Development and characterization of monoclonal antibodies against canine trypsin

Canine cationic trypsin was purified by salting-out, gel filtration and affinity chromatography. Purity was confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The molecular weight was ca. 28 kDa by SDS-PAGE.

Thirty hybridomas were obtained which produced mAb to canine cationic trypsin by the cell fusion technique. Twenty-two of these recognized cationic trypsin only, while eight hybridomas recognized both cationic and anionic trypsin. Several of the anti-canine cationic trypsin mAb were purified by salting-out and DEAE ion-change chromatography using ascites fluid of immunized BALB/c mice. The mAb proved to have very high specificity to canine cationic trypsin as shown by immunoblotting and it may be possible to use them to develop clinical assays.     

Suppression of lipopolysaccharide and galactosamine-induced hepatic inflammation by red grape pomace

Grape pomace is generated in the production process of wine and grape juices and is an industrial waste. This study investigated whether an intake of grape pomace was able to suppress chronic inflammation induced by lipopolysaccharide (LPS) and galactosamine (GalN) in vivo. When Sprague-Dawley rats were orally given methanolic extracts from red and white grape pomace, the extracts inhibited the LPS/GalN-evoked activation of nuclear factor-κB (NF-κB) dose-dependently, and red grape pomace exerted a stronger effect than white grape one. Next, rats were fed an AIN93 M-based diet containing 5% red grape pomace for 7 days, followed by the intraperitoneal injection of LPS and GalN. The intake of the red grape pomace-supplemented diet was found to suppress the LPS/GalN-induced activation of NF-κB and expression of inducible nitric oxide synthase and cyclooxygenase-2 proteins. These results suggest that red grape pomace may contain an abundance of effective compound(s) for anti-inflammatory action.     

Annual changes in macrobenthos abundance in Funka Bay,Japan

The temporal variation in macrobenthos abundance was studied in Funka Bay in the periods 2001–2004 and 2007–2010. Dissolved oxygen just above the sea floor of the basin was lower and ignition loss at the level of the sea floor was higher in 2007–2010 than in 2001–2004. Macrobenthic species were categorized into three groups: long-lived species, sedental polychaeta Notomastus latericeus, and short-lived species. Ignition loss was the most influential factor affecting macrobenthos abundance as this variable restricts the recruitment of macrobenthos. The abundance of both long-lived species and N. latericeus was lower between 2007 and 2010 than between 2001 and 2004. Supplementation of oxygen from outside the bay by the Tsugaru Warm Current may be an important factor impacting the increase in the abundance of long-lived species. The abundance of N. latericeus barely increased between 2007 and 2010, possibly because dissolved organic matter, which is their food item, may have become limited due to a narrow chink between sediments with eutrophication. The abundances of short-lived species did not differ between the periods 2001–2004 and 2007–2010 due to the organisms having a low tolerance to hypoxia. The current conditions in Funka Bay are on the verge of turning the bay into an azoic zone.     

Effect of density of trees on drag exerted on trees in river channels

Recently, riparian forests have attracted attention as they are effective for ecological preservation and landscape enhancement. Uses of such forests in flood prevention, sediment control, and erosion control works have been actively promoted. This study aims to clarify the effect of density of riparian trees on drag exerted on trees along river channels under the regime of bed load transport. Hydraulic model experiments were performed using a straight channel, and the methods to calculate the drag coefficient necessary for deriving drag exerted on trees were studied. Previous studies have stated that the drag coefficient of a cylinder is fairly constant when Reynolds number,R _e, is between 10³ and 10⁵ (e.g., Schlichting, 1979). This study clarified, however, that in cases of relatively dense arrangements of model trees (cylinders), the drag coefficient varies greatly with the density of the trees. As such, correlation between the drag coefficient and Reynolds number was found to be slight. Test results indicated that the drag coefficient of trees,C _d, correlates strongly with the coefficient of velocity,U′/U _* ′, friction factor of the channel bed,f′, and roughness concentration of trees × flow depth,aH′ or ratio of the area occupied by trees,λ. Furthermore, the drag coefficient,C _d, was also found to be slightly correlated with the gradient of the channel bed,I. Equations using either of the following parameters were obtained for deriving the drag coefficient,C _d, based on the experiment results:aH′; λ; U′/U _* ′; and Froude numberF _r. These equations allow fairly accurate calculation of drag exerted on trees.     

