Authentication of flying-fish-meal content of processed food using PCR-RFLP

Nucleotide sequence and polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) analysis of the 3′-portion of the mitochondrial 16S RNA gene (rDNA) coding sequence were used to differentiate between flying fishes and other fishes that are frequently used in ago-noyaki production. In this study, we successfully distinguished between flying fishes and the other fishes by combining amplified DNA fragments with universally designed primers and digesting the PCR products with AfaI and MfeI restriction endonucleases. PCR-RFLP of 15 ago-noyaki, 2 noyaki, and 8 other processed flying-fish products were analyzed to authenticate flying-fish content among processed seafood products. After digestion of the PCR products with both enzymes, we found that all ago-noyaki and processed flying fish products contained either two or three DNA fragments of ~200, 300, and 530 bp, and noyaki samples (which do not contain flying fish) contained only one fragment of ~530 bp. Here, we present a new procedure to confirm the content of flying-fish meal in ago-noyaki.     

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.     

Reactivity and prevalence of neutralizing antibodies against Japanese strains of bovine viral diarrhea virus subgenotypes

Bovine viral diarrhea virus (BVDV) field isolates show genetic and antigenic diversity. At least 14 subgenotypes of BVDV-1 and 4 of BVDV-2 have been identified in Artiodactyla worldwide. Of these, 6 subgenotypes of BVDV-1 and 1 of BVDV-2 have been isolated in Japan. Previously, we reported that each subgenotype virus expresses different antigenic characteristics. Here we investigated the reactivity of neutralizing antibodies against representative strains of Japanese BVDV subgenotypes using sera from 266 beef cattle to estimate the prevalence of this epidemic virus among cattle in Japan. Antibody titers at concentrations at least 4-fold higher than antibodies against other subgenotype viruses were considered subgenotype specific. Subgenotype-specific antibodies were detected from 117 (80.7%) of 145 sera samples (69.7% against BVDV-1a, 1.4% against BVDV-1b, 8.3% against BVDV-1c, and 1.4% against BVDV-2a). The results suggest that neutralization tests are useful in estimating currently epidemic subgenotypes of BVDV in the field.     

Analysis of respiratory quinones in soil for characterization of microbiota

Method for the analysis of respiratory quinones in soil was developed to characterize soil microbiota. The respiratory quinones were extracted with a mixture of chloroform and methanol using a Wahling blender or a sonicator and cleaned-up by a silicagel column cartridge. The quinone species were determined by reverse-phase high performance liquid chromatography. Spectra of peaks were measured with a photodiode array detector to examine the purity. More than 90% of extractable quinones in soil were recovered by three extractions. The recovery of quinones, added to soil as freeze-dried powder of microorganisms, was higher than 96%. This procedure led to higher representative results as follows. Fluctuations within 95% of cumulative frequency were 13% for the extracted amount of quinones and 20% for the dissimilarity, respectively. The quinone profiles of four soils were determined as follows: a soil from the aerobic layer of a paddy field, an ando soil and two yellow upland soils which had received chemical fertilizers with and without farmyard manure. The quinone profiles displayed clear differences in the microbial composition and in the microbial diversity among the soils. It was demonstrated that the analysis of the respiratory quinone profile was useful to characterize the microbial community structure in soil.     

Visual acuity and spectral sensitivity of the elkhorn sculpin <Emphasis Type="Italic">Alcichthys alcicornis</Emphasis>

ABSTRACT:   In this study, the visual acuity, visual accommodation, and spectral sensitivity of the elkhorn sculpin were determined. The elkhorn sculpin Alcichthys alcicornis is a typical groundfish that has conspicuously large eyes. They are caught by bottom gillnets, trawl nets, bottom line angling, and other fishing methods. Although fish behavior arising from visual stimuli is important in these fishing methods, there is little information available regarding the visual sensitivity of the elkhorn sculpin . Three specimens, with total lengths of 300–310 mm, were studied. Visual acuity was calculated, from the maximum cone density in the retina, to be between 0.10 and 0.14. The minimum separable angles were calculated to be 7.14–10.37 min. Measurement of lens movement in five specimens indicated that the near-point distance was between 0.87 and 1.53 times the total length. Lens direction was estimated to be 11–22° in the downward visual axis. Spectral sensitivity was also determined electrophysiologically by recording S-potentials. Three types of response were obtained from 21 specimens, including L responses that showed hyperpolarization at all wavelengths of stimulating light, and two C responses that showed wavelength-dependent hyperpolarization and depolarization. Based on these results, the elkhorn sculpin has color vision and its sensitivity is most acute in light with a wavelength of 554 nm.     

