Effect of tributyltin oxide exposure on the F<Subscript>0</Subscript> and F<Subscript>1</Subscript> generations of a marine teleost,the mummichog <Emphasis Type="Italic">Fundulus heteroclitus</Emphasis>

The effects of organotin compounds on marine organisms, especially on their reproduction, are a continuing source of concern. We performed full-life-cycle exposure tests of tributyltin oxide (TBTO) at sub-lethal concentrations (0.13, 0.25, 0.5 and 1.0 μg/l) using a marine fish, the mummichog Fundulus heteroclitus, and studied the toxic effects of exposure to TBTO at various life-cycle stages of the fish. The effect of TBTO was initially studied on sex differentiation and then on spermatogenesis in maturing fish. A mating experiment was performed to determine the effects of exposure on the fecundity of female fish, fertilization success, and the hatch of the F₁ generation. In all experimental groups, TBTO exposure resulted in a tendency toward a male-biased sex ratio and caused a significant increase in the frequency at which apoptotic cells appeared in the testes. Exposure to 1.0 μg/l TBTO resulted in a significant decrease in fecundity, but no significant effect was found on the fertilization ratio. Time to hatch and hatchability of the F₁ generation were markedly affected in some exposure groups. These results indicate that long-term exposure of the mummichog to TBTO affects not only gonadal sex differentiation and spermatogenesis but also spawning and egg quality.     

Quantification of relative flying fish paste content in the processed seafood ago-noyaki using real-time PCR

Real-time polymerase chain reaction (PCR) analysis of the 3′-portion of the mitochondrial 16S RNA gene (rDNA) coding sequence was used to determine flying fish paste in ago-noyaki. We quantified the amount of flying fish paste in ago-noyaki samples using flying fish-specific primers (Tobi16SF3/Tobi16SR) and universal primers (Univ16SF2/Univ16SR2). Using real-time PCR of standard ago-noyaki, a standard equation was obtained (y = 1.08x − 3.20; R ² = 0.977). This equation was then used to estimate the relative flying fish paste contents of eight commercially available ago-noyaki and two similar products. These results verified that the ago-noyaki products that had already been labeled with the E-mark deserved this status.     

Nitrification Inhibitors from the root tissues of Brachiaria humidicola, a tropical grass

Nitrification inhibitory activity was found in root tissue extracts of Brachiaria humidicola, a tropical pasture grass. Two active inhibitory compounds were isolated by activity-guided fractionation, using recombinant Nitrosomonas europaea containing luxAB genes derived from the bioluminescent marine gram-negative bacterium Vibrio harveyi. The compounds were identified as methyl-p-coumarate and methyl ferulate, respectively. Their nitrification inhibitory properties were confirmed in chemically synthesized preparations of each. The IC50 values of chemically synthesized preparations were 19.5 and 4.4 microM, respectively. The ethyl, propyl, and butyl esters of p-coumaric and ferulic acids inhibited nitrification, whereas the free acid forms did not show inhibitory activity.     

Gingival overgrowth in a dog that received long-term cyclosporine for immune-mediated hemolytic anemia

Gingival mass lesions developed when cyclosporine was administered for 600 days to a female, 7-year-old, longhaired dachshund diagnosed with intractable immune-mediated hemolytic anemia (IMHA). Histopathology indicated hyperplastic suppurative gingivitis. As the anemia improved, the dosage of cyclosporine A (CsA) was markedly decreased, and the mass lesions decreased in size and disappeared, thus suggesting that the mass lesions were an adverse reaction to CsA.     

Clock mutation affects circadian regulation of circulating blood cells

Background  

Although the number of circulating immune cells is subject to high-amplitude circadian rhythms, the underlying mechanisms are not fully understood.     