Role of platelet activating factor in development of thrombocytopenia and neutropenia in dogs with endotoxemia

OBJECTIVE: To determine the role of platelet activating factor (PAF) in lipopolysaccharide (LPS)-induced thrombocytopenia and neutropenia in dogs. ANIMALS: 42 dogs. PROCEDURES: Blood samples were obtained from dogs given LPS (40 microg/kg of body weight; n = 16), PAF (1 microg/kg; 6), PAF (5 microg/kg/h for 90 minutes; 4), or physiologic saline (0.9% NaCl) solution (0.1 ml/kg/h for 90 minutes; 3) IV to monitor changes in blood cell counts, using automated counters and blood smears stained with Giemsa. Blood samples were also obtained from dogs given LPS (40 microg/kg) that had (n = 5) or had not (6) been treated beforehand with TCV-309, a potent PAF antagonist. Concentration of PAF in blood was determined by use of 125I-radioimmunoassay in dogs given LPS at 1 mg/kg (n = 3) and 40 microg/kg (9). RESULTS: Thrombocytopenia and neutropenia were found in all dogs except those given saline solution. The LPS-induced thrombocytopenia was significantly suppressed by prior treatment with TCV-309. The PAF concentrations increased markedly 1 hour after injection of 1 mg/kg of LPS and increased slightly but significantly 10 minutes after injection of 40 microg/kg of LPS. CONCLUSION AND CLINICAL RELEVANCE: PAF plays an important role in the development of LPS-induced thrombocytopenia and neutropenia in dogs. Control of PAF production, PAF-induced effects, or both may be important in the treatment of dogs with gram-negative bacterial infections and associated thrombocytopenia and neutropenia.     

The distribution and early growth of juvenile Pacific bluefin tuna Thunnus orientalis around Sado Island in the eastern Sea of Japan

Fisheries Science - The distribution and early growth of juvenile Pacific bluefin tuna (PBF) Thunnus orientalis were studied based on trawl surveys conducted around Sado Island, eastern Sea of...     

A functional link between RuBisCO-like protein of Bacillus and photosynthetic RuBisCO

The genomes of several nonphotosynthetic bacteria, such as Bacillus subtilis, and some Archaea include genes for proteins with sequence homology to the large subunit of ribulose bisphosphate carboxylase/oxygenase (RuBisCO). We found that such a RuBisCO-like protein (RLP) from B. subtilis catalyzed the 2,3-diketo-5-methylthiopentyl-1-phosphate enolase reaction in the methionine salvage pathway. A growth-defective mutant, in which the gene for this RLP had been disrupted, was rescued by the gene for RuBisCOfrom the photosynthetic bacterium Rhodospirillum rubrum. Thus, the photosynthetic RuBisCO from R. rubrum retains the ability to function in the methionine salvage pathway in B. subtilis.     

Dietary Bacillus subtilis C-3102 Supplementation Enhances the Exclusion of Salmonella enterica from Chickens

Among the reported probiotic Bacillus strains, B. subtilis C-3102 has the unique potential to improve feed uptake under stress conditions in the broilers, piglets, and cows. In this study, we sought to evaluate the protective effect of feed additive probiotic Bacillus subtilis C-3102 against Salmonella enterica infection of specific pathogen-free (SPF) chicks in floor pens in two experiments. In the experiment-1, the chicks in the control group (n=32) were fed a basal diet and those in the C-3102 group (n=32) were fed a basal diet supplemented with 1×10⁶ CFU/g of feed for 28 days. On day 7 post-challenge with S. enterica, there was no significant change in the body weight between both the groups throughout the test period, whereas detection rates of S. enterica in the C-3102 group were significantly lower in the cecum and liver on days 21 and 14 post-challenge, respectively. In the experiment-2, minimum dosage of C-3102 cells required to protect Salmonella infection was evaluated using 3 dosages. Chicks were divided into four groups, fed with different dosages of C-3102 (1×10⁶, 5×10⁵, 3×10⁵, and 0 CFU/g of feed), and challenged with S. enterica (2.8×10⁸ CFU/chicken). S. enterica infection was completed within 7 days post- challenge and was almost excluded from the liver and spleen on day 21 post- challenge in the control group. Average values showed a trend for higher infection rates in the control group >3×10⁵>5×10⁵>1×10⁶ CFU/g on days 14 and 21 post-challenge. These results suggest that B. subtilis C-3102 supplementation has the potential to reduce S. enterica infection rates and/or to accelerate the exclusion of S. enterica from the chicks.     

Effect of Bacillus subtilis C-3102 supplementation in milk replacer on growth and rumen microbiota in preweaned calves

We aimed to assess the effect of feeding Bacillus subtilis C-3102 on the growth and rumen microbiota in the preweaned calves. Twelve newborn Japanese Black calves were randomly allocated to either the control (n = 6) or the treatment (n = 6) groups in the present study. Calves in the treatment group were offered B. subtilis C-3102 supplemented milk replacer throughout the preweaning period. Rumen fermentation during the first 21 days of life seemed to be slightly suppressed by feeding B. subtilis C-3102. This fermentation shift was probably attributed to the lower abundance of the core members of rumen microbiota until 21 days of age in the calves fed B. subtilis C-3102. However, feeding B. subtilis C-3102 did not influence the abundance of the core members of rumen microbiota at 90 days of age. Distribution of Sharpea spp. and Megasphaera spp., which potentially contribute to low methane production and are regarded as beneficial rumen bacteria, was higher in the rumen of calves fed B. subtilis C-3102 at 90 days of age. These results suggest that B. subtilis C-3102 supplementation in milk replacer could potentially contribute to the improvement of feed efficiency after weaning via the establishment of beneficial rumen bacteria.