Accumulation of gamma-oryzanol in teleost

Japanese people consume crop bran, which contains relatively high amounts of gamma-oryzanol (ORZ), as foodstuffs and food materials. We have recently confirmed that ORZ inhibits NF-κB activation, activates peroxisome proliferator-activated receptor gamma (PPARγ), and increases plasma adiponectin levels. ORZ is therefore expected to improve lipid and carbohydrate metabolisms and various inflammatory diseases, i.e., metabolic syndrome including type 2 diabetes. ORZ administration also allowed fish, such as rainbow trout, yellowtail, and red sea bream, to accumulate protein through enhancement of lipid and carbohydrate metabolisms. In the present study, we have investigated ORZ accumulation levels in mouse and rainbow trout administered ORZ-containing feed. Although mouse muscle and liver hardly contained ORZ, muscle tissues of every fish species accumulated higher amounts of ORZ. These findings suggest that rainbow trout, red sea bream, and yellowtail accumulate ORZ in muscle.     

Production and characterization of monoclonal antibodies to leukocytes and serum immunoglobulin in Japanese flounder <Emphasis Type="Italic">Paralichthys olivaceus</Emphasis>

Two monoclonal antibodies (MAbs: JFW1 and JFW10) were produced against peripheral blood leukocytes (PBL) in Japanese flounder. Additionally, MAbs against flounder immunoglobulin (Ig; JFW20 and JFW21) were generated for the surface marker of Ig⁺ leukocytes using purified serum Ig as an antigen. MAb JFW1 recognized the surface marker of granulocytes and monocytes and MAb JFW10 specifically bound to the surface antigen of thrombocytes. Flow cytometric analysis of PBL incubated with JFW1, JFW10, JFW20 and JFW21 revealed that 2.5–7.4, 23.7–50.1, 25.2–26.1 and 5.2–8.3% of all leukocytes were positive for these markers. Analysis of head kidney leukocytes (HKL) showed that JFW1, JFW10, JFW20 and JFW21 bound to 30.5–36.3, 1.9–2.8, 6.4–8.3 and 1.9–3.0% of all leukocytes, respectively. Western blot analysis after SDS-PAGE showed that JFW10 recognizes a protein of 115 kDa from lysed PBL. JFW20 recognized the 70 and 74 kDa proteins of the heavy chain of Ig from serum. No band was observed for either JFW1 or JFW21. These antibodies will be useful for the identification and isolation of Japanese flounder leukocyte subpopulations and will facilitate immunological studies of flounder.     

Oral administration of Lactobacillus plantarum Lq80 and Megasphaera elsdenii iNP‐001 induces efficient recovery from mucosal atrophy in the small and the large intestines of weaning piglets

Weaning causes atrophy of intestinal mucosa and a drop of IgA protection in piglets which increases vulnerability to pathogenic infections. Probiotic lactobacilli may support recovery from such weaning stresses. Butyrate‐produce bacteria may support the growth of colonic mucosa. Megasphaera elsdenii, a lactate‐utilizing butyrate producer, may help butyrate production particularly when combined with lactobacilli. Weaned piglets (Experiment 1: 20 days old, Experiment 2: 28 days old) were orally dosed once a day with either (L) 10¹⁰ (cell/dose) L. plantarum Lq80, or (LM) 10¹⁰ (cell/dose) Lq80 with 10⁹ (cell/dose) M. elsdenii iNP‐001. Lq80 was contained in capsules resistant to gastric digestion. M. elsdenii was contained in capsules resistant to gastric and intestinal digestion. An untreated control (C) was also prepared. After 2 weeks of administration, L. plantarum enhanced the recovery from the villous atrophy in both experiments. The rectal and colonic IgA tended to be higher in L and LM than in C in Experiment 1. Colonic butyrate was higher in LM than in the others in Experiment 1. The thickness of the colonic mucosa was greater in LM than in the others in Experiment 1. In early weaned piglets, the effects of L. plantarum and M. elsdenii were clear.     

