In order to make a toxicological assessment of organic sulfur compounds in heavy oil various conventional biological tests were applied.
(1) Killifish test for median tolerance limit (TLm 24hr). (2) Drosophyla growth test. (3) Chicken embryo assay. (4) Mutagenic activity to Salmonella typhimurium TA 98. (5) Sea urchin egg growth test.
The results were as follows: TLm 24hr of C class heavy oil corresponded to around 80ppm, while A and B class heavy oil gave over 100ppm. TLm 24hr of dibenzothiophene was 17ppm. No significant effects of heavy oils on the developmental stage of Drosophyla were observed.
In the sea urchin egg growth test, the effective concentration of dibenzothiophene was 25ppm, i. e., this compound was more toxic to marine or fresh water organisms than heavy oil itself.
Higher boiling distillate oil such as B and C class heavy oil showed higher toxicity to chicken embryos than A class heavy oil.
In the tests for mutagenic activity using TA 98, B class heavy oil showed slight mutagenicity at a concentration of 500μg/plate. Sulfur compounds (benzothiophene and dibenzothiophene) and sulfur containing oil fraction from heavy oil appeared to exhibit no mutagenicity with respect to TA 98.

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カネミ油症原因油 (1968年2月10日製造) からPCBおよびPCDFをそれぞれ分離しマウスに経口投与した後, 体重, 肝重量および各組織中のPCBパターンの解析を行った.
1. PCBとともにPCDFをマウスに経口投与すると肝/体重比は増加した. また体重抑制の傾向も見られた.
2. PCBとともにPCDFをマウスに経口投与するとピーク25, 28の濃度が減少し油症患者の血中PCBパターンに類似のPCBパターンが得られた. すなわち油症患者の血中PCBパターンにはPCDFが関与していると考えられる.
3. 市販の合成PCDFよりも油症原因油より分離したPCDFの方がピーク25, 28の濃度減衰に及ぼす影響は大きかった.
4. PCBパターンは, 脂肪および肝臓で経日変化が認められた. また脂肪と肝臓の間でもPCBパターンに差異が認められた.

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In 1977, there was an outbreak of photosensitivity dermatitis by among persons who had ingested a certain brand of chlorella tablets. At least 23 patients were identified in and around the Tokyo metropolitan area.
The unconsumed chlorella tablets collected from these patients invariably induced photosensitivity dermatitis in mice. These chlorella tablets contained large amounts of pheophorbide and its ester, which are well-known phototoxic pigments derived from chlorophyll, while chlorella tablets from another maker, which contained less than a tenth as much pheophorbide induced no photosensitivity lesion in mice.
Furthermore, a linear dose response relationship was observed between the severity of photosensitivity dermatitis in experimental animals and the content of pheophorbide in chlorella administered. However, the reaction induced by pheophorbide extracted from the suspect chlorella was weaker than that induced by the original chlorella sample.
It was confimed that ethanol, used in forming granules of chlorella before pelleting, activates chlorophyllase, which in turn hydrolyzes chlorophyll naturally contained in chlorella into pheophorbide and its ester.
We concluded that the photosensitivity dermatitis was caused by the chlorella tablets, which had been treated with ethanol. We believe that pheophorbide and its ester found in large amount in these tablets, have played a major role in inducing the photosensitivity dermatitis.

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Both protein and carbohydrate were necessary for sporulation and enterotoxin production of Clostridium perfringens NCTC 8239 in the ligated intestine of the rabbit. The administration of 10⁸ vegetative cells suspended in 2ml of skim milk was confirmed to induce positive reactions consistently. However, skim milk dialysate, which contained the same amount of protein as the 10% skim milk, had no effect. An addition of 10% lactose to the skim milk dialysate restored the effect. Proteose peptone (Difco), peptone for toxin production (Mikuni), meat extract (Wako) and purified casein (Merck) could replace the skim milk dialysate.
In addition to lactose, raffinose and fructose were effective as carbon sources whereas galactose, sucrose and glucose were less effective. Enterotoxin was not produced in the ligated intestine when the vegetative cells were administered together with any one of these carbohydrate or protein sources alone.

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In September and October 1977, two outbreaks of food poisoning due to Bacillus cereus occurred in Osaka city.
Outbreak 1: On 4th of September 1977, 9 out of a total 13 of employees (69.2%) in two factories became ill 0.5 to 3 hours after taking lunch. Vomiting was the main clinical symptom. B. cereus was isolated (1.4×10⁷ per gram) from the suspect lunches.
Fecal specimens collected from 2 patients on the 3rd day of illness were positive for B. cereus. No other known food-poisoning organism was found either in the lunch or in the feces of patients.
Outbreak 2: On 9th of October 1977, 211 out of 1809 persons (11.7%) who had eaten a packed lunch (‘Onigiri’, ‘Maki-sushi’ and ‘Inari-sushi’) complained of nausea and vomiting after 0.5 to 6 hours. B. cereus were isolated from ‘Onigiri’ (4.7×10⁸/g), ‘Maki-sushi’ (1.3×10⁴/g) and ‘Inari-sushi’ (3.7×10⁴/g), and from 18 out of 21 fecal specimens (85.7%) collected from the patients on the 1st to 3rd day of illness.
All isolates from outbreaks 1 and 2 had the same biological characteristics. Culture supernatants of the two isolates showed heat-sensitive lethal activity to mice on intraperitoneal or intravenous injection.

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アマランスおよびポンソー3Rなどは単離核のRNA合成を促進する作用があり, その機作の解明を試み, 次の結果を得た.
1) 1mMのアマランスおよびポンソー3Rはクロマチンを鋳型としたRNA合成を単離核のRNA合成と同程度促進した. したがって, これらの色素によるRNA合成反応促進は, 核膜が色素により変化して基質の取り込みが増加したためではないと考えられる. しかし, ライトグリーンSFはクロマチンの鋳型活性を促進しなかったことから, 単離核のRNA合成反応促進の機作において, ポンソー3Rおよびアマランスと異なることが推察される.
2) 単離核およびクロマチンにおいて, ポンソー3Rおよびアマランス添加区は色素無添加区およびサンセットイエローFCF添加区よりDNase Iによる分解に対して約2倍程度強い感受性を示した. また, アマランス, ポンソー3RおよびサンセットイエローFCFは仔牛胸腺DNAを基質にしたときのDNase I活性および核の内在性ヌクレアーゼ活性に対して影響を及ぼさなかった.
3) アマランス, ポンソー3RおよびサンセットイエローFCFは単離核のRNaseを1mMで若干阻害した. しかし, サンセットイエローFCFは単離核のRNA合成反応を促進しないことから, 単離核のRNA合成促進にはほとんど関与していないと考えた.

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The determination of bismuth and lead in foods was examined using a flameless atomic absorption spectrometer equipped with a carbon tube. To elminate the matrix effect, which is observed in direct measurements of acid test solutions, the organic layer obtained by extraction of a test solution (pH ca. 8) with a sodium diethyldithiocarbamic acid (DDTC) -methylisobutylketone system was injected into a carbon tube. Examination of ashing and atomizing conditions gave optimum sensitivities of 3 (bismuth) and 5ppb (lead) for 1% absorption. Coefficients of variation obtained by repeated extractions and injections were 1.9 (bismuth) and 3.4% (lead). Linear standard curves were obtained up to 120 (bismuth) and 100ppb (lead). Although the coexistence of chelating metal ions with DDTC caused some interference it is possible to detemine these metals with fairly good accuracy by a calibration method without using standard addition procedures when this method is used for the analysis of bismuth and lead in modified milk powder.

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