Microorganisms which cause infectious diseases are generally called pathogenic microorganism. In this article pathogenic bacteria including those causing zoonosis, especially diarrhogenic bacteria and causative bacteria of legionnaires' disease will be discussed in relation to their contamination in our life-environment. I. Bacterial diarrhea 1. Present status of traveller's diarrhea. 2. Imported cholera cases frequently reported since a cholera outbreak in Arida in June, 1977. 3. Distribution and cotamination in the life-environment of newly recognized bacteria as causative agents of food poisoning. II. Legionnaires' disease 1. Cases of legionnaires' disease in Japan. 2. Contamination of the causative bacteria of legionnaires' disease in the life-environment.

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For the appropriate regulation of natural environment, microbiological treatment of some disinfectants with activated sludge was investigated, especially the degradation and acclimatization of some disinfectants against the activated sludge were examined. Bubbling rate of above 200 ml/min is required for the maintenance of good degradation condition in the treatment system with activated sludge. Glutaraldehyde (GA) was degraded nearly perfect, benzalkonium chloride was more degraded, 8-hydroxyquinoline sulfate was degraded moderately and chlorhexidine digluconate was slightly degraded in the activated sludge. Acclimatization of GA was worse than the other disinfectants. Three kinds of Ps. aeruginosa isolated from the acclimatization treatment system are responsible for the good treatment ability of some disinfectants.

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Gas chromatography-negative ion chemical ionization mass spectrometry (GC-NCIMS) was evaluated as a method for qualitative and quantitative determination of organobromine compounds in water. The negative ion mass spectra of bromine containing trihalomethanes (CHBrCl₂, CHBr₂Cl and CHBr₃) consisted of two peaks of m/z 79 and 81, corresponding to mass numbers of bromine atom. The detection limit for these compounds was 1.0 ppb. The results in this study showed that organobromine compounds in water could be assayed by GC-NCIMS.

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The relative nitration rates of 26 kinds of polycyclic aromatic hydrocarbons by nitric acid were examined to predict the formation of their nitro compounds in the diesel engine. The three nitration reagents used in this experiment were a mixture of nitric acid-acetic acid (1 : 10), (5 : 10), (10 : 10). Each reaction product was determined by gas chromatography with ECD (ECD-GC) and gas chromatography-mass spectrometry (GC-MS). Rates of nitration were in the following order, 3, 4, 8, 9-dibenzopyrene>perylene>1, 12-benzoperylene>3, 4, 9, 10-dibenzopyrene>benzo [a] pyrene>benzo [a] anthracene>1-methylpyrene>pyrene>anthracene>2, 3-(o-phenylene) pyrene>acenaphthene>1, 2, 3, 4-dibenzoanthracene>1, 2, 5, 6-dibenzoanthracene>benzo [k] fluoranthene>benzo [e] pyrene>2, 3-benzofluorene>1, 2-benzofluorene>chrysene>fluorene>fluoranthene>phenanthrene>benzo [b] fluoranthene>naphthalene>triphenylene. Nitro derivatives of naphthacene and picene were not given under these three nitration conditions. The reaction of polycyclic aromatic hydrocarbons with nitrogen dioxide was also examined.

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The mechanism of reaction of glycine with hypochlorite was investigated under neutral conditions in phosphate buffer. At the first stage in this reaction, monochloroglycine was formed at any molar ratio of glycine and hypochlorite. When the amount of hypochlorite exceeded the equimolar of glycine, dichloroglycine was formed from monochloroglycine. Although monochloroglycine was fairly stable, dichloroglycine was labile, so that it underwent decarboxylation and dehydrochlorination to form cyanide, as the corresponding nitrile. Cyanide thus formed was chlorinated rapidly by combined residual chlorine in chloroglycine molecules or by excess hypochlorite, and converted to cyanogen chloride. One mole glycine needed three moles of hypochlorite for the formation of the equivalent cyanogen chloride. Cyanogen chloride was decreased by the subsequent addition of excess hypochlorite. Total chlorine consumed by 1 mol glycine was 4.5 mol.

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Methods for analyzing low concentrations of chlorine in air are discussed. The use of 0.2% sulfamic acid solution for the collection of chlorine made it possible to sample air in large quantities as much as 2.0 l/min for 60 min. Sulfur dioxide and ammonia, which are interfering gases could be completely removed by the use of a chromic acid scrubber. The color that developed was stable for more than 60 min when the barbituric acid method of Asmus, et al. was employed. Quantitative determination was carried out fluorospectrometrically with a detection limit of 0.03 to 0.04 ppb. A good analytical precision was obtained with a coefficient variation of 5.3% for 6.6×10^-6 mol of Cl₂.