Commensal bacteria coated by secretory immunoglobulin A and immunoglobulin G in the gastrointestinal tract of pigs and calves

A large amount of secretory immunoglobulin A (S‐IgA) is secreted in the alimentary tract of mammals. It has been reported that S‐IgA coats a portion of commensal intestinal bacteria in human and mouse. However, S‐IgA‐coated bacteria have not been studied in pigs and calves. In this study, we evaluated the distribution of S‐IgA‐coated commensal intestinal bacteria in each portion of the gastrointestinal tracts of pigs and calves. Immunoglobulin G (IgG)‐coated bacteria were also analyzed because a considerable amount of IgG is secreted in the gastrointestinal tracts of pigs, and in particular, calves. S‐IgA‐ or IgG‐coated bacteria were detected in all the segments of the gastrointestinal tracts of pigs and calves. The proportion of S‐IgA‐coated bacteria to total bacteria (i.e. S‐IgA coating ratio) varied in the segments of the gastrointestinal tract in pigs, whereas those of calves were nearly the same throughout the gastrointestinal tract. The S‐IgA and IgG coating ratios were higher in pigs than in calves for all segments of the gastrointestinal tract.     

Effects of replacing fish oil with canola oil on the growth performance,fatty acid composition and metabolic enzyme activity of juvenile yellowtail Seriola quinqueradiata (Temminck & Schlegel, 1845)

In this study, fish oil (FO) was replaced with canola oil (CO) in juvenile yellowtail diets to establish the optimal replacement levels that ensure adequate feed quality for aquaculture. Juvenile yellowtails (initial body weight: 104.7 g) were fed one of four diets for 10 weeks: 100% FO (FO group) or FO replacement with 25%, 50% or 100% CO (CO25, CO50 and CO100 groups respectively). Body weight, specific growth rate and feed efficiency were significantly affected by the replacement of FO with CO. The final body weight of the CO100 group was significantly lower than that of the FO (control) and CO50 groups. Furthermore, the CO50 group showed significantly higher feed efficiency than the FO group during the first 4 weeks (long photoperiod and high water temperature conditions). Increasing dietary CO increased serum glucose and triglyceride concentrations in a dose‐dependent manner. Regarding the proximate composition of fillet and liver, only crude protein in fillet was affected by the replacement of FO with CO. Fatty acid compositions of fillet and liver were correlated to the amount of CO in each diet. The activities of the metabolic enzymes phosphofructokinase and alanine aminotransferase were lowest in the CO25 and CO50 groups respectively. Thus, FO replacement with CO affected growth performance, serum components, fatty acid composition and the activity of metabolic enzymes. Overall, results obtained in the present study suggest that partial replacement (50%) of FO with CO is effective and might increase juvenile yellowtail growth under long photoperiod and/or high water temperature conditions.     

Development of green onion and cabbage certified reference materials for quantification of organophosphorus and pyrethroid pesticides

Green onion and cabbage certified reference materials for the analysis of pesticide residues were issued by the National Metrology Institute of Japan, part of the National Institute of Advanced Industrial Science and Technology. Green onion and cabbage samples were grown so as to contain several kinds of organophosphorus and pyrethroid pesticides, and those were collected from a field in the Kochi Prefecture in Japan. The certification was carried out by using multiple analytical methods to ensure the reliability of analytical results; the values of target pesticides (diazinon, fenitrothion, cypermethrin, etofenprox, and permethrin for green onion and chlorpyrifos, fenitrothion, and permethrin for cabbage) were obtained by isotope dilution mass spectrometry. Certified values of target pesticides were 0.96-13.9 and 2.41-6.9 mg/kg for green onion and cabbage, respectively. These are the first green onion and cabbage powder certified reference materials in which organophosphorus and pyrethroid pesticides are determined.     

