• Korean Journal of Veterinary Research
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• v.55 no.2
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• pp.117-123
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• 2015

Necrotic enteritis (NE) caused by Clostridium (C.) perfringens commonly occurs in domestic broiler farms since antibiotic supplementation in poultry feed has been banned. We evaluated the antibacterial activities of medicinal plant extracts against C. perfringens isolates to select alternative compounds for preventing NE. We compared antibacterial activities using two methods and evaluated susceptibilities of the isolates based on minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). Two (Fraxinus rhynchophylla Hance [FRH] and Geranium koreanum Kom. [GKK]) of the 30 plant extracts had potent antibacterial activities against C. perfringens ATCC 13124 in two assays. The MIC values for FRH and GKK against 20 C. perfringens isolates were $128{\sim}256{\mu}g/mL$ and $32{\sim}128{\mu}g/mL$, respectively. The geometric MIC mean values for the two extracts were $147.2{\mu}g/mL$ and $68.8{\mu}g/mL$, respectively. The MBCs for the two extracts against the same strains were $1,024{\sim}2,048{\mu}g/mL$ and $256{\sim}1,024{\mu}g/mL$, respectively. The geometric mean MIC and MBC for GKK were about two-fold lower than those of FRH. The modified spot-on-lawn assay may be useful for measuring primary antibacterial potential. FRH and GKK are expected to be used as feed additives to prevent or treat NE in veterinary practice.

• Journal of Food Hygiene and Safety
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• v.36 no.1
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• pp.93-99
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• 2021

This study was undertaken to compare the efficacy of different sample preparation (stomaching, pulsifying, and sonication) and DNA extraction methods (boiling and commercial kit) for detection of enterotoxin-producing Clostridium perfringens from produce by polymerase chain reaction (PCR). Each produce type was inoculated at concentrations of 102, 103, 104, 105, 106, and 107 spores/g. Produce inoculated with spores was treated with three sample preparation methods, and DNA was extracted by boiling method and a commercial kit, followed by PCR. The detection limit of stomached samples was lower than that of pummeled and sonicated samples by 10-100 times. Moreover, the DNA extraction efficiency of the commercial kit was found to be superior to that of boiling. In particular, the PCR efficiency of cherry tomato and perilla leaf samples was greatly affected by sample preparation and DNA extraction method. These data suggest that DNA extraction with a commercial kit after pulsification is an optimum sample preparation method for detection of C. perfringens by PCR.

• Journal of Microbiology and Biotechnology
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• v.13 no.4
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• pp.522-528
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• 2003

The growth responses of Phellodendron amurense root-derived materials against seven intestinal bacteria were examined, using an impregnated paper disk agar diffusion method and spectrometric method under $O_2$-free condition. The biologically active constituent of the P. amurense root extract was characterized as berberine chloride ($C_{20}H_{18}NO_{41}Cl$) using various spectroscopic analyses. The growth responses varied depending on the bacterial strain, chemicals, and dose tested. At 1 mg/disk, berberine chloride strongly inhibited the growth of Clostridium perfringens, and moderately inhibited the growth of Escherichia coli and Streptococcus mutans without any adverse effects on the growth of three lactic acid-bacteria (Bifidobacterium bifidum, B. longum, and Lactobacillus acidophilus). The structure-activity relationship revealed that berberine chloride exhibited more growth-inhibiting activity against C. perfringens, E. coli, and S. mutans than berberine iodide and berberine sulfate. These results, therefore, indicate that the growth-inhibiting activity of the three berberines was much more pronounced as chloridated analogue than iodided and sulphated analogues. As for the morphological effect caused by 1 mg/disk of berberine chloride, most strains of C. perfringens were damaged and killed, indicating that berberine chloride showed a strong inhibition against C. perfringens. As naturally occurring growth-inhibiting agents, the P. amurense root-derived materials described could be useful as a preventive agent against diseases caused by harmful intestinal bacteria such as clostridia.

• Food Science and Biotechnology
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• v.14 no.5
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• pp.685-688
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• 2005

Antibacterial and antiplatelet activities of Acorus gramineus rhizome-derived asaronaldehyde and asaron were analyzed using platelet aggregometer and six human intestinal bacteria. Active constituent of A. gramineus rhizome was isolated and characterized as asaronaldehyde by spectral analyses. At 2 and 1 mg/disk, asaronaldehyde exhibited strong inhibition of Clostridium perfringens and C. difficile without adverse effects on growth of beneficial bacteria such as Bifidobacterium bifidum, Lactobacillus acidophilus, and L. casei. Asaron also revealed moderate growth inhibition against C. perfringens and C. difficile at 2 mg/disk, no growth-inhibiting activity was observed on B. bifidum, L. acidophilus, L. casei, and E. coli. At 50% inhibitory concentration ($IC_{50}$) value, asaronaldehyde was effective in inhibiting platelet aggregation induced by collagen ($IC_{50}$, $27.6\;{\mu}M$) and arachidonic acid ($IC_{50}$, $53.7\;{\mu}M$). These results suggest asaronaldehyde may be useful as lead compound for inhibiting platelet aggregation induced by collagen and arachidonic acid.

