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.     

Intake of Heated Leaf Extract of Coriandrum sativum Contributes to Resistance to Oxidative Stress via Decreases in Heavy Metal Concentrations in the Kidney

Plant Foods for Human Nutrition - Coriandrum sativum (coriander) is an annual herb of the Apiaceae family and has been used as a traditional remedy. Here we examined whether heated leaf extract of...     

Internationally harmonized certified reference materials and proficiency testings for pesticide residue analysis

Matrix certified reference materials (CRMs) and proficiency testings (PTs) are effective for evaluation of the quality of analytical results. The National Metrology Institute of Japan has developed five kinds of CRMs and has provided eight PTs so far for the quantification of pesticide residues in foods. Target pesticides were sprayed on growing crops, and the harvests were used for the preparation of CRMs and PT samples. In most cases, multiple analytical methods based on isotope dilution mass spectrometry were used to ensure the reliability of certified values (for CRMs) and reference values (for PT samples). These activities were carried out with corresponding to the international standards such as ISO 17034 and ISO/IEC 17043. An overview of the development of CRMs and the implementation of PTs is described, with some examples.     

Prevalence of porcine reproductive and respiratory syndrome virus and porcine circovirus type 2 in piglets after weaning on a commercial pig farm in Japan

To investigate the transition in concentration of porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV2) and antibody for these viruses in serum, serum samples were collected from 29 pigs on weaning day and at 7, 14, 21, 28, 53, 84, and 120 days after weaning. The concentration of circulated PRRSV and PCV2 in serum was measured by real-time RT-PCR and real-time PCR, respectively. The specific antibody for PRRSV and PCV2 was measured using ELISA. PRRSV was not detected on 0 days post-weaning (dpw). The specific antibody for PRRSV began to increase as the concentration of PRRSV in serum increased, and the level of PRRSV then tended to decrease. PCV2 was detected in 12 of 28 pigs on 0 dpw. The concentration of PCV2 and the specific antibody for PCV2 showed a similar tendency to those of PRRSV. The correlation analysis suggests that a decline in the daily weight gain coincided with an increase in the PRRSV concentration. Pigs with a higher antibody titer against PRRSV or PCV2 on 0 dpw showed the lower level of PRRSV or PCV2, respectively.     

Classification and Antimicrobial Susceptibilities of Enterococcus Species Isolated from Apparently Healthy Food‐Producing Animals in Japan

Summary A total of 1024 enterococci were recovered from faeces of healthy animals from 178 cattle, 178 pig and 156 broiler farms. Enterococcus faecium was the predominant species recovered (35.8%), followed by E. faecalis (31.3%) and E. hirae (25.6%). Oxytetracycline resistance was most frequently found among E. faecalis (85.9%), E. faecium (58.8%) and E. hirae (48.1%). Resistance rates to almost all antimicrobials were higher in E. faecalis than E. faecium and E. hirae. Isolates from cattle were more susceptible to the antimicrobials studied than those from pigs and broilers. VanA‐ or VanB‐type vancomycin‐resistant enterococci have not been found since the ban of avoparcin use 5 years ago.     

Inclusion of Bacillus amyloliquefaciens strain TOA5001 in the diet of broilers suppresses the symptoms of coccidiosis by modulating intestinal microbiota

Coccidiosis is an intestinal parasitic infection and one of the most prevalent and economically damaging diseases of chickens. Furthermore, coccidia‐induced mucogenesis promotes secondary colonization by Clostridium perfringens, a major pathogen of chickens that causes necrotic enteritis. Our previous work found that supernatant of a culture of Bacillus amyloliquefaciens strain TOA5001 (BA) inhibited the growth of C. perfringens on Gifu anaerobic broth medium. Accordingly, we evaluated the effectiveness of dietary BA administration in inhibiting C. perfringens colonization of the intestine in broilers that were experimentally infected with coccidia. Ten healthy broilers from a BA‐supplemented (2 × 10⁵ colony‐forming units/g of feed) broiler group and 10 from a non‐treated group were challenged with Eimeria tenella and E. maxima (5000 oocysts of each species/chick) at 28 days old. At 36 days old, five chicks from each group were slaughtered, whereas the remaining five in each group were killed at 49 days old. Dietary BA administration into Eimeria‐challenged birds reduced coccidial symptoms such as intestinal lesions. It also modified the cecal microbiota through suppressing C. perfringens and E. coli colonization, and inducing domination of Faecalibacterium prausnitzii, the Lactobacillus group and unknown Lachnospiraceae genera by bacterial DNA‐based metagenome analyses. B. amyloliquefaciens TOA5001 supplementation suppressed the symptoms of coccidiosis by modulating cecal microbiota in Eimeria‐challenged broilers.     

Swimming angle and target strength of larval Japanese anchovy (<Emphasis Type="Italic">Engraulis japonicus</Emphasis>)

The swimming angle of larval Japanese anchovy (Engraulis japonicus) was measured in a tank, and target strength (TS) was calculated using a theoretical scattering model. The mean swimming angle was 12.8° (SD ±22.1). Increased speeds of flow led to increased mean swimming angles. The mean swimming angle at flow of 5 cm s⁻¹ was higher than at other speeds. TS values were estimated using a distorted-wave Born approximation model for two cases. Average values were 1–3 cm s⁻¹ (11.5° ± 22.1) and 5 cm s⁻¹ (16.6° ± 21.7) for cases 1 and 2, respectively. For case 1, TS ranged from −92.0 to −74.7 dB with a mean of −79.4 dB at 120 kHz. For case 2, TS ranged from −92.2 to −75.2 dB with a mean of −79.9 dB. The mean TS in case 2 was lower than that in case 1, with the maximum difference being 1.0 dB at 120 kHz (standard length 22.0 mm). However, there were no significant differences between the regression lines of cases 1 and 2. Thus, changes in flow speed altered the swimming angle of larval Japanese anchovy, but had little influence on TS.     

DNA markers linked to Pga1, an adzuki bean gene that confers resistance to Cadophora gregata race 1

Brown stem rot (BSR) caused by Cadophora gregata f. sp. adzukicola (syn. Phialophora gregata) is a serious soilborne disease of adzuki bean (Vigna angularis) in Japan. Cultivation of resistant cultivars is the most effective disease control method, therefore the selection of resistant lines is a priority for breeders. BSR-resistant adzuki bean lines have been screened in pathogen-infected fields. However, field selection using the pathogen and artificial inoculation methods is time-consuming and labor-intensive. In the present study, we used 105 F₃ lines derived from a cross between a BSR-resistant cultivar ‘Syumari’ and a susceptible cultivar ‘Buchishoryukei-1’ for BSR inoculation tests. Amplified fragment-length polymorphism (AFLP) analyses with 1024 primer sets revealed that six fragments were polymorphic between resistance and susceptible bulked groups. Five DNA markers (Pg77, Pg118, Pg138, Pg139 and Pg126) were developed from the nucleotide sequences of polymorphic AFLP markers and their flanking regions. Pg118, which was derived from E-ACT/M-ACT-118, was tightly linked to the resistance gene Pga1 and was converted into a codominant marker for its easier use in marker-assisted selection for adzuki bean BSR resistance. Finally, the applicability of the developed markers for BSR resistance was tested on 32 adzuki bean accessions or cultivars.