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The lungs or seventy three necropsy cases, most of which were the cases of unexpected sudden deaths, from the Tokyo Metropolitan Medical Examiner Office were used for this study. Materials for culture were taken aseptically from the respiratory parenchyma of the lungs (subpleural region). The six areas of the lungs were examined in each cases, respectively. Peptone glucose chloramphenicol agar media was used for the culture, and isolated filamentous fungi and yeasts were identified by routine procedure. The areas, from which the materials for culture were taken, were examined histopathologically. The results obtained were as follows1) Among the 73 cases, fungi were isolated from 60 cases. Namely, filamentous fungi were detected from 38 cases, yeasts were isolated from 7 cases, and both filamentous fungi and yeasts were obtained from 15 cases. No pulmonary fungus infection was disclosed in the cases examined.2) Filamentous fungi were isolated from 53 cases (72.6%). The number of isolated filamentous fungus colonies was under 5 in the almost all the fungi positive cases which included the cases without any pathological changes in the lungs. There was no definite relation between the presence or grade of inflammatory changes of the lungs and detection rate and genus of filamentous fungi. 272 strains of filamentous fungi were isolated from the 53 cases, and the most frequently isolated fungi was Penicillium, and then Aspergillus, and Basidiomycetous fungi. It was noteworthy finding that the genus or species of the isolated filamentous fungi from the human lung were only the limited genus or species of air borne filamentous fungi.3) Yeasts were isolated from 22 cases (30.1%). The rate of yeasts isolation was higher in the cases with bronchitis or bronchopneumonia than in the cases without any inflammatory changes. Candida albicans and Torulopsis glabrata were isolated most frequently.4) Investigations and discussions were made on relations between the rate of fungus isolation and pleural adhaesion, air contents, and regional incidence of the examined areas, and age, sex, post-mortem hours, body temperature of the examined cases, room temperature and seasonal difference at which the materials for culture were taken.5) From the results obtained, the authors concluded that even a healthy respiratory parenchyma of the human lung can not be assumed as aseptic, but the isolated fungi from the lung without fungus infection were a few in number, and the genus or species of the fungi isolated from the respiratory parenchyma were only the limited genus or species of air borne fungi. Furthermore, an attainability of air borne fungi to the respiratory parenchyma of the human lung was discussed in connection with the pathogenesis of opportunistic fungus infections.

The Corticium foot-rot disease caused by Corticium gramineum Ikata et MATSUURA, occurring particulary on the basal part of wheat stem, is characterized by the dark-brown lesion and later by the decay of stem lesion and infection of young leaves in severe case. The effects of soil types, organic manure, chemical fertilizers such as ammonium sulphate, calcium superphosphate, potassium chloride and lime on the development of the disease were studied by the artificial inoculation during two seasons at Akashi, Pref. Hyogo. The results of these investigations showed that the least injury occurred on the wheat plants grown in the sandy soil in which added organic manure, chemical fertilizers together with lime. The sandy soil seemed to be more unfavorable to the development of the disease than the loam and clay soil. There were marked seasonal changes in the microbial populations with the difference of the soil types and the fertilizers. The severity of the disease generally increased with the number of fungi in soil, while decreased with that of bacteria.

When two groups are present, they are said to form an allometric model, if one group is the extension of the other group along the main axis of variation. The model is widely used in the context of principal component analysis, especially for the description of growth processes of creatures. In this paper, the notion of allometric extension model is applied to conditional distributions. More specifically, we derive a sufficient condition, for which the two conditional distributions given the sign of the first principal component form an allometric extension model.

This paper investigates the efficiencies of several generalized least squares estimators (GLSEs) in terms of the covariance matrix. Two models are analyzed: a seemingly unrelated regression model and a heteroscedastic model. In both models, we define a class of unbiased GLSEs and show that their covariance matrices remain the same even if the distribution of the error term deviates from the normal distributions. The results are applied to the problem of evaluating the lower and upper bounds for the covariance matrices of the GLSEs.

