Genetic diversification of allohexaploid Brassica hybrids (AABBCC) using a fertile octoploid with excessive C genome set (AABBCCCC).

MAIN CONCLUSION: Using octoploid somatic hybrids with excessive C genome sets, AABBCCCC, a diverse allohexaploid, AABBCC, was produced by C genome reduction through subsequent crossing with various AABB cultivars. Even when somatic hybrids are produced, the plants that are produced are rarely in themselves an innovative crop. In this study, we used somatic hybrids of Brassica juncea (AABB) and B. oleracea (CC) as model cases for the genetic diversification of the somatic hybrids. One cell of 'Akaoba Takana' (B. juncea) and two cells of 'Snow Crown' (B. oleracea) were fused to create several somatic hybrids with excessive C genomes, AABBCCCC. Using AABBCCCC somatic hybrids as mother plants and crossing with 'Akaoba Takana', the AABBCC progenies were generated. When these AABBCC plants were self-fertilized, and flow cytometric (FCM) analysis was performed on the next generations, differences in the relative amount of genome size variation were observed, depending on the different AABBCCCC parents used for AABBCC creation. Further self-progeny was obtained for AABBCC plants with a theoretical allohexaploid DNA index by FCM. However, as the DNA indices of the progeny populations varied between plants used and aneuploid individuals still occurred in the progeny populations, it was difficult to say that the allohexaploid genome was fully stabilized. Next, to obtain genetic diversification of the allohexaploid, different cultivars of B. juncea were crossed with AABBCCCC, resulting in diverse AABBCC plants. Genetic diversity can be further expanded by crossbreeding plants with different AABBCC genome sets. Although genetic stability is necessary to ensure in the later generations, the results obtained in this study show that the use of somatic hybrids with excess genomes is an effective strategy for creating innovative crops.

Degenerate oligonucleotide primer MIG-seq: an effective PCR-based method for high-throughput genotyping.

Next-generation sequencing (NGS) library construction often involves using restriction enzymes to decrease genome complexity, enabling versatile polymorphism detection in plants. However, plant leaves frequently contain impurities, such as polyphenols, necessitating DNA purification before enzymatic reactions. To overcome this problem, we developed a PCR-based method for expeditious NGS library preparation, offering flexibility in number of detected polymorphisms. By substituting a segment of the simple sequence repeat sequence in the MIG-seq primer set (MIG-seq being a PCR method enabling library construction with low-quality DNA) with degenerate oligonucleotides, we introduced variability in detectable polymorphisms across various crops. This innovation, named degenerate oligonucleotide primer MIG-seq (dpMIG-seq), enabled a streamlined protocol for constructing dpMIG-seq libraries from unpurified DNA, which was implemented stably in several crop species, including fruit trees. Furthermore, dpMIG-seq facilitated efficient lineage selection in wheat and enabled linkage map construction and quantitative trait loci analysis in tomato, rice, and soybean without necessitating DNA concentration adjustments. These findings underscore the potential of the dpMIG-seq protocol for advancing genetic analyses across diverse plant species.

A simple method for measuring pollen germination rate using machine learning.

The pollen germination rate decreases under various abiotic stresses, such as high-temperature stress, and it is one of the causes of inhibition of plant reproduction. Thus, measuring pollen germination rate is vital for understanding the reproductive ability of plants. However, measuring the pollen germination rate requires much labor when counting pollen. Therefore, we used the Yolov5 machine learning package in order to perform transfer learning and constructed a model that can detect germinated and non-germinated pollen separately. Pollen images of the chili pepper, Capsicum annuum, were used to create this model. Using images with a width of 640 pixels for training constructed a more accurate model than using images with a width of 320 pixels. This model could estimate the pollen germination rate of the F2 population of C. chinense previously studied with high accuracy. In addition, significantly associated gene regions previously detected in genome-wide association studies in this F2 population could again be detected using the pollen germination rate predicted by this model as a trait. Moreover, the model detected rose, tomato, radish, and strawberry pollen grains with similar accuracy to chili pepper. The pollen germination rate could be estimated even for plants other than chili pepper, probably because pollen images were similar among different plant species. We obtained a model that can identify genes related to pollen germination rate through genetic analyses in many plants.

The mutation of CaCKI1 causes seedless fruits in chili pepper (Capsicum annuum).

