Experimental Evidence of the Diarrheal Activity of Sarcocystis sp. in Sika Deer (Cervus nippon).

Recently, the wild deer population has been increasing in Japan, causing serious feeding-related damage to the agricultural and forestry industries. In conjunction with the government's promotion of hunting for population control, the effective utilization of resources and promotion of the game meat industry as a sixth sector of industrialization are desired by local governments. However, several cases in which patients showed intestinal symptoms such as diarrhea due to the consumption of sika deer meat infected with protozoan Sarcocystis spp. have been reported, and the pathogenic microorganisms found in wild deer should be investigated. In this study, Sarcocystis sp. parasitized Kyushu sika deer (Cervus nippon nippon) in Nagasaki Prefecture, Japan, was examined for its enterotoxicity. A phylogenetic analysis based on the sequence of the 18S rRNA gene and cox1 showed that the species was highly homologous to Sarcocystis japonica and/or Sarcocystis sp. HM050622. We attempted to confirm the diarrhea-evoking toxicity of Sarcocystis sp. in sika deer meat, which has been previously reported in human case reports. A mouse ileal loop assay showed that Sarcocystis sp. in sika deer meat induced significant fluid accumulation in the loop at doses of ∼5 × 106 bradyzoites. Western blotting showed that these Sarcocystis parasites possess actin-depolymerizing factor, a diarrhea-evoking factor, similar to Sarcocystis fayeri, which exists in horsemeat. However, the pathogenic conditions of the ileal loop were different from those of similar experiments with S. fayeri. This study suggests that S. japonica parasitizing C. n. nippon may cause diarrhea via a different mechanism from that of S. fayeri.

Characteristics of people seeking consultation after progressing to severe dementia: A mixed-method analysis.

OBJECTIVES: This explores the characteristics of patients with worsening dementia who did not receive a specialized medical examination or care. METHODS: This study utilized a mixed methods analysis. Of the 2712 people who received the Mini Mental State of Examination (MMSE) at the Community Consultation Center for Citizens with MCI and Dementia between December 2007 and December 2019, 1413 people who scored 23 points or less were included. Participants were categorized into mild, moderate, and severe groups, based on their MMSE scores. Participants' characteristics-gender, age, presence or absence of an escort, demographics, family type, and presence or absence of a family doctor-were compared between the groups. To further understand the severe group's characteristics, clinical psychologists recorded consultation forms were categorized. RESULTS: More than 80% of the patients in each group had a family doctor. Moreover, all the severe groups had escorts, and the role of family members and supporters was important for the consultation. In the severe group, 29 patients had never received specialized medical care. Their characteristics were coded "non-existence" (fewer people or opportunities to notice their needs), "connection failure" (a lack of access or connections to consultations), and "evaluation failure" (not recognized as a problem requiring consultation). CONCLUSIONS: It is necessary to improve primary physician education, disseminate knowledge, and raise awareness about dementia, besides building and strengthening networks to alleviate the isolation of dementia patients and their families. The psychological aspects of family members' denial regarding their family members with dementia must be addressed through intervention.

Increased frequency of clindamycin-resistant Cutibacterium acnes strains isolated from Japanese patients with acne vulgaris caused by the prevalence of exogenous resistance genes.

