Measurements of auditory sensitivity in common carp <Emphasis Type="Italic">Cyprinus carpio</Emphasis> by the auditory brainstem response technique and cardiac conditioning method

ABSTRACT:   Recently, the non-invasive auditory brainstem response (ABR) technique has been applied to the determination of hearing sensitivity in fish. The technique has some advantages over, and methodological differences from, the classical techniques. The auditory thresholds of common carp were measured by the ABR technique. The classical method was applied to measure cardiac response by electrocardiogram (ECG) after conditioning with electric shock. The most sensitive frequency obtained by ABR was 505 Hz, and by ECG, 1000 Hz. The shapes of the audiogram obtained by ABR and ECG were similar, though threshold levels in both audiograms differed from each other. Furthermore, audiograms obtained by using behavioral techniques were compared with those taken by the ABR and ECG techniques to assess the validity of the ABR technique for testing fish hearing sensitivity. The measured threshold levels increased in the order of behavioral, ABR, and ECG techniques, in the range below 1000 Hz.     

Estimation of the mode of inheritance of thermal tolerance in the guppy <Emphasis Type="Italic">Poecilia reticulata</Emphasis>

High water temperature influences the survival, growth, and maturation of fish. Genetically characterizing thermal tolerance is one of the most important subjects in fish culture. To identify the genetic characterization of thermal tolerance, this characteristic was compared among strains, and among parents and their offspring, in the guppy Poecilia reticulata. In the strain comparison, significant differences in survival rate were observed among the five strains examined, and between females and males. Females exhibited greater tolerance than males in four of five strains examined. In the comparison between parents and their offspring, stronger influence of female parent than of male parent was observed. Offspring obtained from surviving females exhibited greater tolerance than those from dead females. This tendency was typically observed in male offspring. The survival rate in male offspring obtained from dead female parents was lower than that of those from surviving females. The high-temperature tolerance of male parents did not influence this characteristic in offspring as strongly as that of female parents. These results suggest that the major gene or genes, which has a dominant resistant allele and a recessive sensitive allele, are probably passed on by sex-linked inheritance, located on the X chromosome.     

Immunostimulant effects of nucleotide isolated from yeast RNA on carp, Cyprinus carpio L.

Nucleotides from yeast RNA were evaluated for their ability to enhance the non-specific immune responses in carp, Cyprinus carpio . Oral administration of nucleotides to fish daily for 3 days resulted in enhanced responses of phagocytic and nitroblue tetrazolium (NBT) activities in kidney phagocytic cells. This activation of kidney cells was observed for at least 10 days post-treatment. The serum complement and lysozyme activities also increased in fish treated with nucleotides. Furthermore, the number of Aeromonas hydrophila in nucleotide-treated fish significantly decreased in the blood, kidney and liver after intraperitoneal injection. Thus yeast nucleotides appear to enhance non-specific immune responses in fish.     

Inhibitory effect of 2,4-dibromophenol and 2,4,6-tribromophenol on larval survival and metamorphosis of the sea urchin <Emphasis Type="Italic">Strongylocentrotus nudus</Emphasis>

ABSTRACT:   As a possible factor leading to the low recruitment level of sea urchins in kelp forests, the inhibitory effect of 2,4-dibromophenol (DBP) and 2,4,6-tribromophenol (TBP) released from the large perennial brown algae Ecklonia kurome and Eisenia bicyclis on survival and metamorphosis of eight-armed larvae of the sea urchin Strongylocentrotus nudus was examined. The percentage of larvae that underwent metamorphosis in filtered sea water after 1 h exposure to one-half dilution of saturated dibromomethane solution (∼60 ppm) as a chemical inducer reached approximately 100% after 1 h, while that in filtered sea water containing 1 ppm TBP was reduced to 73%. This was further reduced to less than 40% in the presence of 10 and 20 ppm TBP after 2 h. In filtered sea water containing 1 and 10 ppm DBP, the proportion of metamorphosed larvae was reduced markedly to 43 and 5% after 2 h, respectively. All larvae exposed to 50 ppm TBP and to 20 and 50 ppm DBP died after 1 h. These findings suggest that DBP is more toxic than TBP for sea urchin larvae, strongly inhibiting their metamorphosis.     

