• Title/Summary/Keyword: toxigenicity

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Development of a toxA Gene Knock-out Mutant of Pasteurella multocida and Evaluation of its Protective Effects

  • Kim Tae-Jung;Lee Jae-Il;Lee Bong-Joo
    • Journal of Microbiology
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    • v.44 no.3
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    • pp.320-326
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    • 2006
  • Pasteurella multocida is an important veterinary and opportunistic human pathogen. In particular, strains of P. multocida serogroup D cause progressive atrophic rhinitis, and produce a potent, intracellular, mitogenic toxin known as P. multocida toxin (PMT), which is encoded by the toxA gene. To further investigate the toxigenic and pathogenic effects of PMT, a toxA-deleted mutant was developed by homologous gene recombination. When administrated to mice, the toxigenicity of the toxA mutant P. multocida was drastically reduced, suggesting that the PMT constributes the major part of the toxigenicity of P, multocida. Similar results were obtained in a subsequent experiment, while high mortalities were observed when toxA(+) P. multocida bacterial culture or culture Iysate were administrated. Mice immunized with toxA(-) P. multocida were not protected (none survived) following challenge with toxA(+) P. multocida or bacterial culture Iysate (toxin). These results suggest that the toxigenicity of P. multocida is mainly derived from PMT.

Detection of Toxigenicity of Cyanobacteria by Molecular Method (분자생물학적 방법에 의한 남조류의 독성 생성능의 확인)

  • Lee, Kyung-Lak;Jheong, Weon-Hwa;Kim, Jong-Min;Kim, Han-Soon
    • Korean Journal of Ecology and Environment
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    • v.40 no.1
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    • pp.149-154
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    • 2007
  • In the present study, we performed the PCR assay using TOX2P/TOX2M primer targeting a specific region within mcyB gene to identify potential microcystin-producing cyanobacteria. TOX2P/TOX2M primer set was effective in amplifying mcy gene in the field samples containing Microcystis spp. of 1,000 cells per mL. Moreover, the results from the PCR assay agreed with those of the ELISA analysis. Consequently, this study demonstrated that TOX2P/TOX2M primer set can be used as a genetic probe for the early detection of cyanobacterial toxigenicity in Korean water bodies.

Growth Profile and Toxigenicity of Bacillus cereus in Ready-to-eat Food Products of Animal Origin

  • Oh, Mi-Hwa;Ham, Jun-Sang;Seol, Kuk-Hwan;Jang, Ae-Ra;Lee, Seung-Gyu;Lee, Jong-Moon;Park, Beom-Young;Kang, Eun-Sil;Kwon, Ki-Sung;Hwang, In-Gyun
    • Food Science of Animal Resources
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    • v.31 no.1
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    • pp.40-46
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    • 2011
  • The growth profile of Bacillus cereus in ready-to-eat (RTE) food products of animal origin was examined under different temperature and incubation conditions. In sandwiches and Kimbab, B. cereus did not grow or exhibited only minimal growth at 4 and $10^{\circ}C$, but it grew rapidly at ambient temperature. In sandwiches, B. cereus did not grow efficiently at $25^{\circ}C$, however, in ham, the main ingredient of sandwiches, B. cereus growth was observed at the same temperature, with bacterial levels reaching 7.94 Log CFU/g after incubation for 24 h at $25^{\circ}C$. Toxigenicity of B. cereus was observed only at temperatures above $25^{\circ}C$. In Kimbab, B. cereus produced toxin after 9 h at $30^{\circ}C$ and after 12 h at $25^{\circ}C$. Ingredients of sandwiches and Kimbab were collected from 3 different Korean food-processing companies to investigate the source of contamination by B. cereus. Among the 13 tested food items, 6 items including ham were found to be contaminated with B. cereus. Of these ingredients, B. cereus isolates from 3 items produced enterotoxins. None of these isolates harbored the emetic toxin-producing gene. The findings of the present study can be used for risk assessments of food products, including ham and cheese, contaminated with B. cereus.

