Bacteriological evaluation of composted manure solids prepared from anaerobic digested slurry for hygienic recycled bedding materials for dairy cows

Changes in mastitis‐causing pathogens, pH and water content in composted manure solids (CMS) prepared from digested slurry were evaluated during turning at 2‐day intervals for 8 days (C1–C4). The numbers of streptococci, coagulase‐negative staphylococci and coliforms were 2.6 × 10¹, 1.7 × 10² and 1.0 × 10¹ colony‐forming units (cfu)/g in CMS (C4) (summer), and these counts were markedly lower (P < 0.05) than those in CMS (C0 and C1). The bacterial counts ranged from 10¹ to 1.7 × 10² cfu/g in CMS (C4) (summer) and were within approved levels, <1 × 10⁶ cfu/g, indicating a minimal mastitis risk. The temperatures in CMS (C1–C4) increased to 63°C–74°C in summer and 67°C–70°C in winter. The mean pH values in CMS (C0–C4) were 9.2 in summer and 8.7 in winter, and water contents ranged from 61.7% to 69.6% in summer and 73.2% to 66.2% in winter. The significant decrease of pathogenic bacteria in CMS appears to be closely related to temperature >63°C for 8 days, pH 8.7–9.2, and water content 62% to 73%. This study demonstrates that prepared CMS has value as a recycled material with the potential to alleviate udder health issues in dairy cows.     

Morphological features of the spermatic cord in the musk shrew (Suncus murinus) with special reference to extratesticular Leydig cells

Morphological features of the testicular artery and vein in the spermatic cord of the musk shrew (Suncus murinus) were evaluated by light microscopy, transmission electron microscopy, corrosion cast technique combined with scanning electron microscopy and immunohistochemistry. The vascular architecture in the spermatic cord of the musk shrew was simple. The testicular artery in the musk shrew was straight and accompanied by 1 to 3 branches of testicular vein. The testicular vein was also straight and anastomosed with each other in some points along its length, but it did not form a delicate pampiniform plexus. In the middle and distal portions of the spermatic cord, the tunica adventitia of the artery and vein was joined together to form a single connective tissue septum. Clusters of cells were found in this connective tissue septum in the middle portion of the cord. These cells were located close to the arterial wall and nerve endings, but they did not appear inside of neurium. They showed several typical characteristics similar to Leydig cells, and they were positive for 3beta hydroxysteroid dehydrogenase (HSD) antibody. Ultrastructural and immunohistochemical studies also indicated that the cells in cluster found in the vascular wall of the musk shrew spermatic cord may be equivalent to Leydig cells in testes. These extratesticular Leydig cells had characteristics of the active steroid-producing cell and seemed to be another source of testosterone.     

Density and sound-speed contrasts,and target strength of Japanese sandeel <Emphasis Type="Italic">Ammodytes personatus</Emphasis>

Sound-speed and density contrasts (h and g, respectively), important acoustic material properties, of Japanese sandeel Ammodytes personatus were measured to estimate theoretical target strength (TS). The measured sound-speed contrast of adult fish varied between 1.016 and 1.023 (mean 1.020), and showed temperature dependence. The measured density contrast differed significantly between juvenile and adult. The density contrast of juvenile varied between 1.017 and 1.024 (1.021), and that of adult varied between 1.026 and 1.038 (1.032). Using these results, TS at 38 and 120 kHz in the fishing season were estimated by an empirical sound scattering model. TS of an individual fish varied significantly with change of tilt angle. TS of near-dorsal aspect (TS_max) and tilt-averaged TS (TS_ave) differed by up to 7 dB. At both frequencies, two different TS_ave−length relationships (TS_ave = a log L + b) were obtained for adult and juvenile. The coefficients of log L of adult were close to 20, suggesting that backscattering strength was proportional to square of body length. These values were larger in juvenile (34.0 at 120 kHz, 56.5 at 38 kHz), suggesting that backscattering strength varied drastically with the cube or fifth power of body length.     

