Horizontal and vertical variations in survival rates of juvenile bivalves,Anadara kagoshimensis (Tokunaga, 1906), Ruditapes philippinarum (Adams & Reeve, 1850) and Atrina sp., experimentally caged in the inner area of the Sea of Ariake,Japan

International Aquatic Research - In the inner area of the Sea of Ariake, rearing experiments were conducted to examine how the survival rates of juveniles of three bivalve species (Anadara...     

Detection of Babesia equi (Laveran, 1901) by nested polymerase chain reaction

We describe a nested polymerase chain reaction (PCR) for the detection of Babesia equi in equine infected erythrocytes using oligonucleotides designed on the published sequence of a B. equi merozoite antigen gene (ema-1). A 102bp DNA fragment is specifically amplified from B. equi but not from Babesia caballi, Babesia bovis or Babesia bigemina DNA. In a mock infection we were able to detect down to six infected cells in 10(8) equine erythrocytes or to detect the parasite in blood with an equivalent parasitemia of 0.000006%. Furthermore, gene polymorphism was found by performing a PCR-RFLP (PCR combined with restriction fragment length polymorphism) on both the 102bp and the entire ema-1 gene DNA amplified from two B. equi isolates, Florida (USA) and Pelotas (Southern Brazil) isolates. The polymorphism was confirmed by sequencing the entire ema-1 gene from the B. equi isolate Pelotas. Our results demonstrate that the ema-1 based nested PCR is a valuable technique for routine detection of B. equi in chronically infected horses. It may be used for epidemiological and phylogenetic studies of the parasite as well as monitoring B. equi infected horses in chemotherapeutic trials.     

Effects of fat coated rumen bypass lysine and methionine on performance of dairy cows fed a diet deficient in lysine and methionine

Two experiments were conducted to evaluate the effect of fat coated rumen bypass lysine (RPLys) and methionine (RPMet) on the lactation performance of dairy cows. In Experiment 1, three lactating cows were supplied with RPLys and fat coated DL‐Met which was highly protected (H‐RPMet) as an indigestible marker, and total fecal emission was collected for 72 h following administration. Measuring the proportional difference in fecal excretion of lysine derived from RPLys relative to methionine derived from H‐RPMet, the intestinal availability of RPLys was estimated to be 66.2%. In Experiment 2, 20 multiparous Holstein cows producing approximately 40 kg/day of milk were assigned to two treatments; fed RPLys (16 g/day as lysine) and RPMet (6.5 g/day as methionine) or none (control) from 5 to 21 weeks postpartum. The consumption of dry matter, organic matter, crude protein, neutral detergent fiber and acid detergent fiber were significantly more in the control cows throughout the experimental period. Their milk protein yield, the contents of their milk protein and milk fat were higher by 0.03 kg (P = 0.03), 0.06% (P < 0.001) and 0.11% (P = 0.07), respectively, in the treatment group compared to the control. These results suggest that the RPLys and RPMet used in this study improved the lactation performance of dairy cows.     

Antibodies to Borna disease virus in infected adult rats: an early appearance of anti-p10 antibody and recognition of novel virus-specific proteins in infected animal brain cells

The time course for appearance of antibodies to Borna disease virus (BDV) major antigens, p40, p24, p18 and p10 were investigated in BDV-inoculated adult rats by Western blotting. Anti-p10 antibodies were detected in sera as early as anti-p40 and -p24 antibodies at four or five weeks after inoculation. Furthermore, in addition to these major antigens of BDV, the rat serum could detect additional 80-, 58-, 43-, 20-, and 16-kDa proteins in BDV-infected cultured cells and/or animal brain cells by Western blot analysis. Of these proteins, the 20- and 16-kDa proteins were shown to be related to p24 protein by their reactivity with anti-p24 monoclonal antibody. Interestingly, the 58- and 24-kDa were found only in BDV-infected animal brain cells but not in cultured cells. The results in this study could provide a useful information on the mechanism for the viral replication and pathogenesis.     

