The nucleotide sequence of the genes, fanE and fanF of Escherichia coli K99 fimbriae.

K99 fimbriae of enterotoxigenic Escherichia coli consist of eight different subunits. A major subunit called fimbrillin forms fimbrial structure and a minor subunit called adhesin localizes at the tip of fimbriae and recognizes host receptor ganglioside. Within this eight gene cluster, fanE and fanF have not yet been sequenced. In this study, fanE and fanF genes were sequenced by analyzing several DNA fragments produced by endonuclease or exonuclease digestion. The fanE gene encoded 227 amino acids containing 20 amino acids of signal peptide starting from GTG (valine) and showed a homology to fanA-fanB. The fanF gene encoded 271 amino acids containing 20 amino acids of signal peptide starting from ATG (methionine) and showed homologies to the fanD gene, fimbrillin gene of F41, adhesin gene of P fimbriae (papG) and adhesin gene of Type 1 fimbriae (fimH). E and F subunits had fifteen and fourteen hydrophobic domains, respectively, which periodically appeared possibly forming a hydrophobic region.     

Preparation of latex sensitized with rabbit IgG antibody for slide reversed passive agglutination

A method is described for preparing latex particles sensitized with IgG antibody (IgG-sensitized latex) applicable to the slide reversed passive agglutination (RPLA) test. Soap-free latex (latex) was sensitized with IgG which had been isolated from rabbit anti-bovine lactoferrin serum using protein A Sepharose CL 4B. Unadsorbed protein-binding sites on the surface of latex were blocked with bovine serum albumin (BSA). IgG-sensitized latex that gave better agglutination in RPLA could be selectively obtained by centrifugation at 19 900g for 15 min in 0.01 mol/L glycine buffer (pH 7.3; specific gravity 1.042) containing 3% NaCl, 5% saccharose and 2% choline chloride. By dispersing this IgG-sensitized latex in 0.01 mol/L glycine buffer (pH 7.3) containing 1–2% BSA, a uniformly suspended, highly reactive, readily agglutinable preparation was obtained.     

Isolation, characterization, and quantitative analysis of C-reactive protein from horses

C-reactive protein (CRP) was isolated from equine serum by use of calcium-dependent affinity chromatography conjugated pneumococcal C-polysaccharide, anion exchange chromatography, and gel filtration. It was identified as genuine CRP by its immunochemical cross-reactivity with anti-human CRP, its homology with human CRP in amino acid composition, and its pentameric structure as revealed by electron microscopy. Purified equine CRP had a molecular weight of approximately 118,000 and was composed of 5 identical, nonglycosylated and noncovalently associated subunits with molecular weight of approximately 23,000 each. Equine CRP migrated in the region between beta- and gamma-globulin by results of immunoelectrophoresis, and its isoelectric point was about 7.0. In horses, increased CRP concentration was associated with clinical pneumonitis, enteritis, and arthritis, compared with values obtained in clinically normal horses by use of single radial immunodiffusion method. After IM administration of turpentine oil or castration, serum CRP concentration increased to 6 times higher than baseline values. Results indicate that CRP may be an acute-phase reactant protein in horses.     

Concentrations of C-reactive Protein in Normal Monkeys (Macaca irus) and in Monkeys Inoculated with Bordetella bronchiseptica R-5 and Measles Virus

The concentrations of C-reactive protein (CRP) in serum from normal crab-eating monkeys (Macaca irus) were measured by means of a monkey-specific turbidimetric immunoassay (TIA), and the changes in the serum CRP concentrations in crab-eating monkeys inoculated with Bordetella bronchiseptica R-5 and measles virus (Ichinose or NK 3 strain) were also examined. The CRP concentrations in sera from 54 normal crab-eating monkeys ranged from 0 to 8.3 g/ml (mean 2.2±1.9). No significant difference was found in the CRP concentrations between males and females (p>0.05). The concentrations of CRP in the sera from four crab-eating monkeys inoculated intrabronchially with 10⁹ live B. bronchiseptica increased gradually to a peak at 2 days after inoculation. The peak concentrations of CRP were from 102.4 to 313.2 g/ml, 54–96 times the preinoculative values of 1.9–5.6 g/ml. When the same four crab-eating monkeys were inoculated intrabronchially with measles virus 34 days after inoculation of B. bronchiseptica, the serum CRP concentrations did not increase. Monitoring of CRP is useful for assessing monkeys with acute B. bronchiseptica infection and will probably be of value in the diagnosis of other bacterial infections.     

