• Asian-Australasian Journal of Animal Sciences
• /
• v.16 no.7
• /
• pp.1015-1019
• /
• 2003

A 50 day feeding trial was conducted with White Leghorn (WL) laying hens, 42 weeks old, to determine if feeding of varying levels of aflatoxin (AF), ochratoxin A (OA) or their combinations has any effect on their performance and egg quality parameters. Feeding of $T_4$, $T_7$, $T_8$, $T_9$ and $T_10$ caused significant reduction in feed intake of hens. Hen day egg productions were significantly reduced at all the levels of toxins except 0.5 ppm of AF. Maximum reduction in egg production was noticed at 2 and 4 ppm of AF and OA, respectively. Average body weight and egg weight were not affected by toxin feeding. The feed efficiency in terms of net feed efficiency and feed consumed per dozen egg produced was significantly reduced at higher levels of both the toxins and their combinations. Feed consumption for production of 1 kg egg mass remained uninfluenced due to aflatoxin feeding whereas significant increase in the value of the same was noticed at 4 ppm level of OA and combination of 1 and 2 ppm of AF and 2 and 4 ppm of OA ($T_9$ and $T_10$), respectively. Various levels of OA (1-4 ppm) and all the combination of two toxins ($T_8$, $T_9$ and $T_10$) significantly altered the shape index of eggs in laying hens. The shell thickness was significantly reduced by higher level of AF (2 ppm), OA (2 and 4 ppm) and their combination. Albumen index, Haugh Unit and yolk index remained unchanged due to incorporation of toxins in the diet. It is concluded that AF, OA either singly or in combination at higher levels could depress the performance in terms of egg production and feed efficiency significantly. The egg quality parameters i.e. shape index and shell thickness were also significantly affected.

• Proceedings of the Korean Society of Food Hygiene and Safety Conference
• /
• 1997.06b
• /
• pp.189-189
• /
• 1997

• Korean Journal of Microbiology
• /
• v.40 no.4
• /
• pp.286-294
• /
• 2004

Between 1997 and 1998 in Korea, 56 isolates of Escherichia coli were obtained from pig suffering diarrhea. Among those, 38 isolates that showed the hemolytic activity, antimicrobial resistance, and toxin production were studied. Among 38 isolates, thirty-six isolates $(94.7\%)$ were resistant to tetracycline, 27 isolates $(71.0\%)$ were resistant to ampicillin, 26 isolates $(68.4\%)$ were resistant to chloramphenicol, and 21 isolates $(55.2\%)$ were resistant to trimethoprim, while none was resistant to aztreonam, amikacin, and norfloxacin. Among these iso­lates, 21 isolates $(55.3\%)$ were multiple drug resistant to at least four different class antimicrobial agents. Extended spectrum $\beta-lactamase$ producing isolates were not detected in the double disk synergy test. In these hemolytic Escherichia coli, heat-stable enterotoxin $(89.5\%)$ was the most prevalent toxin, followed by vero­toxins $(47.4\%),$ and then heat-labile enterotoxin $(31.6\%).$ Except 8 isolates $(21.0\%)$ which produced ST only, 12 isolates $(31.6\%)$ produced ST and LT, 13 isolates $(34.2\%)$ produced ST, VT, and VTe, and 5 isolates $(13.2\%)$ produced VT and VTe. However, none produced all 4 types of toxin, simultaneously. The predominant serotype could not be determined by the agglutination method. Sixteen isolates $(42.1\%)$ were strongly adhered to T-24 bladder cell and 17 isolates $(44.7\%)$ were to Caco-2 intestinal cell. Especially, 11 strains $(28.9\%)$ were evaluated as strongly adhesive to both T-24 cells and Caco-2 cells. Genes for toxin and the antimicrobial resistance were transferred to clinical isolates of Escherichia coli from human urine by the filter mating method. Results suggest the possibility that antimicrobial resistance and toxin can be transferred from animals to humans by direct con­tact of resistant bacteria as well as gene transfer, although there was no correlation between toxin production, adherent activity, and antimicrobial resistance among hemolytic E. coli isolated from pig suffering diarrhea.

• Food Science and Biotechnology
• /
• v.17 no.4
• /
• pp.761-765
• /
• 2008

Bacillus cereus causes two different types of food poisoning syndromes: diarrhea and emesis. The diarrheal syndrome is attributed to various enterotoxins, including nonhemolytic enterotoxin, hemolytic enterotoxin, and enterotoxin-T, whereas the emetic syndrome is caused by the dodecadepsipeptide toxin cereulide. A multiplex polymerase chain reaction (PCR) assay was developed to rapidly detect and identify B. cereus strains. Three primer pairs specific to regions within genes encoding nonhemolytic enterotoxin (nheA), molecular chaperonin (groEL), and cereulide synthetase (ces) were used to identify and differentiate between the enterotoxin-producing and emetic toxin-producing B. cereus strains. The cereulide-producing emetic B. cereus showed 3 PCR products of 325, 405, and 685 bp for the groEL, ces, and nheA genes, respectively, whereas the enterotoxin-producing B. cereus showed 2 PCR products without a ces gene specific DNA fragment. Specific amplifications and differentiations by multiplex PCR assay were obtained using 62 B. cereus strains and 13 strains' of other bacterial species. The detection limit of this assay for enterotoxin-producing strain and emetic toxin-producing strain from pure cultures were $2.4{\times}10^1$ and $6.0{\times}10^2\;CFU/tube$, respectively. These results suggest that our multiplex PCR method may be useful for the rapid detection and differentiation of B. cereus strains in foods.

