PCB 118 and Aryl Hydrocarbon Receptor Immunoassays for Screening Dioxins in Retail Fish

The efficacy of a combination of two enzyme-linked immunosorbent assay (ELISA) kits was examined for screening the toxic equivalent (TEQ) concentrations of dioxins in retail fish. The coplanar PCB-EIA system, which is a competitive immunoassay specific for polychlorinated biphenyl (PCB) 118, was tested as a screening method for mono- ortho PCBs. The Ah immunoassay (Ah-I), which is an ELISA-based aryl hydrocarbon receptor binding assay, was analyzed for its screening ability for non- ortho PCBs, polychlorinated dibenzo- p-dioxins (PCDDs), and dibenzofurans (PCDFs). Dilution and recovery tests using purified fish extracts revealed no major interference of the matrix in the PCB-EIA and suggested that the matrix effect was minimized in the Ah-I. Finally, the results for the fish samples ( n = 20) showed a strong correlation between this method and high-resolution gas chromatography coupled to high-resolution mass spectrometry for the determination of the TEQ concentrations of mono- ortho PCBs ( r = 0.99) and non- ortho PCBs and PCDD/Fs ( r = 0.97). These data indicate that our method is suitable for screening retail fish to determine the TEQ concentrations of dioxins.     

Occurrence of tetrodotoxin in three Nassarius gastropod species in Khanh Hoa Province,Vietnam

Fisheries Science - Neurotoxic poisonings with fatal symptoms caused by consumption of Nassarius gastropods have been reported in Vietnam but the causative toxins have not been confirmed. In the...     

Identification of vitamin B12 and pseudovitamin B12 from various edible shellfish using liquid chromatography–electrospray ionization/tandem mass spectrometry

In this study, the vitamin B₁₂ contents were analyzed in the edible portions of various shellfish (bivalves and snails). High vitamin B₁₂ contents (30.5–53.3 μg/100 g wet weight) were detected in mussels, surf clams, bloody clams, and freshwater clams. However, scallops and abalone had extremely low vitamin B₁₂ contents (0.1–1.1 μg/100 g wet weight) which was attributed to only the muscle portions being edible. These results suggest that high levels of vitamin B₁₂ are accumulated in the viscera of shellfish. Vitamin B₁₂ levels were also significantly higher in bivalves than in snails. The corrinoid compounds purified from all bivalves were identified as “true” vitamin B₁₂ using liquid chromatography–electrospray ionization/tandem mass spectrometry. In edible snails, abalone, and pond snails, however, both vitamin B₁₂ and pseudovitamin B₁₂ (an inactive corrinoid) were observed to be the major and minor corrinoid compounds, respectively. Based on these results, we conclude that the whole bodies of these edible bivalves are excellent sources of vitamin B₁₂ for humans.     

Distribution of protein gene product 9.5-immunopositive and NADPH-diaphorase-positive neurons in the common marmoset (Callithrix jacchus) accessory olfactory bulb

The principal center of the accessory olfactory system is the accessory olfactory bulb (AOB). In primates, simians are divided into two groups, New and Old World monkeys, and the AOB is present in only New World monkeys. The common marmoset (Callithrix jacchus) is a species of New World monkey. Although the morphology of the common marmoset AOB has been demonstrated, the distribution patterns of the mitral/tufted and granule cells of the AOB remain unclear. In the present study, therefore, the distribution of the mitral/tufted and granule cells in the common marmoset AOB was examined using two histochemical markers including immuno-staining for protein gene product (PGP) 9.5 and NADPH-diaphorase staining. The vomeronasal nerves, gomeruli and mitral/tufted cells showed PGP 9.5-immunoreactivity. The mitral/tufted cells were arranged in only one or two rows along the margin of the glomerular layer to form the mitral/tufted cell layer (MTL). Since the mitral/tufted cells occurred sparsely in the common marmoset, the MTL was illegible. NADPH-diaphorase reactivity was primarily detected in the rostral and caudal areas of the AOB. In these areas, granule cells showed NADPH-diaphorase reactivity. Since the granule cells were sparse, the common marmoset AOB displayed less-developed granule cell layer. Although the functional significance of the AOB remains to be solved in the common marmoset, small-sized and less-laminated AOB may show that sexual behavior of the common marmoset has lesser dependence on the accessory olfactory system.     

