Bacteriological Survey of Insect Products in Japan.

A microbiological study was conducted on 41 insect product samples (29 raw frozen [21 domestic and 8 imported], 10 powdered, and 2 processed), which were commercially available in Japan. The total aerobic count for raw frozen insects was 5.61 log cfu/g (range: 2.52-8.40), whereas the powdered insect count was 2.89 log cfu/g (range: 1.00-4.57). The bacterial count was significantly higher in raw frozen insects (p < 0.05). The coliform count for the raw frozen insects ranged from <1 to 6.90 log cfu/g, and that for the powdered insects ranged from <1 to 1.00 log cfu/g. The number of samples with values above the detection limit was significantly higher in raw frozen insects (p < 0.05). The detection frequencies of aerobic spores (<1-4.63 log cfu/g), anaerobic spores (<0-4.40 log cfu/g), and Bacillus cereus (<1.7-3.83 log cfu/g) showed no sample type-related significant difference. Listeria spp. was isolated from four samples of raw frozen insects, one of which was Listeria monocytogenes. We did not detect any of the following: Salmonella spp., Shiga toxin-producing E. coli (STEC), Campylobacter jejuni/coli, or pathogenic Yersinia. We isolated insect products retailed in Japan harboring food poisoning bacteria, including L. monocytogenes and B. cereus. In particular, raw frozen products displayed high levels of hygienic indicator bacteria.

Environmental Monitoring of Food Manufacturing Facilities for Listeria: A Case Study.

Environmental monitoring programs (EMPs) for food production facilities are useful for verifying general sanitation controls and are recommended as verification measures to ensure that the Hazard Analysis Critical Control Point plan is working effectively. In this study, EMPs for Listeria were conducted at three food production facilities to assess the efficacy of sanitation control and establish effective sanitation control methods. In Facility A, L. monocytogenes was detected in the clean area although in Zone 3, non-food-contact surfaces. To prevent contamination from dirty areas, the cleaning practices in the preparation room were investigated. Normal cleaning combined with disinfection with carbonated hypochlorite water (chlorine concentration, 150 ppm) proved effective. At Facility B, a salad product and its ingredients (pastrami and salami) were positive for L. monocytogenes serotype 3b. The bacterial count was <10/g in all samples. However, when inoculated with L. monocytogenes isolates, the growth of approximately 2 log cfu/g was observed on pastrami after 48 h of incubation at 10°C. The ingredients were commercially purchased blocks that were sliced in a slicer at Facility B and used as salad toppings. Because both unopened blocks were negative for L. monocytogenes, contamination of the slicer was suspected. Sampling of the slicer revealed that contamination by L. monocytogenes serotype 3b was more extensive after use than before use. Therefore, the slicer was disassembled, cleaned, and disinfected thoroughly. In Facility C, L. monocytogenes serotype 4b (4e) was detected in all the dirty, semiclean, and clean areas. The strain was also isolated from the wheels of a smoking cart transported across the zones. Therefore, efforts were made to frequently clean and disinfect the cart. EMPs revealed the presence of Listeria in each facility and allowed remedial measures to be undertaken. Continued monitoring and Plan-Do-Check-Act cycles were considered desirable.

Bacterial Distribution and Community Structure in Beef Cattle Liver and Bile at Slaughter.

ABSTRACT: In this study, the distribution of hygienic indicator bacteria in cattle livers and bile was examined at slaughterhouses. One hundred twenty-seven cattle livers with gallbladders were carefully eviscerated from carcasses at 10 slaughterhouses. Microbiological examination revealed that nine bile samples (7.1% prevalence) and 19 liver parenchyma samples (15.0% prevalence) were positive for Enterobacteriaceae (EB) with means ± standard deviations of 3.68 ± 4.63 log CFU/mL and 1.59 ± 2.47 log CFU/g, respectively; thus, bacterial contamination was apparent even at the postevisceration stage. Subsequently, 70 cattle livers were obtained at the postprocessing and storage stage from 7 of the 10 slaughterhouses. Microbiological analysis revealed significantly higher levels of EB in the liver parenchyma (3.00 ± 3.89 log CFU/g, P = 0.011) than those at the postevisceration stage, suggesting that bacterial dissemination and/or replication occurred in the liver parenchyma during processing and storage. According to 16S rRNA ion semiconductor sequencing analysis of representative samples from 12 cattle, Proteobacteria, Firmicutes, and Actinobacteria were dominant in both the parenchyma and bile in which EB and Escherichia coli were predominant among livers with higher EB levels. These results suggest that bile plays a role as a vehicle for bacterial transmission to the liver parenchyma. This study is the first to evaluate bacterial distribution and community structure in the liver and biliary microecosystem of cattle at slaughter. Our data support the use of EB testing of bile to screen cattle livers contaminated with high levels of fecal indicator bacteria.

