from a cockroach sample for the first time. Our data suggest that insects (particularly flies), birds, and rodents are potential vectors and reservoirs of abortion, diarrhea, and respiratory infectious agents, and that they may transmit more than one pathogen at the same time.

A seroepidemiological survey for paragonimosis among boar-hunting dogs was conducted throughout central and southern Kyushu, Japan. Among a total of 224 sera collected from boar-hunting dogs kept by 38 owners, 147 (65.6%) were positive for IgG antibodies against Paragonimus westermani antigen by ELISA. In the 147 seropositive dogs, 83 (56.5%) dogs demonstrated high antibody titers (OD>0.700), which appeared to indicate active infection. There were no obvious differences noted in age, sex and the genetic background of the dogs. The seroprevalence was dependent on the feeding manner of the owners and was extremely variable (0-100%). The majority (34/38) of owners fed their dogs uncooked boar meat and/or allowed them free-access to the residues of hunted boars after dissection, resulting in dogs with seropositive results. The dogs that were never fed wild boar meat were entirely seronegative. These results clearly demonstrate that boar-hunting dogs play an important role as a definitive Paragonimus host and that wild boars serve as a reservoir host for the maintenance of the Paragonimus life-cycle, presumably P. westermani, in the mountainous areas of central and southern Kyushu, Japan.

A 5-year-old male mixed-bred boar-hunting dog with a Plott hound background weighing 23 kg was brought to the Veterinary Teaching Hospital, University of Miyazaki, in October 2002. The dog was diagnosed with active infection with the lung fluke Paragonimus westermani by serological testing and also by detection of parasite eggs in his feces. Subsequent examination of four other dogs working with this dog as a boar-hunting team revealed that all five dogs were infected with P. westermani.

For 6 years, 5 Japanese Black cows of the same herd showed anorexia, depression, and dehydration with no feces in the rectum. Biomedical examination of 3 animals showed severe hypokalemia and hypochloremia. Although the first 3 animals died or were slaughtered (causes unknown), necropsy results showed that the cow in case 4 had intestinal obstruction due to phytobezoar derived from napier grass, fed mainly to the cattle as roughage. Therefore, farmers were recommended to avoid the hard root-stem portion of napier grass as roughage. Consequently, less phytobezoar was recovered from the fifth cow, and no similar clinical case of intestinal obstruction was observed thereafter. This is the first report on intestinal obstruction caused by phytobezoars derived from napier grass.

We recently found a group of 5 boar-hunting dogs infected with Paragonimus westermani. As wild boars are known to be the potential paratenic hosts for this parasite, boar-hunting dogs have obviously a high risk of infection by this parasite. In the present study, therefore, we investigated 20 dogs of another group kept by a hunter in Miyazaki Prefecture, Japan, in order to determine whether paragonimosis is a common problem among boar-hunting dogs. The results showed that P. westermani eggs were present in the feces of 10 out of 20 dogs, while 17 dogs were seropositive on ELISA. Taken together with our previous results, it appears that paragonimosis is a serious problem in boar-hunting dogs. The possible risks of infected dogs acting as a source for maintaining the P. westermani life cycle are also discussed.

A coprological survey with detailed clinical observation of naturally occurring haemorrhagic enteritis (HE) cases was conducted to understand the pathophysiology of HE by clarifying the infection status of Eimeria and enteropathogenic bacteria in cattle. Faecal samples from 55 cases of HE and 26 clinically normal animals were collected, and a quantitative examination of Eimeria and potential enteropathogenic bacteria was performed. The number of Eimeria species oocysts per gram of faeces (OPG) exceeded 10,000 in 69.1 per cent of HE cases with a maximum of 1,452,500 OPG and Eimeria zuernii was found to be overwhelmingly dominant. A significant increase in faecal coliform count was observed in HE cases compared with clinically normal animals. Among the animals shedding >10,000 OPG, 42.9 per cent showed a remarkable increase in Clostridium perfringens abundance (>104 CFU/g) in the faeces. In the cases with C. perfringens detected, its abundance was positively correlated with Eimeria OPG and high C. perfringens abundance was always accompanied by high Eimeria OPG. E. zuernii is likely to play a crucial role in massive multiplication of C. perfringens in HE in cattle.

The perinatal transmission of bovine leukaemia virus (BLV) plays a critical role in the spread and persistence of BLV infection in cattle herds. The purpose of this study was to examine the frequency of perinatal infections in an area in Japan and investigate some risk factors associated with infection. Altogether, 129 calves born to BLV-infected cows in a herd in Japan were tested for infection immediately after birth and again at one month of age using nested PCR. Twenty-four calves (18.6 per cent) were infected with BLV, of which 14 (10.8 per cent) and 10 (7.7 per cent) calves were infected via the transplacental and the birth canal routes, respectively. Maternal viral loads, breed, the presence or absence of assistance during parturition and the number of births per dam were evaluated to investigate risk factors associated with infection. Maternal viral load was significantly correlated with the frequency of perinatal infection, and more than 40 per cent of newborn calves born to dams with high viral loads were infected with BLV. The results of this study could contribute towards developing effective eradication programmes by providing necessary data for replacement of breeding cow in the field.

With the rise of the One Health paradigm, ethicists have called for new research approaches, considering the interdependent relationships of humans, animals, and their environment. These relationships can be particularly complex within resource-poor, smallholder livestock systems, necessitating a rigorous informed-consent process. Little has been published on informed consent beyond human-subject research. This paper outlines two studies on informed consent, for research identifying diseases of animal and human importance, within smallholder livestock value chains.Firstly, a randomized independent-group study compared three communication tools (written, cartoons, and photographs) for informing 22 Tanzanian livestock-keepers before seeking their consent. A significant difference in comprehension and engagement in the informed-consent process was found between tools, and cartoons had the highest (i.e. best combined comprehension and engagement) scores. Most (21 out of 22) farmers answered half or more the questions correctly, but none were able to answer all questions. Comprehension testing allowed identification of common misunderstandings, such as immediate benefits the farmers would receive and the process to be used for relaying research results. Dialogue stimulated by cartoons and photographs allowed researchers to determine and respond to participants’ varied relationships with their livestock.The second study assessed preferred methods for indicating consent among informal-sector milk vendors in Nairobi, Kenya. Of consenting participants, 61% (140/230) indicated consent verbally, 39% (90/230) signed consent and none chose thumbprint. There was a significant enumerator-effect on both overall consent and the methods chosen.Several of these findings echo those published in human-medical research. Additionally, highlighted here is the importance of facilitating dialogue during the informed-consent process in One Health research, for a more nuanced understanding of relationships between humans, animals, and their environment. Also discussed is how a requirement to sign consent forms might limit consent among workers in informal markets, which are commonly studied in One Health research. We suggest expansion of these, and development of further, studies towards improving consent processes in One Health research.