Immunogenic properties and mycoplasmal pneumonia of swine (MPS) lung lesions in Large White pigs selected for higher peripheral blood immune capacity

Immunogenic properties and mycoplasmal pneumonia of swine (MPS) lung lesions were compared between the immunity‐selected Large White line and the non‐selected Large White line. The selected Large White line showed a higher level of pulmonary MPS lesions compared with the non‐selected Large White line. Subsequent to vaccination, the percentage of natural killer cells and T cells (CD3⁺CD4⁺CD8^‐ and CD3⁺CD4^?CD8⁺ T cells) were significantly increased in the non‐selected line but remained unchanged in the immunity‐selected Large White line. Secretion of Mycoplasma hyopneumoniae vaccine‐specific immunoblogulin G and phagocyte activity in peripheral blood were significantly higher in the immunity‐selected Large White line than in the non‐selected line. Expression of interleukin (IL)‐4 and IL‐6 messenger RNA in hilar lymph nodes was significantly lower in the immunity‐selected Large White line than in the non‐selected line. However, expression of IL‐10 in all immune tissues was significantly higher in the immunity‐selected Large White line. These results suggest that the selection for high immunity was not effective in increasing resistance to MPS lung lesions.     

Pathogenicity of emerging Japanese type 1 porcine reproductive and respiratory syndrome virus in experimentally infected pigs

To clarify the pathogenicity of Japanese type 1 porcine reproductive and respiratory syndrome virus (PRRSV) isolate in experimentally infected pigs, we evaluated clinical signs and monitored viremia for 21 days post-inoculation (dpi). Lungs were mottled, tanned and reddish in appearance; had lesions predominantly in the cranial, middle and accessory lobes; and failed to collapse at 10 dpi. Although microscopic lesions of lungs were reproduced using the Japanese emerging type 1 PRRSV isolate under experimental conditions, no significant differences were noted between the challenge and control groups regarding mean rectal temperature and daily weight gain. These results provide useful insights into the limited pathogenicity of single infection with the Japanese type 1 PRRSV isolate in piglets, which differ from findings in reported field cases.     

Census and effective population sizes of white‐spotted charr (Salvelinus leucomaenis) in a fragmented landscape

Several habitat models have been proposed to predict population size for stream fishes and to guide habitat assessment and monitoring techniques. However, most models do not incorporate the potential advantage of molecular genetic markers. We conducted a field survey and microsatellite DNA analyses to quantify the relationships among genetic diversity, census/effective population size and habitat variables in fragmented populations of white‐spotted charr (Salvelinus leucomaenis). The census population size significantly increased with the stream length, the number of pools and a pool‐riffle sequence index, a proxy for channel‐unit habitat type complexity within reaches. Population density was correlated with the pool‐riffle sequence index only. Genetic diversity and effective population size were not correlated with the habitat variables or census population size. There was a lack of isolation‐by‐distance population structure in the studied populations. Our results suggest that stream length and the number of pools within reaches associated with habitat complexity are the habitat variables that explain the majority of variation in population size of white‐spotted charr. Our findings provide further evidence that census population size per se is a poor indicator of the inclusive genetic diversity within populations in a fragmented landscape.     

Effect of retinoic acid on gene expression profiles of bovine intramuscular preadipocytes during adipogenesis

To investigate genes involved in intramuscular adipogenesis in ruminants, 16 genes with dramatic variable expression were selected. These were selected from the differentiation‐ and proliferation‐phase libraries of our previous serial analysis of gene expression (SAGE) studies of a clonal bovine intramuscular preadipocyte (BIP) cell line. We harvested the BIP cells over 12 days after adipogenic stimulation with all‐trans retinoic acid (ATRA). Quantitative real‐time PCR confirmed the earlier SAGE study results of the expression patterns of 15 of the genes. On day 6, TG accumulation increased significantly in the BIP cells but was completely inhibited in the 3T3‐L1 cells (the monogastric reference). ATRA enhanced expression levels of six genes whereas it suppressed expression of eight genes on day 3 of adipogenesis in the BIP cells. Forty‐eight hours after transfection, the messenger RNA expression level of the adipose differentiation‐related protein (ADFP), encoded by one of the upregulated genes, in the ADFP small interference RNA (siRNA)‐transfected cells was 3.5% of that in negative control‐transfected cells. Also, 6 days after induction the TG level in the ADFP siRNA‐transfected cells was 21.8% lower than that in negative control‐transfected cells. This analysis of gene expression profiles after ATRA treatment will contribute to our understanding of the molecular mechanisms involved in bovine intramuscular adipogenesis.