Studies on Pasteurella multocida isolated from pneumonic lungs of slaughter pigs (도축돈의 폐렴병소에서 분리한 Pasteurella multocida 에 대한 연구)

  • Ahn, Byung-chul;Cho, Kwang-hyun;Kim, Bong-hwan
    • Korean Journal of Veterinary Research
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    • v.34 no.3
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    • pp.511-516
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    • 1994
  • P multocida was isolated from 80(17.7%) of 450 pneumonic lungs of slaughter pigs. The majority of the biochemical and cultural characteristics of P multocida isolates were identical to those of the reference strains employed. Seventy seven strains(96.3%) among 80 isolates were capsular serotype A while the remaining 3(3.8%) were serotype D. All isolates were very susceptible to ampicillin, ceftiofur, cephalothin, ciprofloxacin and penicillin-G although some of them were resistant to sulfamethoxin and/or streptomycin. Sixty one(76.3%) of all 80 P multocida isolates were dermonecrotic toxin producers. Out of 77 isolates of serotype A and 3 isolates of serotype D, 59(76.6%) and 2(66.7%) were toxigenic, respectively. No difference was noted in dermonecrotic toxigenicity of the isolates in relation to capsular serotypes.

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Response of Growth and Toxigenicity to Varying Temperature and Nutrient Conditions in Aphanizomenon flos-aquae (Cyanophyceae) (환경조건에 따른 Aphanizomenon flos-aquae (Cyanophyceae) 균주의 성장 반응 및 독소 생성)

  • Ryu, Hui-Seong;Shin, Ra-Young;Lee, Jung-Ho
    • Journal of Korean Society on Water Environment
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    • v.33 no.5
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    • pp.538-545
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    • 2017
  • The purpose of this study is to investigate growth response and toxigenicity under various temperature and nutritional conditions, in order to understand the physioecological characteristics of Aphanizomenon flos-aquae, which is a bloom-forming cyanobacterium in the Nakdong River. The strain was inoculated into media under combinations of four temperatures (4, 12, 21, $30^{\circ}C$) and three nutrients (modified CB medium, P-depleted CB medium, N-depleted CB medium) for 28 days. The algae-inhibition tests were performed to assess the potential allelopathic effects of the strains' filtrates on the growth of four algae strains (Microcystis aeruginosa, Aulacoseria ambigua f. spiralis, Aphanizomenon flos-aquae, Scenedesmus obliquus). Toxin production of a strain was measured by Enzyme-Linked ImmunoSolbent Assay (ELISA). The optimal growth temperature (Topt) of strains was $19.9^{\circ}C$ ($18.3-21.2^{\circ}C$), and the temperature range for growth was from $-0.3^{\circ}C$ to $34.3^{\circ}C$. Specific growth rate (${\mu}$) in modified CB medium varied from 0.10 to $0.16day^{-1}$, and the maximum growth rate (${\mu}_{max}$) was $0.17day^{-1}$. Although growth curves under N-existed and N-depleted conditions were almost the same, growth under N-depleted condition was relatively slowed (${\mu}=0.09$ to $0.14day^{-1}$), with a decreased maximum cell density. However, growth under the P-depleted condition was restricted for all temperatures, Two stains of Aphanizomenon flos-aquae were confirmed as not producing toxins, because saxitoxin and cylindrospermopsin were not detected by ELISA. The exudates or filtrates from the Aphanizomenon flos-aquae (DGUC003) resulted in significant inhibition of algal growth on the Aulacoseira ambigua f. spiralis (DGUD001) and Aphanizomenon flos-aquae (DGUC001) (p < 0.01).

Clinical and bacteriological studies on infectious atrophic rhinitis of swine (돼지 전염성 위축성 비염의 임상학적 및 세균학적 연구)

  • Kim, Bong-hwan;Tak, Ryun-bin;Cho, Gil-jae;Jang, Hee-kyung
    • Korean Journal of Veterinary Research
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    • v.31 no.4
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    • pp.457-469
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    • 1991
  • Clinical and bacteriological observations on infectious atrophic rhinitis of swine were conducted in order to obtain some basic information for the clinical and immnnological control of the disease prevailing in the republic. Samples were collected from nasal cavities of 135 4~12 week old pigs from 12 herds and from turbinates of 199 slaughtering pigs from 14 swine herds to investigate the prevalence of Bordetella bronchiseptica and/or Pasteurella multocida in the nasal cavities of the pigs. On the examination of nasal swabs by cultural techniques and of gross lesion of snouts of slaughtering pigs, all the swine herds investigated were found to be affected by atrophic rhinitis and a total of 84 B bronchiseptica and 139 P multocida cultures were isolated from the nasal cavities of the pigs. Of the 199 slaughtering pigs, some 48% revealed gross pathological lesion typical of atrophic rhinitis and the prevalence of B bronchiseptica and P multocida were 27.6% and 46.7%, respectively. Biochemical properties, antimicrobial susceptibilities, serological characteristics and toxigenicity of the isolates of B bronchiseptica and P multocida were investigated.

