Biocontrol science

Publisher: Nihon Bōkin Bōbai Gakkai


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    ABSTRACT: A commercial aflatoxin detection ELISA kit, "RIDASCREEN FAST Aflatoxin", was validated with corn samples naturally contaminated with aflatoxin and non-contaminated reference corn samples according to the Japanese Notification Method ShokuAnHatsu 0816-7. The trueness, intra-laboratory repeatability, intermediate precision, limit of detection and limit of quantitaion were found to be 91%, 10%, 6.4%, 0.6 ug/kg and 2 ug/kg, respectively, and the performance of the kit was recognized as complying with all criteria in the Supplement Table of the Notification. These data suggest that this kit is useful as a simplified device to screen out negative corn samples contaminated with less than 4 ug/kg.
    Biocontrol science 08/2013; 19(1):39-43.
  • Biocontrol science 01/2013;
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    ABSTRACT: The proposed approach to validation of steam sterilization in autoclaves follows the basic life cycle concepts applicable to all validation programs. Understand the function of sterilization process, develop and understand the cycles to carry out the process, and define a suitable test or series of tests to confirm that the function of the process is suitably ensured by the structure provided. Sterilization of product and components and parts that come in direct contact with sterilized product is the most critical of pharmaceutical processes. Consequently, this process requires a most rigorous and detailed approach to validation. An understanding of the process requires a basic understanding of microbial death, the parameters that facilitate that death, the accepted definition of sterility, and the relationship between the definition and sterilization parameters. Autoclaves and support systems need to be designed, installed, and qualified in a manner that ensures their continued reliability. Lastly, the test program must be complete and definitive. In this paper, in addition to validation study, documentation of IQ, OQ and PQ concretely were described.
    Biocontrol science 06/2012; 17(2):57-67.
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    ABSTRACT: We analyzed the production of neutral lipids by the marine hydrocarbonoclastic bacteria Marinobacter sp. strain PAD-2 using hexadecane or succinate as the sole carbon source. Results showed that strain PAD-2 was able to grow and reduce the surface tension to 33±1.5 mN m(-1) and 58±1.5 mN m(-1) when n-hexadecane or succinate was used as the sole carbon source, respectively. The lipophilic compounds produced by Marinobacter sp. strain PAD-2 were extracted, and then crude lipophilic compounds, expected to be wax ester-like lipids, were analyzed by thin layer chromatography (TLC) . Furthermore, the lipophilic compound demonstrating surface activity was purified and subjected to gas chromatography/mass spectrometry (GC/MS) analysis. Although these did not give definite structural information due to the weak molecular ion peak (M(+)) , one component Ma-1 showed almost the same mass spectrum as that of component Fa-2, which represented a biosurfactant derived from Dietzia maris reported previously. Cell hydrophobicity was measured by a test of bacterial adhesion to hydrocarbons. A higher hydrophobic cell surface was observed in strain PAD-2. Extracellular wax ester-like compounds seem to be one type of the surface active compounds when bacteria grow on hexadecane or succinate as the sole carbon source.
    Biocontrol science 06/2012; 17(2):69-75.
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    ABSTRACT: In May 2011, strain HYNE-20 (=JCM 17837) was isolated from a sample of hot spring water from a foot spa in Niigata, Japan, by a plating method using glycine vancomycin polymyxin B cycloheximide α-ketoglutarate (GVPCα) medium at 36°C for 7 d. The 16S rDNA sequences (1,469bp) of this strain (accession number: AB638719) had high (99.7%) similarity to Legionella rubrilucens, and we identified that this strain was indeed Legionella rubrilucens. When this strain was cultured on buffered charcoal yeast extract α-ketoglutarate (BCYEα) agar at 36°C for 7 d, it exhibited red autofluorescence under UV light (365 nm) . The dominant cellular fatty acids of the strain HYNE-20 were 16:1ω7c (29.9%) , and the guanine-plus-cytosine (G+C) content of DNA was 49.0 mol%. This is the first report that Legionella rubrilucens was isolated from a hot spring for foot soaking.
    Biocontrol science 06/2012; 17(2):101-5.