Tumour necrosis factor‐related apoptosis‐inducing ligand induces apoptosis in canine hemangiosarcoma cells in vitro

Tumour necrosis factor‐related apoptosis‐inducing ligand (TRAIL) is an apoptosis‐inducing cytokine that shows potential therapeutic value for human neoplasms, and is effective in some canine tumours; however, its potential for killing canine hemangiosarcoma (HSA) cells is unknown. Thus, we evaluated the proapoptotic effect of TRAIL in nine canine HSA cell lines. Cells (JuA1, JuB2, JuB2‐1, JuB4, Re11, Re12, Re21, Ud2 and Ud6) were cultured with three recombinant human TRAILs (rhTRAILs): TRAIL‐TEC derived from Escherichia coli, TRAIL‐TL derived from mammalian cells and isoleucine zipper recombinant human TRAIL (izTRAIL) containing an isoleucine‐zippered structure that facilitates trimerization. TRAIL‐TEC did not decrease the cell viability in any of the cell lines tested, whereas the other two rhTRAILs effectively decreased the viability of all cell lines as assessed by the WST‐1 assay. In canine HSA cells, izTRAIL induced apoptosis more effectively than TRAIL‐TL. In JuB4, Re12, and Ud6 cells, izTRAIL increased the activation of caspase‐3 and caspase‐8 and caused poly (ADP‐ribose) polymerase degradation. Moreover, izTRAIL treatment increased the proportion of Annexin V+/ Propidium iodide (PI)? apoptotic cells and nuclear fragmentation in izTRAIL‐sensitive cells. These results show that rhTRAIL can induce apoptosis in canine HSA cells, but the sensitivity of TRAIL was different depending on the cell lines. Therefore, TRAIL could be an effective therapeutic agent against canine HSA, but the specific mechanism of resistance should be determined to clarify under what conditions this treatment would be most effective.     

Effect of setting conditions on mechanical properties of acid-induced Kamaboko gel from squid <Emphasis Type="Italic">Todarodes pacificus</Emphasis> mantle muscle meat

Squid Todarodes pacificus suwari gels, set at various temperatures and times, and acid-induced kamaboko gel, which was prepared by soaking suwari gel in 5% acetic acid for 20 h, were studied to evaluate the mechanical properties that are affected by setting conditions. Unset squid meat paste did not form a gel when soaked in acetic acid. The breaking strength of both suwari gel and acid-induced kamaboko gel showed a tendency to increase with setting temperature and time. SDS-PAGE analysis of suwari gel and acid-induced kamaboko gel, which were set at various temperatures and times, showed that myosin heavy chain (MHC) was observed at 30°C only for the first hour. The intensity of the MHC band at 30°C gradually decreased with setting time, while the intensity of the polymer band gradually increased with setting time. These results suggest that the protein-protein bonds in suwari gel affect the final texture of acid-induced kamaboko gel. Based on the analysis of the mechanical properties, and in consideration of the fact that the purpose of this experiment was to reduce energy usage, the best setting condition was determined to be 40°C for 3 h.     

Comparison of extractive components in muscle and liver of three Loliginidae squids with those of one Ommastrephidae species

Extractive components were determined for the mantle muscle and liver of three species of Loliginidae squid, Sepioteuthis lessoniana, Loligo bleekeri, and L. edulis, and one species of Ommastrephidae, Todarodes pacificus, as a control. Total free amino acid levels and the major free amino acids in muscle, taurine, proline, glycine, alanine, and arginine, were significantly higher in Loliginidae squids than in T. pacificus. The main nucleotide was adenosine 5′-monophosphate, which did not differ significantly between species. Malate was the organic acid found in muscle in highest concentrations. The muscle of these species contained a large amount of trimethylamine oxide. A large amount of glycine betaine was also detected in the muscle, but showed no large species difference. From these results, the muscle of Loliginidae species is considered to have a much sweeter taste than that of T. pacificus. Compared with muscle, squid liver was characteristic with high contents of taurine, glutamate, bitter amino acids, succinate, propionate, trimethylamine, and glycine betaine, and with low contents of sweet amino acids, arginine, nucleotides, malate, and trimethylamine oxide. These results suggest that squid liver is characterized by a complicated taste containing umami, bitterness, sourness, fishy flavor, and less sweetness.     

