Expression of alpha-hemoglobin stabilizing protein and cellular prion protein in a subclone of murine erythroleukemia cell line MEL

Alpha-Hemoglobin stabilizing protein (AHSP) functions as the erythroid-specific molecular chaperon for alpha-globin. AHSP gene expression has been reported to be downregulated in hematopoietic tissues of animals suffering from prion diseases though the mechanism remains to be clarified. Herein, we demonstrate that MELhipod8 cells, a subclone of murine erythroleukemia (MEL) cells, have prion protein (PrPc) on the cell surface and have highly inducible expression of the AHSP and alpha- and beta-globin genes, resembling the expression pattern of the PrP and AHSP genes in bipotential erythroid- and megakaryocyte-lineage cells followed by erythroid differentiation in normal erythropoiesis. Moreover, MELhipod8 cells exhibit greater effective erythroid differentiation with a population of hemoglobinized normoblast-like cells than that observed for the parental MEL cells. These findings suggest that MELhipod8 cells could provide a mechanism for downregulation of the AHSP gene in prion diseases.     

FCV-VBS isolated from cats with typical symptoms caused VSD in experimental cats

Commercially available vaccines have been used widely to prevent feline calicivirus infection (FCI). However, with their widespread use, field strains, which are weakly cross-reactive with the live-virus vaccine strain F9, have posed the problem of vaccine breakdown. Recently the existence of FCV—associated virulent systemic disease (VSD) has been published. But their molecular diversity, antigenic mutations and physicochemical property have not been sufficiently clarified. Thus, we experimentally gave the vaccine breakdown strain (VBS) H10 to cats that had been inoculated with an F9 live vaccine. After the administration of strain H10, vaccinated cats (1 through 4) had no respiratory symptoms, whereas the non-vaccinated cat 5 showed clinical symptoms such as a fever of over 40°C, loss of vitality, decreased appetite, diarrhea, and nasal discharge after receiving strain H10, and died. Lethal FCV is rare, and may be a virulent systemic disease (VSD)—inducing strain. This is the initial report on VSD in Japan. It has been reported that symptoms of VSD were similar in vaccinated and nonvaccinated cats on experimental infection. However, no VSD-like symptoms developed, and the incidence of the disease varied depending on the presence or absence of vaccination, suggesting that there are two mechanisms of vaccine breakdown: one is associated with the vaccine immunity level, and the other is not. The characteristics of the VBS revealed were: (1) the duration of virus excretion was short when the originally carried antibody titer before virus challenge was high, (2) the excreted viral molecular species varied daily, not being limited to a specific species with time, and (3) the acquired physicochemical properties did not persist, and altered daily. FCV-VBS alters the molecular species and physicochemical properties daily due to the reduction of host immunity, which may lead to VSD.     

Pharmacokinetics and toxicity of zonisamide in cats

With the eventual goal of making zonisamide (ZNS), a relatively new antiepileptic drug, available for the treatment of epilepsy in cats, the pharmacokinetics after a single oral administration at 10mg/kg and the toxicity after 9-week daily administration of 20mg/kg/day of ZNS were studied in healthy cats. Pharmacokinetic parameters obtained with a single administration of ZNS at 10mg/day were as follows: Cmax=13.1microg/ml; Tmax=4.0h; T(1/2)=33.0h; areas under the curves (AUCs)=720.3microg/mlh (values represent the medians). The study with daily administrations revealed that the toxicity of ZNS was comparatively low in cats, suggesting that it may be an available drug for cats. However, half of the cats that were administered 20mg/kg/day daily showed adverse reactions such as anorexia, diarrhoea, vomiting, somnolence and locomotor ataxia.     

The effect of approach direction and pace on flight distance of beef breeding cows

The aim of this study is to determine the effect of approach direction and pace on flight distance of beef breeding cows. Five Japanese Black breeding cows were used as experimental cattle. Three approach directions were tested; that were the approach toward the head of the test cow from the front direction (FRONT), the approach toward to the foreleg of the test cow from the right or left (SIDE) and the approach toward to hind‐leg of the test cow diagonally from the right or left rear (BACK). Three levels of approach pace were also tested; that were the approach at a pace of 7 km/h, the approach at a pace of 4 km/h, and the approach at a pace of 1 km/h. The shortest distance between observer and cow's at the moment of the cow's withdrawal was defined as the flight distance. The flight distance in FRONT was significantly shorter than that in SIDE. The effect of the approach pace was significant. The faster an observer approached to cow, the greater the flight distance shown by the cow. The manner of approach to cattle markedly affects the flight distance. Therefore, the manner of approach to cattle must be standardized to measure flight distance for sharing results obtained by several researchers under various conditions.     