KEY MESSAGE: The seedless mutant tn-1 in chili pepper is caused by a mutation in CaCKI1 (CA12g21620), which encodes histidine kinase involving female gametophyte development. An amino acid insertion in the receiver domain of CaCKI1 may be the mutation responsible for tn-1. Seedlessness is a desirable trait in fruit crops because the removal of seeds is a troublesome step for consumers and processing industries. However, little knowledge is available to develop seedless chili peppers. In a previous study, a chili pepper mutant tn-1, which stably produces seedless fruits, was isolated. In this study, we report characterization of tn-1 and identification of the causative gene. Although pollen germination was normal, confocal laser microscopy observations revealed deficiency in embryo sac development in tn-1. By marker analysis, the tn-1 locus was narrowed down to a 313 kb region on chromosome 12. Further analysis combined with mapping-by-sequencing identified CA12g21620, which encodes histidine kinase as a candidate gene. Phylogenetic analysis revealed CA12g21620 was the homolog of Arabidopsis CKI1 (Cytokinin Independent 1), which plays an important role in female gametophyte development, and CA12g21620 was designated as CaCKI1. Sequence analysis revealed that tn-1 has a 3-bp insertion in the 6th exon resulting in one lysine (K) residue insertion in receiver domain of CaCKI1, and the sequence nearby the insertion is widely conserved among CKI1 orthologs in various plants. This suggested that one K residue insertion may reduce the phosphorylation relay downstream of CaCKI1 and impair normal development of female gametophyte, resulting in seedless fruits production in tn-1. Furthermore, we demonstrated that virus-induced gene silencing of CaCKI1 reduced normally developed female gametophyte in chili pepper. This study describes the significant role of CaCKI1 in seed development in chili pepper and the possibility of developing seedless cultivars using its mutation.

MIG-seq is an effective method for high-throughput genotyping in wheat (Triticum spp.).

MIG-seq (Multiplexed inter-simple sequence repeats genotyping by sequencing) has been developed as a low cost genotyping technology, although the number of polymorphisms obtained is assumed to be minimal, resulting in the low application of this technique to analyses of agricultural plants. We applied MIG-seq to 12 plant species that include various crops and investigated the relationship between genome size and the number of bases that can be stably sequenced. The genome size and the number of loci, which can be sequenced by MIG-seq, are positively correlated. This is due to the linkage between genome size and the number of simple sequence repeats (SSRs) through the genome. The applicability of MIG-seq to population structure analysis, linkage mapping, and quantitative trait loci (QTL) analysis in wheat, which has a relatively large genome, was further evaluated. The results of population structure analysis for tetraploid wheat showed the differences among collection sites and subspecies, which agreed with previous findings. Additionally, in wheat biparental mapping populations, over 3,000 SNPs/indels with low deficiency were detected using MIG-seq, and the QTL analysis was able to detect recognized flowering-related genes. These results revealed the effectiveness of MIG-seq for genomic analysis of agricultural plants with large genomes, including wheat.

UV excited fluorescence image-based non-destructive method for early detection of strawberry (Fragaria × ananassa) spoilage.

The soon spoiled strawberries need to be classified from healthy fruits in an early stage. In this research, a machine vision system is proposed for inspecting the quality of strawberries using ultraviolet (UV) light based on the excitation-emission matrix (EEM) results. Among the 100 fruits which were harvested and stored under 10 °C condition for 7 days, 7 fruits were confirmed to be spoiled by using a firmness meter. The EEM results show the fluorescence compound contributes to a whitish surface on the spoiled fruits. Based on the EEM results, UV fluorescence images from the bottom view of strawberries were used to classify the spoiled fruits and healthy fruits within 1 day after harvest. These results demonstrate the UV fluorescence imaging can be a fast, non-destructive, and low-cost method for inspecting the soon spoiled fruits. The proposed index related to the spoiling time can be a new indicator for qualifying strawberry.

A Fragment of S38AA is a Novel Plasma Biomarker of Alzheimer's Disease.