• BMB Reports
• /
• v.30 no.2
• /
• pp.144-149
• /
• 1997

An expression vector was constructed containing the gene encoding diphtheria toxin A (DTA) which was placed after a T7 promoter. Cytoplasmic expression of the DTA gene resulted in the formation of an insoluble inclusion body. The inclusion body was collected after the complete lysis of the cell, and subsequent washing with 0.1% Triton X-100 released 16~30% of DTA protein from the inclusion body along with other contaminating proteins. The released DTA protein was purified by dialysis. The remaining pellet was dissolved in 8 M urea containing 5% ${\beta}-mercaptoethanol$, and the denatured DTA was renatured by the dilution-dialysis method. The total yield was 35%, and about 5 mg DTA was obtained from 1 L culture. The DTA protein has a free sulfhydryl group exposed to the protein surface, and was shown to have a tendency to dimerize through disulfide formation in the absence of ${\beta}-mercaptoethanol$. The utility of the sulfhydryl group was tested for the construction of recombinant toxins.

• Biomedical Science Letters
• /
• v.23 no.3
• /
• pp.175-184
• /
• 2017

Cholera toxin (CT) is an ADP-ribosylating bacterial exotoxin that has been used as an adjuvant in animal studies of oral immunization. The mechanisms of mucosal immunogenicity and adjuvanticity of CT remain to be established. In this study, we investigated the role of indoleamine 2,3-dioxygenase (IDO), which participates in the induction of immune tolerance, in CT-mediated breakdown of oral tolerance. When IDO-deficient ($IDO^{-/-}$) mice and their littermates were given oral ovalbumin, significant changes in antibody responses, footpad swelling and $CD4^+$ T cell proliferation were not observed in $IDO^{-/-}$ mice. Feeding of CT decreased IDO expression in mesenteric lymph nodes (MLN) and Peyer's patch (PP). CT-induced downregulation of IDO expression was reversed by inhibitors of nuclear factor-kappa B (NF-${\kappa}B$), pyrrolidine dithiocarbamate and p50 small interfering RNA. IDO expression was downregulated by the NF-${\kappa}B$ inducers lipopolysaccharide and tumor necrosis factor-${\alpha}$. CT dampened IDO activity and mRNA expression in dendritic cells from MLN and PP. These data indicate that CT disrupts oral tolerance by activating NF-${\kappa}B$, which in turn downregulates IDO expression. This study betters the understanding of the molecular mechanism underlying CT-mediated abrogation of oral tolerance.

• Journal of Periodontal and Implant Science
• /
• v.37 no.sup2
• /
• pp.339-351
• /
• 2007

Cytolethal distending toxin(CDT)은 세포 주기 중 G2에서 M 기로의 전환을 막아 세포의 증식을 억제할 수 있는 세균 단백 독소의 일종이다. 구강 미생물 중 유일하게 Actinobacillus actinomycetemcomitans (A. actinomycetemcomitans)만이 이 CDT를 생성 할 수 있는 것으로 알려져 있다. A. actinomycetemcomitans는 localized aggressive periodontitis (LAP)의 원인균으로 여겨지며 비 운동성의 그람 음성 구간균이고 $37^{\circ}C$, 5% $CO_2$ 하에 성장이 왕성하다. A. actinomycetemcomitans의 CDT는 3개의 인접한 유전자인 cdtA, cdtB, cdtC에 의해 형성 되며 각각의 유전자에 대한 단백질의 기능은 아직 완전히 밝혀지지 않았다. 현재까지 연구에 의하면 cdtA는 CDT의 세포부착과 관련이 있는 것으로 여겨지며 이 유전자의 기능 이상 시 CDT의 독성 효과가 현저히 감소한다고 알려져 있다. 따라서 본 연구는 A. actinomycetemcomitans의 cdtA 유전자를 클로닝, 단백질 발현하여 향후 치주질환의 발병 과정에서 CdtA의 역할을 규명하고 질환의 예방 및 치료법에 도움을 주고자 하였다. A. actinomycetemcomitans Y4균주를 cdtA 유전자 클로닝을 위해 사용하였다. A. actinomycetemcomitans의 genomic DNA는 genomic DNA 추출 kit를 사용하여 분리하고 cdtA에 특이적인 primer를 이용하여 PCR을 통해 cdtA 유전자를 증폭하였다. 증폭된 cdtA 유전자를 T-vector에 클로닝 하였으며, 클로닝 된 cdtA 유전자는 단백질 발현을 위해 pRSET Avector에 서브클로닝 한 후 발현 균주인 BL21(DE3)를 이용하여 발현시켰다. 발현 후 Ni-NTA AP conjugate를 이용한 Western blot을 통해 pRSET-CDTA를 확인하였다.