Cutibacterium acnes, a resident bacterium of the skin, is a target for antimicrobial treatment of acne vulgaris, because it exacerbates inflammation. Recently, antimicrobial-resistant C. acnes strains have been isolated worldwide, and their prevalence has led to failure of antimicrobial treatment. This study aimed to analyze the antimicrobial resistance of C. acnes strains isolated from Japanese patients with acne vulgaris who visited the hospital and dermatological clinics between 2019 and 2020. Resistance rates to roxithromycin and clindamycin increased during 2019 to 2020 compared with those during 2013 to 2018. Additionally, the proportion of doxycycline-resistant and low-susceptibility strains (minimum inhibitory concentration [MIC] ≥8 µg/mL) increased. No difference in clindamycin resistance rates between patients with and without a history of antimicrobial use was observed during 2019 to 2020, which were significantly higher for patients with a history than for patients without a history during 2016 to 2018. The proportion of high-level clindamycin-resistant strains (MIC ≥256 µg/mL) gradually increased; particularly, the resistance rate was 2.5 times higher in 2020 than that in 2013. The proportion of strains showing high-level clindamycin resistance that also have the exogenous resistance genes erm(X) or erm(50), which confer high resistance, showed a strong positive correlation (r = 0.82). Strains with the multidrug resistance plasmid pTZC1 encoding erm(50) and tet(W) genes were frequent in clinic patients. Notably, most strains with erm(X) or erm(50) were classified as single-locus sequence types A and F (traditional types IA1 and IA2). Our data show that the prevalence of antimicrobial-resistant C. acnes is increasing in patients with acne vulgaris attributable to acquisition of exogenous genes in specific strains. To control the increasing prevalence of antimicrobial-resistant strains, it is important to select the appropriate antimicrobials while taking into consideration the latest information on resistant strains.

The Relationship Between Epidemiological Factors and the Parasite Load of Sarcocystis in Yezo-Deer (Cervus nippon yesoensis) in Hokkaido, Japan.

Several cases of gastrointestinal symptoms including diarrhea and vomiting due to the consumption of Sarcocystis-infected venison have been reported in Japan. However, the control of case incidence is difficult, as epidemiological information concerning Sarcocystis in venison in Japan is insufficient. We examined the prevalence and parasite load of Sarcocystis in 89 samples of Yezo-deer (Cervus nippon yesoensis) venison in Hokkaido by quantifying the copy numbers of the 18S rRNA gene of Sarcocystis, followed by a statistical analysis that considered the sampling area, age, and sex to clarify the parameters related to the parasite load. The copy numbers per gram of venison in samples ranged from 4.8 to 8.8 log. Wilcoxon rank-sum test, the one-way factorial analysis of variance (ANOVA), Steel-Dwass test, and a two-way factorial ANOVA showed significant differences in the copy numbers among sampling areas, not by age or sex, suggesting that the load of Sarcocystis in wild deer depended on the sampling area in Hokkaido. Notably, more than 80% of Hokkaido venison has a higher gene copy number than the meat that caused Sarcocystis fayeri-food poisoning. This information is expected to contribute to the establishment of hygiene standards for safe venison consumption and the control of gastrointestinal symptom cases due to consumption of Sarcocystis-infected venison.

Mite allergen avoidance decreases allergic symptoms in children in Ishinomaki city of Japan after natural disasters.

OBJECTIVE: We investigated the prevalence of asthma, rhinitis, and atopic dermatitis in children, evaluated the mite allergen levels in their bedding after the Great East Japan Earthquake, and assessed changes in allergic symptoms in children and their families after allergen avoidance practices. METHODS: We performed a survey for the International Study of Asthma and Allergies in Childhood (ISAAC) comprising 1109 children, aged 7-8 years, living in Ishinomaki, Japan. We collected responses from 464 children, and in 2016, measured the level of Dermatophagoides group 1 (Der 1) in the bedding of 202 of these children. The intervention group of children attended at least one allergen avoidance seminar. The levels of Der 1 in their bedding were measured, along with changes in allergic symptoms, in 17 children in 2017 and 14 children in 2018. The nonintervention group comprised children who did not attend an allergen avoidance seminar at any time. RESULTS: Of the 464 children who participated in the ISAAC, 50 (10.8%) reported having asthma, 179 (38.8%) allergic rhinitis, and 126 (27.3%) atopic dermatitis. The average level of Der 1 measured in the bedding of the 202 children in 2016 was 295.8 ng/m2. The levels of Der 1 in the intervention group-but not in the nonintervention group-significantly decreased in 2017 and 2018. The symptoms of asthma, allergic rhinitis, and atopic dermatitis in the children of intervention group and their families decreased after allergen avoidance practices. CONCLUSIONS: Allergen avoidance practices relieved allergic symptoms in school children after the Great East Japan Earthquake.