Neurovascular Interactions in the Development of the Vasculature.

Vertebrates have developed a network of blood vessels and nerves throughout the body that enables them to perform complex higher-order functions and maintain homeostasis. The 16th-century anatomical text 'De humani corporis fabrica' describes the networks of blood vessels and nerves as having a branching pattern in which they are closely aligned and run parallel one to another. This close interaction between adjacent blood vessels and nerves is essential not only for organogenesis during development and repair at the time of tissue damage but also for homeostasis and functional expression of blood vessels and nerves. Furthermore, it is now evident that disruptions in neurovascular interactions contribute to the progression of various diseases including cancer. Therefore, we highlight recent advances in vascular biology research, with a particular emphasis on neurovascular interactions.

Anterior segment dysmorphogenesis of the eye and glaucoma in MG-W gerbils.

Unilaterally swollen eyes were histopathologically characterized in four MG-W gerbils. The primary lesions resided in the anterior segment of the eye where neural crest cells play a critical role in embryonic development. They included indistinct filtration angle, unformed canal of Schlemm, hypoplastic iris, and ciliary body. The findings noted in the retina, optic nerve, optic tract, and lateral geniculate nucleus were consistent with the lesions induced following the persistent elevation of intraocular pressure as a result of insufficient drainage of aqueous humor. Thus, the present cases observed in the eyes of MG-W gerbils exemplified the anterior segment dysmorphogenesis associated with inadequate neural crest migration or differentiation, leading to subsequent glaucoma.

Meningitis and brain abscess formation caused by Cronobacter malonaticus sequence type 440 in a full-term neonate.

Cronobacter spp. cause serious diseases, such as necrotizing enterocolitis, bacteremia, and meningitis in neonates and infants. Most Cronobacter-associated meningitis is reportedly due to C. sakazakii and the majority of infections caused by C. malonaticus occur in adults and are less severe. We report the case of meningitis and brain abscess caused by C. malonaticus Sequence Type (ST) 440 in a healthy full-term neonate. We should consider the possibility that full-term neonates may develop meningitis due to C. malonaticus and treat appropriately because its mortality rate is very high, and survivors are usually left with severe neurologic impairment. In addition, C. malonaticus ST440 may have virulence factors that cause neonatal meningitis akin to the previous report of meningitic ST307 strain.

Advanced paternal age alone does not adversely affect pregnancy or live-birth rates or sperm parameters following intrauterine insemination.

PURPOSE: This study aimed to evaluate the effect of advanced paternal age on pregnancy outcomes and sperm parameters following intrauterine insemination (IUI). We used IUI data rather than assisted reproductive technology data, which might mask the effects of sperm impairments. METHODS: We retrospectively analyzed 1576 IUI cycles in women under 40 years old between April 2012 and May 2016 at the National Center for Child Health and Development in Japan. The main outcomes were clinical pregnancy and live birth. RESULTS: The mean male age was significantly lower in cycles that resulted in pregnancy compared with those without pregnancy (38.0 vs 39.1 years; P < 0.001), with a similar trend for live-birth cycles. However, there was no relationship between advanced paternal age and pregnancy outcomes after adjusting for confounding factors and correlations within patients using generalized estimating equations, and the age of the female partner was the only factor affecting pregnancy rate. Furthermore, advanced paternal age had no effect on sperm parameters. CONCLUSIONS: Advanced paternal age alone does not adversely affect pregnancy or live-birth rates or sperm parameters following IUI.

