New fermentation technique to process fish waste

The traditional method of processing fish waste is costly and usually overheats the waste, making it less digestible. In the present study, a fermentation technique using Aspergillus awamori was used to dry minced sardine fish to find a new model for processing fish waste and to evaluate its digestibility. Wheat bran (700 g), minced fresh sardines (2638 g) and A. awamori spores (0.7 g) were mixed together and kept in a ventilated incubator for 5 days at 40°C. At the end of the fermentation, a sample of fishmeal was taken to measure the nutrient content, enzyme activity and digestibility of crude protein (CP) and dry matter (DM) using an in vitro method. Furthermore, the effect of enzymes produced during the fermentation of DM and CP digestibilities of corn–soybean mixture in vitro were also examined. The percentage of moisture, CP, crude fat and nitrogen‐free extract in the fermented fish waste were 11, 39, 23 and 25%, respectively. The in vitro digestibilities of DM (69%) and CP (84%) of the fishmeal were almost the same as those of the heat‐dried sardine powder. During the fermentation, glucoamylase, α‐glucosidase, α‐amylase and acidic protease were produced, which may improve the DM digestibility of the diet. In conclusion, fish waste can be dried successfully using the fermentation technique described in the present study. Furthermore, the DM digestibility of the corn–soybean meal mixture is improved by the enzymes produced during the fermentation.     

Nucleotide sequence polymorphisms of beta1‐, beta2‐, and beta3‐adrenergic receptor genes on Jinhua,Meishan, Duroc and Landrace pigs

The full amino acid coding sequences of adrenergic receptor genes beta1, beta2, and beta3 (ADRB1, ADRB2, and ADRB3)were determined for Jinhua, Meishan, Duroc and Landrace pigs. Non‐synonymous substitution of Arg458Pro was found in the porcine ADRB1 gene, resulting in a 469 amino acid sequence. Continuous substitutions of Asn29Asp and Glu30Gln were found in the porcine ADRB2 gene, resulting in a 418 amino acid sequence. Additionally, a Lys30 polymorphism of the ADRB2 gene was found in the Jinhua pigs. There were three non‐synonymous substitutions of Asn24Thr, Arg264Gln and Asn398Asp on the porcine ADRB3 gene. A thymine insertion in the ADRB3 gene, resulting in a protein with two fewer amino acids, was found in the Jinhua and Meishan pigs. To assess the effect of ADRB polymorphisms on porcine subcutaneous fat layer thickness, we calculated the genetic frequency of the variants in fatty and lean groups, each consisting of 24 pigs that were crossbreds of Duroc and Jinhua pigs. The effect of the ADRB3 gene polymorphism was not evaluated, because there was insufficient variation on the ADRB3 gene in the examined groups. Although Fisher's exact test showed no significant difference in the frequency of ADRB1 and ADBR2 variants between the two groups, the Arg458 variant of ADRB1 was higher (P = 0.11) in the lean group, and pigs in that group had a thinner fat layer than did those with the Pro458 variant. These results imply a possibility of ADRB1 polymorphism as a minor factor in porcine fat layer thickness. The Asp29 variant of ADRB2 was higher in the lean group (P = 0.11), and the Glu30 variant was higher in the fatty group (P = 0.15), but the Asp29 variant was found only in the Chinese pigs. Thus, the effect of ADRB2 polymorphisms was not clear in this study.     

Mouse in vivo-derived late 2-cell embryos have higher developmental competence after high osmolality vitrification and −80°C preservation than IVF or ICSI embryos

Mammalian embryos are most commonly cryopreserved in liquid nitrogen; however, liquid nitrogen is not available in special environments, such as the International Space Station (ISS), and vitrified embryos must be stored at −80°C. Recently, the high osmolarity vitrification (HOV) method was developed to cryopreserve mouse 2-cell stage embryos at −80°C; however, the appropriate embryo is currently unknown. In this study, we compared the vitrification resistance of in vivo-derived, in vitro fertilization (IVF)-derived, and intracytoplasmic sperm injection (ICSI)-derived mouse 2-cell embryos against cryopreservation at −80°C. The ICSI embryos had lower survival rates after warming and significantly lower developmental rates than the in vivo and IVF embryos. Further, IVF embryos had a lower survival rate after warming, but a similar rate to the in vivo embryos to full-term development. This result was confirmed by simultaneous vitrification of in vivo and IVF embryos in the same cryotube using identifiable green fluorescent protein-expressing embryos. We also evaluated the collection timing of the in vivo embryos from the oviduct and found that late 2-cell embryos had higher survival and developmental rates to full-term than early 2-cell embryos. Some early 2-cell embryos remained in the S-phase, whereas most late 2-cell embryos were in the G2-phase, which may have affected the tolerance to embryo vitrification. In conclusion, when embryos must be cryopreserved under restricted conditions, such as the ISS, in vivo fertilized embryos collected at the late 2-cell stage without long culture should be employed.     

