Effects of high potassium chloride supplementation on water intake and bodyweight gains in pregnant and lactating mice

Thirty‐one ICR pregnant mice were assigned to a control or a potassium chloride (KCl) diet group to clarify the effects of KCl supplementation on water intake, bodyweight gains and serum components in pregnant and lactating mice, and 5% KCl was supplemented in KCl diets from 6.5 days post coitus to 1 or 14 days after parturition. Feed intake was not affected by treatment, but supplemental KCl decreased bodyweight gains of lactating mice and their neonatal mice. Water intake and urine volume of KCl supplemented mice were significantly higher than those of control mice during pregnancy and supplemental KCl decreased serum urea N in pregnant mice. Supplemental KCl increased water intake drastically in lactating mice immediately after parturition and increased serum K at 14 days after parturition. Histological alteration using hematoxylin–eosin was not found in the kidney of each mouse at 1 or 14 days after parturition. These results indicate that high KCl supplementation accelerates water intake in lactating mice and prevents bodyweight gains of maternal and neonatal mice during lactation.     

Production of transgenic cloned pigs expressing the far-red fluorescent protein monomeric Plum

Monomeric Plum (Plum), a far-red fluorescent protein with photostability and photopermeability, is potentially suitable for in vivo imaging and detection of fluorescence in body tissues. The aim of this study was to generate transgenic cloned pigs exhibiting systemic expression of Plum using somatic cell nuclear transfer (SCNT) technology. Nuclear donor cells for SCNT were obtained by introducing a Plum-expression vector driven by a combination of the cytomegalovirus early enhancer and chicken beta-actin promoter into porcine fetal fibroblasts (PFFs). The cleavage and blastocyst formation rates of reconstructed SCNT embryos were 81.0% (34/42) and 78.6% (33/42), respectively. At 36–37 days of gestation, three fetuses systemically expressing Plum were obtained from one recipient to which 103 SCNT embryos were transferred (3/103, 2.9%). For generation of offspring expressing Plum, rejuvenated PFFs were established from one cloned fetus and used as nuclear donor cells. Four cloned offspring and one stillborn cloned offspring were produced from one recipient to which 117 SCNT embryos were transferred (5/117, 4.3%). All offspring exhibited high levels of Plum fluorescence in blood cells, such as lymphocytes, monocytes and granulocytes. In addition, the skin, heart, kidney, pancreas, liver and spleen also exhibited Plum expression. These observations demonstrated that transfer of the Plum gene did not interfere with the development of porcine SCNT embryos and resulted in the successful generation of transgenic cloned pigs that systemically expressed Plum. This is the first report of the generation and characterization of transgenic cloned pigs expressing the far-red fluorescent protein Plum.     

Comparison of ultrasound- and electrostimulation-guided nerve blocks of brachial plexus in dogs

Objective

To compare the effectiveness of ultrasound- and electrostimulation-guided nerve blocks of the brachial plexus and to determine whether ultrasound guidance is feasible in conscious dogs.

Percutaneous transcatheter coil embolization of a ventricular septal defect in a dog

A 4-month-old male French Bulldog weighing 5.0 kg (11 lb) was referred for a heart murmur. A grade 3/6 systolic murmur was detected at the left heart base and a grade 4/6 systolic murmur was detected at the right heart base. By use of color-flow Doppler ultrasonography and cardiac catheterization, a diagnosis of supracristal ventricular septal defect (VSD) with accompanying aortic regurgitation was made. Percutaneous transcatheter coil embolization was used to close the VSD. Because residual shunt was detected via echocardiography after coil implantation, the residual shunt was followed periodically via echocardiography to detect spontaneous closure of the VSD. Volume overload in the left ventricle was detected in the dog 131 days after admission. Additional coils were placed 137 days after admission. Hemolysis resulting in hemoglobinuria was detected, but this complication was mild. In the dog of this report, results of coil occlusion for correction of VSD were promising. Thus, coil occlusion should be considered as an alternative treatment for VSD in dogs.     

Single-locus EST-SSR markers for characterization of population genetic diversity and structure across ploidy levels in switchgrass (<Emphasis Type="Italic">Panicum virgatum</Emphasis> L.)