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    ABSTRACT: To determine the cytotoxicity of antibiotic eyedrops to ocular surface cells using a semi-quantitative method, a range of commercially available antibiotic eyedrops were assessed by using three corneal cell lines and one conjunctival cell line. All antibiotic solutions were free of benzalkonium chloride. Cell viability was determined by the MTT assay and neutral red assay following the exposure of cells to the undiluted, 2- and 10-fold diluted drugs for 10, 30, and 60 min. Toxicity was compared using % cell viability score (%CVS) . The tested eyedrops and values of %CVS50 and %CVS40/80 were Bestron(®) (cefmenoxime, 100, 94) , Panimycin(®) (dibekacin, 86, 58) , Noflo(®) (norfloxacin, 90, 50) , Cravit(®) (levofloxacin, 86, 46) , Tosfulo(®) (tosufloxacin, 57, -3) , and Vigamox(®) (moxifloxacin, 57, -6) . Cell viability markedly increased after dilution. For instance, cell viability assayed by MTT was > 80% for all the measurements in antibiotics diluted 10-fold, and the rate of the measurements showing > 80% cell viability decreased to 43% (31 out of 72 measurements) in the solutions diluted 2-fold. Of the drugs tested, Bestron(®) containing cefmenoxime showed the weakest toxicity. Vigamox(®) containing moxifloxacin and Tosuflo(®) containing tosufloxacin were more toxic when compared with the other antibiotics. CVS was useful for the comparison of the cytotoxicity of the drugs.
    Biocontrol science 06/2012; 17(2):93-9.
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    ABSTRACT: Feline calicivirus (FCV) is a pathogenic microorganism that causes upper respiratory diseases in cats. Recently, an FCV infection with a high mortality rate has been confirmed, and there is need to develop a treatment for cases of acute infection. We evaluated whether the replication of FCV could be prevented by RNA interference. For this study, we designed an siRNA targeted to the polymerase region of the strain FCV-B isolated from a cat that died after exhibiting neurological symptoms. Cells transfected with siR-pol dose-dependently suppressed the replication of FCV-B. siR-pol suppressed its replication by suppressing the target viral RNA.
    Biocontrol science 06/2012; 17(2):87-91.
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    ABSTRACT: We tried to discriminate 16 strains of the Bacillus cereus group including B. cereus, B. thuringiensis, B. mycoides, B. pseudomycoides, and B. weihenstephanensis strains by the pattern analysis of Random Amplified Polymorphic DNA (RAPD) -PCR. Eight oligonucleotides primers were prepared and the polymorphic patterns of the DNA of each strain were compared with those of others. The primers E and F gave different patterns of RAPD-PCR products in all strains of the B. cereus group, so these primers are effective tools for the discrimination of closely related strains. All eight primers showed different polymorphic patterns of DNA for the four strains of B. cereus isolated from the kitchen of a private home, which verifies the advantage of the RAPD-PCR analysis for the discrimination of isolated strains of B. cereus from the environment.
    Biocontrol science 06/2012; 17(2):83-6.
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    ABSTRACT: We found that an additive for a resin, which was comprised of collagen and aluminum (Al), showed a strong and stable antibacterial effect against various bacterium under certain conditions. We tried to clarify its mechanism of action, and investigated optimum conditions for its effects. This additive (Al cross-linked collagen powder: Al-COL) absorbed phosphorus in LB medium, gradually released aluminum in the phosphorus-reduced LB medium, and exhibited a bactericidal effect. Allophane was very suitable as the control subject, because it did not release Al in the medium, decreased phosphorus levels in the medium, and the phosphorus decrease led to a reduction in bacterial growth, though not to a bactericidal effect. On the other hand, the addition of Al to the phosphorus-reduced solution led to a bactericidal effect. These results suggested that Al can exert a strong antibacterial effect in the absence of phosphorus. This phenomenon was confirmed using film-shaped test items mixed with Al-COL powder. Furthermore, the reduction of phosphorus also synergistically led to the enhancement of the antibacterial effect of silver (Ag). The phosphorous absorption promoted the antibacterial action of Al and Ag, and Al, which has seldom been used as an antimicrobial agent, is available as an antibacterial agent in the absence of phosphorus.
    Biocontrol science 03/2012; 17(1):37-44.
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    ABSTRACT: We investigated the virucidal activity of commercially available alcohol-based hand rub products against coxsackievirus A7, B5, feline calicivirus F9, and human adenovirus type 3, type 7, type 8 using susceptible cell lines, Vero cells, CRFK cells, and A549 cells. Fifteen tested hand rub products were ethanol (EtOH) for disinfection (Japanese Pharmacopoeia Grade), two EtOH-based products, one povidone iode-containing product, one alkyldiaminoethylglycine hydrochloride-containing product, six benzalkonium chloride (BAK)-containing products, and four chlorohexidine gluconate (CHG)-containing products. Some active ingredients (BAK, benzetonium chloride, and CHG) were diluted with EtOH to make 0.5% and 0.2% solutions. Virus inactivation rates were calculated after contact with each hand rub product for 10 or 60 seconds. Of the hand rub products tested, only the povidone iode-based product showed antiviral activity superior to that of EtOH against all the strains. EtOH solutions of active ingredients (0.2% and 0.5%) also showed decreased antiviral activity. In conclusion, antiviral activity of all the commercially available alcohol-based hand rub products except that containing povidone idode was dependent on their active ingredients. The povidone idode-containing hand rub product kept its effectiveness even after the dilution with EtOH. Although alcohol-based hand rub products are convenient and suitable for the control of some microbes, they are not generally recommended for the control of viral infections.