This paper investigates some properties of Euclidean distance matrices (EDMs) with focus on their ordering structure. The ordering treated here is the group majorization ordering induced by the group of permutation matrices. By using this notion, we establish two monotonicity results for EDMs: (i) The radius of a spherical Euclidean distance matrix (spherical EDM) is increasing with respect to the group majorization ordering. (ii) The larger an EDM is in terms of the group majorization ordering, the more spread out its eigenvalues are. Minimal elements with respect to this ordering are also described.

Five selenium compounds, Na 2 SeO 4 , H 2 SeO 4 , Na 2 SeO 3 , H 2 SeO 3 and SeO 2 , were tested for their capacity to induce chromosome aberrations in cultured human leukocytes and for their reactivity with DNA by a rec-assay system and inactivation of transforming activity in Bacillus subtilis . Chromosome-breaking activity was significantly higher for the compounds with four-valent than with six-valent selenium, the efficiency being in the decreasing order H 2 SeO 3 > Na 2 SeO 3 > SeO 2 ⪢ H 2 SeO 4 > Na 2 SeO 4 . Rec assay using B. subtilis with different recombination capacities suggested that damage to DNA was produced by selenites but not by selenates. The reactivity of selenites with DNA was also indicated by a significant loss of transformation of the tryptophan marker of B. subtilis DNA treated with H 2 SeO 3 and SeO 2 .

Microsporum canis was isolated from the inflammatory skin lesions of a Bengal tiger in April, 1979. In clinical findings rounded area of alopecia, 3 to 8 cm in size was observed on the right haunch and the tail. The skin lesions were covered with thin scales, but the inflammatory changes were not severe. Findings, on physical examination, were unremarkable and the animal appeared healthy except for the skin lesions. The infected hairs were invaded by the fungal elements and arthroconidia were around the hair shaft. Cuticles and cortex of the infected hairs were brittle. They were evidently digested and were easily pulled out from the hair follicles. In mycological findings numerous and typical macroconidia were observed. By cross mating of a isolate from tiger and Nannizzia otae(-) strains, cleistothecia were produced. Both organic iodide and undecylenic acid ointment were effective for therapy. In this paper the dermatophytosis of tiger caused by M. canis is described.

The culture filtrates of Bacillus cereus strains isolated from diarrheal-type and vomiting-type food poisoning outbreaks were examined for their enterotoxigenicity, vascular permeability-enhancing (VP) activity and mouse lethality. Seven of nine diarrheal strains were starch-hydrolysis positive; all vomiting ones were negative. All diarrheal strains elicited VP reaction; the vomiting strains did not. The mouse lethal activity of the diarrheal strains paralleled the VP activity; such was not the case with the vomiting strains. The culture filtrates of four diarrheal strains among eight and none of vomiting strains caused fluid accumulation in the mouse ileal loop. The concentrated (50-fold) culture filtrates of five diarrheal strains of eight caused fluid accumulation in rabbit ileal loops; only three of 21 vomiting strains did so.

Human hairs infected with Microsporum ferrugineum were examined by light and scanning electron microscopy to obtain information on the parasitic form. The fungus invaded the hair filaments and developed hyphae between the hair cuticles and cortex. Spheroids of arthroconidia were frequently observed to invade the hair follicle. A mosaic sheath of massive spheroids was observed around the hair filaments. The smooth-walled arthroconidia with chitinous surfaces were single, spherical or subspherical particles of 2.0-2.5 X 2.5-3.5 microns, and 2-celled, gourd-shaped (constricted at the septum) particles of 2.5-3.0 X 5.0-7.0 microns.

Summary A statistical survey of visceral aspergillosis in Japanese pathological autopsies (1972–1981) was carried out. It was found that the infection has increased remarkably in recent years, and the results obtained were briefly described. Pathogenicity of several strains of Aspergillus fumigatus in rabbits was studied on three occasions in the past 31 years. The pathogenicity appeared to be unchanged even after long subcultures for 13 years, while lowered pathogenicity was noted with long-term subcultures over 30 years. However, the lowered pathogenicity of the fungus seemed to be recovered by animal passage.

