Expression dynamics of bovine MX genes in the endometrium and placenta during early to mid pregnancy

MX belongs to a family of type I interferon (IFN)-stimulated genes, and the MX protein has antiviral activity. MX has at least two isoforms, known as MX1 and MX2, in mammals. Moreover, bovine MX1 has been found to have alternative splice variants—namely, MX1-a and MX1B. In ruminants, IFN-τ—a type I IFN—is temporarily produced from the conceptus before implantation and induces MX expression in the endometrium. However, the expression dynamics of MX after implantation are not clear. In the present study, we investigated the expression of MX1-a, MX1B and MX2 in the endometrium and placenta before and after implantation along with the expression of IFN-α, type I receptors (IFNAR1 and IFNAR2) and interferon regulatory factors (IRF3 and IRF9). Pregnant uterine samples were divided into five groups according to pregnancy days 14–18, 25–40, 50–70, 80–100, and 130–150. Tissue samples were collected from the intercaruncular endometrium (IC), caruncular endometrium (C) and fetal placenta (P). Although all the MX expressions were significantly higher in the IC and C at days 14–18, presumably caused by embryo-secreted IFN-τ stimulation, their expressions were also detectable in the IC, C and P after implantation. Furthermore, IFN-α expression was significantly higher in the IC. RT-PCR indicated IFNAR1, IFNAR2, IRF3 and IRF9 mRNA in all the tissues during pregnancy. These results suggest that all the MX genes are affected by the type I IFN pathway during pregnancy and are involved in an immune response to protect the mother and fetus.     

Feeding habits of albacore Thunnus alalunga in the transition region of the central North Pacific

ABSTRACT:   The feeding habits of albacore Thunnus alalunga (fork length: 48.9–76.2 cm, n  = 132) were examined from late spring to early autumn in relation to its northward migration in the transition region between the subtropical and subarctic fronts in the central North Pacific. Samples were collected at night using surface gill nets or during daytime pole-and-line surveys in 2001 and 2002. During May and June, albacore fed mainly on Japanese anchovy Engraulis japonicus , which accounted for 27.2%, 67.0%, and 45.5% of the total stomach contents by number ( Cn ), wet weight ( WW ), and frequency of occurrence ( F ), respectively, and secondarily on the subarctic gonatid squid Gonatopsis borealis ( Cn , 15.8%; WW , 10.8%; F , 28.8%). From July to September, albacore continued to depend on Japanese anchovy ( Cn , 48.2–52.8%; WW , 79.9–95.2%; F , 27.8–85.4%). These results corresponded well with the remarkable rebound of the Japanese anchovy stock since the 1990s. Gonatopsis borealis , the main squid prey from May to June, almost disappeared from the stomachs of albacore from July to September, probably due to the northward migration of this squid to subarctic waters in summer. The feeding impact of albacore on the Japanese anchovy stock in the transition region was conservatively estimated to be from 1400 to 2100 tons per day from late spring to early autumn.     

Prey selection of common minke (Balaenoptera acutorostrata) and Bryde's (Balaenoptera edeni) whales in the western North Pacific in 2000 and 2001

A study of common minke and Bryde's whales was conducted in the western North Pacific in the 2000 and 2001 summer seasons to estimate prey selection of cetaceans as this is an important parameter in ecosystem models. Whale sighting and sampling surveys and prey surveys using quantitative echosounder and mid‐water trawl were carried out concurrently in the study. Biomasses of Japanese anchovy, walleye pollock and krill, which were major prey species of common minke and Bryde's whales, were estimated using an echosounder. The results suggested that common minke whale showed prey selection for Japanese anchovy while they seemed to avoid krill in both the offshore and coastal regions and walleye pollock in the continental shelf region. Selection for shoaling pelagic fish was similar to that in the eastern North Atlantic. Bryde's whale showed selection for Japanese anchovy in August 2000 and July 2001, while it showed prey selection for krill in May and June in 2001.     

Early metaphase II oocytes treated with dibutyryl cyclic adenosine monophosphate provide suitable recipient cytoplasm for the production of miniature pig somatic cell nuclear transfer embryos

We investigated the effects of in vitro maturation duration and treatment with dibutyryl cyclic adenosine monophosphate (dbcAMP) on the blind enucleation efficiency and developmental competence of miniature pig somatic cell nuclear transfer (SCNT) embryos. Oocytes were cultured for 22 h in NCSU-23 medium with or without 1 mM dbcAMP and then additionally cultured in dbcAMP-free NCSU-23 for 14, 18, or 22 h. Regardless of dbcAMP treatment, the rate of nuclear maturation reached a plateau at 36 and 40 h. However, mitochondrial distribution, a marker for cytoplasmic maturation, differed between the dbcAMP-untreated oocytes at 36 h and dbcAMP-treated oocytes at 40 h. The metaphase II chromosomes were adjacent to the first polar body in 68.8% and 63.5% of the dbcAMP-untreated oocytes at 36 h and dbcAMP-treated oocytes at 40 h, respectively. Furthermore, the blind enucleation efficiency by removing a small volume of cytoplasm was significantly higher in the dbcAMP-untreated oocytes at 36 h (82.9%) and dbcAMP-treated oocytes at 40 h (89.9%) than other groups. The rate of blastocyst formation was highest in the dbcAMP-treated oocytes at 40 h. Hence, this study demonstrated that dbcAMP-treated early metaphase II oocytes are suitable for the production of miniature pig SCNT embryos.     