Comparison of Chromogenic Selective Media for the Detection of Cronobacter spp. (Enterobacter sakazakii).

　The four types of chromogenic selective media that are commercially available in Japan were compared for establishing a Japanese standard method for detecting Cronobacter spp. based on ISO/TS 22964:2006. When assessed using 9 standard Cronobacter spp. strains and 29 non-Cronobacter strains, Enterobacter sakazakii isolation agar, ChromocultTM Enterobacter sakazakii agar, CHROMagarTM E. sakazakii, and XM-sakazakii agar demonstrated excellent inclusivity and exclusivity. Using the ISO/TS 22964:2006 method, the recovered numbers of 38 Cronobacter spp. strains, including 29 C. sakazakii isolates obtained from each medium, were equivalent, indicating that there was no significant difference (p > 0.05) among the four types of chromogenic selective media. Thus, we demonstrated that these four chromogenic selective media are suitable alternatives when using the standard method for detecting Cronobacter spp. in Japan, based on the ISO/TS 22964:2006.

Comparative toxicity of dinophysistoxin-1 and okadaic acid in mice.

The mouse bioassay for diarrhetic shellfish poisoning toxins has been used worldwide. In this study, dinophysistoxin-1 (DTX-1) and okadaic acid (OA) were compared for toxicity. The lethality rate increased and the median survival time decreased in a dose-dependent manner in both DTX-1 and OA. The median lethal dose value was 150.4 µg/kg (95% confidence interval=130.1-171.2 µg/kg) for DTX-1 and 185.6 µg/kg (95% confidence interval=161.2-209.6 µg/kg) for OA. The toxicity equivalent factor 1:1 has been used for OA and DTX-1 in the EU and Japan. Thus, it may be considered that toxicity potential of DTX-1 has remained underestimated as compared to that of OA and DTX-1 might be more toxic than OA.

Extracranial internal carotid artery dissection caused by compression from a giant osteophyte due to atlantoaxial osteoarthritis: case report.

Deformed osseous structures have been reported as rare causes of extracranial internal carotid artery (ICA) dissection, including the styloid process and the hyoid bone. Here, the authors describe the first known case of symptomatic ICA dissection caused by a giant osteophyte due to atlantoaxial osteoarthritis. The left ICA was fixed at the skull base and at the ICA portion compressed by the osteophyte, and it was highly stretched and injured between the two portions during neck rotation. The patient was successfully treated with ligation of the affected ICA following balloon test occlusion. Atlantoaxial osteoarthritis should be considered in the differential diagnosis of ICA dissection in patients with a severely deformed cervical spine.

Intraoperative Transcranial Motor-Evoked Potentials Predict Motor Function Outcome in Intracerebral Hemorrhage Surgery.

OBJECTIVE: Prediction of motor function after intracerebral hemorrhage (ICH) often poses a diagnostic challenge. This study was performed to investigate whether intraoperative monitoring of motor-evoked potentials (MEPs) could predict postoperative motor function recovery. METHODS: We reviewed 16 consecutive patients undergoing evacuation of supratentorial ICH with hemiplegia between June 2011 and October 2014. Patients were categorized according to the results of MEPs before and after evacuation of hematoma. The correlation between detection of MEPs and prognosis of motor function was analyzed. RESULTS: In 10 of 16 cases (62%), stable MEPs were detected before and after evacuation of hematoma, and postoperative motor function was improved in all cases, including 3 cases with severe preoperative motor impairment on manual muscle test (1-2). In 3 cases (19%) in which MEPs were not detected throughout the procedure, motor function was not improved. In the other 3 cases (19%), MEPs were not measured before evacuation of ICH but were detected after evacuation despite poor prognosis of motor function. The results of postevacuation MEPs were considered false-negative results. Predictions using pre-evacuation MEP results were completely consistent with prognosis for recovery, whereas MEPs obtained during and after evaluation were useful for monitoring. CONCLUSIONS: Intraoperative MEPs may indicate preservation of pyramidal tracts, and pre-evacuation MEPs can predict motor function outcome after ICH surgery.

