Influence of death struggle on the structural changes in chub mackerel muscle during chilled storage

ABSTRACT: Chub mackerel (34–35 cm, approximately 500 g), which were caught by fishing with a rod and line at the Bungo Channel, Oita prefecture, were rested overnight in a fish preserve and either killed by decapitation (control group) or allowed to struggle in air for 30 min (struggled group). Muscle samples were excised every 4 h, and measurements on breaking strength and histological observations were done for both groups. The breaking strength of muscle in the control group was significantly higher than that in the struggled group, whereby a decrease in breaking strength was delayed for 12 h compared to the struggled group. Light microscopy showed space extension among muscle cells in association with a decrease in breaking strength. Especially in the struggled group, the extended area was larger and the difference in area was significant at the time when breaking strength showed a significant difference. Using electron microscopy, the extended area showed cut and/or disappeared collagen fibrils. From these results, it was demonstrated that struggling to death promoted the degradation of collagen fibrils and the weakening of connective tissue and, resultantly, led to the faster softening of muscle of chub mackerel.     

Movement of Xanthomonas campestris pv. vitians in the stems of lettuce and seed contamination

Xanthomonas campestris pv. vitians , the causal agent of bacterial leaf spot of lettuce (BLS), can be seedborne, but the mechanism by which the bacteria contaminates and/or infects lettuce seed is not known. In this study, the capacity of X. campestris pv. vitians to enter and translocate within the vascular system of lettuce plants was examined. The stems of 8- to 11-week-old lettuce plants were stab-inoculated, and movement of X. campestris pv. vitians was monitored at various intervals. At 4, 8, 12 and 16 h post-inoculation (hpi), X. campestris pv. vitians was recovered from 2 to 10 cm above (depending on stem length) and 2 cm below the inoculation site. Xanthomonas campestris pv. vitians was also recovered from surface-disinfested stem sections of spray-inoculated plants. Together, these results are consistent with X. campestris pv. vitians invading and moving systemically within the vascular system of lettuce plants. To investigate the mechanism of seed contamination, lettuce plants at the vegetative stage of growth were spray-inoculated with X. campestris pv. vitians and allowed to develop BLS. Seed collected from these plants had a 2% incidence of X. campestris pv. vitians external colonization, but no bacteria were recovered from within the seed.     

Growth and morphologic response of rumen methanogenic archaea and bacteria to cashew nut shell liquid and its alkylphenol components

The growth and morphology of rumen methanogenic archaea (15 strains of 10 species in 5 genera, including 7 strains newly isolated in the present study) and bacteria (14 species in 12 genera) were investigated using unsupplemented in vitro pure cultures and cultures supplemented with cashew nut shell liquid (CNSL) and its phenolic compound components, anti-methanogenic agents for ruminant animals. Growth of most of the methanogens tested was inhibited by CNSL and alkylphenols at different concentrations ranging from 1.56 to 12.5 μg/ml. Of the alkylphenols tested, anacardic acid exhibited the most potent growth inhibition. Three gram-negative bacterial species involved in propionate production were resistant to CNSL and alkylphenols (>50 μg/ml). All the methanogens and bacteria that were sensitive to CNSL and alkylphenols exhibited altered morphology; disruption of the cell surface was notable, possibly due to surfactant activity of the tested materials. Cells division was inhibited in some organisms, with cell elongation and unclear septum formation observed. These results indicate that CNSL and alkylphenols, particularly anacardic acid, inhibit both rumen bacteria and methanogens in a selective manner, which could help mitigate rumen methane generation.     

A direct enzyme immunoassay for the measurement of furosemide in horse plasma

A new enzyme immunoassay (EIA) for the measurement of furosemide in horse plasma is described. The lower limit of detection of this EIA method was 7.8 ng/ml. The intra-and inter-assay coefficients of variation ranged from 2.5% to 4.9% and 7.5% to 9.8%, respectively. Cross-reactivity with other compounds was not observed. There was a high correlation (r2=0.987) between the high-performance liquid chromatography and EIA results obtained for furosemide concentrations in horse plasma. These results indicate that the newly developed EIA method is useful for the quantitative analysis of furosemide in horse plasma.     

Phytoplankton surveys off the southern Myanmar coast of the Andaman Sea: an emphasis on dinoflagellates including potentially harmful species

A detailed list of dinoflagellate species for the Mergui Archipelago, off southern Myanmar, was compiled for the first time. This was based on surveys conducted in two seasons in pre- and post-monsoons, and on observations for both plankton and cyst assemblages. In the plankton samples, 57 and 26 dinoflagellate species were recorded, respectively, in the pre- and post-monsoons. Among the list from plankton and cyst samples, harmful species were found: red-tide species including Prorocentrum spp. and Alexandrium affine, and causative species of paralytic shellfish poisoning, e.g., Alexandrium tamiyavanichii and Gymnodinium catenatum, and of diarrhetic shellfish poisoning, e.g., Dinophysis spp. Therefore, although no incidences have been reported yet, attention must be paid to these harmful algal bloom (HAB) events in Myanmar where exploitation of fishery resources is drastically increasing. Other dinoflagellate compositions were also unique and may relate to the oceanographic system in this region. In the pre-monsoon, rather rare oceanic species of Ornithocercus spp. were found concurrently with neritic species, probably due to the extensive southwesterly current from the Indian Ocean. Throughout the seasons, diverse species of Protoperidinium were predominately recorded in both planktonic and sediment samples, indicating richness of prey organisms probably sustained by upwelling systems.     