Purification and characterization of an angiotensin I‐converting enzyme inhibitory peptide derived from porcine troponin C

To search for a novel angiotensin I‐converting enzyme (ACE) inhibitory peptide, porcine skeletal troponin was hydrolyzed with pepsin. This hydrolysate showed ACE inhibitory activity, and was applied to various kinds of chromatography to separate an active peptide. Analysis using a protein sequencer identified this peptide as RMLGQTPTK (9mer). This sequence was estimated to occur at the 44–52 position of troponin C, and its 50% inhibitory protein concentration (IC₅₀) was 34 µM. RMLGQTP (7mer), a partial peptide of 9mer, showed activity with an IC₅₀ of 503 µM. RP‐HPLC analysis of a reaction mixture of 9mer and ACE showed that 9mer was slowly hydrolyzed by ACE. On the other hand, 7mer was rapidly hydrolyzed by ACE. Activity of 9mer was reduced as its hydrolysis by ACE proceeded. To estimate the resistance of 9mer to digestive proteases after oral administration, it was reacted with pepsin, α‐chymotrypsin, or trypsin. In each of these reaction mixtures, a significant amount of 9mer remained as a substrate after digestion. Remaining ACE inhibitory activity was close to that of 9mer. These results suggest that 9mer might not be digested after oral administration, because of its relatively high resistance to digestive proteases. Therefore, 9mer might be expected to work well in vivo as an ACE inhibitor.     

Lipid peroxidation-derived hepatotoxic aldehydes, 4-hydroxy-2E-hexenal in smoked fish meat products

ABSTRACT:   Contents of 4-hydroxy-2 E -hexenal (HHE), hepatotoxic aldehydes, in smoked fish meat products (smoked salmon and fish meat sausage) were analyzed. Large differences in these contents between the different samples were observed. Very low levels of HHE were detected in fish meat sausage samples. However, a high level of HHE was observed in one batch of smoked salmon. Changes of HHE contents in yellowtail meat containing cherry and sugi wood vinegar stored at 0°C were also analyzed for 7 days. Malonaldehyde (MA) was also analyzed in these samples as an index of the lipid peroxidation level. After 3 or 7 days of storage, HHE contents in both wood vinegar-added samples were significantly higher, but MA contents were significantly lower than those of the control.     

Toxicity and immunogenicity of Aeromonas salmonicida extracellular products to salmonids

Abstract. Aeromonas salmonicida extracellular products (ECP) were fractionated by gel filtration chromatography. Three protein fractions were found. The first and second fractions showed low and high haemolytic activity, respectively, whereas the third fraction showed protcolytic activity. By intramuscular injection into juvenile rainbow trout, the median lethal doses of the first, second and third fractions were calculated at >669, 152 and <1514μg/g body weight, respectively. Cytopathic effects of the fractionated ECP to coho salmon lymphocytes were observed in vitro. The first and second fractions caused cell deformation, nuclear granulation and cytoplasmic streaming after 3h. No cytologic effects with the third fraction were observed. A mixture of the first and third fractions, a mixture of the second and third fractions, and non-fractionated ECP caused nuclear granulation and cytoplasmic streaming after 3Qmin. Using immunodiffusion analysis, the first and second fractions formed a single precipitating line each against white-spotted char anti-ECP sera, but the third fraction did not formed a precipitating line against the antisera.     

The investigation of probiotic potential of lactic acid bacteria isolated from traditional Mongolian dairy products

The aims of this study were to investigate the diversity of lactic acid bacteria (LAB) isolated from traditional Mongolian dairy products, and to estimate the probiotic potential of the isolated strains. We collected 66 samples of the traditional Mongolian dairy products tarag (n = 45), airag (n = 7), aaruul (n = 8), byasulag (n = 1) and eezgii (n = 5), from which 543 LAB strains were isolated and identified based on 16S ribosomal DNA sequence. The predominant species of those products were Lactobacillus (L.) delbrueckii ssp. bulgaricus, L. helveticus, L. fermentum, L. delbrueckii ssp. lactis and Lactococcus lactis ssp. lactis. However, we could not detect any LAB strains from eezgii. All LAB isolates were screened for tolerance to low pH and to bile acid, gas production from glucose, and adherence to Caco‐2 cells. In vitro, we found 10 strains possess probiotic properties, and almost identified them as L. plantarum or L. paracasei subspecies, based on 16S ribosomal DNA and carbohydrate fermentation pattern. These strains were differentiated from each other individually by randomly amplified polymorphic DNA analysis. Additionally, it was notable that 6/10 strains were isolated from camel milk tarag from the Dornogovi province.