[Collaboration between public health nurses and the private sector].

OBJECTIVES: We clarified collaborations between public health nurses (PHNs) and the private sector, such as nonprofit organizations. METHODS: Semi-structured interviews were conducted with 11 private sector organizations and 13 PHNs who collaborate with them between December 2012 to October 2013. Interview guides were: overall suicide preventive measurements, details of collaboration between private sector organizations and PHNs, and suicide prevention outcomes/issues. Data from private sector organizations and PHNs were separately analyzed and categories created using qualitative and inductive design. Private sector organizations' and PHNs' categories were compared and separated into core categories by similarities. RESULTS: Six categories were created: 1. establishing a base of mutual understanding; 2. raising public awareness of each aim/characteristic; 3. competently helping high suicidal risk persons detected during each activity; 4. guarding lives and rehabilitating livelihoods after intervention; 5. restoring suicide attempters/bereaved met in each activity; and 6. continuing/expanding activities with reciprocal cohesion/evaluation. CONCLUSION: PHNs are required to have the following suicide prevention tasks when collaborating with private sector organizations: understanding the private sector civilization, sharing PHN experiences, improving social determinants of health, meeting basic needs, supporting foundation/difficulties each other (Dear editor. Thank you for kind comments. I was going to explain that PHNs and NPOs support each other their foundation of activity and difficulties in their activities. The foundations include knowledge, information, budgets, manpower etc. The difficulties mean like suffering faced with suicide during activities.), and enhancing local governments' flexibilities/ promptness.

Ability of NT-pro-BNP to Diagnose Cardioembolic Etiology in Patients with Acute Ischemic Stroke.

Background Cardioembolic stroke (CE) is usually associated with a larger ischemic area leading to higher morbidity and mortality rates. No biomarkers for CE are available, which causes difficulty in differential diagnosis of CE from other subtypes of acute ischemic stroke. Methods: We prospectively evaluated consecutive patients with acute ischemic stroke to identify biomarkers that could distinguish between CE and other subtypes of acute ischemic stroke. Etiological diagnoses were identified according to the National Institute of Neurological Disorders and Stroke (NINDS) 111 classification using clinical examinations, computed tomography (CT), magnetic resonance imaging (MRI), cardiac evaluations, and other tests. The biomarkers N-terminal pro-brain natriuretic peptide (NT-pro-BNP), Thrombin-Antithrombin III Complex (TAT), and D-dimer were determined in blood samples collected within 48 hours of onset and compared between groups with and without CE. Non- CE consisted of atherothrombotic brain infarction (ATBI), lacunar infarction (LI), and other stroke subtypes of unknown cause (other). Results: This study included 279 patients diagnosed with acute ischemic stroke. Serum levels of NT-pro- BNP were significantly higher in those with than in those without CE stroke (p<0.0001). Analysis of receiver operating characteristics (ROC) curves indicated that an NT-pro-BNP cutoff of 332 pg/mL provided optimal sensitivity (98.3%) and specificity (75.8%) for distinguishing CE from non-CE. Conclusions: Serum levels of NT-pro-BNP may help in diagnosis of CE during the acute phase and thus allow appropriate therapy t6 prevent subsequent cardiogenic stroke.

Fournier's gangrene caused by Listeria monocytogenes as the primary organism.

A 70-year-old man with a history of tongue cancer presented with Fournier's gangrene caused by Listeria monocytogenes serotype 4b. Surgical debridement revealed undiagnosed rectal adenocarcinoma. The patient did not have an apparent dietary or travel history but reported daily consumption of sashimi (raw fish). Old age and immunodeficiency due to rectal adenocarcinoma may have supported the direct invasion of L monocytogenes from the tumour. The present article describes the first reported case of Fournier's gangrene caused by L monocytogenes. The authors suggest that raw ready-to-eat seafood consumption be recognized as a risk factor for listeriosis, especially in cases of skin and soft tissue infection.