Polyploidy, ploidal variation between populations, and aneuploidy within some populations complicate population genetic analyses in switchgrass. We report 21 genic-simple sequence repeat marker loci with single-locus disomic segregation in tetraploids and apparently tetrasomic inheritance in octoploids, thus allowing population genetic analyses across ploidy levels. Based on 472 individuals sampled over four tetraploid and eight octoploid cultivars, six to 55 alleles were detected per locus with an average of 24.1. Genetic diversity was greater in octoploids than tetraploids, as expected from polysomic inheritance. One tetraploid cultivar displayed comparable diversity to the least diverse octoploid cultivars, suggesting breeding history or population history in the native stands of origin may have also affected within-cultivar diversity. Amplicon number at each locus and population relationships suggest autopolyploid origin of octoploids within upland tetraploids with significant cultivar differentiation. However, model-based Bayesian clustering of individuals indicated that closely related octoploid cultivars are difficult to identify, possibly due to slowed differentiation by polysomic inheritance. The analysis of the sampling effect indicated addition of loci is more effective for cultivar identification than more individuals sampled per cultivar. Discriminating power of loci tended to correlate with their variability. The eight loci with greatest discriminatory power within tetraploids were equally successful as 20 loci at identifying the four tetraploid cultivars. The set of markers reported in this study are useful for characterization of switchgrass germplasm and identifying population structure for association studies.     

cDNA cloning and expression analysis of two heat shock protein genes,Hsp90 and Hsp60, from a sterile Ulva pertusa (Ulvales,Chlorophyta)

Molecular chaperones are synthesized and accumulated under a variety of unfavorable conditions in all organisms. Heat shock protein 90 (Hsp90) and Hsp60, which are classified into the major classes of molecular chaperones, play important roles in cellular stress responses. In this study, we characterized sterile Ulva pertusa Hsp90 (UpHsp90) and UpHsp60 genes which may be involved in tolerance to thermal and heavy metal stresses in this alga. The UpHsp90 cDNA consisting of 2,118 nucleotides encoded a polypeptide of 705 amino acids (AA). On the other hand, the UpHsp60 cDNA consisting of 1,722 nucleotides encoded a protein whose predicted length was 573 AA. The AA sequence alignment and phylogenetic analyses showed that the UpHsp90 and UpHsp60 proteins were more similar to cytoplasmic Hsp90s and mitochondrial Hsp60s, respectively, than to other types of the respective Hsps. Southern blot analysis indicated that the sterile U. pertusa genome had at least two cytoplasmic Hsp90-encoding genes and two mitochondrial Hsp60-encoding genes. The UpHsp90 and UpHsp60 mRNA levels were significantly affected by diurnal and temperature changes, and slightly affected by exposure to heavy metals. These results suggest that UpHsp90 and UpHsp60 genes play particularly important roles in adaptation to diurnal and temperature changes.     

Purification and functional characterization of complement C3 and a novel zymosan-binding protein in tilapia serum

Zymosan, a yeast cell wall preparation that binds activated forms of complement C3, is a useful model target to activate the complement system. In our trial to analyze C3 diversity in Nile tilapia at the protein level using zymosan, we found that a novel 240-kDa serum protein (ZBP-240) also bound to zymosan in addition to C3-derived fragments. In the present study, we aimed to characterize tilapia C3 and ZBP-240, focusing on their immune-related functions. Four distinct C3 isoforms were purified from tilapia serum and shown to possess an intrachain thioester bond. ZBP-240 was also isolated from tilapia serum and examined for its binding properties to various microbial targets. As a result, ZBP-240 showed a wide spectrum of binding to Gram-positive and Gram-negative bacteria and yeasts. Amino acid sequence analysis of CNBr fragments of ZBP-240 suggested that this is a novel protein with no homologous sequence in protein databases. It was also suggested that the binding of ZBP-240 to microbes largely depends on hydrophobic interactions in a divalent-cation-independent manner, and that there may be a divalent-cation-dependent factor that enhances the binding of ZBP-240 in tilapia serum. Interestingly, ZBP-240 showed opsonic activity for tilapia kidney phagocytes at a level comparable to that of C3, implying that ZBP-240 is a novel teleost opsonic serum protein.     

Aggressive behaviour and cannibalism in greater amberjack,Seriola dumerili : effects of stocking density,feeding conditions and size differences

Aggressive behaviour, assessed by 24‐h observations, and survival rates, determined in a 2‐week rearing experiment, were examined in greater amberjack, Seriola dumerili, subjected to single and multiple factors (high density, restricted feeding and diverse size). Aggressive behaviour frequency in the group subjected to both restricted feeding and size diversity (FS) was significantly higher than that in the other groups (Tukey, n=4, P<0.05). In the FS group, aggressive behaviour increased with the starvation time from 19 days post hatching (dph) to 23 dph (two‐way analysis of variance, Tukey, n=4, P<0.05), but decreased gradually thereafter to 46 dph. In the rearing experiment, the survival rates for the diverse size and FS groups were lower than those of the control or restricted feeding groups (Kaplan–Meier, log‐rank test, n=100, P<0.01). Daily mortality in all groups was the highest at 24 dph; although dead fish were usually small, disappearance due to cannibalism was not observed during the rearing period. The results indicate that aggressive behaviour in greater amberjack is induced by hunger and size diversity in the early juvenile stages (23 dph). Mass mortality during seedling production was mainly caused by injury and the death of small fish due to aggressive behaviour/cannibalism.