• 제목/요약/키워드: Inoculation method

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Development of an IoT Device for Detecting Escherichia coli from Various Agri-Foods and Production Environments (IoT 적용 대장균 검출기 개발과 농식품 및 생산환경에 적용)

  • Nguyen, Bao Hung;Chu, Hyeonjin;Kim, Won-Il;Hwang, Injun;Kim, Hyun-Ju;Kim, Hwangyong;Ryu, Kyoungyul;Kim, Se-Ri
    • Journal of Food Hygiene and Safety
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    • 제34권6호
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    • pp.542-550
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    • 2019
  • To detect Escherichia coli from agri-food and production environments, a device based on IoT (internet of things) technology that can check test results in real time on a mobile phone has been developed. The efficiency of the developed device, which combines an incubator equipped with a UV lamp, a high-resolution camera and software to detect E. coli in the field, was evaluated by measuring the device's temperature, detection limit, and detection time. The device showed a difference between its programmed temperature setting and actual temperature of about 1.0℃. In a detection limit test performed with a single-colony inoculation, a color change to yellow and a florescent signal were detected after 12 and 15 h incubations, respectively. The incubation time also decreased along with increasing bacteria levels. When applying the developed method and device to various samples, including utensils, gloves, irrigation water, seeds, and vegetables, detection rates of E. coli using the device were higher than those of the Korean Food Code method. These results show that the developed protocol and device can efficiently detect E. coli from agri-food production environments and vegetables.

Purtification of Parasporal Protein Crystals of Bacillus thuringiensis (Bacillus thuringiensis의 내독소 단백질의 분리1)

  • Kim, Yeong-Hun;Kim, Sang-Hyeon;Gang, Seok-Gwon
    • Journal of Sericultural and Entomological Science
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    • 제33권1호
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    • pp.32-36
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    • 1991
  • The study has been carried out to acquire some basic informations about Bacillus thuringiensis for developing the microbial pesticide. Three strains of Bacillus thuringiensis var. Kurstaki, dendrolimus and aizawai, were used in the experiments as follows. Growth characteristics of each strain were examined and parasporal protein crystals were isolated from the mixtures of spores and protein crystals by the new method of centrifugation in two-layer cushion of Renograffin using a fixed angle rotor. The results are as follows. 1. No difference was shown in growth characterestics among three strains of B. thuringiensis. In growth curves, all strains reached to exponential phase by 2 hr and stationary phase by 7-8 hr after inoculation. 2. The pH of the culture media during exponential growth stage decreased about 1.4 of a pH unit at the beginning of sporulation, but recovered during the early stage of sporulation and then remained nearly constant during the later stage. 3. As 10$m\ell$ sample was applied to two-layer cushion of Renograffin and then centrifuged for 1hr at 27,000g a fixed angle rotor, the purity and recovery ratio was 99.9% and 5.8%, respectively. It has been shown that the new method for the isolation of parasporal protein crystals was more efficient than any from the estabilished methods.

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A Study on the Method of Culture for Paecilomyces japonica Using an Egg (계란을 이용한 눈꽃동충하초 재배방법에 관한 연구)

  • 강한석;손장호;이길왕;김선구;조병욱;신택순;전해열
    • Korean Journal of Organic Agriculture
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    • 제11권1호
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    • pp.67-77
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    • 2003
  • This study was conducted to established method of culture for Paecilomyces japonica using an egg. Mycelia grew favorably at the temperature of 22~26$^{\circ}C$ on eggs. 5.1g of dry matter basis(average 7.2cm of longer and 199.6 of numbers) of artificial fruiting bodies were harvested at 60 days after inoculation from one of egg. Commercial fruiting bodies of Paecilomyces japonica from silkworms was used for comparative nutriental contents. Cordycepin contents of fruit bodies of Paecilomyces japonica cultivated on eggs and silkworms were not significantly different. Crude fat contents of fruiting bodies of Paeilomyces japonica cultivated from eggs was significantly higher than from silkworms(P<0.05). Mn and Cu contents of fruiting bodies of Paecilomyces japonica cultivated from silkworms were significantly higher than from eggs(P<0.05), but Na, Mg, Fe and Zn contents were significantly higher from eggs(P<0.05). Glycine, Arginine and Proline contents in the fruiting bodies of Paecilomyces japonica cultivated from silkworms were tend to higher than from eggs, but Serine, cystein. methionine, isoleucine and phenylalanin were tend to higher from eggs. These results were made possible that possible mass production of artificial fruiting bodies of Paecilomyces Japonica cultivated on eggs.