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Electron capturing detector-gas chromatography was applied to study the behavior of trace amounts of chromium dissolved from a stainless-steel tableware which was examined according to The Standard Methods of Analysis for Hygienic Chemists authorized by the Pharmaceutical Society of Japan. Three kinds of solution were prepared by 1. heating with 4% acetic acid in the tableware to boil for 10 min, 2. standing with 4% acetic acid in the tableware at room temperature for 24 h, 3. standing with 4% acetic acid in the tableware at room temperature for 10 min, by considering the use of each tableware. After the treatments, each test solution was determined by gas chromatography. The present study showed that i) chromium dissolved from tableware was trivalent since gas chromatography detected only the chelate which was formed from trivalent chromium and trifluoroacetylacetone, ii) amounts of chromium dissolved repeatedly from the identical tableware was reduced gradually, iii) relationship between amounts of chromium and iron in test solution obtained from dissolution test was linear (y=19.8×-23.7, r=0.956), and amounts of iron was about 50 to 100 times than that of chromium, iv) it was found that the range of amounts of chromium dissolved was 0.04-29 ng/ml (or 0.09-34 ng/cm²) from the results obtained from various new stainless-steel tablewares and the amounts of chromium dissolved from dissolution test 1, 2, and 3 was in the order 1>2≨3.

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To investigate when the changes of lipoperoxide contents in early stage of the inflammation are induced in the testis of rats after CdCl₂ injection, we measured the lipoperoxide contents in plasma and tissues (liver, kidney and testis) at 1, 3, 5, 10, 15, 24 and 48 h after CdCl₂ (5.0 mg/kg) injection. Lipoperoxide value in the testis increased significantly from 15 h after CdCl₂ injection. Although no change of lipoperoxide value was observed in the liver through the experimental period, the value in the kidney increased significantly at 10 and 48 h after CdCl₂ treatment. Heme protein value which represents the degree of hemorrhage in the testis also increased from 15 h after CdCl₂ injection. This suggests that hemorrhage in the testis has a good correlation with an increase in lipid peroxidation in situ. The fact that significant increases in lipoperoxide values and in the degree of hemorrhage were observed from 15 h after CdCl₂ injection, reveals the possibility that oxidative reactions occur in the testis of rats nearly 15 h after the treatment. Assay of enzyme activities in serum was performed simultaneously. The activity of β-glucuronidase which is considered to be one of parameters showing the occurrence of an inflammation, increased significantly at an early time (from 1 to 5 h) after CdCl₂ treatment. This implies that some biochemical reactions occur in the testis before the time when the increase of lipoperoxide formation occurs. In the liver where no fluctuation of lipoperoxide concentration was observed during the experimental period, the Zn contents increased as the time elapsed after CdCl₂ administration. On the contrary, a significant reduction of the Zn content was observed in the testis in which a considerable increase in lipoperoxide concentration was noticed, at 3-5 h after CdCl₂ administration. The inflammation of testis by Cd is supposed to be initiated at this time stage. The difference of the Zn contents in these tissues may be considered to be one of factors which induce the difference among tissues in lipoperoxide concentration.

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A rapid and reproducible method for the determination of aspartame (AT) in foods by high-performance liquid chromatography (HPLC) was investigated. A food sample was extracted with water. Phosphate buffer, pH 4.0, and cetyltrimethyl ammonium chloride (CTA) were added to the water extract and the solution was passed through SEP-PAK C₁₈ cartridge (Waters Associates). After washing the cartridge with water, AT was eluted with a mixture of methanol-0.027 M phosphate buffer pH 5.0 (1 : 3). The eluate was injected on a Li Chrosorb RP-8 column by using a mixture of methanol-0.025 M phosphate buffer, pH 5.0, (1 : 4) as a mobile phase. The effluent was monitored with a UV detector at a wavelength of 210 nm. The recoveries of AT from liquid foods, solid foods and fatty foods were 96.9-100.9%, 91.1-96.9% and 85.7-98.0%, respectively.

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Impurities in methamphetamine (MA) synthesized from ephedrine were identified by thin-layer chromatography, gas liquid chromatography, infrared spectroscopy, ultraviolet spectroscopy, proton magnetic resonance spectroscopy, gas chromatography/mass spectrometry and neutron activation analysis. Benzyl methyl ketone and iodine were detected as impurities in MA prepared by reducing ephedrine with hydrogen iodide and red phosphorus. Ephedrine, 1-phenyl-2-methylamino-1-propanone, palladium and barium were detected as impurities in MA prepared by reducing ephedrine with hydrogen and Pd-BaSO₄. Ephedrine, 1, 2-dimethyl-3-phenyl aziridine, palladium and barium were detected as impurities in MA prepared by reducing chloroephedrine with hydrogen and Pd-BaSO₄. Knowledge of impurities contained in MA provides important information regarding synthetic methods. Impurities contained in seized MA were analyzed by the method described above, and synthetic methods were assumed.