Muscle fiber conduction velocity in human biceps brachii muscle, produced by voluntary contraction and by contraction owing to microstimulation of a single motor unit, was measured with surface array electrodes. The conduction velocity of the fibers in the motor unit was calculated from the conduction time of the motor unit action potential along the electrode array and the electrode separation. With voluntary contraction, a conduction velocity of 4.25 +/- 0.43 m/sec (mean +/- S.D., n = 68) was obtained. In recording the surface EMG, the mean firing rate of the motor unit was 15.8 imp/sec (range 6-24 imp/sec). Significantly slower conduction velocity of 3.69 +/- 0.33 m/sec (mean +/- S.D., n = 56) was found after microstimulation (P less than 0.001). The higher the stimulation rate the higher was the conduction velocity. With increasing stimulus rates of 5, 10, 20 and 40 c/sec, the mean and S.D. of the conduction velocity were 3.74 +/- 0.33 m/sec (2.1% increase in the mean value to 1 c/sec stimulus rate), 4.16 +/- 0.37 m/sec (13.6%), 4.35 +/- 0.54 m/sec (18.8%) and 4.80 +/- 0.49 m/sec (31.1%), respectively. The firing rate for voluntary contraction was in the same range of the one obtained with 10-20 c/sec electrical stimulation, conduction velocity was the same in the two conditions. We conclude that measurement of muscle fiber conduction velocity should also be standardized with muscle fiber firing rate.

Larvae of the xanthid mud crab Eurytlum Umosum were reared in the laboratory from hatching to first crab stage. Four zoeal stages and one megalops stage were obtained and are described. Complete larval development required about 15 days under culture conditions of 26.0 to 28.0 C and 25 ppt sea water. A long antenna and short antennal exopods consign E. Umosum to the Group I xanthid zoeae of Rice (1980). The first stage zoea closely resembles that of Panopeus herbstii and is distinguished by having the dorsal spine strongly recurved at the extremity. Stages 2 to 4 are dis­ tinguished from Rhithropanopeus harrisii and Neopanope sayi larvae by having two lateral spines on the telson.

An ecological study was carried out on the fungal flora in the human lung of 159 autopsy cases. Fungi were isolated from 129 cases (81.1%). Filamentous fungi consisting of 918 strains were isolated from 113 cases, and the dominant genera were Aspergillus and Penicillium. Yeasts were isolated from 58 cases, and the dominant genus appeared to be Candida. With morbid anatomical study, the incidence of the fungus isolation was examined from various points of view. It was concluded that even a healthy respiratory parenchyma of the human lung cannot be assumed as aseptic. However, only a few numbers of a few genera of air-born fungi were isolated. Special stress was laid on the exposure of the respiratory parenchyma of the human lung to air-born fungi in connection with the pathogenesis of opportunistic fungus infections.

SUMMARY: A highly radioresistant bacterium was isolated from the faeces of a giant panda (Ailuropoda melanoleuca). When the organism was subjected to gamma irradiation in phosphate buffer, the induction dose and D10 values were 846 and 345 krad, respectively, for cells grown on PCNZ agar, and 700 and 460 krad, respectively, for the enlarged cells grown on 5 % (v/v) horse blood brain heart infusion agar. The D10 value of the former cells was about 1.8 times higher than that of Micrococcus radiodurans grown on PCNZ agar.

Forty-nine powdered herbal drugs were analyzed for their mold profile and for the potential presence of three mycotoxins (aflatoxin, sterigmatocystin, ochratoxin A). Aspergillus and Penicillium species were predominant, but Rhizopus, Mucor, Cladosporium, and Aureobasidium spp. were also found in a few samples. Mycotoxins were not detected in any samples, and only one isolated culture was found to be a mycotoxin producer on laboratory media.

Fifteen strains of Phoma species were examined for their production of toxic metabolite (s). Although three of them showed the toxicity for cultured HeLa cells and mice and the metabolites were not identical with the known metabolites, attempts to isolate toxic principle (s) were abandoned due to their instability. In the course of the work cynodontin (3), 6-methylsalicylic acid, an unidentified phenolic of mp 191-193°, and a chromanone (4) were isolated. The chromanone (4) was proved to be (2R)-5-hydroxy-6-(2'-hydroxyethyl)-7-methoxy-2-methylchromanone.