Alzheimer's disease (AD) is a progressive neurodegenerative disease without a cure. The pathological process starts decades before clinical onset, and thus clinical trials of drugs aimed at treating AD should start at a presymptomatic stage. Therefore, it is critical to diagnose AD at an early stage. Tau, phosphorylated tau, and amyloid-ß peptide (Aß) in cerebrospinal fluid (CSF), and positron emission tomography (PET) imaging of Aß or tau accumulation are supportive biomarkers for AD diagnosis, but there is no reliable presymptomatic diagnostic marker. Since CSF sampling is invasive, and PET imaging is expensive and available only at specialized centers, a reliable blood biomarker has long been sought for. Here we describe a novel extramembrane fragment from solute carrier family 38 member 10 (SLC38A10, S38AA) that we found to be decreased in pyramidal neurons in AD cases by proteomics and immunohistochemical analysis. We detected a S38AA fragment in CSF and found the levels to correlate with severity of AD and APOE genotype. Importantly, the plasma levels of the fragment also showed a significant correlation with Mini-Mental State Examination scores in AD. Moreover, plasma from other neurodegenerative disease was analyzed and the fragment was found to be increased specifically in AD. Interestingly, the fragment is detected in mouse, rat, and monkey, and increases in amyloid precursor protein transgenic mice as the AD-like pathology progresses. We propose that the S38AA fragment in plasma could be a novel quantitative diagnostic marker for AD and potentially a marker of disease progression in AD.

Impact on predictability of tropical and mid-latitude cyclones by extra Arctic observations.

Recent research has demonstrated that additional winter radiosonde observations in Arctic regions enhance the predictability of mid-latitude weather extremes by reducing uncertainty in the flow of localised tropopause polar vortices. The impacts of additional Arctic observations during summer are usually confined to high latitudes and they are difficult to realize at mid-latitudes because of the limited scale of localised tropopause polar vortices. However, in certain climatic states, the jet stream can intrude remarkably into the mid-latitudes, even in summer; thus, additional Arctic observations might improve analysis validity and forecast skill for summer atmospheric circulations over the Northern Hemisphere. This study examined such cases that occurred in 2016 by focusing on the prediction of the intensity and track of tropical cyclones (TCs) over the North Atlantic and North Pacific, because TCs are representative of extreme weather in summer. The predictabilities of three TCs were found influenced by additional Arctic observations. Comparisons with ensemble reanalysis data revealed that large errors propagate from the data-sparse Arctic into the mid-latitudes, together with high-potential-vorticity air. Ensemble forecast experiments with different reanalysis data confirmed that additional Arctic observations sometimes improve the initial conditions of upper-level troposphere circulations.

Development of an ensemble Kalman filter data assimilation system for the Venusian atmosphere.

The size and mass of Venus is similar to those of the Earth; however, its atmospheric dynamics are considerably different and they are poorly understood due to limited observations and computational difficulties. Here, we developed a data assimilation system based on the local ensemble transform Kalman filter (LETKF) for a Venusian Atmospheric GCM for the Earth Simulator (VAFES), to make full use of the observational data. To examine the validity of the system, two datasets were assimilated separately into the VAFES forecasts forced with solar heating that excludes the diurnal component Qz; one was created from a VAFES run forced with solar heating that includes the diurnal component Qt, whereas the other was based on observations made by the Venus Monitoring Camera (VMC) onboard the Venus Express. The VAFES-LETKF system rapidly reduced the errors between the analysis and forecasts. In addition, the VAFES-LETKF system successfully reproduced the thermal tide excited by the diurnal component of solar heating, even though the second datasets only included horizontal winds at a single altitude on the dayside with a long interval of approximately one Earth day. This advanced system could be useful in the analysis of future datasets from the Venus Climate Orbiter 'Akatsuki'.

Direct high-pressure NMR observation of dipicolinic acid leaking from bacterial spore: A crucial step for thermal inactivation.

A bacterial spore protects itself with an unusually high concentration (~10% in dry weight of spore) of dipicolinic acid (DPA), the release of which is considered the crucial step for inactivating it under mild pressure and temperature conditions. However, the process of how the spore releases DPA in response to pressure remains obscure. Here we apply 1H high-resolution high-pressure NMR spectroscopy, for the first time, to the spore suspension of Bacillus subtilis natto and monitor directly and in real-time the leaking process of DPA in response to pressure of 200MPa at 20°C. We find that about one third of the total DPA leaks immediately upon applying pressure, but that the rest leaks slowly in hrs upon decreasing the pressure. Once DPA is fully released from the spore, the proteins of the spore become easily denatured at a mild temperature, e.g., 80°C, much below the temperature commonly used to inactivate spores (121°C). The success of the present experiment opens a new avenue for studying bacterial spores and cells at the molecular level in response to pressure, temperature and other perturbations.

Effects of super-hard rice bread blended with black rice bran on amyloid ß peptide production and abrupt increase in postprandial blood glucose levels in mice.