Lumenal localization in the endoplasmic reticulum of the C-terminal tail of an AE1 mutant responsible for hereditary spherocytosis in cattle

An R664X nonsense mutant AE1 is responsible for dominant hereditary spherocytosis in cattle and is degraded by the proteasomal endoplasmic reticulum-associated degradation. The present study demonstrated that R664X AE1 translated in vitro had the trypsin-sensitve site identical to that of the wild-type AE1. The P661S/R664X mutant containing a possible N-glycosylation site at Asn660 showed an increase in size by 3 kDa both in the cell-free translation system and in transfected HEK293 cells. Moreover, steady state levels of R664X and P661S/R664X in HEK293 cells were markedly increased in the presence of a proteasome inhibitior. These findings indicate that the truncated C-terminal region of R664X AE1 has lumenal localization in the endoplasmic reticulum and is not accessible to proteasomal machineries in the cytosol.     

Codend selectivity for jack mackerel and whitefin jack and unequal split parameter estimates observed in trouser trawl experiments

Codend selectivity for the jack mackerel Trachurus japonicus and the whitefin jack Kaiwarinus equula were evaluated based on data from trouser trawl experiments carried out in the East China Sea, using a test codend of 60 mm diamond mesh and a control codend made of minnow net with a square mesh of 9 mm bar length. Between-haul variations in parameters and the mean selection curves were tested with the catch data in the SELECT approach, and then the model of between-haul variation in the split parameter with the mean selection curve was chosen as the best fit using Akaike’s information criterion model selection. The 50% retention lengths and the selection ranges were 11.4 and 3.36 cm for jack mackerel and 8.83 and 0.93 cm for whitefin jack, respectively. The selection curve for whitefin jack was sharp, whereas that for jack mackerel was relatively wide. As the estimated split parameters indicated, about 80% of the whitefin jack entered the control codend, but 85 and 90% of the jack mackerel entered the control codend in the second and third hauls, respectively. The inequality in the split parameter is discussed from the viewpoint of the animal’s swimming behavior and water movement based on underwater video observations.     

Dual effects of reversed winter–spring temperatures on year‐to‐year variation in the recruitment of chub mackerel (Scomber japonicus)

To clarify the effects of temperature on the recruitment of chub mackerel (Scomber japonicus) in the North Pacific, we investigated the influence of winter surface temperature (WST) on spawners at the time of maturity around the spawning grounds and the influence of ambient spring temperature on larvae using estimated temperature (ET) obtained from particle tracking experiments. We found a significant positive correlation between ET approximately 10 days following hatching and the recruitment per spawning stock biomass (RPS) after 2000. The closer (more meandering) the Kuroshio Current (KC) axis was in relation to the spawning ground, the higher (lower) the spring surface temperature and the higher (lower) RPS was in the spawning ground. In contrast, WST inside KC near the maturity/spawning ground was significantly negatively correlated with RPS. A significant negative correlation between the temperatures in winter and spring was detected in the area after 2000, when the conditions of the Pacific decadal oscillation index and the stability of the Kuroshio Extension were synchronous, indicating that KC shifted northward during this time. The reversed temperature pattern was consistent with the winter–spring movement of KC axis in the offshore direction and was correlated with the winter–spring difference in the intensity of the Aleutian low. These results suggest that the annual variation in chub mackerel recruitment after 2000 was strongly affected by the combined effects of ambient temperature because of the reversal of conditions that occurred between winter and spring around the maturity/spawning ground, which was related to the KC path.     