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Toxicity and Fumonisin B1 Production by Fusarium Isolates from Chines Corn Samples (중국산 옥수수로부터 분리한 Fusarium균주의 독성 및 Fumonisin B1의 생성)

  • 이인원;강효중
    • Korean Journal Plant Pathology
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    • v.10 no.2
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    • pp.129-135
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    • 1994
  • Ninety-two isolates of Fusarium species were obtained from Chinese corn samples. The predominant Fusarium species isolated from corn seeds were F. moniliforme, F. proliferatum, F. oxysporum and F. subglutinans, and all 13 species were identified. Each isolate was grown on autoclaved wheat grains and wheat cultures were fed by twenty-one-day-old female rats for the toxicity test. Twenty-six out of 92 isolates caused the death accompanying feed refusal, severe weight loss, liver damage, and hemorrhages in the stomach and intestines. Of the toxigenic isolates, 17 isolates of F. moniliforme, 4 of F. oxysporum, 3 of F. proliferatum, and one of each F. sporotrichioides and unknown species were lethal to rats. The analyses of fumonisin B1 production of the 26 toxigenic Fusarium isolates were carried out by thin layer chromatography and high-performance liquid chromatography, and fumonisin B1 was confirmed by mass spectrometry. Fumonisin B1 was produced in wheat culture at levels ranging from 280 $\mu\textrm{g}$/g to 3,952 $\mu\textrm{g}$/g by all of toxigenic F. moniliforme and F. proliferatum, but by none of the other toxigenic Fusarium species. The present results suggest the high possibility of natural occurrence of fumonisin B1 in corn samples imported from China.

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Taxonomic Characterization, Evaluation of Toxigenicity, and Saccharification Capability of Aspergillus Section Flavi Isolates from Korean Traditional Wheat-Based Fermentation Starter Nuruk

  • Bal, Jyotiranjan;Yun, Suk-Hyun;Chun, Jeesun;Kim, Beom-Tae;Kim, Dae-Hyuk
    • Mycobiology
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    • v.44 no.3
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    • pp.155-161
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    • 2016
  • The most economically important species used in a wide range of fermentation industries throughout Asia belong to Aspergillus section Flavi, which are morphologically and phylogenetically indistinguishable, with a few being toxigenic and therefore a major concern. They are frequently isolated from Korean fermentation starters, such as nuruk and meju. The growing popularity of traditional Korean alcoholic beverages has led to a demand for their quality enhancement, therefore requiring selection of efficient non-toxigenic strains to assist effective fermentation. This study was performed to classify the most efficient strains of Aspergillus section Flavi isolated from various types of traditional wheat nuruk, based on a polyphasic approach involving molecular and biochemical evaluation. A total of 69 strains were isolated based on colony morphology and identified as Aspergillus oryzae/flavus based on internal transcribed spacer and calmodulin gene sequencing. Interestingly, none were toxigenic based on PCR amplification of intergenic regions of the aflatoxin cluster genes norB-cypA and the absence of aflatoxin in the culture supernatants by thin-layer chromatography analysis. Saccharification capability of the isolates, assessed through ${\alpha}-amylase$ and glucoamylase activities, revealed that two isolates, TNA24 and TNA15, showed the highest levels of activity. Although the degrees of variation in ${\alpha}-amylase$ and glucoamylase activities among the isolates were higher, there were only slight differences in acid protease activity among the isolates with two, TNA28 and TNA36, showing the highest activities. Furthermore, statistical analyses showed that ${\alpha}-amylase$ activity was positively correlated with glucoamylase activity (p < 0.001), and therefore screening for either was sufficient to predict the saccharifying capacity of the Aspergillus strain.

