Effect of temperature and cell density on ethanol fermentation by a thermotolerant aquatic yeast strain isolated from a hot spring environment

ABSTRACT: A thermotolerant, fermentative yeast strain named RND13 from a hot spring drainage was evaluated for its ethanol-producing ability at elevated temperatures at a high substrate concentration [15% (w/v) glucose] close to the level reflecting industrial practice. The RND13 was capable of utilizing glucose almost completely at 40°C with increasing inoculum size, producing ethanol up to 6.6% (w/v), which is comparable to levels (7.0–7.2%) at 30°C. The maximum rate of ethanol production by the RND13 was found to be 9.0 g/L per h at 40°C in an inoculum sized 5% (w/v). At 43°C, however, the RND13 could not utilize glucose to completion and showed a slight drop in the extent of produced ethanol [6.0% (w/v)]. Thus, the culture at 40°C with a 5% cell inoculum was considered to be the optimal condition for ethanol production at higher temperatures in terms of batch fermentation. In the phylogenetic analysis based on the small-subunit rDNA sequence, the strain was grouped together with both Candida glabrata and Kluyveromyces delphensis , which are relatively close to Saccharomyces cerevisiae .     

How do visual and auditory cues influence calf stress reactions at weaning?

We investigated the effects of visual and auditory information related to cow and calf stress responses at weaning. Three weaning conditions were established with physical separation only (PS), physical and visual separation (VS), and physical, visual and auditory separation (VAS). Stress responses such as vocalization and self‐grooming, and changes of three maintenance behaviors (lying, walking and grazing) were recorded for experimental cows and calves in the respective conditions. Comparison of the data revealed that the cow responses differed slightly among groups. However, the peak frequency of vocalization was higher in the PS calves than in those of either VS or VAS. Self‐grooming frequency was significantly different in calves and was high in the order of VAS, VS and PS calves. Furthermore, the percentage of time spent lying down was greater, and that of walking was less, in VAS calves than in either VS or PS calves. These results demonstrate that different information at weaning might strongly affect the stress responses of calves but not those of cows. Relative impacts of different types sensory information on stressors must be considered to plan animal‐friendly weaning.     

Serological assessment of chlamydial infection in the koala by a slide EIA technique

A rapid and simplified slide enzyme immunosorbent assay (EIA) was developed for the diagnosis of chlamydial infection in the koala. HeLa 229 cells infected with koala strain Chlamydia psittaci were fixed on the surface of multiwell slides and used as the antigen. The assay consisted of first reacting koala antiserum with the fixed C psittaci antigen, followed by reaction with biotinylated rabbit anti-koala IgG, ABC reagent and substrate. The chlamydial EIA antibody titres obtained were compared with those of a complement fixation (CF) test using koala strain C psittaci as antigen. Of 35 koala sera tested, 16 CF positive sera (greater than or equal to 1:8) also had a positive titre (greater than or equal to 1:200) in the slide EIA test (sensitivity 93.8%, 15/16). Nineteen CF negative sera were also negative in the slide EIA (specificity 100%, 19/19). Sixty-eight samples of koala blood were collected by ear-prick using a sampling paper method and were assayed by both tests. Sensitivity of the slide EIA was 100% (15/15) and specificity of the test was 96.2% (51/53). To simplify the slide EIA for use as a practical screening test, a 3-point serum dilution series (1:100, 1:200, 1:400) was used. This 3-point slide EIA was compared with the CF test using sheep strain chlamydial antigen. Thirty-nine sera were assayed by both tests. The sensitivity of the 3-point method was 85.7% (6/7) and the specificity was 71.9% (23/32) as compared with the sheep antigen CF test.(ABSTRACT TRUNCATED AT 250 WORDS)     

Magmatic dike resonances inferred from very-long-period seismic signals

Very-long-period (VLP) signals showing simple decaying harmonic oscillations with periods near 10 seconds and lasting for about 300 seconds were observed in association with an earthquake swarm that occurred beneath Hachijo Island, Japan. Results from the source-mechanism analysis and waveform simulation based on a fluid-filled crack model consistently point to the resonance of a dike filled with a basaltic magma as the source of the VLP signals. Thus, VLP signals can be used to probe the state of the fluid and dynamic processes within a volcanic system.     

A planar rhombic charge-separated tetrasilacyclobutadiene

The cyclobutadiene (CBD) molecule C(4)H(4) deviates from a high-symmetry square geometry to compensate for its antiaromatic electronic structure. Here, we report a CBD silicon analog, Si(4)(EMind)(4) (1), stabilized by the bulky 1,1,7,7-tetraethyl-3,3,5,5-tetramethyl-s-hydrindacen-4-yl (EMind) groups, obtained as air- and moisture-sensitive orange crystals by the reduction of (EMind)SiBr(3) with three equivalents of lithium naphthalenide. X-ray crystallography reveals a planar and rhombic structure of the Si(4) four-membered ring, with alternating pyramidal and planar configurations at the silicon atoms. The large (29)Si chemical shift differences (Δδ > 350 parts per million) in the solid-state nuclear magnetic resonance spectra suggest a contribution of an alternately charge-separated structure. The rhombic-shaped charge-separated singlet state of compound 1 thus stabilizes its cyclic 4π-electron antiaromaticity in a manner that contrasts sharply with the bond-length alternation, characterizing the rectangular distortion of carbon-based CBD.     

Detection of the EGFP sequence in breast muscle of 3‐year‐old chicken after transfection using sonoporation

In our continuing effort to generate transgenic chickens, sonoporation was chosen to insert an exogenous gene into the chicken genome. An EGFP expression vector (pCAG‐EGFPac) and microbubbles were injected into the central disc of stage‐X blastoderm or the germinal crescent of stage‐4 embryos, followed by ultrasonic vibration. Nineteen chicks out of 108 treated embryos hatched, six females and six males out of these 19 chicks grew to sexual maturity and two females and three males lived for 3 years. Genomic DNA from 17 out of 35 gonads from embryos and chicks that died before sexual maturity was EGFP‐positive by PCR. No EGFP sequence was detected in the genomic DNA of 322 embryos from six sexually mature females and the semen from four sexually mature males by PCR. When genomic DNA was obtained from various tissues of five 3‐year‐old chickens, the EGFP sequence was amplified from the genomic DNA of the breast muscle of a female (No. 85). The above sequence was subjected to DNA sequencing and verified to be the EGFP sequence. These results showed that sonoporation is an effective tool for the transduction of exogenous genes into chicken embryos for the generation of transgenic chickens.     

Analysis of strigolactones,germination stimulants for striga and orobanche,by high-performance liquid chromatography/tandem mass spectrometry

A simple and rapid analytical method for strigolactones, germination stimulants for the root parasitic weeds witchweed (Striga spp.) and broomrape (Orobanche spp.), has been developed using high-performance liquid chromatography connected to tandem mass spectrometry (LC/MS/MS). The natural strigolactones (strigol, sorgolactone, orobanchol, and alectrol) were clearly separated and identified by LC/MS/MS. As low as 0.1 pg/microL of strigol and 0.5 pg/microL of sorgolactone could be quantified, whereas 1 pg/microL was needed for the quantification of orobanchol (S/N > 10). Using this method, it was found that red clover produces orobanchol and alectrol but not strigol. The roots of red clover seedlings were found to produce 13, 70, 58, and 65 pg of orobanchol/plant 1, 2, 3, and 4 weeks after germination, respectively.