Mite allergen avoidance decreases allergic symptoms in children in Ishinomaki city of Japan after natural disasters

Objective We investigated the prevalence of asthma, rhinitis, and atopic dermatitis in children, evaluated the mite allergen levels in their bedding after the Great East Japan Earthquake, and assessed changes in allergic symptoms in children and their families after allergen avoidance practices.Methods We performed a survey for the International Study of Asthma and Allergies in Childhood (ISAAC) comprising 1109 children, aged 7–8 years, living in Ishinomaki, Japan. We collected responses from 464 children, and in 2016, measured the level of Dermatophagoides group 1 (Der 1) in the bedding of 202 of these children. The intervention group of children attended at least one allergen avoidance seminar. The levels of Der 1 in their bedding were measured, along with changes in allergic symptoms, in 17 children in 2017 and 14 children in 2018. The nonintervention group comprised children who did not attend an allergen avoidance seminar at any time.Results Of the 464 children who participated in the ISAAC, 50 (10.8%) reported having asthma, 179 (38.8%) allergic rhinitis, and 126 (27.3%) atopic dermatitis. The average level of Der 1 measured in the bedding of the 202 children in 2016 was 295.8 ng/m2. The levels of Der 1 in the intervention group—but not in the nonintervention group—significantly decreased in 2017 and 2018. The symptoms of asthma, allergic rhinitis, and atopic dermatitis in the children of intervention group and their families decreased after allergen avoidance practices (AU)

Mite avoidance decreased mite-specific IgE levels and ameliorated asthma symptoms in subjects who lived in temporary housing after natural disasters

Background: We previously reported an increased prevalence of asthma among patients who had lived in temporary housing after the 2011 Great East Japan Earthquake. We investigated the prognosis of asthma in former residents of temporary housing after allergen avoidance. Methods: Asthma was diagnosed in adults ≥ 15 years from 2014 to 2019 who had lived in temporary housing in Ishinomaki City for at least 1 year. The disease prognosis after the intervention of allergen avoidance in cases that were followed for more than 3 years during the 6-year study period was analyzed. We measured the Dermatophagoides farinae -specific immunoglobulin E (IgE) levels in serum, and the amount of Dermatophagoides group 1 (Der 1) antigen on their futons or mattresses. We instructed residents in an allergen avoidance strategy that included 32 tasks, including using microfiber bedding covers. Results: Of the 202 examinees who were followed for at least 3 years during the 6-year study period, 72(35.6%) were asthmatic during at least one examination. Of these 72 asthmatics, 55(76.4%) developed the disease after the earthquake, and more than half of the cases that we diagnosed at the examination were mild intermittent asthma. After the allergen-avoidance intervention, both the Der 1 level on the futons or mattresses of residents who were diagnosed with asthma but who were nonasthmatic at the final screening and their serum Der f-specific IgE levels significantly decreased (P < 0.01) at the final examination. Conclusion: Antigen avoidance ameliorated mild asthma that was prevalent among residents of temporary housing after the earthquake (AU)

Association between glycemic control and cardiovascular events in older Japanese adults with diabetes mellitus: An analysis of the Japanese medical administrative database.