• Korean Journal of Veterinary Research
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• v.37 no.1
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• pp.137-146
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• 1997

About Clostridium perfringens causing clinically necrotic enteritis or isolated from the intestinal contens of healthy chicken, We examined the usefulness of a rapid identification method by gas-liquid chromatography as well as the types of toxins. For this study, there were used 169 chickens including 116 broilers, 27 layers and 26 breeders which collected from 9 healty flock and 21 diseased flock showing necrotic enteritis. Among them, Cl perfringens was isolated from 30 chickens(17.8%) including 7 breeders(26.9%), 5 layers(18.5%) and 18 broilers(15.5%). Isolation of Cl perfringens was mainly from ceca (100%) and followed by small intestines(70.0%) and livers(16.7%), respectively. Average concentration of the pathogen in intestinal contents was $10^{3.8}CFU/g$ in cases occuring necrotic enteritis and on the contrary $10^{3.8}CFU/g$ in healthy cases. All isolates tested showed the same characterstics in biochemical tests compared to those in standard strain. Analysis of gas-liquid chromatography to volatile fatty acids produced by Cl perfringens in PYG broth showed the typical peaks of acetic and butyric acids compatible with the standard chromogram and was confirmed as a effective and reliable tool for rapid identification of the bacteria. Toxin types of 30 strains were mostly classified in A type(26 isolates) and the rest in C type(2 isolates) and unidentifed type(2 isolates). All the isolates were highly susceptible to amphicillin, amoxicillin and cephalothin.

• Journal of the East Asian Society of Dietary Life
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• v.19 no.5
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• pp.798-802
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• 2009

This study was performed to assess the presence of contaminated microorganisms of Escherichia coli, Clostridium perfringens, and Bacillus cereus in the 112 commercial Saengshik products. E. coli was not detected in all the samples, but C. perfringens was detected in 11 products (9.8%). The number of the bacteria was less than 100 CFU/g, which was satisfactory to KFDA microbiological requirement. B. cereus was detected less than $10^2{\sim}10^3$ CFU/g in 7 products and $10^3{\sim}10^4$CFU/g in 13 products out of 25 products. Those detected bacteria from tryptose sulphite cycloserine agar and mannitol egg yolk polymyxin agar showed the typical characteristics of Gram positive and contained lecithinase, which can decompose egg-yolks layers in the biochemical test. Therefore, much more attention must be applied to satisfy the B. cereus requirement for Saengshik products sold in the market.

• Food Science and Biotechnology
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• v.14 no.3
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• pp.420-423
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• 2005

Effects of Dasom Valley and Bora Valley on fecal microflora, fecal moisture, and fecal pH of twelve healthy human volunteers were investigated. Numbers of Bifidobacterium, Clostridium perfringens, Escherichia coli, and Lactobacillus of control group were $9.24{\pm}0.63$, $4.44{\pm}1.21$, $7.75{\pm}0.38$, and $6.98{\pm}0.81$ (Log CFU/g wet feces), respectively. During administration of Dasom Valley, numbers of Bifidobacterium and Lactobacillus were $10.70{\pm}0.44$ and $8.84{\pm}0.77$, whereas those of C. perfringens and E. coli were $2.96{\pm}1.50$ and $6.69{\pm}0.29$, respectively. Administration of Dasom Valley significantly increased growth responses of beneficial bacteria, Bifidobacterium and Lactobacillus, whereas those of harmful bacteria, C. perfringens and E. coli, significantly decreased. Moisture content of feces increased and fecal pH decreased with intake of Dasom Valley. Intake of Bora Valley slightly increased numbers of Bifidbacterium and Lactobacillus and slightly decreased those of C. perfringens and E. coli. Results indicate Dasom Valley has greater intestinal-modulating effect than Bora Valley and Atlantic. Daily intake of Dasom Valley may normalize disturbed physiological functions, resulting in improvement of growth and composition of microbial community within intestinal tract.

• Journal of Veterinary Clinics
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• v.34 no.2
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• pp.112-114
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• 2017

Two one-month-old dairy calves which have Eimeria oocysts in their bloody diarrhea died acutely a few days after showing the first clinical signs. At necropsy, hemorrhagic and congestive gastrointestinal organs were observed in both calves, and abomasal ulcerations existed. As a prevalent agent in all of the collected intra-intestinal specimens, Clostridium perfringens was isolated and the strain was identified as type A possessing alpha and beta2-toxins. In these clinical cases, intercurrent infection by C. perfringens type A and Eimeria through contaminated environment may be responsible for acute hemorrhagic enteritis.

• Journal of Veterinary Clinics
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• v.32 no.2
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• pp.154-157
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• 2015

A major cause of diarrhea in a dog is an infection with bacteria which include Salmonella spp., Campylobacter (C.) spp., and Clostridium (Cl.) spp.. It is fastidious to identify these bacteria by the culture. The purpose of this experiment is to devise the method for detecting Cl. perfringens, C. jejuni, C. coli, and Salmonella spp. with rapid and high sensitivity. The fecal samples collected from 71 normal and 66 diarrheic dog feces were used to compare the prevalence of the enteric pathogens and to develop a multiplex real-time polymerase chain reaction (PCR) assay for clinical use. Detection of Cl. perfringens, C. coli, and C. jejuni in diarrhea feces was higher than normal feces. A developed multiplex real-time PCR is useful for determining the presence and quantity of pathogen-specific or other unique sequences with in a fecal sample.

• Journal of Microbiology and Biotechnology
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• v.26 no.1
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• pp.38-43
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• 2016

Novel and specific recognition elements are of central importance in the development of a pathogen detection method. Here, we describe a simple method for identifying the cell-wall binding domain (CBD) from a sequenced bacterial genome employing homology search for phage lysin genes. A putative CBD (CPF369_CBD) was identified from a genome of Clostridium perfringens type strain ATCC 13124, and its function was studied with the CBD-GFP fusion protein recombinantly expressed in Escherichia coli. Fluorescence microscopy showed the specific binding of the fusion protein to C. perfringens cells, which demonstrates the potential of this method for the identification of novel bioprobes for specific detection of pathogenic bacteria.