Distinguishing local growth from immigration-based size shifts for juvenile chum salmon communities in coastal Hokkaido, northern Japan

The body size of juvenile fish is often used as an index of growth rate, which in turn is influenced by local habitat conditions. We evaluated the size and origin of juvenile Chum salmon Oncorhynchus keta in the coastal areas of three regions (Atsuta, Shari, and Konbumori) of Hokkaido, northern Japan. The origin of the individuals in these communities differed between Konbumori and the other two regions. The former contained juveniles that originated from both the nearest stocked river and other rivers that were outside the area of interest. Conversely, the communities in Atsuta and Shari consisted exclusively of fish from the nearest stocked river. Moreover, the juveniles in Konbumori were larger than those in Atsuta and Shari. The results of our otolith analysis suggest that the larger size of the fish in Konbumori was due to the immigration of large individuals from natal rivers at distant locations. Thus, immigrants were likely to enlarge their body size composition in the area of interest. In summary, if the distance from a natal river is adjusted and daily growth is taken into account, body size can be used as an indicator of growth performance in coastal juvenile chum salmon.     

Isolation of ligament protein-1 from shell ligament of the scallop Patinopecten yessoensis

Elastomeric proteins occur in the shell ligament of bivalve molluscan shellfish. A newly identified protein named ligament protein-1, with a molecular mass of approximately 12 kDa, was extracted from the shell ligament of the scallop, Patinopecten yessoensis, with 70 % formic acid, and purified by gel filtration column chromatography and reverse-phase C18 column chromatography. Although the amino acid composition of this protein differs from that of the known shell ligament protein abductin, the partial amino acid sequences are similar to those of abductin. CD spectra and FT–IR spectra analyses showed that ligament protein-1 retains its secondary structures of α-helix, β-turn, and β-sheets. To the best of our knowledge, this is the first study on protein purification from the shell ligament of a bivalve molluscan shell.     

Density estimation of the giant jellyfish Nemopilema nomurai around Japan using an alternative modified detection function for left truncation in a line transect survey

Information on the distribution and bloom scale of the giant jellyfish Nemopilema nomurai is necessary to understand the impact on fisheries. In October 2009, we had an opportunity for visual counting of N. nomurai around Japan from a research/training vessel. To take into account possibly severe missed detection of N. nomurai in the close vicinity of the vessel, detection functions were modified by introducing an additional parameter δ expressing the proportion of detectable N. nomurai within a threshold perpendicular distance. Some covariate effects in the detection process were also investigated. The δ-modified hazard-rate detection function with δ variation parameter of jellyfish body size was chosen as the best-fit one by AIC model selection. The results suggested that a higher proportion (>30 % or more) of N. nomurai were undetected in the vicinity of vessel. The δ-modified model had an advantage of no loss of left-truncated data and provided better estimation of density with the smaller coefficient of variation than the conventional left truncation approach. It is applicable to other line transect surveys, e.g. aerial surveys. High densities of N. nomurai were found in the central part of the Sea of Japan and off Iwate Prefecture.     

Low water exchange Gracilariopsis bailiniae Zhang & B.M. Xia culture in intensive milkfish culture effluents: effects of seaweed density on seaweed production and effluent treatment

This study tested the feasibility of a low-cost seaweed biofiltration system for pond-based aquaculture through an indoor-integrated fish-seaweed culture experiment using weekly nutrient supply regime and different seaweed stocking densities. The culture experiment was conducted in glass aquaria that were stocked with Gracilariopsis bailiniae at 3 densities (low = 0.5 kg m^?2, middle = 2 kg m^?2 and high = 3.5 kg m^?2) and provided either with effluents from intensive milkfish (Chanos chanos) culture or with effluent-free seawater (control) as nutrient source. Stocking density was used as a factor in optimizing nutrient availability for growth and nutrient removal under such low water exchange conditions. Our results showed that G. bailiniae cultivated in milkfish effluents had higher growth, biomass and nitrogen yields than those cultivated in effluent-free seawater. Among the different stocking densities tested, highest growth rate (1.03 % day^?1) was obtained in the middle density. Increasing biomass and nitrogen yields were also obtained at this density until the end of the culture period. Poorer growth rates at low and high stocking densities were attributed to light limitation from phytoplankton and self-shading, respectively. Due to seaweed treatment, average outflow concentration of NH₄ ⁺ was reduced from half of its pretreated level. This study showed that a weekly effluent supply at 2 kg m^?2 seaweed stocking density can sustainably support the growth of G. bailiniae as long as the dissolved nutrients are present at high levels.     

Comparisons among MRI signs,apparent diffusion coefficient,and fractional anisotropy in dogs with a solitary intracranial meningioma or histiocytic sarcoma

Although MRI has become widely used in small animal practice, little is known about the validity of advanced MRI techniques such as diffusion‐weighted imaging and diffusion tensor imaging. The aim of this retrospective analytical observational study was to investigate the characteristics of diffusion parameters, that is the apparent diffusion coefficient and fractional anisotropy, in dogs with a solitary intracranial meningioma or histiocytic sarcoma. Dogs were included based on the performance of diffusion MRI and histological confirmation. Statistical analyses were performed to compare apparent diffusion coefficient and fractional anisotropy for the two types of tumor in the intra‐ and peritumoral regions. Eleven cases with meningioma and six with histiocytic sarcoma satisfied the inclusion criteria. Significant differences in apparent diffusion coefficient value (× 10^?3 mm²/s) between meningioma vs. histiocytic sarcoma were recognized in intratumoral small (1.07 vs. 0.76) and large (1.04 vs. 0.77) regions of interest, in the peritumoral margin (0.93 vs. 1.08), and in the T2 high region (1.21 vs. 1.41). Significant differences in fractional anisotropy values were found in the peritumoral margin (0.29 vs. 0.24) and the T2 high region (0.24 vs. 0.17). The current study identified differences in measurements of apparent diffusion coefficient and fractional anisotropy for meningioma and histiocytic sarcoma in a small sample of dogs. In addition, we observed that all cases of intracranial histiocytic sarcoma showed leptomeningeal enhancement and/or mass formation invading into the sulci in the contrast study. Future studies are needed to determine the sensitivity of these imaging characteristics for differentiating between these tumor types.     