Studies on mycotoxin-producing fungi contaminating foodstuffs have been carried out with the aim of disclosing the possible causative agents of human diseases, particularly of human cancer, in Japan. Several areas showing relatively high incidence of liver and stomach cancers were selected for this project, including one rural city in central Japan (Honshu district) and three towns or villages in southern Japan (Kyushu district). Nutritional and epidemiological investigations were also performed in addition to the collection of the foodstuffs in these areas. Final conclusions concerning this mycotoxin-epidemiological study have not yet been obtained but the mycological examination and some of the chemical assays for mycotoxins are nearly completed. The results of these studies will be presented here as the first report.

Culture filtrates and chloroform extracts from Penicillium purpurogenum isolated from foodstuffs were subjected to preliminary survey for toxic effects on HeLa cells and mice. The toxic metabolite was isolated in a crystalline form from P. purpurogenum and was identified as rubratoxin B. Pathological findings observed in HeLa cells and mice treated with the metabolite are briefly described.

Aflatoxin B 1 (0.05 μM per larvae) was detected in silkworm larvae artificially attacked by an Aspergillus flavus isolate from a sericultural farm in Japan.

In the previous paper of this series, we reported that three strains of bacteria, i. e. Clostridium sordellii, Propionibacterium acnes and Campylobacter sp. were identified as being able to form cyclohexylamine (CHA) from sodium cyclamate (CHS-Na) in in vitro experiments. These strains have been isolated from the gastrointestinal flora of CHS-Na converter guinea pigs. To confirm their ability to convert CBS-Na to CHA in vivo, these were given alone or in combination to germfree and conventional mice in drinking water containing CBS-Na, and excretion of CHA in the urine and the stools was examined. The growth of the bacteria was also checked. Campylobacter sp. seemed unable to establish itself in the intestine, because the bacterial count decreased from 104 at 3 days to below 102 at several weeks when this bacterium was administered alone. On the other hand, the mixed cultures of Campylobacter sp. with each of the other two strains and of all three strains combined were found to establish themselves well in the intestine of the germfree mice as indicated by bacterial counts of 107-1010. C. sordellii and P. acnes were also able to colonize the intestine alone in or combination; the bacterial counts were always 109-10. As regards excretion of CHA in the urine and the stools, positive results were obtained with the combination of Campylobacter sp. and C. sordellii, and of all three strains. In the former case, the excretion of CHA gradually increased for 4 weeks and reached 2mg/day/mouse. The results were similar in germfree and conventional mice. The results of this experiment suggest that the converting bacteria were Campylobacter sp. and C. sordellii. However, it is not clear which bacterium is the main converting bacterium at present, so a more detailed investigation of the CH-Na conversion mechanism by both bacteria will be carried out in the next phase of this study.

The formation of nitrosodimethylamine (NDMA) from dimethylamine (DMA) and sodium nitrate in the stomach of monkeys was investigated.Five monkeys maintained on commercial monkey diet were force-fed with 20ml/kg of brain heart infusion broth containing 500ppm of DMA and 1200ppm of sodium nitrate. Three monkeys, used as control animals, were starved for one day before the force-feeding with the two compounds. The concentrations of NO2- and NDMA, the pH and the types of microorganisms in the sampled stomach contents were chemically and microbiologically determined at 0, 0.5, 1, 3, 5 and 7 hours after the administration of DMA and sodium nitrate. The blood concentration of NDMA was also measured.The results were as follows:1) After the administration of DMA and sodium nitrate, the peak concentrations of NO2- and NDMA in stomach contents of the three one-day starved monkeys were 2.7, 89 and 16.2ppm, and 41.1, 144.4 and 5.4ppb, respectively, while the peaks of the two compounds in the normal diet monkeys were 75.6, 69.9, 5.7, 1.0 and 201.5ppm of NO2- and 833.6, 63.9, 6.1, not detected and 4.7ppb of NDMA. However, NDMA was not detected (detection limit 0.2ng) in the blood of any monkey investigated.2) The good NDMA converter monkeys generally had a higher pH value of the stomach contents than the poor converter monkeys.3) No remarkable variation of microbial counts was observed in the stomach contents of the monkeys tested.