Alzheimer's disease and type 2 diabetes are very serious diseases with the latter having been suggested to cause the former. We prepared super-hard rice bread blended with black rice bran (SRBBB), which contained a high amount of resistant starch that showed strong inhibitory activities against ß-secretase and acetylcholinesterase even after heating. Black rice bran showed greater ß-secretase inhibitory activity (3.6-fold) than Koshihikari rice. The bran contained more oleic acid and anthocyanin, meaning that it is potentially a biofunctional food with a high antioxidant capacity. Furthermore, aged mice, which were fed a SRBBB diet for four weeks, showed lower amyloid ß 40 peptide in the blood than mice fed a commercial diet (p < 0.01). Additionally, their initial blood glucose levels (BGLs) after 12 weeks of being fed SRBBB were significantly lower than those in the control group. Taken together, our results indicate SRBBB shows promise for inhibiting not only amyloid ß production, but also abrupt increases in postprandial BGLs.

Additional Arctic observations improve weather and sea-ice forecasts for the Northern Sea Route.

During ice-free periods, the Northern Sea Route (NSR) could be an attractive shipping route. The decline in Arctic sea-ice extent, however, could be associated with an increase in the frequency of the causes of severe weather phenomena, and high wind-driven waves and the advection of sea ice could make ship navigation along the NSR difficult. Accurate forecasts of weather and sea ice are desirable for safe navigation, but large uncertainties exist in current forecasts, partly owing to the sparse observational network over the Arctic Ocean. Here, we show that the incorporation of additional Arctic observations improves the initial analysis and enhances the skill of weather and sea-ice forecasts, the application of which has socioeconomic benefits. Comparison of 63-member ensemble atmospheric forecasts, using different initial data sets, revealed that additional Arctic radiosonde observations were useful for predicting a persistent strong wind event. The sea-ice forecast, initialised by the wind fields that included the effects of the observations, skilfully predicted rapid wind-driven sea-ice advection along the NSR.

Application of High-Pressure Treatment to Enhancement of Functional Components in Agricultural Products and Development of Sterilized Foods.

In 1988, the late Prof. Rikimaru Hayashi had first proposed "Use of High Pressure in Food", introducing his views, i.e., "heat and pressure are independently capable of transforming the state of a substance, and such state transforming factors are only heat and pressure in nature." Sc. D. Masaru Nakahara stated in his note that he had been impressed by the unique starting point of Prof. Hayashi's idea. Prof. Hayashi had explored some good method for food processing without using heat, so he alternatively thought of high-pressure treatment (Hayashi R (1989) Use of high pressure to food processing and preservation. In: Hayashi R (ed) Use of high pressure in food. San-Ei Publishing Co, Kyoto, pp 1-30; Nakahara M (1990) Water and ions at high pressure: their fundamental properties relevant to the pressure treatment to food. In: Hayashi R (ed) Pressure-processed food--research and development. San-Ei Publishing Co, Kyoto, pp 3-21). Since the start-up of Japanese research group of high pressure in biological field (the present "Japanese Research Group of High Pressure Bioscience and Biotechnology (JHPBB)") and "International Association of High Pressure Bioscience and Biotechnology (IAHPBB)", we have continued to research into the industrial use of high-pressure treatment over a period of 25 years to realize our dream, that is, the same as Prof. Hayashi's dream. Although heat and pressure were found to be independent factors capable of transforming the state of a substance, use of heat has been overwhelmingly more frequent in food processing up to now. However, the pressure treatment has the advantages of instantaneous transmission, uniform distribution in vessels, and ability of inducing uniform change in quality. The high-pressure treatment does not cause cleavage of the covalent bond in the substance, thereby lessening the decomposition of nutrients, the generation of offensive smell, and the production of abnormal materials when compared with the heat application. In addition, energy consumption in the high-pressure treatment is less than that in the heat treatment. For the reasons mentioned above, the high-pressure treatment has thus been regarded as suitable for future food processing, and much attention has been paid to the researches of high-pressure treatment again. Then, we reviewed the previous researches in which little interest had been taken because of imperfectness of non-heat sterilization. Surprisingly, we discovered some novel findings about the effect of high-pressure treatment, that is, pressure history on the subsequent event. Then, we decided to present two theses on the themes, "Application of High-pressure Treatment to Enhancement of Functional Components in Agricultural Products" and "Application of High-pressure Treatment to Development of Sterilized Foods".

The combination of perception of other individuals and exogenous manipulation of arousal enhances social facilitation as an aftereffect: re-examination of Zajonc's drive theory.