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The following results were obtained from the experiments by use of mice, to which calcium was given in advance, in order to clarify the role of metallothionein (MT) in acute toxicity by cadmium. 1) With an increase in the amount of Calcium (Ca) administered, the rate of death by the administration of Cd decreased. Between MT concentration in the liver induced by Ca and the rate of death induced by Cd, a negative correlation was found. 2) By studying the time-course of Cd after its administration, it was found that MT concentration and the binding rate of Cd to MT in the group to which only Cd was given were lower than those in the group to which Ca was administered in advance. However, no significant difference was detected in the Cd concentration in the livers of both groups. 3) MT concentration in the liver after the use of more than 6 mg/kg of Cd was lower than that after the administration of 3 mg/kg of Cd. These results show that the Cd acute toxicity was defended by replacing Cd with zinc (Zn) which was bound to MT in the liver.

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Stability of trifluoroacetyl (TFA), pentafluoropropionyl (PFP) and heptafluorobutyryl (HFB) derivatives of methamphetamine and its metabolites in man were investigated. TFA derivatives in solution were more stable than those in solid state, and solution of TFA derivatives in ethyl acetate or n-hexane was stable at 5°C for a few hours. Although PFP and HFB derivatives were more stable than the corresponding TFA derivatives, it took long time for the removal of excess reagent after preparation of the derivatives. TFA, PFP and HFB derivatives of methamphetamine, amphetamine and norephedrine were stable, and those of aromatic hydroxylated metabolites were relatively unstable, especially TFA derivatives of p-hydroxynorephedrine was most unstable in these derivatives. Taking into account the facility in preparation of the derivatives, stability of the derivatives and cost of the reagents, it was practically most useful to prepare TFA derivatives and to apply ethyl acetate solution of the derivatives to gas chromatography within a few hours after preparation.

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In order to investigate mercury pollution in humans and animals, a simple and sensitive method for the determination of nanogram levels of total mercury (T-Hg) in extremely small amounts of urine, blood, hair, internal organs and other specimens was established. The proposed method includes furnace combustion under optimum conditions, capture of mercury as gold amalgam and detection of vaporized mercury. The method was successfully applied to the determination of ppb levels of T-Hg in μl or mg amounts of biological samples.

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Malathion in the blood was determined by gas liquid chromatography with a flame thermionic detector on 1.0 m glass column packed with 10% DC-200 at 225°C. The blood was haemolyzed by the addition of 9 parts of water, subjected to modified Extrelut[○!R] column and malathion was eluted from the column with n-hexane. Determination was carried out successfully by the internal standard method using cyanophenphos. The calibration curve of malathion in ethyl acetate solution (0.5-4.0 μg/ml) showed a straight line. The recovery of malathion added to the blood (4.0 μg/ml) was 100.32±2.07%. Malathion in the blood of a suicide was identified by gas liquid chromatography-chemical ionization mass spectrometry and the content was 1.03 μg/ml.

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The residual contents of pesticides in 13 commercial crude drugs, their decoctions and 15 commercial pharmaceutical preparations which contain crude drugs were determined by using gas chromatography. Eleven kinds of organochlorine pesticides and 9 kinds of organophosphorus pesticides were examined. From the several crude drugs, 8 kinds of organochlorine pesticides were detected, but no organophosphorus pesticides were found from the crude drugs. The maximum values of BHC isomers (total BHC), DDT and related compounds (total DDT), dieldrin and heptachlor epoxide were 0.452 ppm, 0.089 ppm, 0.010 ppm and 0.039 ppm, respectively. The organochlorine pesticides in decoctions prepared from the crude drugs were found to decrease to 0-9.4% of those in the raw crude drugs. The tablets and pills containing powdered crude drugs were found to include higher amounts of organochlorine pesticides than the tablets containing the extracts of crude drugs. More than 1 ppm of total DDT was determined in the soft capsules with fish-liver oil. More than 1 ppm of total BHC and heptachlor epoxide were determined in the soft extract of Ginseng. The potential daily intakes of these pesticides from all samples were calculated. The values obtained were under 5% of acceptable daily intake (ADI) recommended by WHO.

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