Un homme de 70 ans ayant des antécédents de cancer de la langue s'est présenté avec une gangrène de Fournier causée par un Listeria monocytogenes de sérotype 4b. Le débridement chirurgical a révélé un adénocarcinome rectal non diagnostiqué. Le patient n'avait pas d'antécédents alimentaires ou de voyage apparents, mais a déclaré consommer des sashimis (poisson cru) tous les jours.L'âge avancé et l'immunodéficience causée par l'adénocarcinome rectal ont peut-être favorisé l'invasion directe du L monocytogenes par la tumeur. Il s'agit du premier cas déclaré de gangrène de Fournier attribuable au L monocytogenes. Les auteurs proposent d'inclure la consommation de fruits de mer crus prêt-à-manger dans les facteurs de risque de listériose, notamment en cas d'infections de la peau et des tissus mous.

Food-borne botulism in Japan in March 2012.

In March 2012, two patients were transported urgently to the hospital in Tottori Prefecture, Japan, because of symptoms suggestive of botulism. Botulinum neurotoxin type A was detected in the clinical specimens and the food consumed by the two patients (vacuum packed adzuki-batto, a sweet adzuki bean soup containing noodles). We were able to make a prompt diagnosis of food botulism associated with the consumption of adzuki-batto, from which the causative pathogen Clostridium botulinum Ab was cultured.

Bacterial translocation in alymphoplasia (aly/aly) mice.

Bacterial translocation (BTL) is defined as the passage of viable bacteria from the gastrointestinal tract to the organs. This study was to elucidate the roles of Peyer's patches (PPs) and/or mesenteric lymph nodes (MLNs) in BTL. Alymphoplastic mutant mice and phenotypically normal heterozygous micewere dominantly colonized with streptomycin-resistant Escherichia coli and BTL was examined. In PP- and MLN-competent mice, BTL to MLNs was detected in 100% of mice, but BTL to organs was rare (25%). On the other hand, in PP- and MLN-deficientmice, BTL to organs was detected in 91% of mice. The results clearly indicate that PPs are not the only site for bacterial entry.

Clinical use of (11)C-methionine and (18)F-FDG-PET for germinoma in central nervous system.

OBJECTIVE: The purpose of this study was to examine the (11)C-methionine (MET) and (18)F-fluorodeoxyglucose (FDG) positron emission tomography (PET) findings of central nervous system (CNS) germinoma and the diagnostic utility of these findings. METHODS: We retrospectively evaluated the cases of 10 patients who were diagnosed with CNS germinoma according to their histopathological or clinical findings. All the patients underwent pretreatment MET and/or FDG-PET scans, and the resultant images were assessed qualitatively and quantitatively. In the qualitative assessments, we used 3- and 5-grade visual scoring systems for the MET- and FDG-PET images, respectively. In the quantitative assessments, the maximal standardized uptake value (SUVmax) and the ratio of the SUVmax of the tumor (T) divided by the mean SUV for the normal white or gray matter [T/N (WM), T/N (GM)], was calculated. RESULTS: The mean and SD values of SUVmax, T/N (WM), and T/N (GM) were 1.9 ± 1.4, 2.5 ± 1.3, and 1.7 ± 0.9 on MET-PET and 5.8 ± 2.2, 1.6 ± 0.5, and 0.8 ± 0.2 on FDG-PET, respectively. On MET-PET, only one lesion was not detected. On the other hand, on FDG-PET all of the lesions exhibited uptake values that were intermediate between those of the normal white matter and gray matter. CONCLUSION: In terms of its tumor-contouring ability, MET is a good tracer for diagnosing CNS germinomas; therefore, MET-PET is considered to be useful for planning biopsies or surgery. Although FDG-PET is capable of detecting CNS germinomas, it produced insufficient image contrast in the present study. Further studies are needed before FDG-PET can be used in clinical examinations of CNS germinoma.

Evaluation of the culture method NIHSJ-02 alternative to ISO 10272-1:2006 for the detection of campylobacter jejuni and campylobacter coli in chicken: collaborative study.