Zajonc's drive theory postulates that arousal enhanced through the perception of the presence of other individuals plays a crucial role in social facilitation (Zajonc, 1965). Here, we conducted two experiments to examine whether the elevation of arousal through a stepping exercise performed in front of others as an exogenous factor causes social facilitation of a cognitive task in a condition where the presence of others does not elevate the arousal level. In the main experiment, as an "aftereffect of social stimulus," we manipulated the presence or absence of others and arousal enhancement before participants conducted the primary cognitive task. The results showed that the strongest social facilitation was induced by the combination of the perception of others and arousal enhancement. In a supplementary experiment, we manipulated these factors by adding the presence of another person during the task. The results showed that the effect of the presence of the other during the primary task is enough on its own to produce facilitation of task performance regardless of the arousal enhancement as an aftereffect of social stimulus. Our study therefore extends the framework of Zajonc's drive theory in that the combination of the perception of others and enhanced arousal as an "aftereffect" was found to induce social facilitation especially when participants did not experience the presence of others while conducting the primary task.

Long-term safety and efficacy of treatment with subcutaneous abatacept in Japanese patients with rheumatoid arthritis who are methotrexate inadequate responders.

OBJECTIVE: To assess the long-term safety, immunogenicity, and efficacy of subcutaneous (SC) abatacept in combination with methotrexate (MTX) in Japanese patients with rheumatoid arthritis who were MTX inadequate responders, in a long-term extension (LTE) to a double-dummy, double-blind study (NCT01001832). METHODS: Patients, who had previously received SC or intravenous (IV) abatacept with MTX (6-8 mg/week) for 24 weeks, received SC abatacept (125 mg/week) with MTX for an additional 52 weeks. Safety, immunogenicity, and efficacy were assessed. RESULTS: The LTE included 112 patients. SC abatacept was generally well tolerated in the LTE, with no new safety signals. American College of Rheumatology 20, 50, and 70 response rates, disease activity score 28 (C-reactive protein) remission rates (< 2.6), and Health Assessment Questionnaire-Disability Index response rates (≥ 0.3 improvement from baseline) achieved at the end of the double-blind period were maintained over the LTE and were comparable in patients who received SC or IV abatacept in the double-blind period. Seropositivity for immunogenicity occurred in 4 (3.6%) patients. Self-injection of SC abatacept was well controlled and not associated with additional safety events. CONCLUSIONS: SC abatacept had acceptable safety and was well tolerated and effective over the LTE (76 weeks in total), with low rates of immunogenicity in Japanese patients.

Efficacy, safety, pharmacokinetics and immunogenicity of abatacept administered subcutaneously or intravenously in Japanese patients with rheumatoid arthritis and inadequate response to methotrexate: a Phase II/III, randomized study.

OBJECTIVE: To evaluate efficacy and safety of subcutaneous (SC) and intravenous (IV) abatacept and background methotrexate (MTX) in Japanese patients with rheumatoid arthritis (RA) and inadequate response to MTX (MTX-IR). METHODS: Double-dummy, double-blind study (NCT01001832); 118 adults with ≥ 10 swollen joints, ≥ 12 tender joints and C-reactive protein (CRP) ≥ 0.8 mg/dL randomized 1:1 to SC abatacept (125 mg weekly) with IV loading (∼10 mg/kg on Day 1), or IV abatacept (∼10 mg/kg monthly) for 169 days, both also receiving MTX (6-8 mg/week). Primary endpoint was Day 169 American College of Rheumatology (ACR)20 response; other efficacy endpoints, safety and immunogenicity were assessed. RESULTS: Similar proportions of patients achieved ACR20 responses at Day 169 with SC (91.5% [95% CI 81.3, 97.2]) and IV abatacept (83.1% [71.0, 91.6]). ACR50/70 responses, adjusted mean changes from baseline in Health Assessment Questionnaire-Disability Index scores and remission rates (28-joint Disease Activity Score [CRP] < 2.6) were also comparable between groups. Serious adverse event frequencies (5.1% vs. 3.4%) were similar with both formulations. One patient per group tested seropositive for immunogenicity. Weekly SC abatacept dosing achieved mean serum concentrations > 10 µg/mL (minimum therapeutic target). CONCLUSIONS: SC abatacept demonstrated comparable efficacy and safety to IV abatacept, with low immunogenicity rates, in MTX-IR Japanese patients with RA.

Semisynthesis of salviandulin E analogues and their antitrypanosomal activity.