AIMS/INTRODUCTION: The relationship between glycated hemoglobin (HbA1c) and cardiovascular events in older adults was investigated using a Japanese administrative medical database. MATERIALS AND METHODS: Anonymized medical data on patients with diabetes mellitus aged ≥65 years for the period from January 2010 to December 2019 were extracted from the EBM Provider database. The primary end-point was a composite of cardiovascular events, whereas the other end-points included severe hypoglycemia and fracture. The association between cardiovascular events and HbA1c at the index date (i.e., approximately 10 months after initial diabetes mellitus diagnosis) was evaluated using the Cox proportional hazards model. RESULTS: Among the 3,186,751 patients in the database, 3,946 older adults with diabetes mellitus were eligible for inclusion and were subsequently grouped according to HbA1c quartiles at the index date. Cardiovascular events occurred in 142 patients. Patients with HbA1c in the highest quartile had significantly higher risk of hospitalization for cardiovascular disease than those with HbA1c in the lowest quartile (hazard ratio 1.948; 95% confidence interval 1.252-3.031, P = 0.003). However, the events risk was similar across subgroups with HbA1c <7.2%. The incidence of hypoglycemia and fracture was not significantly associated with the level of glycemic control. CONCLUSIONS: Among older adults with diabetes mellitus, those with poor glycemic control were at higher risk for cardiovascular events compared with those with better glycemic control. However, strict glycemic control had no effect on cardiovascular risk in patients with HbA1c <7.2%.

Indoor Fungal Contamination in Temporary Housing after the East Japan Great Earthquake Disaster.

To understand fungal contamination in the indoor environment of the disaster region, a field survey was performed to measure the number of fungal counts and identify isolates in the indoor air of prefabricated temporary housing, privately independent-housing, and rented apartments flooded by the East Japan Great Earthquake disaster tsunami. As a result, the period with the highest detected fungal count was from the rainy season to summer in independent-housing and rented apartments. Moreover, in the temporary housing, the fungal number increased further in winter as indicated by the maximum fungal-number throughout the measurement period. The detection frequency of Aspergillus species was relatively higher in the indoor air of temporary housing than in typical housing in the non-disaster area. Since Aspergillus is known as an allergenic genus, it requires careful attention to the health risk for residents. The extremely high level of fungal condensation in indoor air possibly occurred due to high relative humidity and loss of heat insulation in the building attics. It is suggested that this problem commonly happened in the cold region including the entire disaster region of the East Japan Great Earthquake.

Association between Temporary Housing Habitation after the 2011 Japan Earthquake and Mite Allergen Sensitization and Asthma Development.

INTRODUCTION: We previously reported an increased prevalence of asthma in adults who lived in temporary housing after the 2011 Great East Japan Earthquake. The goal of the current study was to investigate changes in asthma prevalence and mite-specific immunoglobulin E (IgE) titers in temporary housing residents during 2014-2019. METHODS: By using the Global Initiative for Asthma guidelines, we diagnosed asthma in Ishinomaki city temporary housing residents aged 15 years or older. We then analyzed serum antigen-specific IgE levels to Dermatophagoides farinae (Der f), Dermatophagoides pteronyssinus (Der p), and Aspergillus fumigatus. RESULTS: The prevalence of asthma exceeded 20% across all age-groups throughout the study period. The proportion of study participants with a "positive" antigen-specific IgE titer (i.e., ≥0.35 IUA/mL) was higher in asthmatics than in nonasthmatics for Der f and Der p but not for Aspergillus fumigatus. Among residents ≥50 years old who were diagnosed with asthma, the percentage with a Der f-specific IgE titer ≥0.10 IUA/mL was higher than the proportion with ≥0.35 IUA/mL. Among study participants, asthma onset occurred before the earthquake, during residence in shelters or temporary housing, and (starting in 2016) after moving out of temporary housing. The Der p-specific IgE level was positively correlated with the duration of temporary housing (p < 0.05, r = 0.41) and inversely correlated with the time elapsed since moving out of temporary housing (p < 0.05, r = -0.35). CONCLUSION: Mite allergen sensitization was found in both asthmatic and nonasthmatic temporary housing residents after the 2011 Japan earthquake and tsunami; asthma developed even after subjects moved out of temporary housing.

Establishment of a Novel Molecular Detection Method for Sarcocystis in Venison.