    Biocontrol science 03/2012; 17(1):45-9.
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    ABSTRACT: Bacteriocins are ribosomally synthesized antibacterial peptides produced by bacteria that inhibit the growth of similar or closely related bacterial strains. A number of bacteriocins from a wide variety of bacteria have been discovered, and their diverse structures have been reported. Growing evidence suggests that bacteriocins have diverse structures, modes of action, mechanisms of biosynthesis and self-immunity, and gene regulation. Bacteriocins are considered as an attractive compound in food and pharmaceutical industries to prevent food spoilage and pathogenic bacterial growth. Furthermore, elucidation of their biosynthesis has led to the use of bacteriocin-controlled gene-expression systems and the biosynthetic enzymes of lantibiotics, a class of bacteriocins, as tools to design novel peptides. In this review, we summarize and discuss currently known information on bacteriocins produced by Gram-positive bacteria and their applications.
    Biocontrol science 03/2012; 17(1):1-16.
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    ABSTRACT: The bacterial communities associated with rotifers (Brachionus plicatilis sp. complex) and their culture water were determined using culture-dependent and -independent methods (16S rRNA gene clone library). The bacterial communities determined by the culture-independent method were more diverse than those determined by the culture-dependent method. Although the culture-dependent method indicated the bacterial community of rotifers was relatively similar to that of the culture water, 16S rRNA gene clone library analyses revealed a great difference between the two microbiotas. Our results suggest that most bacteria associated with rotifers are not easily cultured using conventional methods, and that the microbiota of rotifers do not correspond with that of the culture water completely.
    Biocontrol science 03/2012; 17(1):51-6.
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    ABSTRACT: The antimicrobial activity, toxicity and antimicrobial mechanism of a new type of tris(4-alkylphenyl)sulfonium which has sterically bulky alkyl substituents (bTAPS), were estimated and compared with those of other sulfoniums which we reported previously. Concerning tris {4-(iso-propyl)phenyl}sulfonium (bTAPS-iso3) and tris{4-(tert-butyl)phenyl}sulfonium (bTAPS-tert4), the antimicrobial activity of these compounds tended to be lower than both tri(n-alkyl)sulfoniums (TASs) and tris{4-(n-alkylphenyl)}sulfoniums (TAPSs) at similar ClogP values. However, the activities of tris{4-(cyclohexyl)phenyl}sulfonium (bTAPS-cyclo6) were clearly higher than those of TAS and were almost similar to those of TAPS at similar ClogP values. The mutagenicities of tested bTAPSs were judged to be all negative. Both the acute oral toxicity strength and the acute skin irritation/corrosion toxicity strength tended to follow the order of TAPSs > bTAPSs > TASs. However, only the acute skin irritation/corrosion toxicity strength of bTAPS-cyclo6 was almost as low as that of TAS which has a similar ClogP value to bTAPS-cyclo6. Because bTAPS-cyclo6 has both high antimicrobial activity and low toxicity, this compound might become to be an alternative antimicrobial compound to relatively hazardous antimicrobials which have been widely used in many fields.
    Biocontrol science 03/2012; 17(1):27-35.
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    ABSTRACT: The anti-influenza virus activity of fossilized marine coral powder (sango mineral powder, SMP) was studied. SMP is composed in terms of mass of around 25 % of calcium and 10 % of magnesium, respectively, principally as dolomite (CaMg(CO(3))(2)) but not as calcium oxide (CaO) or magnesium oxide (MgO). By mixing the influenza virus with SMP, the infectivity of the virus substantially decreased and there was more than a 10(4) reduction on the 3rd d of infection. The antiviral effect was observed against all the type A and B strains of the influenza virus examined including the H1N1 2009 pandemic and H5N1 avian viruses. The surface structure of SMP was highly porous and the anti-influenza activity was explained by the adsorption of the viral particles onto its surface. The binding of viruses to SMP was strong and stable in the physiological condition, and the attached viruses detached only in the presence of a high concentration of phosphate. This was similar to the binding of protein to hydroxyapatite, suggesting an ionic interaction between SMP and the viral proteins. SMP maintained its activity to capture influenza viruses even after being immobilized on a non-woven textile. SMP would be useful as a practical anti-influenza tool especially in preparation for the next pandemic virus.
    Biocontrol science 03/2012; 17(1):17-25.
