Protozoan predation of Escherichia coli in hydroponic media of leafy vegetables

Multiple outbreaks of food poisoning associated with fresh vegetable consumptions have occurred in many countries. Numerous reports have described human pathogenic bacteria, such as Escherichia coli O157:H7 and Salmonella spp., that can internalize into fresh vegetables via root or leaf surfaces. While attempting to obtain the threshold concentration of internalization of E. coli inoculated into hydroponic medium during vegetable cultivation, we observed a rapid decrease in E. coli numbers. In the present study, we determined that the rapid decline in E. coli was not due to a physiological change into a viable but non-culturable (VNC) state. The population crash was instead caused by true bacterial death, as the rapid descent was also confirmed by micro-colony fluorescence in situ hybridization, a culture-independent method that can detect VNC cells. We next monitored the number of E. coli inoculated into intact or filter-sterilized hydroponic medium after cultivation of various types of plants. We found that the number of E. coli in intact hydroponic medium decreased markedly, whereas the level in filter-sterilized hydroponic medium was completely unchanged. This result suggests that biotic factors were present that could be eliminated by filtering. Robust predation of E. coli by protozoa (ciliates and flagellates) was observed using fluorescently labeled bacteria incorporated into the hydroponic medium. Finally, morphological identification of flagellates by scanning electron microscopy revealed the presence of a species of Stramenopiles. These findings suggest the importance of protozoa as bacterial feeders in hydroponic systems and hence the use of these organisms as potential control agents of human pathogenic bacteria.     

Stimulatory effect of aluminum on the growth of tea pollen tubes

It is well known that when inorganic elements such as boron (B), calcium, cobalt, and manganese are added to media in combination or alone, they stimulate the germination and/or tube growth of various kinds of pollen (Schmücker 1933; Loo and Hwang 1944; Yamada 1958; Brewbaker and Kwack 1963). Among these elements, the stimulatory effect ofB is the most effective and generally recognized. Aluminum (Al) belongs along with B to Group IIIa in the periodic table, and Fähnrich and Ultlich (1964) reported that Al inhibited the germination and growth of the pollen tube of Petunia hybrida and Antirrhinum majus. However, stimulatory effects of Al on pollen germination or pollen tube growth had not been previously reported.     

Selective staining and visualization of hyphal sheath of a white-rot fungus <Emphasis Type="Italic">Phanerochaete crassa</Emphasis> WD1694 with phloxine B

The hyphal sheath is a morphological feature of many kinds of fungi. Although the fine structures of the sheath have been studied in detail by a number of electron microscopy techniques, the function and physiology of the hyphal sheath are not yet clarified. One reason for this is that the hyphal sheath is a colorless, mucilaginous, and delicate material so that it is not easily identified. We developed a simple method to visualize and identify the hyphal sheath of the white-rot fungus Phanerochaete crassa WD1694. The small mycelial pellets in shaken liquid cultures of P. crassa WD1694 were stained directly with phloxine B. Both the hyphae and the hyphal sheath that filled the gaps between each of the hyphae were visualized and observed by light microscopy. The stained hyphae were further studied by transmission electron microscopy, atomic force microscopy, and fl uorescence microscopy. Based on these observations, we confirmed that the staining of the hyphae was also due to the presence of the hyphal sheath that closely covered the fungal cell wall. These results clearly showed that the hyphal sheath was selectively stained with phloxine B and could be observed and identified by conventional light microscopy. Part of this report was presented at the 50th Lignin Symposium, Nagoya, October 2005     

New conjugated metabolites of 3-phenoxybenzoic acid in plants

The metabolism of 3-phenoxybenzoic acid, a common plant metabolite of deltamethrin, cypermethrin and fenvalerate, has been studied in abscised leaves of cabbage, cotton, cucumber, kidney bean and tomato plants. The [¹⁴C]-acid was readily converted into more polar conjugates by esterification with glucose, 6-O-malonylglucose, gentiobiose, cellobiose, glucosylxylose and two types of triglucose with different isomerism. Other metabolites identified were the glucosyl ether of 3-(4-hydroxyphenoxy)benzoic acid, and a 3-(2-hydroxyphenoxy)benzoic acid derivative with a total of two molar equivalents of glucose linked to the carboxyl and phenolic -OH groups. The conjugation pathways were somewhat plant-specific. The glucosylxylose ester was found only in cotton, and the cellobiose and triglucose esters were found only in tomato. All of the conjugates except the glucose and glucosylxylose esters were plant metabolites that had not been identified previously. Furthermore, this is the first report to show the presence of cellobiose and triglucose conjugates in plants. However, neither of the acetyl derivatives of the [¹⁴C]-triglucoside was identical with the synthetic deca-acetyl derivative of [1→6]-triglucoside.     