An improved transformation system for Lombardy poplar (Populus nigra var. italica)

We report an improved transformation system for Lombardy poplar (Populus nigra var. italica). A new binary vector, pBF2, with 11 unique restriction enzyme sites and the normal neomycin phosphotransferase II (NPTII) gene was constructed for the transformation of Lombardy poplar. Genetically transformed adventitious shoots were directly regenerated after cocultivation of stem segments with Agrobacterium tumefaciens EHA105 that harbored a binary vector with genes for the NPTII and enhanced green fluorescent protein. Successful transformation was confirmed by fluorescence microscopy, immunoblotting and Southern blotting analyses, and resistance to kanamycin and geneticin. This transformation system requires less time than our previous method for the regeneration of transgenic shoots. When explants were incubated on a smedium containing dithiothreitol, the transformation frequency increased to approximately 20%.     

PDE4 and PDE5 regulate cyclic nucleotide contents and relaxing effects on carbachol-induced contraction in the bovine abomasum

The effects of various selective phosphodiesterase (PDE) inhibitors on carbachol (CCh)-induced contraction in the bovine abomasum were investigated. Various selective PDE inhibitors, vinpocetine (type 1), erythro-9-(2-hydroxy-3-nonyl) adenine (EHNA, type 2), milrinone (type 3), Ro20-1724 (type 4), vardenafil (type 5), BRL-50481 (type 7) and BAY73-6691 (type 9), inhibited CCh-induced contractions in a concentration-dependent manner. Among the PDE inhibitors, Ro20-1724 and vardenafil induced more relaxation than the other inhibitors based on the data for the IC₅₀ or maximum relaxation. In smooth muscle of the bovine abomasum, we showed the expression of PDE4B, 4C, 4D and 5 by RT-PCR analysis. In the presence of CCh, Ro20-1724 increased the cAMP content, but not the cGMP content. By contrast, vardenafil increased the cGMP content, but not the cAMP content. These results suggest that Ro20-1724-induced relaxation was correlated with cAMP and that vardenafil-induced relaxation was correlated with cGMP in the bovine abomasum. In conclusion, PDE4 and PDE5 are the enzymes involved in regulation of the relaxation associated with cAMP and cGMP, respectively, in the bovine abomasum.     

Aquagrams of Raw Milk for Oestrus Detection in Dairy Cows

The purpose of this research was to develop rapid and cost‐effective method for oestrus detection in dairy cows by means of near infrared spectroscopy and aquaphotomics, using raw milk from individual cows. We found that aquaphotomics approach showed consistent specific water spectral pattern of milk at the oestrus periods of the investigated Holstein cows. Characteristic changes were detected especially in foremilk collected at morning milking. They were reflected in calculated aquagrams of milk spectra where distinctive spectral pattern of oestrus showed increased light absorbance of strongly hydrogen‐bonded water. Results showed that monitoring of raw milk near infrared spectra provides an opportunity for analysing hormone levels indirectly, through the changes of water spectral pattern caused by complex physiological changes related to fertile periods.     

Effects of various selective phosphodiesterase inhibitors on carbachol-induced contraction and cyclic nucleotide contents in the guinea pig gall bladder

The effects of various selective phosphodiesterase (PDE) inhibitors on muscle contractility and cyclic nucleotide contents in the guinea pig gall bladder were investigated. Various selective PDE inhibitors, vinpocetine (type 1), erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA, type 2), milrinone (type 3), Ro20-1724 (type 4), and zaprinast (type 5), inhibited CCh-induced contractions in a concentration-dependent manner. The rank order of potency for the gall bladder was Ro20-1724 > vinpocetine > EHNA > milrinone > zaprinast, which was different from that of the trachea, taenia coli, and aorta. In the presence of CCh (0.3 muM), vinpocetine, milrinone, and Ro20-1724 each increased cAMP content, but not cGMP. By contrast, zaprinast increased cGMP content, but not cAMP, and EHNA increased both cAMP and cGMP contents. These results suggest that vinpocetine-, milrinone-, and Ro20-1724-induced relaxation was correlated with cAMP, zaprinast-induced relaxation was correlated with cGMP, and that EHNA-induced relaxation was correlated with cAMP and cGMP in the guinea pig gall bladder. In conclusion, the effect of PDE inhibitors in the guinea pig gall bladder was different from those in smooth muscles, such as the trachea, taenia coli, and aorta.