Studies on the properties of organic matfer in buried humic horizon derived from volcanic ash

To investigate the relationship between the change of humus property and time factor, the humus composition in the buried humic horizons with the age from 28,000 years B.P. to the present was studied, according to the method presented by Kumada et al. (7).

The fH decreased with the elapse of time and in case of most of humus in the buried humic horizons with the age over 10,000 years, the fH became lower than 40 and were in the form of so-called ‘combined’ humus.

When 3,000-4,000 years have passed since the surface soil buried, the humic acid first moved . to ‘combined form.’ Second, after over 10,000 years, the fulvic acid moved to ‘combined form.’

The humification of ‘free’ humic acid proceeded with the elapse of time till around 5,000 years but of 'combined' humic acid till around 7,000 years. Both the humification-degree of ‘free’ humic acid after around 5,000 years and of ‘combined’ humic acid after around 7,000 years decreased. The humification-degree of ‘free’ and ‘combined’ humic acid from the samples with the age about 15,000 years B.P. to 28,000 years B.P. may be the same.     

Effect of bensulfuron-methyl (a sulfonylurea herbicide) on the soil bacterial community of a paddy soil microcosm

The effect of bensulfuron-methyl (BSM) on a soil microbial community in a model paddy microcosm was studied. Total bacterial numbers in the overlying water and surface soil were monitored for 2 months after the application of BSM at the field rate and a ten-fold field rate. Pentachlorophenol (PCP) was used for comparison. Neither chemical affected the total bacterial numbers remarkably, either in the overlying water or in the surface soil. In contrast, the nitrification potential was significantly suppressed by the BSM application. The bacterial community structure, as evaluated by the denaturing gradient gel electrophoresis (DGGE) of PCR amplification products from bacterial 16S rDNA, was unaffected by the BSM treatments over 8 weeks in the surface soil, compared with the control (no pesticide). In contrast, the surface soil exposed to PCP at a ten-fold field rate showed different patterns from the controls at 4 weeks and 8 weeks after application. The DGGE patterns of the overlying water were much more variable than those of the surface soil in any treatments. Cluster analysis showed that the BSM plots were classified within the same group as the control at 1 week after application and that the BSM and PCP plots from 2 weeks onward after application were grouped differently from the control. Of 22 clones excised from the DGGE gels, 20 clones belonged to the Proteobacteria and two belonged to the Verrucomicrobia. It was considered that the impact of BSM on the overall microbial community (total numbers, community structure of soil) was negligible, although BSM had an impact on some specific functions of the soil microbial community (nitrification) and a part of the community (overlying water).     

Induction of Systemic Resistance in Cucumber against Several Diseases by Plant Growth-promoting Fungi: lignification and Superoxide Generation

Five fungal isolates (Trichoderma, Fusarium, Penicillium, Phoma and a sterile fungus) from zoysiagrass rhizosphere that promote plant growth were tested for their ability to induce systemic resistance in cucumber plants against Colletotrichum orbiculare. Roots of cucumber plants were treated with these fungal isolates using barley grain inocula (BGI), mycelial inocula (MI) or culture filtrate (CF). Most isolate/inoculum form combinations significantly reduced the disease except BGI of Trichoderma. These fungal isolates were also evaluated for induction of systemic resistance against bacterial angular leaf spot and Fusarium wilt by treatment with BGI. Penicillium, Phoma and the sterile fungus significantly reduced the disease incidence of bacterial angular leaf spot. Phoma and sterile fungus protected plants significantly against Fusarium wilt. Roots treated with CFs of these fungal isolates induced lignification at Colletotrichum penetration points indicating the presence of an elicitor in the CFs. The elicitor activity of CFs was evaluated by the chemiluminescence assay using tobacco callus and cucumber fruit disks. The CFs of all isolates elicited conspicuous superoxide generation. The chemiluminescence activity of the CF of Penicillium was extremely high, and its intensity was almost 100-fold higher than that of other isolates. The chemiluminescence activity was not lost following treatment with protease or autoclaving or after removal of lipid. The MW 12,000 dialyzed CF fraction was highly effective in eliciting chemiluminescence activity. Chemiluminescence emission from cucumber fruit disks treated with Penicillium was the same as that obtained from tobacco callus, except that the lipid fraction also showed a high activity. Both the MW 12,000 fraction and the lipid fraction induced lignification in the epidermal tissues of cucumber hypocotyls.     

An Improved Method for Isolating Violet Root Rot Fungus, Helicobasidium mompa, from Basidiocarps

Basidiocarps of the violet root rot fungus, Helicobasidium mompa, are less frequently used for isolation than are mycelial strands and sclerotia even though the basidiocarps are conspicuously produced at the trunk base of diseased plants. Basidiocarps are also more suitable for storage. This paper describes an improved method for obtaining pure cultures from basidiocarps using microcentrifuge tubes to facilitate the awkward steps of rinsing fungal materials under a dissecting microscope. Received 28 June 2000/ Accepted in revised form 4 August 2000