Purification and properties of β-galactosidase from Tilapia intestine: Digestive enzyme of Tilapia-X

ABSTRACT:   β-galactosidase of the intestine of Tilapia nilotica was purified by ammonium sulfate precipitation, followed by PAPTG-Sepharose 4B affinity chromatography, ethylenediamineetetraacetic acid ion-exchange chromatography, polyexchanger PBE 94 chromatofocusing, and Sephadex G-100 gel filtration. β-galactosidase was found to be a single band when examined by poly-acrylamide gel electrophoresis. The purifications of β-galactosidase were 27-fold from the crude extract. β-galactosidase showed optimum activity at pH 5.0 at 40°C, and was specifically found to be able to hydrolyze p -nitrophenyl β-galactopyranoside. It degrades galactan and agarose, and produces galactose. β-galactosidase was strongly inhibited by Hg²⁺ and PCMB. β-galactosidase is considered to be secreted by the upper and middle parts of the intestine and most of the activity was detected in the intestinal juice.     

Tree-to-tree and clone-to-clone variations of monoterpenes emitted from needles of hinoki (Chamaecyparis obtusa)

Variations in the composition of low boiling point (LBP) monoterpenes emitted from needle samples of 150 hinoki (Chamaecyparis obtuse) trees (30 strains, each with five clones) native to Shimane Prefecture, Japan, were investigated using a headspace technique. The assays revealed considerable proportional variations especially in the amount of sabinene, which ranged from 24% to 78% of the total LBP monoterpenes. The proportions of α-pinene, myrcene, and limonene negatively correlated with that of sabinene overall. In particular, the proportion of limonene showed clear negative correlation with that of sabinene (r = −0.98). Differences in the proportion of sabinene among five clones in each strain were less than 15% in 22 out of 30 strains, indicating that monoterpene composition is constitutively steady in most strains. In a few strains, however, considerable variation in the composition was observed among clones.     

Heat budget and statistical analysis of the relationship between stream temperature and riparian forest in the Toikanbetsu River basin, northern Japan

Riparian forests greatly influence aquatic ecosystems by providing shade cover, which controls water temperature and limits primary production. We examined the relationship between forest cover and summer stream temperature in northernmost Japan. Heat budget and statistical analyses were employed and the results were compared. Heat budget analysis revealed that the water temperature would decrease almost linearly from 29°C to 25°C with an increase in forested reaches along a 3.2 km stretch of the river. Multiple regression analysis by the stepwise method chose only open channel length as a variable to explain the variation in maximum stream temperature. A sharp increase in stream temperature was noted when riparian forest cover was removed in short lengths, of up to 1.0km; this increasing trend gradually flattened as the length of open stretch increased. Thus, even small openings in the riparian canopy resulted in drastic rises in summer stream temperature. The maximum summer temperatures estimated by the two methods were coincided, and can therefore be accurately estimated by regression analysis. Retrospective analysis based on the regression equation showed that the maximum summer temperature in 1947 was 6°C lower than at present, and that a sharp increase occurred from 1947 to 1960, a period of rapid expansion of agricultural land development in the watershed.     

Isolation and antimicrobial susceptibility of Plesiomonas shigelloides from great cormorants (Phalacrocorax carbo hanedae) in Gifu and Shiga Prefectures,Japan

Plesiomonas shigelloides is a causal agent of gastroenteritis, sepsis and meningitis in humans. We examined the prevalence of P. shigelloides among great cormorants (Phalacrocorax carbo hanedae) in Japan and the antimicrobial susceptibility of isolates. P. shigelloides was isolated from 33 (47.8%) of 69 fecal samples from great cormorants in 2014. All 33 isolates were subjected to antimicrobial susceptibility testing using broth microdilution methods, which showed resistance to ampicillin (31 isolates, 93.9%), tetracycline (two isolates, 6.1%) and trimethoprim (one isolate, 3.0%). The high prevalence of P. shigelloides in the great cormorants implicates the possible microbiological risk to public health.     