Samples of wheat and barley grains in the domestic marketing channel from 1971 to 1972, were obtained and examined for their fungal contamination by means of the mycological method. Fungi which emerged from surface-sterilized grains, plated on Potato-Dextrose Agar containing chloramphenicol and on PDA containing 25% dextrose, were isolated and identified.Aspergillus, Penicillium, and Alternaria were predominantly found in 70-80% of the all samples and Cladosporium, Rhizopus and Mucor were commonly isolated, but Fusarium was not frequently detected. Eighteen point eight percent of the wheat samples and 39.3% of the barley samples were recognized as a highly contaminated sample carring the genus Aspergillus. Aspergillus flavus appeared in 44% of the wheat and 31% of the barley, and 8% of the wheat samples and 10% of the barley samples were found as a highly contaminated sample. A. versicolor occured in 38% of the barley samples and 16% of the wheat samples and its highly contaminated barley samples were accounted for about 7%. A. ochraceus and A. clavatus were isolated from 14 to 25% of the wheat and barley samples, but only 2% of these positive samples showed a high contamination with the both fungi.Representative strains of the genus Aspergillus isolated from all the sample grains were determined chemically for their mycotoxin producibility on either liquid or solid media.Mycotoxin-producing strains were: nine Aflatoxins-producing strains from 58 strains of A. flavus (15.5%); 32 Sterigmatocystin-producing strains from 48 strains of A. versicolor (66.7%); 5 Ochratoxin A-producing strains (23.6%) and 17 Penicillic acid-producing strains (77.3%) from 22 strains of A. ochraceus; 10 Patulin-producing strains from 13 strains of A. clavatus (77%).

In Vitro culture of 49 strains of anaerobic bacteria isolated from the stomach of monkeys was carried out to identify those with the ability to form nitrosodimethylamine (NDMA) in GAM broth containing sodium nitrite (1000ppm) and dimethylamine (500ppm). The medium was adjusted to pH 6.The results were as follows:1) Of the 49 strains of anaerobic bacteria, including the family Bacteroidaceae (dominant strains in the stomach), the following 18 strains were recognized as NDMA-forming bacteria: Bacteroides coagulans (2 strains), B. melaninogenieus ss. melaninogenicus (2 strains), B. corrodens (1 strain), Fusobacterium nucleatum (5 strains), Bacteroidaceae spp. (5 strains), Eubacterium lentum (2 strains), and Peptostreptococcus sp. (1 strain).2) Comparative studies of the rate of NDMA formation and the pH dependence indicated that the rate of NDMA formation by the anaerobic NDMA-forming bacteria was (1.1 to 1.5 times) higher than that in the blank culture and the pH was unchanged, while the aerobic strains, except for Neisseria sicca and Streptococcus equi showed a rate two to three times higher than that of the blank culture and the pH apparently decreased to a final pH of about 5.5 (5.1 to 5.9).3) Studies on NDMA formation in GAM broth with 0.3% glucose or in brain heart infusion with 0.2% glucose showed that anaerobic NDMA-forming strains were non-glucose fermenters, whereas aerobic ones were glucose fermenters and oxidizers. It was concluded that there was no specific relationship between NDMA formation and the utilization of glucose.