Predation dynamics of mackerel on larval and juvenile anchovy: is capture success linked to prey condition?

We tested whether the predation dynamics of chub mackerel Scomber japonicus and spotted mackerel S. australasicus on young anchovy Engraulis japonicus relates to individual growth characteristics of the prey and could account for the growth-selective survival predicted by recruitment hypotheses. Juvenile and adult mackerel were sampled along with their young anchovy prey field in 2004 (juvenile mackerel and larval anchovy) and 2005 (adult mackerel and juvenile anchovy) off the Pacific coast of Honshu, Japan. The recent 5-day mean growth rate of larval and juvenile survivors and prey found in the stomach of mackerel was estimated from the otolith microstructure. No significant difference was found between the recent growth of larval or juvenile survivors and that of preyed individuals. We conclude that despite a relatively small body size, the high activity level and predation skills displayed by mackerel prevent fast-growing larvae and early juveniles from benefitting in terms of the expected survival advantage over slow-growers. Hence, growth-selective predation mortality of larval fish would depend on the feeding ecology of the predator rather than predator size. Selection for fast growth is more likely to occur under predation pressure from invertebrate organisms and small pelagic fish specialized on zooplankton, such as herring and anchovy.     

A method for predicting the occurrence of paralytic shellfish poisoning along the coast of Hokkaido in the Okhotsk Sea in summer

To design a method for predicting outbreaks of paralytic shellfish poisoning (PSP) in scallop fishing grounds, the relationship between the distribution of the toxic dinoflagellate Alexandrium tamarense and the dynamics of the Soya Warm Current (SWC) was examined in the Okhotsk Sea off Hokkaido. Surveys were conducted from May to June to clarify the transportation mechanism of A.?tamarense from the oceanic area to the coastal area. The sea-level difference (SLD) between Wakkanai and Abashiri was monitored as an index of the strength of the SWC southeastward flow in an alongshore belt to examine the possible occurrence of A.?tamarense in the coastal area during temporal weakening of the SWC. A bottom-mounted acoustic Doppler current profiler (ADCP) was used for direct observations of the SWC. The results indicated that PSP occurred when low-salinity water contaminated with A.?tamarense extended to the coast during temporal weakening of the SWC due to a decrease of the SLD. Our results strongly indicate that predictions can be realized by monitoring the decrease of SLD as an index of temporal weakening of the SWC after surveys of the distribution of A.?tamarense in the oceanic area before the period of PSP occurrence.     

Estimation of the outcrossing rate for annual Asian wild rice under field conditions

The Asian wild rice, Oryza rufipogon, has partial outcrossing behavior and shows high levels of genetic variation. To estimate an accurate outcrossing rate of annual form of O. rufipogon, two backcross lines (Lines R1 and R2) between Oryza sativa Nipponbare and O. rufipogon W630 were examined under field conditions. A chromosome survey confirmed that these lines had wild chromosomal segments at more than 92% of marker loci. As for the traits of glume, stamen and pistil, Line R1 showed similar floral morphology as that of O. rufipogon W630, whereas Line R2 had larger glumes. In 2005, 2006 and 2008, a total of 22 backcross plants were planted in the middle of wild rice plots. The successive progenies of each plant were examined using microsatellite markers that could clearly detect self-pollination and outcrossing. The outcrossing rates of Line R1 plants ranged from 4.04% to 25.50% with an average of 10.20%. This indicates that cross-pollination of wild rice is a chance event affected by many environmental factors. The outcrossing rates of Line R2 plants also varied, however, no significant difference was observed between the averages of Lines R1 and R2, suggesting that the glume sizes are not critical for outcrossing ability.