Population structure of pigs determined by single nucleotide polymorphisms observed in assembled expressed sequence tags

We have collected more than 190 000 porcine expressed sequence tags (ESTs) from full‐length complementary DNA (cDNA) libraries and identified more than 2800 single nucleotide polymorphisms (SNPs). In this study, we tentatively chose 222 SNPs observed in assembled ESTs to study pigs of different breeds; 104 were selected by comparing the cDNA sequences of a Meishan pig and samples of three‐way cross pigs (Landrace, Large White, and Duroc: LWD), and 118 were selected from LWD samples. To evaluate the genetic variation between the chosen SNPs from pig breeds, we determined the genotypes for 192 pig samples (11 pig groups) from our DNA reference panel with matrix‐assisted laser desorption ionization time‐of‐flight mass spectrometry. Of the 222 reference SNPs, 186 were successfully genotyped. A neighbor‐joining tree showed that the pig groups were classified into two large clusters, namely, Euro‐American and East Asian pig populations. F‐statistics and the analysis of molecular variance of Euro‐American pig groups revealed that approximately 25% of the genetic variations occurred because of intergroup differences. As the F_IS values were less than the F_ST values_, the clustering, based on the Bayesian inference, implied that there was strong genetic differentiation among pig groups and less divergence within the groups in our samples.     

Elevation of serum surfactant protein-A with exacerbation in canine eosinophilic pneumonia

A 7-year-old female spayed Labrador Retriever was admitted to our hospital, because ofcough with sputum. She was diagnosed as having canine eosinophilic pneumonia (CEP) basedon blood eosinophilia, bronchial pattern and infiltrative shadow observed on thoracicradiography, bronchiolar obstruction and air-space consolidation predominantly affectingthe right caudal lung lobe, as revealed by computed tomography (CT), predominanteosinophils in CT-guided fine needle aspiration and the clinical course. She exhibited agood response to steroid therapy, and the cough disappeared. The serum surfactant protein(SP)-A level increased with the aggravated symptom and decreased markedly with improvementcompared with the C-reactive protein level and the number of eosinophils. We propose thatserum SP-A level is a good biomarker in CEP.     

Direct effects of linoleic and linolenic acids on bovine uterine function using in vivo and in vitro studies

The aim of the present study was to evaluate the effects of continuous administration of linoleic acid or linolenic acid into the intra-uterine horn, ipsilateral to the corpus luteum, on the duration of the estrous cycle and plasma progesterone (P4) concentration. The effects of linoleic and linolenic acids on bovine uterine and luteal functions were also studied using a tissue culture system. Intra-uterine administration of linoleic or linolenic acid (5 mg/10 ml of each per day) in cows, between days 12 and 21, resulted in a prolonged estrous cycle compared to the average duration of the last one to three estrous cycles before administration in each group (P < 0.05). Moreover, plasma P4 concentration in cows treated with linoleic or linolenic acid was high between days 19 and 21 (linoleic acid), or on day 20 (linolenic acid), compared to that of the control cows (saline administration; P < 0.05 or lower). Both linoleic (500 µg/ml) and linolenic (5 and 500 µg/ml) acids stimulated prostaglandin (PG) E2 but inhibited PGF2α production by cultured endometrial tissue (P < 0.01), while P4 production by cultured luteal tissue was not affected. These findings suggest that both linoleic and linolenic acids support luteal P4 production by regulating endometrial PG production and, subsequently, prolonging the duration of the estrous cycle in cows.     

Lethal limits of high temperature for two crayfishes, the native species Cambaroides japonicus and the alien species Pacifastacus leniusculus in Japan

ABSTRACT: Lethal limits of high temperature were studied to clarify the effects on the survival of the endangered Japanese crayfish species Cambaroides japonicus and the alien species Pacifastacus leniusculus . After the acclimation period for 2 weeks at 16°C, the temperature was raised at a rate of 1°C per week. As a result, the ultimate upper lethal temperatures of C.   japonicus and P.   leniusculus were 27.0 and 31.1°C, respectively, and the lethal temperature for P.   leniusculus was significantly higher than for C.   japonicus . The natural distributions of these two species are discussed in terms of the temperature tolerance.