A series of analogues of salviandulin E, a rearranged neoclerodane diterpene originally isolated from Salvia leucantha (Lamiaceae), were prepared and their in vitro activity against Trypanosoma brucei brucei was evaluated with currently used therapeutic drugs as positive controls. One of the 19 compounds prepared and assayed in the present study, butanoyl 3,4-dihydrosalviandulin E analogue was found to be a possible candidate for an antitrypanosomal drug with fairly strong antitrypanosomal activity and lower cytotoxicity.

Symptoms related to glaucomatous visual field abnormalities among male Japanese workers in a population-based setting.

PURPOSE: To identify symptoms potentially related to glaucomatous visual field abnormalities (VFAs) in a population-based setting, and to assess the applicability of using these symptoms to identify persons at risk of developing glaucoma. METHODS: In this study, 10,214 Japanese male general workers (mean age, 45.3±8.8 years) filled out a self-administered questionnaire and underwent frequency doubling technology (FDT) perimetry testing. The questionnaire inquired about whether the participant was suffering from any of nine symptoms, with scores for each response graded on a four-item Likert scale: 0 (none), 1 (rarely), 2 (sometimes), and 3 (always). Results from the questionnaire were compared among three groups: participants without any VFAs (9767), participants with VFAs as determined by the FDT test (FDT-VFA; 447), and 227 participants (of the 447 FDT-VFA participants) with glaucomatous VFAs who were newly diagnosed by ophthalmologic examinations. RESULTS: The mean summed scores for the total items were significantly (p<0.01) higher in FDT- and glaucomatous VFA groups than in normal subjects. In particular, responses citing the symptoms 'feeling of something in the front of the eye' and 'feeling of hardness to see in dark places' were significantly (p<0.05) more frequent in subjects with FDT- and glaucomatous VFAs than among normal participants. However, the respective areas under the receiver operating characteristic curve of summed scores for the nine total items and for the two items which showed significant differences for the glaucoma groups were 0.57 (95% confidence interval=0.53-0.60) and 0.58 (95% confidence interval=0.54-0.61). CONCLUSION: Although the symptoms 'feeling of hardness to see in dark places' and 'feeling of something in the front of the eye' could be associated with glaucomatous VFA in a population-based setting, inquiring about symptoms is of little aid in identifying subjects with glaucomatous VFA as a strategy for public health.

Effect of a group health education program in the workplace at the age of 35: a population approach.

Using a population approach, we investigated whether a group health education program exerted a preventive effect on checkup items at five years later. Workers turning 35 years old before the initial implementation of the program were entered in the control group (n = 422) and those turning 35 years on this date or after were entered in the intervention group (n = 206). These groups were compared using data obtained from routine health checkups at 35 and 40 years of age. In the intervention group, self-management ability prior to and after completion of the program was compared using a 2 item questionnaire based on the General Self-Efficacy Scale (GSES) and the Health Locus of Control (HLC). In males, the intervention group showed significant inhibition of increases in body mass index (BMI), systolic and diastolic blood pressure (SBP and DBP), and gamma-glutamyl transpeptidase (gamma-GTP) compared to the control group. In females, however, a similar outcome was seen for gamma-GTP only. The evaluation index of self-management ability for both GSES and HLC significantly improved among males but did not significantly change among females at five years post-completion of the program. In particular, the group with a high GSES evaluation index experienced significant inhibition of weight gain. As a population approach, adoption of this program in the workplace for males aged 35 years may have an inhibitory effect on five-year weight gain. Further, programs which improve GSES appear effective in inhibiting weight gain.

Change of thermal inactivation of Clostridium botulinum spores during rice cooking.

Cooked and packed rice (CaPR), a popular rice product in Japan, is distributed with oxygen-absorbing agents and without refrigeration. When the final product was inoculated with spores of several strains of proteolytic Clostridium botulinum at a dose of 10(3) spores per g (2 x 10(5) spores per package) and incubated at 30 degrees C, the bacteria grew and produced neurotoxins in 40 days. To simulate more realistic cases of contamination, the same dose of spores was inoculated before the cooking process. When cooked at 100 degrees C for 30 min, a small number of spores survived and the toxins were detected in some of the samples after incubation for 180 days. However, when cooked at 100 degrees C for 15 min immediately followed by 105 degrees C for 15 min, neither survivors nor the toxins were detected during incubation for 270 days after cooking. Even when inoculated with 10(5) spores per g of one of the most heat-resistant strains, 213B, viable spores were not detected after cooking. The inactivation by these heating conditions in different media indicated that the spores were inactivated >1,000-fold more in rice suspension than in cooked meat medium or phosphate buffer. It was therefore suggested that rice contains component(s) that facilitates thermal inactivation of C. botulinum.