Recently, horse meat (basashi) contaminated with Sarcocystis spp. caused food poisoning in Japan. An official detection method provided by the Ministry of Health, Labour and Welfare (MHLW), Japan, was designed to detect Sarcocystis fayeri to diagnose and control outbreaks of basashi food poisoning. In 2011, Sarcocystis-contaminated venison also caused food poisoning. However, the official MHLW detection method was not adequate for detecting Sarcocystis spp. in venison. In this study, we established a novel PCR-based detection method that amplifies 18S rRNA gene based on the conserved region of the sequence in 32 species of Sarcocystis for screening and quantification. Fifty venison samples from three areas in Hokkaido were examined by the MHLW method and the novel detection method. All samples were Sarcocystis spp.-positive. A sequence analysis indicated the presence of a species of Sarcocystis specific to sika deer (Cervus nippon), and not to horses. Another primer pair was designed for a quantitative real-time PCR assay to determine the copy number of the Sarcocystis-18S rRNA gene in parasitized venison. The melting curve analysis revealed high specificity of this assay. The calculated curve demonstrated that this quantitative PCR assay showed R2 value of 0.993 with 10-106 copies. Using this quantitative real-time PCR assay, the gene copy numbers were determined in 50 venison samples. The copy numbers of each sample ranged from 104 to 107 per gram. The copy numbers differed according to the area in Hokkaido. This indicates that the density of Sarcocystis spp. that infect Sika deer in Hokkaido is affected by the area. The novel screening and quantitative PCR method for Sarcocystis in venison was useful for collecting epidemiological information on Sarcocystis in wild Japanese sika deer, which will contribute to improve the safety of venison products in Japan.

Seroprevalence of fasciolosis in Hokkaido sika deer (Cervus nippon yesoensis) from Hokkaido Prefecture, Japan revealed by ELISA using recombinant cathepsin L1.

Fasciolosis, a zoonotic disease caused by liver flukes of the genus Fasciola, has been reported in Hokkaido (Yezo) sika deer (Cervus nippon yesoensis) in Hokkaido Prefecture, Japan; however, the actual seroprevalence in the animal has not been adequately evaluated. The objective of the present study was to analyze the seroprevalence of the disease among Hokkaido sika deer. Recombinant cathepsin L1 (rCatL1) was used as an antigen for an indirect enzyme-linked immunosorbent assay (ELISA) to detect antibodies against Fasciola flukes. The sensitivity and specificity of the ELISA were 84.6% and 100%, respectively. The average seroprevalence in 1109 Hokkaido sika deer from 20 locations in Hokkaido Prefecture was 43.9%. Mature deer showed higher seroprevalence than younger individuals; however, even younger animals may act as a reservoir for the disease. Monitoring infection levels in the Hokkaido sika deer population is important not only for the livestock industry, but also for preventing human fasciolosis.

Comprehensive Fungal Community Analysis of House Dust Using Next-Generation Sequencing.

Fungal community analyses in homes have been attracting attention because fungi are now generally considered to be allergens. Currently, these analyses are generally conducted using the culture method, although fungal communities in households often contain species that are difficult to culture. In contrast, next-generation sequencing (NGS) represents a comprehensive, labor- and time-saving approach that can facilitate species identification. However, the reliability of the NGS method has not been compared to that of the culture method. In this study, in an attempt to demonstrate the reliability of this application, we used the NGS method to target the internal transcribed spacer 1 (ITS1) in the fungal genome, conducted fungal community analyses for 18 house-dust samples and analyzed fungal community structures. The NGS method positively correlated with the culture method regarding the relative abundance of Aspergillus, Penicillium, Cladosporium and yeasts, which represent the major fungal components found in houses. Furthermore, several genera, such as Malassezia, could be sensitively detected. Our results imply that the reliability of the NGS method is comparable to that of the culture method and indicates that easily available databases may require modifications, including the removal of registrations that have not been sufficiently classified at the genus level.

Presence of Sarcocystis sybillensis in Japanese sika deer (Cervus nippon) captured in its native territory and its phylogenetic relationship with Sarcocystis nipponi.