Exclusion of cytoplasmic fragments in flow cytometric analysis of lymph node samples from dogs with lymphoma using membrane-permeable violet laser-excitable DNA-binding fluorescent dye (DyeCycle Violet)

Background: Cytoplasmic fragments derived from fragile neoplastic lymphocytes are common in samples of lymph nodes collected from dogs with lymphoma. These cytoplasmic fragments interfere with accurate gating of target cells and quantification protocols used for flow cytometry because of their variable size and expression of lymphoid cell surface antigens on their membranes. Objective: The aim of this study was to develop a method to efficiently exclude cytoplasmic fragments from flow cytometric analysis of canine lymph nodes in which lymphoma was present. Methods: Single‐cell suspensions of neoplastic cells were prepared from biopsy samples and fine‐needle aspirates of lymph nodes from 23 dogs with lymphoma. Suspensions were stained using a violet laser‐excitable (405 nm) membrane‐permeable DNA‐binding fluorescent dye (DyeCycle Violet [DCV]), incubated with antibodies against CD3, CD5, CD21, CD22, and CD45, and then stained with 7‐amino‐actinomycin D (7‐AAD), an argon‐excitable (488 nm) membrane‐impermeable DNA‐binding fluorescent dye. Multiparameter flow cytometry was used for analysis based on selective uptake and laser‐activated fluorescence of these dyes. Results: Cytoplasmic fragments, which were DCV‐negative and CD45‐positive, and dead cells, which were positive for 7‐AAD, were efficiently separated from neoplastic cells. Conclusion: Staining with DCV is a useful method to improve flow cytometric gating methods and quantitative analyses of lymph node samples from dogs with lymphoma.     

Treatment of canine otitis externa using video otoscopy

Otitis externa in 27 toy poodles and 40 miniature dachshunds was treated using a video otoscope. A distinct concavity (external tympanic concavity) was observed at the junction between the ventral part of the external surface of the tympanum and the ear canal to which a considerable amount of hair and debris had adhered. All hair and debris adhering to the external tympanic concavity were removed, and systemic antibiotic and antifungal agents were administered, after which all of the dogs recovered. The pattern of hair growth observed in the external tympanic concavity could be characterized according to the breed of dog. All of the toy poodles presented with curly hairs, while the miniature dachshunds had upright or flat-lying hairs.     

Localization of eNOS in the olfactory epithelium of the rat

Nitric oxide (NO) is a free radical and produced from L-arginine by nitric oxide synthase (NOS). Since NO is recently suggested to be involved in olfactory perception, the expression of eNOS, an isoform of NOS, was examined in the rat olfactory epithelium. The activity of NADPH-diaphorase was also examined as a marker of NOS. In the dorsomedial region of the nasal cavity, intensely positive reactions for NADPH-diaphorase were observed in the entire cytoplasm of sensory cells (olfactory cells). By immunohistochemistry, intensely positive reactions for eNOS were also found in the dorsomedial region of the nasal cavity. These reactions were observed on the free border of the olfactory epithelium. By immunoelectron microscopy, positive reactions for eNOS were found in the cilia of olfactory cells. In addition, in situ hybridization analysis of the olfactory epithelium revealed the expression of eNOS mRNA in the olfactory cells. These results indicate the presence of eNOS in the olfactory cells of the rat, and differential expression of eNOS in the olfactory epithelium depending on the regions of the nasal cavity. In addition, NO produced by eNOS may be involved in olfactory perception in the cilia of olfactory cells.     

The antagonist activity of lipid IVa on the stimulation by lipid A of TNF-alpha production from canine blood mononuclear cells

Lipid A, the active component of lipopolysaccharide (LPS), exists in the outer membrane of Gram-negative bacteria and binds to the Toll-like receptor 4 (TLR4) and MD-2 complex. On the other hand, the synthetic precursor of Escherichia coli lipid A, tetraacylated lipid IVa, is an agonist for TLR4 and MD-2 complex in murine, equine and feline cells but is an antagonist for lipid A in human cells. The aim of the study was to examine the function of canine Toll-like receptor 4 (TLR4) and MD-2 complex on canine blood mononuclear cells (BMC), by analyzing lipid A- or lipid IVa-induction of TNF-α production from these cells in order to understand canine innate immune system. After 5-h culture of canine BMC with lipid A (lipid A culture) or lipid IVa (lipid IVa culture), the TNF-α, as determined by ELISA, had increased in the supernatants of the lipid A cultures in a dose-dependent manner, whereas the TNF-α was undetectable in supernatant of lipid IVa-treated cultures. The TNF-α was statistically significantly different between the lipid A and lipid IVa cultures (100 and 1000 ng/ml). TNF-α production from canine BMC was inhibited, in a lipid IVa-dose-dependent manner, when the BMC were pre-cultured with lipid IVa for 60 min and then cultured with lipid A for 5h, while in control BMC cultures production if TNF-α was unchanged. These results indicate that the TNF-α production stimulated by lipid A was competed out by pre-exposing the BMC to lipid IVa. Thus, lipid A is an agonist for TNF-α production in canine BMC, whereas lipid IVa appears to be an antagonist against this lipid A stimulation of canine BMC.