For the surveillance of the prevalence of Campylobacter jejuni and Campylobacter coli in raw chicken products in Japan, a qualitative method, National Institute of Health Sciences Japan (NIHSJ)-02, was developed as an alternative to International Organization for Standardization (ISO) 10272-1:2006. In the NIHSJ-02 culture method, the enrichment step is carried out in a reduced volume of Preston broth at 42 +/- 1 degrees C to reduce cost and space, and to prevent the overgrowth of background bacteria. To evaluate the performance of NIHSJ-02, a collaborative study was conducted, and the results obtained by NIHSJ-02 were compared with those obtained using the reference method, ISO 10272-1:2006. Fifteen laboratories participated; each examined 48 minced chicken samples consisting of test samples uninoculated, inoculated with C. jejuni at a low or high level, and inoculated with C. coli at a low level. The average probabilities of detection by NIHSJ-02 across laboratories were 0.033, 0.222, 0.678, and 0.267 in samples uninoculated, inoculated with C. jejuni at a low and high level, and with C. coli at a low level, respectively. Those by ISO 10272-1:2006 were 0.051, 0.128, 0.551, and 0.090. Significantly higher probabilities of detection were determined by NIHSJ-02 compared to ISO 10272-1:2006, except for uninoculated samples. On the other hand, significantly lower frequency of occurrence of background bacteria was observed by NIHSJ-02 (43.1%) compared with ISO 10272-1:2006 (92.6%). NIHSJ-02 showed better performance than ISO 10272-1:2006 with regard to the selective detection of C. jejuni and C. coli in chicken.

Comparison of liquid crystal display monitors calibrated with gray-scale standard display function and with γ 2.2 and iPad: observer performance in detection of cerebral infarction on brain CT.

OBJECTIVE: The purpose of the study was to compare observer performance in the detection of cerebral infarction on a brain CT using medical-grade liquid crystal display (LCD) monitors calibrated with the gray-scale standard display function and with γ 2.2 and using an iPad with a simulated screen setting. MATERIALS AND METHODS: We amassed 97 sample sets, from 47 patients with proven cerebral infarction and 50 healthy control subjects. Nine radiologists independently assessed brain CT on a gray-scale standard display function LCD, a γ 2.2 LCD, and an iPad in random order over 4-week intervals. Receiver operating characteristic (ROC) analysis was performed by using the continuous scale, and the area under the ROC curve (A(z)) was calculated for each monitor. RESULTS: The A(z) values for gray-scale standard display function LCD, γ 2.2 LCD, and iPad were 0.875, 0.884, and 0.839, respectively. The difference among the three monitors was very small. There was no significant difference between gray-scale standard display function LCD and γ 2.2 LCD. However, the A(z) value was statistically significantly smaller for the iPad than the γ 2.2 LCD (p < 0.05). CONCLUSION: Observer performance for detecting cerebral infarction on the LCD with γ 2.2 calibration was found to be similar to the LCD with gray-scale standard display function calibration. Although observer performance using the iPad was poorer than that using the other LCDs, the difference was small. Therefore, the iPad could not substitute for other LCD monitors. However, owing to the promising potential advantages of tablet PCs, such as portability, further examination is needed into the clinical use of tablet PCs.

Growth of Listeria monocytogenes in refrigerated ready-to-eat foods in Japan.

The ability of L. monocytogenes to grow in a series of Japanese ready-to-eat (RTE) foods, including boiled baby sardine and Japanese pickle, was tested at two different refrigeration temperatures. In RTE foods in which L. monocytogenes can grow, growth was significantly higher at 10°C than that at 4°C during their shelf lives and growth patterns varied extensively among the different types of foods. However, growth did not occur at 4°C within the shelf life of certain RTE foods, such as broiled squid. The patterns of growth were varied extensively with different sample types. These results suggest that some types of traditional Japanese RTE foods stored at 10°C may be potential sources of listeriosis. To reduce the risk of food-borne listeriosis, studies to determine the contamination levels in RTE foods and the effects of storage temperature on their shelf lives are needed.