In vitro activity of 24 antimicrobial agents against Staphylococcus and Streptococcus isolated from diseased animals in Japan

A total of 88 Staphylococcus and 61 Streptococcus isolates from diseased animals throughout Japan were examined in 2000 for the minimum inhibitory concentrations of 24 different antimicrobials by the agar dilution method standardized by the Japanese Society of Chemotherapy. The resistance rates to aminobenzylpenicillin (36.4%) and benzylpenicillin (35.2%) were high in Staphylococcus isolates, and those to oxytetracycline (45.9%) and kanamycin (21.3%) were high in Streptococcus isolates. Two isolates resistant to oxacillin harbored the mecA gene. One was Staphylococcus epidermidis derived from a pig with arthritis, and the other Staphylococcus cohnii from a head of cattle with mastitis.     

Gene expression profiles of CD4/CD8 double‐positive T cells in porcine peripheral blood

A characteristic subset of T cells, known as double positive T cells (DPTC) and expressing both cluster of differentiation 4 (CD4) and CD8, is observed in porcine peripheral blood. Previous studies suggested that DPTC might be memory cells. However, detailed phenotypes and functions of DPTC are yet to be fully elucidated and thus, the relatedness of DPTC with memory phenotypes remains unclear. In this study, DPTC gene expression profiles in peripheral blood were analyzed by DNA microarray in Experiment 1 and compared with those of CD4 single positive T cells (4SPTC) and CD8 single positive T cells (8SPTC). Expressions of IFNG, CCL5, NCK2, CCR2 and ITGB1 were higher than that of 4SPTC and 8SPTC. In contrast, expressions of CCR7 and SELL were lower than that of 4SPTC and 8SPTC. These results suggested that DPTC were either effector T cells or effector memory T cells (T_EM). Next, to determine whether DPTC were effector T cells or T_EM, differences in the response of DPTC and 8SPTC against immunized/primed antigens were compared (Experiment 2). While DPTC showed quick elevation of IL2 and CD25 gene expressions against in vitro stimulation of primed/immunized antigens, 8SPTC did not. These results suggest that at least some DPTC likely belong to T_EM.     

Shiga toxin 2eB‐transgenic lettuce vaccine is effective in protecting weaned piglets from edema disease caused by Shiga toxin‐producing Escherichia coli infection

Porcine edema disease (ED) is a toxemia that is caused by enteric infection with Shiga toxin 2e (Stx2e)‐producing Escherichia coli (STEC) and is associated with high mortality. Since ED occurs most frequently during the weaning period, preweaning vaccination of newborn piglets is required. We developed stx2eB‐transgenic lettuce as an oral vaccine candidate against ED and examined its protective efficacy using a piglet STEC infection model. Two serially developed Stx2eB‐lettuce strains, 2BN containing ingredient Stx2eB constituting a concentration level of 0.53 mg Stx2eB/g of powdered lettuce dry weight (DW) and 2BH containing ingredient Stx2eB constituting a concentration level of 2.3 mg of Stx2eB/g of powdered lettuce DW, were evaluated in three sequential experiments. Taken the results together, oral administration of Stx2eB‐lettuce vaccine was suggested to relieve the pathogenic symptoms of ED in piglets challenged with virulent STEC strain. Our data suggested that Stx2eB‐lettuce is a promising first oral vaccine candidate against ED.     

Antimicrobial susceptibility of Mannheimia haemolytica isolates from cattle in Japan from 2001 to 2002

A total of 27 clinical isolates of Mannheimia haemolytica from cattle in Japan from 2001 to 2002 were examined for antimicrobial susceptibility to 25 antimicrobial agents. The minimum inhibitory concentrations of 25 different antimicrobials were determined by an agar dilution method according to the guidelines of the National Committee for Clinical Laboratory Standards. Of the 27 isolates, seven isolates (26.9%) were resistant to at least one of the 25 drugs and resistance rates ranged from 3.7 to 18.5%. Resistance rates to dihydrostreptomycin (18.5%), oxytetracycline (11.1%), and doxycycline (11.1%) were relatively high and those to the remaining drugs were less than 10%.