Sodium alginate supplementation modulates gut microbiota,health parameters,growth performance and growth‐related gene expression in Malaysian Mahseer Tor tambroides

This study investigated the effects of sodium alginate supplementation on gut microbiota composition, health parameters, growth performances and growth‐related gene expression of Malaysian mahseer. Five test diets were formulated by supplementing 0%, 0.1%, 0.2%, 0.4% and 0.8% sodium alginate. Triplicate groups of juvenile Tor tambroides (2.19 ± 0.05 g) were stocked in 15 aquaria (20 individuals per aquarium) and fed at 3.0% body weight per day for 60 days. PCoA and UPGMA analysis showed that gut bacterial community were more convergence in higher sodium alginate‐supplemented diets. The percentage of Porphyromonadaceae, Bacteroides, Plesiomonas and Shewanella were substantially higher and Aeromonas, Entomoplasmatales and Prevotellaceae were drastically lower in higher sodium alginate (0.2%–0.8%) diets. Sodium alginate supplementation (≥0.2%) significantly improved the haematocrit value and respiratory burst activity of T. tambroides. Growth performances and feed utilization were significantly higher in 0.2%–0.4% sodium alginate‐supplemented diets. The increased growth rate of T. tambroides was governed by both hyperplastic and hypertrophic muscle growth. Real‐time PCR data demonstrated that most of the growth‐related genes were significantly upregulated in 0.2%–0.4% sodium alginate‐supplemented diet. Finally, it can be concluded that sodium alginate should be supplemented at 2 g/kg in practical fish feed formulation.     

Metagenomic analysis using 16S ribosomal RNA genes of a bacterial community in an urban stream,the Tama River,Tokyo

In an effort to determine genus- or species-level taxonomic profiles and diversity of bacterial consortia in the Tama River around urban Tokyo, next-generation sequencing technology targeting a 16S ribosomal RNA (rRNA) gene amplicon was employed. Metagenomic analysis performed by an Ion Personal Genome Machine after sequentially filtering samples through 5-, 0.8- and 0.2-μm filters yielded 1.48 Gb of 16S sequences (average 2.38 M reads/sample). The results indicated that half of the bacterial sequences belonged to Proteobacteria, followed by Bacteroidetes, Actinobacteria and Cyanobacteria. Flavobacterium (Bacteroidetes), possibly including a potential fish pathogen, was the most numerous genera in the Tama River metagenome, and accounted for?~?16% of assigned 16S reads, followed by Mycobacterium. Other dominant bacterial genera including Zoogloea, Sediminibacterium, Hyphomicrobium, Sphingopyxis, Thiothrix and Lysobacter, were thought to be associated with waste water and sludge. MiSeq metagenomic analysis revealed that environmental factors, particularly water temperature, influenced the bacterial composition throughout the year, with a strong negative correlation observed for Proteobacteria and a positive correlation for Bacteroidetes. In terms of bacterial genera, Flavobacterium was positively correlated with temperature, while Polaromonas, Pseudomonas and Bradyrhizobium were negatively correlated with this, suggesting dynamic change in the free-living bacterial population throughout the year and versatile adaptation strategies in relation to environmental factors.     

Quantifying growth in maricultured corals using photogrammetry

In coral mariculture, growth is one of the most common ways to track success. However, numerous methods of monitoring coral growth make comparative studies challenging. A literature review of 39 studies from 1982 to 2017 indicated that the most predominant non‐invasive methods used were linear and areal measurement and these were evaluated for their accuracy using nursery‐reared corals. The monthly change in linear and areal growth rates of six coral species (n = 215), Pocillopora acuta, Hydnophora rigida, Merulina ampliata, Podabacia crustacea, Echinopora lamellosa and Platygyra sinensis were measured via photogrammetry. We tested whether the planar area of coral colonies can be estimated using three geometric formulas of linear measurements. Based on the literature review, the six types of measurement methods were namely linear, volume, weight, area, count of polyps/branches and calcification, in decreasing order of application. Our results showed that the change in area calculated by geometric mean diameter (GMD) formulas provided the most accurate estimation among the three formulas and was strongly correlated with planar area (R² ≥ .60; p < .05) for all coral species, except E. lamellosa and Pl. sinensis. However, our findings suggest that it is not ideal to use geometric formulas to estimate the change in area. Instead, we posit that areal photogrammetry represents the simplest yet accurate non‐invasive method for rapid monitoring of extensive areas of corals in situ. Lastly, we discuss the recommendations and limitations for areal photogrammetry.