Genomics Reveals Traces of Fungal Phenylpropanoid-flavonoid Metabolic Pathway in the Filamentous Fungus Aspergillus oryzae

  • Juvvadi Praveen Rao;Seshime Yasuyo;Kitamoto Katsuhiko
    • Journal of Microbiology
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    • v.43 no.6
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    • pp.475-486
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    • 2005
  • Fungal secondary metabolites constitute a wide variety of compounds which either playa vital role in agricultural, pharmaceutical and industrial contexts, or have devastating effects on agriculture, animal and human affairs by virtue of their toxigenicity. Owing to their beneficial and deleterious characteristics, these complex compounds and the genes responsible for their synthesis have been the subjects of extensive investigation by microbiologists and pharmacologists. A majority of the fungal secondary metabolic genes are classified as type I polyketide synthases (PKS) which are often clustered with other secondary metabolism related genes. In this review we discuss on the significance of our recent discovery of chalcone synthase (CHS) genes belonging to the type III PKS superfamily in an industrially important fungus, Aspergillus oryzae. CHS genes are known to playa vital role in the biosynthesis of flavonoids in plants. A comparative genome analyses revealed the unique character of A. oryzae with four CHS-like genes (csyA, csyB, csyC and csyD) amongst other Aspergilli (Aspergillus nidulans and Aspergillus fumigatus) which contained none of the CHS-like genes. Some other fungi such as Neurospora crassa, Fusarium graminearum, Magnaporthe grisea, Podospora anserina and Phanerochaete chrysosporium also contained putative type III PKSs, with a phylogenic distinction from bacteria and plants. The enzymatically active nature of these newly discovered homologues is expected owing to the conservation in the catalytic residues across the different species of plants and fungi, and also by the fact that a majority of these genes (csyA, csyB and csyD) were expressed in A. oryzae. While this finding brings filamentous fungi closer to plants and bacteria which until recently were the only ones considered to possess the type III PKSs, the presence of putative genes encoding other principal enzymes involved in the phenylpropanoid and flavonoid biosynthesis (viz., phenylalanine ammonia-lyase, cinnamic acid hydroxylase and p-coumarate CoA ligase) in the A. oryzae genome undoubtedly prove the extent of its metabolic diversity. Since many of these genes have not been identified earlier, knowledge on their corresponding products or activities remain undeciphered. In future, it is anticipated that these enzymes may be reasonable targets for metabolic engineering in fungi to produce agriculturally and nutritionally important metabolites.

Analysis of Potential Toxigenicity and Phylogeny using Target Genes in Aphanizomenon flos-aquae (Cyanophyceae) strains isolated from the Nakdong River (낙동강에서 분리된 Aphanizomenon flos-aquae (Cyanophyceae) 균주의 목표 유전자를 이용한 잠재적 독소 생성능 및 계통학적 분석)

  • Ryu, Hui-Seong;An, Sung-Min;Lim, Chang-Kun;Shin, Ra-Young;Park, Jong-Guen;Lee, Jung-Ho
    • Korean Journal of Ecology and Environment
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    • v.50 no.1
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    • pp.137-147
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    • 2017
  • The identity of toxin producers remains only hypothesis unless there were identified by strain isolation and analytical confirmation of both the cyanotoxin production and the genetic identity of the monoculture. The purposes of this study were to identify a morphologic and phylogenetic classification in Aphanizomenon flos-aquae strains isolated from the Nakdong River and to investigate the potential ability of the strains to produce toxins such as saxitoxin and cylindrospermopsin using target genes. The 16S rRNA and sxtA, sxtI, cyrA, cyrJ genes were analyzed on two strains (DGUC001, DGUC003) isolated from the Nakdong River. Morphological features of the strains were observed a shape of aggregated trichomes in parallel fascicles which can reach up to macroscopic size and a hyaline terminal cell without aerotope. In addition, the 16S rRNA phylogenetic analyses showed that the strains were identified as the same species with high genetic similarity of 98.4% and grouped within a monospecific andsupported cluster I of Aphanizomenon flos-aquae selected from GenBank of the NCBI. The cyrA and cyrJ genes encoding for the cylindrospermopsin-biosynthesis were not detected in the present study. The sxtA gene was in detected both the two strains, whereas the sxtI gene which had been suggested as a suitable molecular marker to detect saxitoxin-producing cyanobacteria was not found both the strains. Thus, the two strains isolated from Nakdong River were identified as the same species of Aphanizomenon flos-aquae Ralfs ex Bornet et Flahault 1888, the two strains were confirmed as potential non-producing strains of the saxitoxin and cylindrospermopsin.