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    ABSTRACT: To establish rapid methods to detect Shiga toxin (Stx)-producing Escherichia coli (STEC) in ground beef samples by using an immunochromatography kit, results of 8-h enrichment in various types of broth with shaking were compared. In pure culture, Stx was detected in the culture of trypticase soy broth (TSB) at 42°C and modified EC broth (mEC) at 36°C from all or most serogroups of O26, O111, O128, O157 and OUT. Ground beef samples inoculated with each serogroup were enriched in TSB at 42°C, mEC at 36°C and mEC with novobiocin (NmEC) at 42°C. Although all conditions led to the successful recovery of each serogroup by the plating method, enrichment in NmEC was relatively superior to the other conditions in the detection of Stx by an immunochromatography kit. These results indicated that the growth of STEC and the release of Stx from cells were different in pure cultures and in culture with ground beef. In addition, polymyxin B treatment for 10 min at 37°C and homogenizing with glass beads enhanced the detection of Stx. From the results, it was suggested that an immunochromatography kit in a combination with enrichment in NmEC at 42°C for 8 h, and treatment with polymyxin B or homogenizing would be a rapid method to detect STEC contamination in ground beef.
    Biocontrol science 12/2011; 16(4):159-64.
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    ABSTRACT: We investigated the bactericidal effects and cytotoxicity of an ortho-phthalaldehyde product in comparison with those of its predecessor glutaraldehyde products. Bactericidal effects ware examined on Mycobacterium terrae, a standard organism used for investigating the bactericidal effect of high-level disinfectants. Cytotoxicity as determined by the MTT assay was examined by using four cell lines. The colony forming test, a method to examine residual toxicity, and the evaporation test, a newly developed method to examine the toxicity of the evaporated ingredients, were performed. Test solutions were 2.25% and 3.5% glutaraldehyde (GA) products and a 0.55% ortho-phthalaldehyde (OPA) product, and glutaraldehyde itself. All the disinfectants showed sufficient bactericidal effects on M. terrae. Meanwhile, the OPA product was less toxic than GA products and GA itself to all the cell lines tested. The colony forming test showed that GA products and GA itself exerted residual cytotoxicity more potently than did the OPA product. The evaporation test showed that GA products and GA itself exerted cytotoxicity via evaporation more potently than did the OPA product. In conclusion, OPA appears to be less cytotoxic than GA even though bactericidal effects were comparable. This may be due to the lower concentration of the active ingredient (ortho-phthalaldehyde) in the OPA product.
    Biocontrol science 12/2011; 16(4):165-70.
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    ABSTRACT: Scolecobasidium, generally found in outdoor samples, were isolated from detergent-rich indoor environments. The isolates from bathrooms and washing machines, because of their exposure to detergents, might be genetically and biologically distinct from outdoor isolates. In this study, 11 Scolecobasidium isolates from detergent-rich indoor environments were examined to find the genetic and biological differences between the indoor and outdoor isolates. One isolate from a wall of a soap factory, showing similar conidia morphology with S. constricta, was phylogenetically distinct from the other Scolecobasidium spp. The 10 isolates from washing machines and bathrooms were identified as S. humicola, but these were classified into 2 groups that differed from the reference strain of S. humicola from leaves. All 11 isolates and the 4 reference strains of S. constricta and S. humicola grew on the medium containing sodium oleate and polyoxyethylene-(9)-lauryl ether, but the reference strains of the other Scolecobasidium spp. grew only on the medium containing sodium oleate. The results showed that S. humicola and S. constricta could utilize both surfactants generally included in soaps or synthetic detergents as nutrients. A further implication is that the genetic variation found in the S. humicola isolates from detergent-rich indoor environments can occur as a result of adaptation to such an environment.
    Biocontrol science 12/2011; 16(4):139-47.
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    ABSTRACT: In August, 2010, strain HYMO-6 was isolated from a sample of hot spring water in Aomori, Japan. The 16S rDNA sequences (1,496bp) of this strain (accession number: AB597175) had a similarity of less than 96.6% to other Legionella species, prompting us to hypothesize that this strain might be a novel species belonging to the genus Legionella. However, in March of 2011, it was became clear that the HYMO-6 strain (=JCM 17450 =KCTC 23560 =DSM 24727) was Legionella nagasakiensis CDC-1796-JAP-E(T) (=ATCC BAA-1557(T) =JCM 15315(T)). When this strain was cultured on BCYEα agar at 36°C for 7 d, no long cells were observed. The dominant fatty acids of strain HYMO-6 were 16:1ω7c (32.4%), and the DNA G+C content was 42.0 mol%.
    Biocontrol science 12/2011; 16(4):171-6.

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