The first study reporting the morphological characterization of Sarcocystis sybillensis was performed in 1983; however, without any molecular analysis. Sarcocystis nipponi has been recently described as a species synonymic to S. sybillensis. We reconfirmed the presence of S. sybillensis in Japanese sika deer (Cervus nippon) captured in its native territory; and performed its molecular and phylogenetic characterization. The morphological characteristics of the sarcocysts were consistent with those of S. nipponi and S. sybillensis described in the first report. However, the nucleotide sequence of 18S rRNA gene of S. sybillensis showed only 91.9% identity to that of S. nipponi, suggesting low homology among the concerned Sarcocystis spp. Accordingly, S. sybillensis was found to occupy a clade distinct from that of S. nipponi in a phylogenetic tree of Sarcocystis. Therefore, the present study provides essential information on 18S rRNA-based molecular characterization of S. sybillensis and disproves the existing notion of morphology-based species synonymity of S. sibillensis and S. nipponi. These results also suggest that S. sybillensis belongs to type 2 Sarcocystis.

PREVALENCE OF YERSINIA AMONG WILD SIKA DEER (CERVUS NIPPON) AND BOARS (SUS SCROFA) IN JAPAN.

We examined the prevalence of Yersinia, including pathogenic species such as Yersinia enterocolitica and Yersinia pseudotuberculosis, among wild sika deer (Cervus nippon) and boars (Sus scrofa) captured in Japan. The prevalence of Yersinia in the wild deer was 75% (207/277) and in the boars was 74% (40/54). A total of 417 isolates of nine Yersinia species were isolated from the animals examined: the largest number of isolates (48%, 200/417) were Y. enterocolitica biotype 1A. Pathogenic Y. enterocolitica 1B/O:8 were also isolated from two deer, and Y. pseudotuberculosis serogroups 3 and 4 were isolated from two boars and a deer, respectively. The pathogenic Y. enterocolitica 1B/O:8 isolates carried four virulence genes (ail, ystA, yadA, and virF), and Y. pseudotuberculosis serogroups 3 and 4 isolates carried three virulence genes (inv, yadA, and lcrF). Although the Y. enterocolitica 1B/O:8 and Y. pseudotuberculosis isolates were sensitive to almost all the antimicrobials tested, the two Y. enterocolitica 1B/O:8 isolates were resistant to azithromycin and ampicillin, and the three Y. pseudotuberculosis isolates were resistant only to azithromycin. These findings suggested that wild deer and boars might be important reservoirs for the agent causing human yersiniosis.

First report of Sarcocystis pilosa sporocysts in feces from red fox, Vulpes vulpes schrencki, in Hokkaido, Japan.

Sarcocysts of various Sarcocystis spp. are highly prevalent in wild sika deer, Cervus nippon yesoensis, in Hokkaido, Japan, and four species have been identified based on morphological and molecular characteristics: S. ovalis, S. pilosa, S. tarandi-like, and S. truncata-like. The definitive hosts of S. ovalis are corvids, but the hosts of the other species have not yet been identified. Aiming to determine the definitive hosts of these species, we collected 65 red fox (Vulpes vulpes schrencki) fecal samples in eastern Hokkaido and examined them for fecal sporocysts using a modified sucrose flotation method. One fecal sample contained typical Sarcocystis sporocysts, which were identified as S. pilosa based on 18S ribosomal RNA and cytochrome c oxidase subunit I gene sequences. This is the first identification of S. pilosa sporocysts in the wild. These findings indicate that red foxes serve as a definitive host of S. pilosa, and that red foxes constitute a source of S. pilosa infection for deer in Hokkaido.

Molecular characterization of three Sarcocystis spp. from wild sika deer (Cervus nippon yesoensis) in Hokkaido, Japan.