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Comparative Study on the Characteristics of Microalgae as Standard Species for Marine Ecotoxicity Tests (Skeletonema sp., Dunaliella tertiolecta) (해양생태독성시험 표준생물로서 미세조류의 특성 비교 연구(Skeletonema sp., Dunaliella tertiolecta))

  • Kim, Tae Won;Moon, Chang Ho;Lee, Su Jin
    • Journal of the Korean Society of Marine Environment & Safety
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    • 제26권5호
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    • pp.514-522
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    • 2020
  • To understand the ecotoxicological differences between representative Skeletonema sp. and Dunaliella tertiolecta, both producers as international standard test species for marine ecotoxicity testing, we compared each standard test method, and comparatively analyzed the suitability of the species for environmental assessment and their sensitivity to various test substances. Although most of the test conditions were the same in each method, there were differences in limitation of pH changing and the initial inoculation density in the validation criteria, which is supposed to originate from the low growth rate of D. tertiolecta. In terms of suitability, both species showed consistency in test performance by repeatedly meeting the validation criteria required by the standard test methods. The salinity ranges available for testing were 20 and 10 psu for Skeletonema sp. and D. tertioelecta, respectively. Finally, regarding sensitivity, the toxicity sensitivity of Skeletonema sp. was relatively higher than that of D. tertiolecta for the reference toxicant, actual polluted water discharged (ballast water), and other chemicals. This implies that using at least two species of microalgae from different classification groups could help increase the reliability and objectivity of test results in the performance of marine ecotoxicity tests using producers.

Blackeye Cowpea Mosaic Virus and Cucumber Mosaic Virus Causing Mosaic Disease on Asparagus Bean (Vigna sesquipedalis) in Korea (동부(Vigna sesquipedalis)에 발생하는 Blackeye Cowpea Mosaic Virus와 Cucumber Mosaic Virus에 관한 연구)

  • Yoon Tae Kyu
    • Korean Journal Plant Pathology
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    • 제3권4호
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    • pp.291-298
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    • 1987
  • Samples showing mosaic symptom of cowpea (Vigna sesquipedalis) with vein banding, chlorotic spot, vein yellow were collected from Chinju areas in Korea, Two viruses were distinguishable by stability in sap, host range, and relations with cells and tissues were examined under an electron microscope, Blackeye cowpea mosaic(BICMV) was sap-transmissible to 7 plant species in 2 families, Of the plants, only leguminous species were systemically infected. This virus was inactivated by heating at $50-65^{\circ}C$ for 10 min, by diluting at $10^{-4}-10^{-5}$, and aging at room temperature for 1-6 days. Preparations examined under the electron microscope by direct negative staining method(DN -method) always showed particles of flexuous filament bout 750nm in length and cytopasmic inclusions. Cytoplasmic inclusions and virus particles were also confirmed to present in the cytoplasm of a mesophyll cell by ultrathin sections of BICMV infected cowpea leaves. Cucumber mosaic virus (CMV) was transmitted by sap- inoculation on inoculated leaves of Chenopodium amaranticolor, C. quinoa producing local lesions, but non-inoculated upper leaves of Nicotiana glutinosa, Cucurbita pepo and Vigna sesquipedalis producting systemic mosaic symptoms. Electron microscopic examination of virus preparation by direct negative staining showed spherical particles of about 30nm in diameter. In ultrathin sections of CMV infected tissues, virus particles of crystalline array were found in the vacuole and a large number of virus particles were found in the cytoplasm and the plasmodesmata of mesophyll cells.

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Fine mapping of qBK1, a major QTL for bakanae disease resistance in rice

  • Ham, Jeong-Gwan;Cho, Soo-Min;Kim, Tae Heon;Lee, Jong-Hee;Shin, Dongjin;Cho, Jun-Hyun;Lee, Ji-Yoon;Yoon, Young-Nam;Song, You-Chun;Oh, Myeong-Kyu;Park, Dong-Soo
    • Proceedings of the Korean Society of Crop Science Conference
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    • 한국작물학회 2017년도 9th Asian Crop Science Association conference
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    • pp.92-92
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    • 2017
  • Bakanae disease is one of the most serious and oldest problems of rice production, which was first described in 1828 in Japan. This disease has also been identified in Asia, Africa, North America, and Italy. Germinating rice seeds in seed boxes for mechanical transplantation has caused many problems associated with diseases, including bakanae disease. Bakanae disease has become a serious problem in the breeding of hybrid rice, which involves the increased use of raising plants in seed beds. The indica rice variety Shingwang was selected as resistant donor to bakanae disease. One hundred sixty nine NILs, YR28297 ($BC_6F_4$) generated by five backcrosses of Shingwang with the genetic background of susceptible japonica variety, Ilpum were used for QTL analysis. Rice bakanae disease pathogen, CF283, was mainly used in this study and inoculation and evaluation of bakanae disease was performed with the method of the large-scale screening method developed by Kim et al. (2014). SSR markers evenly distributed in the entire rice chromosomes were selected from the Gramene database (http://www.gramene.org), and the polymorphic markers were used for frame mapping of a $BC_5F_5$ resistant line. Here, we developed 168 near-isogenic rice lines (NILs, $BC_6F_4$) to locate a QTL for resistance against bakanae disease. The lines were derived from a cross between Shingwang, a highly resistant variety (indica), and Ilpum, a highly susceptible variety (japonica). The 24 markers representing the Shingwang allele in a bakanae disease-resistant NIL, YR24982-9-1 (parental line of the $BC_6F_4$ NILs), were located on chromosome 1, 2, 7, 8, 10, 11, and 12. Single marker analysis using an SSR marker, RM9, showed that a major QTL was located on chromosome 1. The QTL explained 65 % of the total phenotype variation in $BC_6F_4$ NILs. The major QTL designated qBK1 was mapped in 91 kb region between InDel15 and InDel21. The identification of qBK1 and the closely linked SSR marker, InDel18, could be useful for improving rice bakanae disease resistance in marker-assisted breeding.