Improvement in the feeding activity, early growth and survival of Pacific bluefin tuna Thunnus orientalis larvae fed a casein peptide-based microdiet supplemented with inosine monophosphate

Two feeding experiments were conducted to determine the effect of a 5′-inosine monophosphate (IMP)-supplemented casein peptide-based microdiet (MD) on the feeding activity, growth and survival of Pacific bluefin tuna (PBT) larvae. PBT larvae [total length: 14.0 mm (Exp. 1) or 13.3 mm (Exp. 2)] were fed a casein peptide-based MD supplemented with or without 3% IMP for 12 days. As a negative control, a limited number of yolk-sac larvae of spangled emperor Lethrinus nebulosus were fed only for the first 7 days and starved for the next 5 days. A sufficient amount of yolk-sac larvae of spangled emperor was fed in the Exp. 2. All fish in the control group had died by 9 or 10 days after the experiment. Extensive mortality was observed in fish fed the MD. The MD was only found in the stomach contents of PBT fed the MD + IMP. Of the fish fed an IMP-supplemented diet, 50% survived for 12 days after the initiation of both feeding experiments. In addition, significantly higher growth than that observed during the first 7 days was seen in the PBT in the IMP treatment after 12 days of the feeding experiment. These results suggest that the supplementation of casein peptide-based MD with IMP could improve the feeding activity, survival and growth performance of PBT.     

Canine C-reactive protein (CRP) does not share common antigenicity with human CRP

Differences in antigenicity between human and canine C-reactive proteins were investigated by Western blotting analysis. It was confirmed that several commercial anti-human CRP sera reacted with canine CRP. However, 34 anti-canine CRP sera prepared by immunization of rabbits and goats with canine CRP all reacted with canine CRP but not with human CRP in either immunoelectrophoresis or Western blotting.Immunization with human CRP produced a cross-reacting antibody that reacted with canine CRP. Conversely, immunization with canine CRP did not produce a cross-reacting antibody that reacted with human CRP. These findings may be interpreted as showing that, while canine and human CRPs do not share common antigenicity, they do contain structurally similar antigenic determinants.Abbreviations CRP C-reactive protein - PBS phosphate-buffered saline - SDS-PAGE sodium dodecyl sulphate-polyacrylamide gel electrophoresis     

Projections of nutrient supply to a wakame (Undaria pinnatifida) seaweed farm on the Sanriku Coast of Japan

Wakame (Undaria pinnatifida) aquaculture is one of the major fishery industries on the Sanriku Coast of Japan. Low concentrations of nutrients in the autumn, when the germinated wakame sporophytes are installed at aquaculture facilities, induce defective growth, resulting in withering. To determine the characteristics of the nutrient variations in this region, we conducted hydrographic measurements off the coast of Iwate Prefecture. The measured results showed that in the autumn, vertical mixing pumped nutrients from the deeper layers to the mixed layer, where nutrients had been depleted during the summer. Based on these results, we developed a one‐dimensional model incorporating vertical mixing by atmospheric cooling to project the vertical distribution of the nutrient concentrations. By conducting ensemble calculations, we created a probability projection calendar for the nutrient supply up to 50 days ahead. This information was provided to wakame farmers before they installed the germinated sporophytes. The projected probability calculated from the initial conditions on 11 October 2016 was 0% until 25 October and gradually increased to 85% on 30 November. The nitrate concentration detected by an ultraviolet nitrate sensor moored at a wakame farm revealed that the nutrient concentrations reached a critical threshold on 18 November. The projected probability on that date was 75%; therefore, the projection was successful. The observed temporal evolution of the temperature, salinity, and nutrient concentrations was well reproduced by our model. The information obtained using this model is expected to reduce the risk of wakame withering and improve the efficiency of wakame production.     

Population and distribution of sclerotia of Rhizoctonia solani kühn in sugar beet field soil

The population and distribution of sclerotia of Rhizoctonia solani Kühn in two sugar beet field soils was determined at harvest by a sieving-flotation method. In rhizosphere soil (RS) and non-rhizosphere soil (NRS) from the most heavily infected roots of sugar beets, 1.43–2.5 and 0.83–1.0 sclerotia g^?1 dry soil were detected, respectively. In the soil around healthy sugar beet, these values were 0.04–0.12 and 0.03–0.04 sclerotia g^?1 dry soil. More sclerotia were always obtained from RS than from NRS. More than 80% of the sclerotia were in the upper 10 cm of soil and within 10 cm of diseased roots. Therefore, there is a non-uniform distribution of sclerotia of R. solani in soil.The sclerotial population in soil increased significantly with disease severity and a good correlation was obtained between the number of sclerotia and the disease severity on infected plants. Most of the sclerotia collected from the field soil ranged in size from 0.5 to 2.0 mm diameter.Viability of sclerotia increased as severity of crown rot increased and as the size of the sclerotia increased. Conversely, there was a progressive decrease in sclerotial germination with increasing depth in soil and increasing distance from the infected root.