Immunolocalization of steroidogenic enzymes P450scc, 3betaHSD, P450c17, and P450arom in Göttingen miniature pig testes

The objective of this study was to investigate immunolocalization of steroidogenic enzymes in G?ttingen miniature (GM) pig testes. Testes of 6 adult GM pigs were obtained in September 1996 (n=2), February (n=2) and June (n=2), 1997. Steroidogenic enzymes were immunolocalized using polyclonal antisera raised against bovine adrenal cholesterol side-chain cleavage cytochrome P450 (P450scc), human placental 3beta-hydroxysteroid dehydrogenase (3betaHSD), porcine testicular 17alpha-hydroxylase cytochrome P450 (P450c17), and human placental aromatase cytochrome P450 (P450arom). Histologically, all types of spermatogenic cells including mature-phase spermatozoa in seminiferous tubules were observed in all testes throughout the year. Moreover, P450scc, 3betaHSD, P450c17and P450arom were identified in Leydig cells but not in Sertoli cells of all testes. These results suggested that adult GM pig testes have the ability to produce germ cells throughout the year, and the synthesis of progestin, androgen and estrogen occurs in the Leydig cells of GM pig testes.     

Purification and characterization of a corrinoid-compound in an edible cyanobacterium Aphanizomenon flos-aquae as a nutritional supplementary food

The vitamin B12 concentration of the dried cells of Aphanizomenon flos-aquae was determined by both microbiological method with Lactobacillus delbrueckeii ATCC7830 and chemiluminescence method with intrinsic factor. The Aphanizomenon cells contained 616.3 +/- 30.3 micro g (n = 4) of vitamin B12 per 100 g of the dried cells by the microbiological method. The values determined with the chemiluminescence method, however, were only about 5.3% of the values determined by the microbiological method. A corrinoid-compound was purified from the dried cells and characterized. The purified corrinoid-compound was identified as pseudovitamin B12 (an inactive corrinoid-compound for humans) by silica gel 60 TLC, C18 reversed-phase HPLC, ultraviolet-visible spectroscopy, and 1H NMR spectroscopy. The results suggest that the Aphanizomenon cells are not suitable for use as a vitamin B12 source, especially in vegans.     

Characterization of vitamin B<Subscript>12</Subscript> compounds from marine foods

Vitamin B₁₂ is synthesized by only certain bacteria and archaea but not by animals or plants. In marine environments, bacterial vitamin B₁₂ is transferred and concentrated into fish and shellfish bodies by plankton in the marine food chain. Moreover, marine macrophytic red algae, Porphyra spp. specifically contain substantial amounts of vitamin B₁₂, due to microbial interaction. Although some meats or viscera of edible fish and shellfish are excellent sources of biologically active vitamin B₁₂, an inactive corrinoid, pseudovitamin B₁₂, was found in some edible shellfish using liquid chromatography/electrospray ionization–tandem mass spectrometry. To prevent elderly people from developing vitamin B₁₂ deficiency due to food protein-bound vitamin B₁₂ malabsorption, we present a survey of marine foods containing free vitamin B₁₂. The results of our study suggest that bonito and clam extracts (or soup stocks), which contain considerable amounts of free vitamin B₁₂ are useful not only as seasonings and flavorings but also as excellent sources of free vitamin B₁₂.     

Ocular Toxicity of Benzalkonium Chloride Homologs Compared with Their Mixtures

This study was performed to assess the in vivo ocular toxicity of benzalkonium chloride (BAK) homologs compared with commercially available BAK (BAK mixture) and to assess the ocular toxicity of BAK homolog after repeated ocular application. Rabbit eyes were examined by ophthalmology and scanning electron microscopy (SEM) after 10 applications of BAK homologs with C12 (C12-BAK) and C14 (C14-BAK) alkyl chain lengths and a BAK mixture at concentrations of 0.001% (w/v), 0.003% (w/v), 0.005% (w/v), 0.01% (w/v) and 0.03% (w/v). The ocular toxicity of C12-BAK to rabbit eyes was examined by ophthalmology and histopathology after repeated ocular application for 39 weeks. In addition, the antimicrobial activities of C12-BAK and C14-BAK against A. niger, S. aureus and P. aeruginosa were assessed. Ocular toxicity of C12-BAK was less than those of the BAK mixture and C14-BAK. No ocular toxicity was noted after ocular application of 0.01% C12-BAK to rabbits for 39 weeks. C12-BAK showed antimicrobial activities at a concentration of 0.003%. These results suggest that the use of C12-BAK to replace BAK mixture as a preservative in ophthalmic solutions should be considered in order to reduce the incidence of the corneal epithelial cell injury induced clinically by BAK.