Genetic analysis of fat‐to‐protein ratio,milk yield and somatic cell score of Holstein cows in Japan in the first three lactations by using a random regression model

We estimated the genetic parameters of fat‐to‐protein ratio (FPR) and the genetic correlations between FPR and milk yield or somatic cell score in the first three lactations in dairy cows. Data included 3 079 517 test‐day records of 201 138 Holstein cows in Japan from 2006 to 2011. Genetic parameters were estimated with a multiple‐trait random regression model in which the records within and between parities were treated as separate traits. The phenotypic values of FPR increased soon after parturition and peaked at 10 to 20 days in milk, then decreased slowly in mid‐ and late lactation. Heritability estimates for FPR yielded moderate values. Genetic correlations of FPR among parities were low in early lactation. Genetic correlations between FPR and milk yield were positive and low in early lactation, but only in the first lactation. Genetic correlations between FPR and somatic cell score were positive in early lactation and decreased to become negative in mid‐ to late lactation. By using these results for genetic evaluation it should be possible to improve energy balance in dairy cows.     

Determination of theaflavins including methylated theaflavins in black tea leaves by solid-phase extraction and HPLC analysis

A quantitative method for four theaflavins and two methylated theaflavin derivatives in black tea leaves was developed by solid-phase extraction and a high-performance liquid chromatographic method with photodiode array detection. The theaflavins in black tea leaves were extracted three times with 40 vol 50% aqueous ethanol (mg dry tea powder/mL) containing 2% ascorbic acid. The ethanol extracts were diluted 4-fold with distilled water. All diluted extracts were directly applied to the solid-phase C18 cartridge column without concentration. The fraction of theaflavins was obtained by 40% ethanol extraction after rinsing with water followed with 15% ethanol extraction. An aliquot of theaflavins after concentration was injected onto an ODS C18 reversed-phase column, and four theaflavins and two methylated theaflavins were sufficiently separated by a linear gradient system using distilled water and acetonitrile with 0.5% acetic acid. This analytical method is sensitive for the determination of a small amount of methylated theaflavins, since various interfering substances produced during the fermentation process were eliminated in advance by solid-phase extraction. Using this analytical method, we also demonstrated that methylated theaflavins were easily produced during the manufacture of black tea.     

Monetary costs of dietary energy reported by young Japanese women: association with food and nutrient intake and body mass index

OBJECTIVE: Little is known about the relationship of monetary diet costs to dietary intake and obesity, particularly in non-Western populations. This study examined monetary cost of dietary energy in relation to diet quality and body mass index (BMI) among young Japanese women. DESIGN: Dietary intake was assessed by a validated, self-administered, diet history questionnaire. Diet costs were estimated using retail food prices. Monetary cost of dietary energy (Japanese yen 1000 kcal-1) was then calculated. BMI was computed from self-reported body weight and height. SUBJECTS: A total of 3931 female Japanese dietetic students aged 18-20 years. RESULTS: Monetary cost of dietary energy was positively associated with intakes of fruits, vegetables, fish and shellfish, and pulses; however, higher monetary cost of dietary energy was also associated with higher consumption of fat and oil, meat and energy-containing beverages, and lower consumption of cereals (rice, bread and noodles) (all P for trend <0.01). At the nutrient level, monetary cost of dietary energy was positively associated with intakes of dietary fibre and key vitamins and minerals, but also associated positively with intakes of fat, saturated fatty acids, cholesterol and sodium, and negatively with carbohydrate intake (all P for trend <0.0001). After adjustment for possible confounders, monetary cost of dietary energy was quite weakly but significantly negatively associated with BMI (P for trend = 0.0197). CONCLUSIONS: Increasing monetary cost of dietary energy was associated with both favourable and unfavourable dietary intake patterns and a quite small decrease in BMI in young Japanese women.