An Efficient Inoculation Method for Colonization of Chinese Cabbage Seedlings by the Root Endophytic Fungus Heteroconium chaetospira

To develop an efficient method to inoculate Chinese cabbage seedlings with the root endophytic fungus Heteroconium chaetospira, an appropriate nursery soil and glucose concentration for the nutrient medium for fungal colonization were determined. A grid-sheet method was established for estimating the degree of colonization of entire roots by the fungus. The fungus colonized at high frequencies when peat moss was used as the rooting medium. Colonization was highest (75%) when peat moss was amended with 0.1% glucose. Under these conditions, fungal hyphae developed intracellularly in root cortical cells. In contrast, under high glucose conditions, fungal colonization was restricted mostly to intercellular regions of epidermal or cortical root tissues. Here, hyphae formed inter- or intracellular microsclerotia. Received 19 August 2002/ Accepted in revised form 5 December 2002     

Molecular identification of Echinochloa oryzicola Vasing. and E. crus-galli (L.) Beauv. using a polymerase chain reaction–restriction fragment length polymorphism technique

Polymerase chain reaction (PCR) and polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) techniques were applied for establishing the reliable practice in identification of Echinochloa oryzicola Vasing. and E. crus-galli (L.) Beauv. (barnyardgrass). Total DNA was extracted from 18 accessions and 86 individuals of E. oryzicola , 33 accessions and 140 individuals of E. crus-galli var. crus-galli , 23 individuals of E. crus-galli var. praticola , and six individuals of E. crus-galli var. formosensis that were collected from Japan. A partial region of intergenetic spacer between trn T and trn L, and an intron of trn L were amplified separately using a trn-a and trn-b1 primer set, and a trn-c and trn-d primer set, respectively. All individuals of E. oryzicola showed the same fragment amplified by the trn-a and trn-b1 primer set. The fragment was 481 bp in length, and was undigested by Eco R I, whereas all individuals of E. crus-galli , including three botanical varieties, showed the same fragment with a 449-bp length. The fragment was digested by Eco R I into two fragments (178 and 271 bp). The fragment amplified by the trn-c and trn-d primer set in all individuals of E. oryzicola was digested by Alu I into two fragments (174 and 452 bp), but undigested by Dra I. In contrast, the fragment amplified by the trn-c and trn-d primer set in all individuals of E. crus-galli was digested by Dra I into two fragments (134 and 487 bp), but undigested by Alu I. There was no intraspecific variation in these regions; thus, these two species are easily identifiable by using our method.     

Natural hybridization in wild soybean (Glycine max ssp. soja) by pollen flow from cultivated soybean (Glycine max ssp. max) in a designed population

The frequency of hybridization through pollen flow from the cultivated soybean to the wild soybean was evaluated for the purpose of assessment of the ecological risk of genetically modified crops. The flowering habits of three soybean cultivars and one wild soybean accession were monitored on an experimental farm. A cultivar and a wild accession, both of which flowered at a similar period, were then planted alternately in 5 12 arrays with 50 cm spacing on the farm. The seedlings of progeny seeds gathered from individual plants of the wild accession were used for an isozyme analysis to identify whether they were hybrid or not. In 23 plants of the wild accession, four plants produced hybrids (the incidence of hybridization = 17.4%). There was no directionality in hybridization. The hybridization rate per maternal plant varied from 0 to 5.89% with a mean of 0.73% for all maternal plants. The results indicate that natural hybrids are easily produced in a certain frequency by pollen flow from the cultivated soybean to the wild soybean under their simultaneous flowering with adequate pollinators.     

Role of the α subunit of nicotonic acetylcholine receptor in the selective action of imidacloprid

Examination of agonist interactions of imidacloprid on recombinant chicken α4β2 and Drosophila SAD/Chicken β2 hybrid receptors, expressed in Xenopus oocytes by nuclear injection of the cDNAs, indicates that imidacloprid is a partial agonist. Replacement of the α4 subunit for the Drosophila SAD subunit lowered the imidacloprid EC₅₀ 37-fold, whereas EC₅₀s for other agonists increased 4-50 fold, suggesting that the α subunit contributes to the high affinity of insect nicotonic receptors for imidacloprid. ©1999 Society of Chemical Industry