Eighty-nine representative yeast cultures isolated from 5 kinds of chilled foods such as potato or macaroni salad, Chinese dumplings and raw seafoods (sea urchin roe, sliced tuna, hen clam feet), were taxonomically studied. These yeast isolates were identified as the following 36 species belonging to 8 genera: Candida beechii Buckley et van Uden, Candida guilliermondii (Castellani) Langeron et Guerra, Candida humicola (Daszewska) Diddens et Lodder, Candida intermedia (Cifferri et Ashford) Langeron et Guerra, Candida lambica (Lindner et Genoud) van Uden et Buckley, Candida lipolytica (Harrison) Diddens et Lodder, Candida membranaefaciens (Lodder et Kreger-van Rij) Wickerham et Burton, Candida parapsilosis (Ashford) Langeron et Talice, Candida pelliculosa Redaelli, Candida sake (Saito et Ota) van Uden et Buckley, Candida santamariae Montrocher, Candida solani Lodder et Kreger-van Rij, Candida valida (Leberle) van Uden et Buckley, Candida zeylanoides (Castellani) Langeron et Guerra, Cryptococcus albidus (Saito) Skinner, Cryptococcus infirmo-miniatus (Okunuki) Phaff et Fell, Cryptococcus laurentii (Kufferath) Skinner, Cryptococcus luteolus (Saito) Skinner, Cryptococcus macerans (Frederiksen) Phaff et Fell, Debaryomyces hansenii (Zopf) Lodder et Kreger-van Rij, Kloeckera apiculata (Reess emend. Klocker) Janke, Leucosporidium gelidum Fell, Statzell, Hunter et Phaff, Rhodotorula aurantiaca (Saito) Lodder, Rhodotorula glutinis (Fres.) Harrison, Rhodotorula graminis di Menna, Rhodotorula minuta (Saito) Harrison, Rhodotorula pallida Lodder, Rhodotorula rubra (Demme) Lodder, Torulopsis candida (Saito) Lodder, Torulopsis halonitratophila Onishi ex van Uden et Vidal-Leiria, Torulopsis holmii (Jorgensen) Lodder, Torulopsis magnoliae Lodder et Kreger-van Rij, Trichosporon cutaneum (de Beurm., Gougerot et Vaucher) Ota, Trichosporon penicillatum do Carmo-Sousa, Trichosporon pullulans (Lindner) Diddens et Lodder, Trichosporon variabile (Lindner) Delitsch.

Mycological survey of Salami sausages in retail markets, 13 samples of mold-fermented type and 18 of natural fermented type, was carried out from viewpoints of food sanitation. Most important genus of fungi detected from the mold-fermented type Salamis was Penicillium, particularly representative of P. cyclopium, P. miczynskii, and P. viridicatum. These isolates might be brought from the artificial inocula during a manufacturing process of the Salamis. In the natural fermented type, more broader range of fungal groups were distributed: Cephalosporium sp., Mucor mucedo, M. racemosus, Aspergillus versicolor, Penicillium cyclopium, and others. Furthermore, when the populations of individual groups of fungi were considered, great differences were noted in the total amounts of Penicillium in the natural fermented Salamis as compared with those from the mold-fermented. Totally 23 strains of Penicillium, belonging in P. cyclopium, P. expansum, P. miczynskii, P. viridicatum, and other 3 species, were screened for production of penicillic acid (PA). None of the examined Penicillium strains yielded detectable amounts of PA in liquid shaking culture. It may be concluded from the above results and data of Ciegler, et al. (1972), that consumption of mold-fermented sausage does not represent a potential health hazard.

Proteolytic and lipolytic activities of 2 psychrophilic yeast species and 31 psychrotrophic yeast species isolated from 5 kinds of chilled foods, such as potato or macaroni salad, Chinese dumplings and raw seafoods (sea urchin roe, sliced tuna, hen clam feet), were investigated under various incubation conditions. Proteolytic activity was determined by use of YM agar plus 1% skim milk. Lipolytic activity was determined by use of butterfat in Crossley's agar and beef fat in Gorodkowa agar (Eijkman's method). These 3 kinds of agar plates were incubated for 2 weeks at 5, 15 and 25°C, respectively.The yeasts which demonstrated proteolytic activity under the conditions of incubation for one week at 5, 15 and 25°C, respectively, were 21.2, 18.2 and 15.2% of the 33 species of yeasts tested. The yeasts which demonstrated lipolytic (butterfat) activity under the same conditions of incubation amounted to 30.3, 60.6 and 54.5% of the total, respectively. It was generally found that lipolytic activity was higher than proteolytic activity.Proteolytic and/or lipolyic yeast species were widely distributed among the genera Candida, Cryptococcus, Debaryomyces, Leucosporidium, Rhodotorula, and Trichosporon. Candida sake, C. guilliermondii, C. lambica and C. solani exhibited proteolytic activity when incubated at 5°C within one week. Candida parapsilosis, C. santamariae and C. zeylanoides showed lipolytic activity under the incubation condition. Leucosporidium gelidum, Candida lipolytica and Trichosporon pullulans showed both proteolytic and lipolytic activities at 5°C within 4 days. Strain U1, a yeastlike organism resembling Aureobasidium pullulans showed both activities at 15°C within 2 days, and Rhodotorula aurantiaca, Rh. graminis and Trichosporon cutaneum showed both activities at 15°C within 2 weeks.