Behavioral responses of arabesque greenling to trap entrance design

ABSTRACT:   Fish catch efficiency can vary with the design of the trap entrance. To clarify how trap entrance design affects catch efficiency, the responses of fish to different trap entrance designs with different inclination angles or funnel lengths were observed. The passing ratio (i.e. catch) increased and then decreased with increasing inclination angle. An inclination angle of 34° had the highest passing ratio of 43%; the lowest passing ratio was 8% at an angle of 53°. A significant difference in passing ratio was found between two entrance designs ( P  = 0.006). There were significant differences in passing ratio for entrance designs with different funnel lengths ( P  = 0.01). The highest passing ratio was 38% for a funnel length of 29 cm. The approach and passing durations were also compared among the trap entrance designs. The results suggest that modifying the entrance shape can significantly improve the catch efficiency.     

Feeding behavior and food consumption of Japanese whelk, <Emphasis Type="Italic">Buccinum isaotakii</Emphasis> (Neogastropoda: Buccinidae)

Abilities of detection and preference of food by a Japanese whelk, Buccinum isaotakii, were investigated under laboratory conditions. Consumption per food item and the corresponding assimilation efficiency were also de termined. Whelks were more sensitive to fresh foods than boiled foods, detecting them in a short period of time varying between 3.8 (±0.8) and 23.1 (±3.1) minutes. B. isaotakii preferred fish, polychate and bivalves and the time spent on these foods was more than 10 min as compared with those of shrimp (2.0 min) and squid (1.0 min). The highest daily consumption rate was observed on sardine, followed by scallop, polychaete and oyster. The relationship between shell length and consumption of different food items was positive. The coefficients of determination were relatively high for sardine, scallop, polychaete and oyster (r ², between 0.46 and 0.51) but not for clam, mussel shrimp and squid (r ², between 0.003 and 0.176), suggesting that the former could be predators of consumption for B. isaotakii. Observed were relatively high assimilation efficiencies on sardine, scallop, polychaete, and oyster, ranging from 30 to 70%. The present results are the intial information pertaining to the feeding biology of this whelk, thus, in-depth studies like an energy budget are recommended to deepen the understanding about seed production and resource management.     

Effects of acquiring FSC forest management certification for Japanese enterprises using SmartWood Audits

We explore the organizational, environmental, and economic effects of sustainable forest management (FM) certification by Smart-Wood (SW), one of the certification bodies accredited by the Forest Stewardship Council (FSC) to carry out certification in Japan. We closely investigated three enterprises: the Yusuhara Forest Owners’ Cooperative, Yamanashi Prefectural Forest, and Ryujin-mura Forest Owners’ Cooperative. Interviews with representatives of these entities provided crucial empirical information regarding the influence of certification on environmental assessment, rare-species protection, landscape-management planning, management objectives, and forest supervision. Attainment of FM status improved environmental awareness and engendered positive changes in assessing and managing forests and other natural resources but simultaneously imposed heavier economic costs.     

Histiocytic ulcerative colitis in a French bulldog

A 9-month-old French bulldog was referred for signs of chronic large bowel diarrhea. The dog had an increased frequency of defecation, tenesmus and hematochezia. Flexible colonoscopy showed hyperemia, irregularities and ulcerations with multifocal hemorrhages in the mucosa from the descending colon to the proximal rectum. Multiple colonic biopsies were characterized by infiltrations of PAS positive histiocytes in the lamina propria. A diagnosis of histiocytic ulcerative colitis (HUC) was made, and the animal showed only minimal improvement, although it was treated with nutritional and medical therapies. This is the second case of HUC in French bulldog, a breed which has ancestral relations to Boxer dogs.     

Angiotensin converting enzyme and chymase activity in the feline heart and serum

The feline cardiac and serum angiotensin converting enzyme (ACE) and chymase activities were determined and compared in dogs, and hamsters. In all three species, cardiac chymase activity exceeded ACE activity; however, there were some differences. In cats, left ventricular ACE and chymase activities (0.15 +/- 0.01 and 0.59 +/- 0.1 mU/mg-protein, respectively) were lower than in dogs (0.42 +/- 0.05: p<0.01 and 2.0 +/- 0.4 mU/mg-protein: p<0.01) and hamsters (0.93 +/- 0.06: p<0.001 and 2.1 +/- 0.2 mU/mg-protein: p<0.01); in contrast, serum ACE activities was higher in cats (12.7 +/- 1.0 mU/ml) than in dogs (5.9 +/- 0.6 mU/ml: p<0.001). The relative contribution of chymase (cats: 84.0 +/- 5.1%, dogs: 81.4 +/- 3.4%, and hamsters: 72.6 +/- 5.6 %) to ANG-II formation in the heart was greater than that of ACE in these animals (cats: 10.9 +/- 4.1%, dogs: 11.5 +/- 3.6%, and hamsters: 17.2 +/- 0.8%). These species-specific differences suggest that the efficacy of renin-angiotensin system modulating agents may differ among species.