Diaphragm samples from 65 hunted sika deer (Cervus nippon yesoensis) from Hokkaido, Japan were examined for the presence of sarcocysts based on histological sections. Morphologically, the detected sarcocysts grouped into three types: (Type 1) 108.0-305.0 µm in width, thick-walled (4.3-7.0 µm) with tombstone-like protrusions; (Type 2) 25.0-69.5 µm in width, thick-walled (3.8-8.0 µm) with finger-like protrusions; and (Type 3) 22.5-55.0 µm in width, thin-walled (under 1 µm) with no visible protrusions under light microscopy. All samples contained at least one sarcocyst type, and multiparasitism was apparent in 58 samples. Morphologically, Type 1 sarcocysts were found in 19 (29.2%) samples, Type 2 in 62 (95.4%) samples, and Type 3 in 60 (92.3%) samples. The sarcocysts were collected using laser microdissection, the DNA extracted from them was PCR-amplified, and their 18S ribosomal RNA and cytochrome c oxidase subunit 1 genes were sequenced. Phylogenetic analysis showed that, for both genes, each morphological sarcocyst type (Types 1, 2, and 3) aligned most closely with S. silva/S. truncata, S. tarandi/S. elongata, and S. pilosa, respectively. Based on the sequence identities between taxa and the molecular information for sarcocysts in C. nippon centralis, the sarcocyst types were presumed to be S. truncata-like (Type 1), S. tarandi-like (Type 2), and S. pilosa (Type 3). The phylogenetic analyses based on the present comprehensive molecular characterization of three Sarcocystis spp. from C. nippon yesoensis in Hokkaido suggest that canids (e.g., wild foxes) may be the definitive hosts for S. pilosa, and felids (or unknown species) the definitive hosts for the other two species.

Yersinia infection in two captive guereza colobus monkeys (Colobus guereza).

Two guereza colobus monkeys (Colobus guereza) reared in a zoological garden in Japan suddenly died of multifocal fibrinonecrotic gastroenteritis and septicemia associated with infection by Yersinia spp. It was necessary to microbiologically differentiate Yersinia frederiksenii and Y. enterocolitica. We described the pathological findings and discuss the causal agent to emphasize the need to revert to using a combination of multiple examinations for diagnosis.

Effects of temperature, pH and curing on the viability of Sarcocystis, a Japanese sika deer (Cervus Nippon centralis) parasite, and the inactivation of their diarrheal toxin.

Recently, the Sarcocystis parasite in horse and deer meat has been reported to be a causative agent of acute food poisoning, inducing nausea, vomiting and diarrhea. Compared with other causative agents, such as bacteria, viruses and other parasites, in deer meat, the Sarcocystis species parasite, including its stability under various conditions, is poorly understood. In this study, we assessed the viability of Sarcocystis spp. and the activity of their diarrhea toxin (a 15-kDa protein) in deer meat under conditions of freezing, cold storage, pH change and curing. In addition, the heat tolerance was assayed using purified bradyzoites. The results showed that the species lost viability by freezing at -20, -30 and -80°C for <1 hr, heating at 70°C for 1 min, alkaline treatment (pH 10.0) for 4 days and addition of salt at 2.0% for <1 day. Immunoblot assays showed that the diarrhea toxin disappeared together with the loss of viability. However, the parasite survived cooling at 0 and 4°C and acidification (pH 3.0 and 5.0) for more than 7 days with the diarrhea toxin intact. These results provide useful information for developing practical applications for the prevention of food poisoning induced by diarrheal toxin of Sarcocystis spp. in deer meat during cooking and preservation.

The sensitivity of commercial kits in detecting the genes of pathogenic bacteria in venison.

The expansion of the wild deer population is a major problem for the Japanese farm and forestry industries because their damage to farm products and vegetation results in huge economic loss. To promote game meat consumption, hygiene inspections should be performed to detect main bacterial pathogens before products are shipped. In this study, we aimed to evaluate the ability of commercial test kits to genetically detect EHEC, Salmonella and Listeria monocytogenes in venison. Our results demonstrated that the kits for three pathogens could be useful for venison as well as other domestic meat products. Our comparative study showed that the LAMP kits were more sensitive than the RT-qPCR kits in the detection of all of these pathogens.