Mycotoxin production was investigated in 27 kinds of commercial foods artificially inoculated with following representative toxigenic fungi, i. e., afiatoxin-producing strain of Aspergillus parasiticus NRRL-3000, sterigmatocystin-producing strain of A. versicolor I-20-b, ochratoxin A-producing strain of A. ochraceus 10-21-N, and Fusarium toxinproducing strain of Fusarium solani F-135.It was observed that among vegetables and food products tested, dried sweet potato and salami sausage were favorable substrates for the production of several kinds of mycotoxins and dried persimon was favorable for sterigmatocystin production. Pepper, roasted coffee bean and cinnamon bark were not susceptible to mycotoxin contamination under the moistend condition.Substances contained in a chloroform extract of cinnamon and pepper inhibited the growth of the four toxigenic fungi. These results indicate that cinnamon and pepper might contain inhibitory materials.

Cyclohexylamine (CHA) is a metabolite of the artificial sweetening agent Cyclamate which was rejected from the food additives. The notice about cyclamate is subsiding, but studies with this substance have been continued. To determine sodium cyclamate (CHS-Na) assimilating bacteria inhabiting gastrointestinal tract of animals, a quantitative and qualitative bacteria analysis for faecal materials have been conducted in a monkey administered orally with 20% CHS-Na solution (1.25ml/kg daily), and in untreated monkey. The faecal flora of the normal monkeys predominently constituted Catenabacteria and Peptostreptococci, and Clostridia, Bacteroidaceae, Bifidobacteria, and Veillonella, all of which belong to the obigate anaerobe, were commonly found in order. Facultative anaerobes like Lactobacilli and Streptococci were abundantly found, and Enteroba cteria and Staphylococci could be detected in the faecal homogenates. On the other hand, the constitution of faecal flora in the monkey excreting CHA in the urine was not quite different from that of the normal monkeys. In addition, any marked alteration of the flora in the treated monkey was not observed throughout this analysis. In parallel with this, in vitro, culture study was carried out by using the modified BC broth containing 0.25% of CHS-Na. Initial mass cultures obtained from the faecal homogenates were inoculated into the BC broth and incubated at 37°C for 14 days under anaerobic condition, and production of CHA periodically examined by the chemical procedure. The production of CHA by the initial mass culture was gradually increasing in progressively. After 14 days culture, bacteria survived were Clostridia, Bacteroidaceae, Lactobacilli, and Streptococci, those which may have CHA productivity. These bacteria and the initial mass culture were subcultured again on the BC broth but no production of CHA was observed in these subcultures. After the cultivation on differential media for identification, these organisms exposed to the air (O2) may have lost their CHA productivity. From this study, it is indicated that although CHA converting bacteria was not definitely detected in vitro, some anaerobic bacteria such as Clostridia, Bacteroidaceae, Lactobacilli and Streptococci seemed to have sodium cyclamate metabolizing activity in gastrointestinal tract of the monkeys.