• The Korean Journal of Pain
• /
• v.18 no.1
• /
• pp.29-33
• /
• 2005

Background: Chronic headache (CH) constitutes a significant public health problem, impacting on both the individual sufferer and society. Patients with CH, unresponsive to drug therapy or nerve block, suffer considerable disability due to the frequency and severity of attacks; therefore, they should be considered for novel therapy. Botulinum toxin type A (BoNT-A) has shown significant promise in the management of CH. In this paper, we review recent evidence on the efficacy of BoNT-A, and also report our experience with this treatment in CH patients. Methods: BoNT-A was used to treat 69 CH patients, including 47 in a chronic migraine group and 22 in a non-migraine CH group, who showed therapy-resistance to palliative drug or nerve block. We investigated the demography, dosage and site of BoNT-A injection, and used a visual analogue scale (VAS) for pain and the degree of satisfaction. The data were analyzed using t-tests and a Friedman repeated measures analysis of variance on ranks. Results: Significant decreases in the VAS for pain were found in both the chronic migraine and non-migraine CH groups, from 2, 4 and 12 weeks and from 4 and 12 weeks, respectively, after BoNT-A administration (P < 0.05). The chronic migraine group showed significantly lower VAS scores for pain than the non-migraine CH group from 2, 4 and 12 weeks after the BoNT-A administration (P < 0.05). Twenty eight patients (59.2%) in the chronic migraine group and eight (36.4%) in the non-migraine CH were satisfied with the BoNT-A treatment. Conclusions: This clinical study revealed that the use of BoNT-A demonstrated efficacy for CH patients resistant to drug therapy or nerve block. Moreover, BoNT-A proved itself more effective in the chronic migraine than non-migraine CH group.

• Toxicological Research
• /
• v.1 no.1
• /
• pp.1-15
• /
• 1985

The trichothecenes are a chemically related sesquiterpenoid fungal metabolites of Fusarium, Trichoderma, Stachybotrys and others, and at moment more than 70 kinds of derivatives are identified. Historically, they are identified as antifungal and phytotoxic compounds, but after the finding of T-2 toxin from Fusarium tricinctum, several trichothecenes are now considered to be natural toxicants in foodstuffs and feeds.

• Microbiology and Biotechnology Letters
• /
• v.11 no.3
• /
• pp.223-231
• /
• 1983

The productions of beta-exotoxin from sixteen Bacillus thuringiensis strains were examined by Micrococus flava primarily, and then measured by spectrophotometer during culturing in Conner and Hansen mineral salts medium at 28$^{\circ}C$. Also the toxic effects of the toxin to mice were checked. The growth of Bacillus thuringiensis K2 and BTK2-T1, -T13, -T33 and -T40 got into stationary phase at 6 hour culture and then maintained it up to 48 hours without severe fluctuation. The production of beta-exotoxin from the strains, BTK2, BTK2-T1, -T13, -T17 and -T33 appeared at 6 hour culture and the amounts of the toxin were about 40 $\mu\textrm{g}$/$m\ell$ at 6 hour culture, approximately 70 $\mu\textrm{g}$/$m\ell$ at 12 hours, approximately 85$\mu\textrm{g}$/$m\ell$ from 24 hours to 48 hours. At 48 hour-culture, BTK2 produced 80 $\mu\textrm{g}$/$m\ell$ of beta-exotoxin (5.5$\times$10$^{8}$ cells/$m\ell$, BTK2-T13 produced 84 $\mu\textrm{g}$/$m\ell$ (4.3$\times$10$^{8}$ cells/$m\ell$), BTK2-T17 produced 87$\mu\textrm{g}$/$m\ell$ (1.4$\times$10$^{8}$ cells/$m\ell$), and BTK2-T33 produced 84 $\mu\textrm{g}$/$m\ell$ (4.9$\times$10$^{8}$ cells/$m\ell$). All other serotypes also produced beta-exotoxin. At 48 hour culture, BTK-37 produced 88$\mu\textrm{g}$/$m\ell$ (6.1$\times$10$^{8}$ cells/$m\ell$), BTK-35 produced 81 $\mu\textrm{g}$/$m\ell$), and the rest of them produced less than 70 $\mu\textrm{g}$/$m\ell$. To check the toxicity of beta-exotoxin and B. thuringiensis, the cultured media with microorganisms were inoculated to mice by per os, intraperiloneal, subcutaneous and intracerebral injection, and nasal cavity inoculation for 30 days. However, the toxin did not kill all of the treated mice.