Walnut hoarding by the Japanese wood mouse,Apodemus speciosus Temminck

Walnuts,Juglans ailantifolia Carriere, are large and the shell is difficult to open. Thus, seed predators in Japan are restricted mainly to the Japanese squirrel (Sciurus lis Temminck) and the wood mouse (Apodemus speciosus Temminck), which can shave away the hard shells with their sharp incisors. Previous work indicated that the squirrels are not only predators, but also disperse walnuts. In this study, 95 walnuts in Period1 (September 1996–February 1997) and 80 walnuts in Period 2 (September 1999–February 2000) each attached to a miniature radio transmitter, were placed in a feeding box, and 93 and 53 of them were transported by free-living wood mice in the respective periods. Most of the walnuts provided from September to November were scatter-hoarded in the litter layer, while from December to February, the mice fed immediately and seldom scatter-hoarded on the ground surface. In all, 66 walnuts were hoarded and 80 were eaten soon after discovery, although walnuts were eaten more frequently in Period 1 than in Period 2, which may be related to annual differences in seed crop size in the study site. The mean transport distance of hoarded seeds was 6.2 ± 5.0 m (mean ± SD,n=66). The fate of hoarded walnuts was followed from September 1996 to May 1997; 21 (75%) of the scatter-hoarded walnuts were recovered, but 7 (25%) were left uneaten. Consequently, the Japanese wood mouse also plays a role in seed-dispersion of the walnut, but the transport distance for mice was shorter than that for squirrels.     

The effect of 7.2% hypertonic saline solution on the duration of sodium gradient between the cerebrospinal fluid and the venous circulation in the dog

To determine the duration of water movement from cerebrospinal fluid (CSF) into venous blood by the infusion of 7.2% hypertonic saline solution (HSS), the sodium gradient between venous blood and CSF were examined. Venous sodium concentrations remained higher than that in CSF for duration of 60 min following HSS infusion. By 90 min, the CSF sodium concentration reached the equilibrium with venous sodium concentration. Those data suggests that the duration of time during which water moved from CSF into capillaries in brain by the gradient of sodium concentration was less than 90 min.     

A case of protein-losing enteropathy treated with methotrexate in a dog

A 9-year-old female Pug was presented to us with chronic diarrhea. Hematologic findings indicated severe hypoproteinemia and hypoalbuminemia, and endoscopy revealed severe edema of the duodenal mucosa. Based on these results and on additional histopathological findings, we made a diagnosis of protein-losing enteropathy caused by lymphocytic-plasmacytic enteritis with lymphangiectasia. The dog was initially treated with prednisolone and cyclosporine. This treatment regimen was not effective. However, when methotrexate was substituted for cyclosporine, progress was obtained and the diagnosis was confirmed.     

Effects of feeding calves genetically modified corn bt11: a clinico-biochemical study

Genetically modified corn Bt11 is insect-resistant and expresses Cry1Ab toxin, an insecticidal protein, in kernels. Although Bt11 corn is considered safe based on animal performance, there are no reports available on the clinico-biochemical effects of feeding it to cattle. In this study, we evaluated the effects of feeding Bt11 to calves, using blood and ruminal clinico-biochemical parameters. Our three-month-long feeding experiment demonstrated that calves (n=6), fed with a ration containing 43.3% of Bt11 corn kernels as dry matter, did not develop any discernible clinical, hematological, biochemical, or ruminal abnormalities as compared with control calves (n=6) fed non-Bt11 corn. The results suggest that the transgenic Bt11 has no negative clinico-biochemical effects on calves.     

Economic contribution of participatory agroforestry program to poverty alleviation: a case from Sal forests, Bangladesh

In the Forest Department of Bangladesh, a Participatory Agroforestry Program (PAP) was initiated at a denuded Sal forests area to protect the forest resources and to alleviate poverty amongst the local poor population. We explored whether the PAP reduced poverty and what factors might be responsible for poverty alleviation. We used three poverty measurement methods: the Head Count Index, the Poverty Gap Index and the Foster-Greer-Thorbecke index to determine the extent poverty reduction. We used a linear regression model to determine the possible differences among factors in poverty reduction. Data were collected through semi-structured questionnaires and face to face interviews within the study area. PAP proved effective at poverty alleviation, considerably improving the local situation. The linear regression model showed that PAP output explained the income differences in poverty reduction. Participants identified bureaucracy and illegal money demands by forest department officials, an uncontrolled market system, and underdeveloped road infrastructure as the main obstacles to reduction of poverty. Overall, PAP is quite successful in alleviating poverty. So this program might be ofinterest at other degraded forest areas as a tool to alleviate poverty.     

Isolation and characterization of eight microsatellite loci in the Otton frog, Babina subaspera

The Otton frog, Babina subaspera, is an endangered forest frog species with unusual morphological and ecological features compared with other frogs. Eight microsatellite markers were isolated from B. subaspera and characterized with 24 individuals from Amami Islands, Kagoshima prefecture, Japan. The number of alleles observed for each locus ranged from 9 to 19 (average 13.9). The observed heterozygosity (H _O) and the expected heterozygosity (H _E) ranged from 0.71 to 0.92 and 0.83 to 0.92, respectively. These microsatellites will be useful for future genetic studies of B. subaspera, which will be essential in the conservation of this species.