Microbiological examination of chilled roe of sea urchin (Namauni; Heriocidaris spp., etc., 20 samples), sliced raw tuna (Maguro sashimi; Thunnus spp., etc., 20 samples) and the foot of the hen clam (Bakagai sashimi; Mactra spp., 17 samples) was carried out to examine the microflora, especially the yeasts. The samples were collected from retail markets by official food examiners and investigated by the official microbiological methods. Counts of bacteria, yeasts and fungi in the samples of roe of sea urchin ranged from 103 to 107, 102 to 107 and below 104 per gram, respectively. Counts of bacteria, yeasts and fungi in the samples of sliced raw tuna and the foot of the hen clam were found to range from 103 to 106, 101 to 104 and 104 per gram, respectively.In the three kinds of seafood samples, yeasts were found to be the second largest microbial group and the predominant species were asporogenous yeasts such as Rhodotorula (R. glutinis, R. rubra, R. pallida), Candida (C. guilliermondii, C. humicola, C. lipolytica, C. parapsilosis, C. zeylanoides), Trichosporon (Tr. cutaneum, Tr. pullulans, Tr. variabile) and Cryptococcus (Cr. albidus, Cr. infirmo-miniatus, Cr. laurentii, Cr. macerans). Trichosporon cutaneum, a species of opportunistic pathogen, was isolated from 45% of the samples of sliced raw tuna. In all the samples tested, the main bacterial genera were Pseudomonas, Corynebacterium, Micrococcus, Staphylococcus, etc., and representative fungal genera were Penicillium, Phoma, Fusarium, Mucor, etc.Psychrotrophic organisms grown within 2 weeks at 5°C accounted for 94.6% of the total yeast cultures and 55.0% of the total fungal cultures isolated from the seafood samples.

Fate of nitrate and nitrite in saliva of monkey was investigated by use of five monkeys (2.5-5.0kg) which were force-fed by a stomach tube with 4, 20, 40mg/kg of body weight of sodium nitrate solution at the ten-days intervals. As the results of this preliminary investigation, the monkeys could be grouped three different converters such as very good (M1), good (M2 and M3) and poor converter (M4 and M5).To obtain more detail information about the reduction of ingested nitrate, further investigation was carried out on the good converter (M1), and the two poor converters (M4 and M5) of monkey as used for control. All monkeys used in this experiment were force-fed with 100mg/kg of sodium nitrate solutionby the stomach tube. Concentration of nitrate and nitrite in blood and saliva were successively calculated by the chemical procedure. Paralleled with this experiment, comparative survey for microflora of oral cavity of the monkeys was also conducted. Results of these studies were summarized as follows:1) Nitrate concentration of blood in all the monkey tested peaked at 7hr after ingestion showing 180ppm. In the good monkey (M1), about 70ppm of the concentration was maintained for 48hr after administration.2) Salivary concentration of nitrate in the good monkey showed in maximum concentration of 800ppm after 7hr, and the salivary concentration of nitrate was always kept in higher than that in blood. In contrary, the poor monkeys indicating rather lower concentration of nitrate in blood was observed.3) Salivary concentration of nitrite in the good monkey indicated that a maximum concentration was 30ppm and 10-20ppm of nitrite was maintained in saliva for over 24hr. In the poor monkeys, however, no apparent correlation between ingestion of nitrate and formation of nitrite in saliva was observed.4) A total count of bacteria and molds in oral cavity of the good monkey always showed a greater number of microbes than that of the poor monkeys. Streptococcaceae, Peptococcaceae and Bacteroidaceae were predominantly isolated from oral cavity of the good monkey. Of which, Bacteroides melaninogenicus was mostly common species among the microflora. Nitrate reducing species of bacteria belonging anaerobic bacteria, i. e., Bacteroidaceae and Veillonella seemed to be indigenous species in mouth of the good monkey.

The qualitative and quantitative analytical methods for sterigmatocystin in grains were studied. The mycotoxin was extracted with a mixture of methanol and 1% NaCl solution (55:45) from grains and was reextracted with chloroform. After cleaned up by silica gel column chromatography, it was detected and estimated by thin layer chromatography. The quantitative step of the presented procedure was based on conversion of sterigmatocystin to a more highly fluorescent derivative by spraying a 20% AlCl3 ethanol solution on a developed thin layer plate. The measurement of its fluorescence was carried out by using a fluorodensitometer. For the chemical confirmation of sterigmatocystin, it was suitable to treat a spot of the mycotoxin with trifluoroacetic anhydride to produce a fluorescent derivative of the mycotoxin, before the second development of the procedure of two-dimensional thin layer chromatography. Furthermore, analytical method for sterigmatocystin by gas chromatography was investigated.