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Studies on the Applicability of Furazolidone to the Silkworm Rearing Industry as a Useful Remedy for Certain Silkworm Diseases (Report I) (푸라졸리돈의 누에병치료약으로서의 응용가능성에 관한 연구 (제1보))

  • 이장락
    • Journal of Sericultural and Entomological Science
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    • 제15권1호
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    • pp.15-25
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    • 1973
  • The author studied the applicability of Furazolidone to the silkworm rearing industry as a useful remedy for certain silkworm diseases, at the silkworm rearing house of the college of agriculture, Seoul national university, during both the spring and the autumn silkworm rearing season of 1972. Discovering the fact that Furazolidone, when put on the mulberry leaves in a powdered form, is eaten along with the leaves by silkworms and thus the systematic administration of Furazolidone to silkworms is possible, the experimenter carried on a series of experiments (1. determining the in vitro antibacterial activity of Furazolidone to four pathogens of silkworm diseases-Bacillus thuringiensis, Aspergillus oryzae, Aspergillus flavus, and Isaria farinosa, 2. observing the prophylactic and therapeutic effect of Furazolidone against the experimental flacherie caused by inoculation of B. thuringiensis, and 3. examining the toxicity of Furazolidone to silkworm larvae). As the results of the experiments the investigator found out the fundamental fact that Furazolidone exerts a good prophylactic and therapeutic effect against flacherie which is the most common and important silkworm disease: Furazolidone, in in vitro test, inhibited completely the growth of B. thuringiensis, the pathogen of bacterial flacherie, at the concentration of 1 ${\mu}$g/mι. with the tube method and at the concentration of 5 ${\mu}$g/mι. with the plate method, and the drug showed an excellent prophylactic effect and a considerably good therapeutic effect, depending on the time of administration, on the 5th instar silkworms inoculated B. thuringiensis, at the tentative dose of 150mg. per 10 silkworms administered once a day for 2 days. For the practical administration of Furazolidone against flacherie, the dose, the time and duration of administration, and the form of preparation, will be investigated more closely.

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Experimental Studies on Differential Diagnosis of Non-pathogenic Lesion Dairy Cattle with Positive Tuberculin Reaction (Tuberculin 양성(陽性) 무병소유우(無病巢乳牛)의 감별진단(鑑別診斷)에 관한 실험적연구(實驗的硏究))

  • Kim, Jong Myeon
    • Korean Journal of Veterinary Research
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    • 제16권2호
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    • pp.151-158
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    • 1976
  • The tuberculin test have been widely utilized to diagnose tuberculosis of the dairy cattle. It was reported that approximately 70% of the dairy cattle with positive tuberculin reaction in Korea had been non-pathogenic lesion. So the studies on the specific method to diagnose tuberculosis of them is really need in Korea. Therefore this study investigated upon the diagnostic method for cattle tuberculosis in the aspect of cellular-immunology. The results obtained in this investigation are summarized as follows: 1. All the tuberculin tests to the swine inoculated with BCG.(B group), Mycobacterium avium (A group) and Mycobacterium smegmatis (S group), respectively, were represented high positive reaction and were no differences in the species of inoculated bacteria. 2. In the migration inhibition test using Iymphocyte in circulating blood of the swine inoculated with three species of acid-fast bacteria respectively, the test to A group was represented slight positive for migration in the presence of $15{\mu}g/ml$-PPD and the other reaction were clear and total positive for migration inhibition in the presence of $25{\mu}g/ml$-PPD or more, and the test to B group was the same results as to A group approximately. The test to S group was represented slight positive for migration inhibition in the presence of $15{\mu}g/ml$-PPD, but the results were the same in the presence of $25{\mu}g/ml$-PPD and $35{\mu}g/ml$-PPD. These results showed that there were remarkable differences between group A, B and group S in the presence of $25{\mu}g/ml$-PPD and $35{\mu}g/ml$-PPD. 3. The transformation rates of lymphocyte in PPD treated tissue culture had no significance without any relation between PPD treatment and non-treatment but A group and B group showed significance. And A group and B group showed high significance in comparison with N group and S group in the LSD examination. 4. The infection rated in lymphocyte of BCG inoculated after 3 days tissue culturing were represented those of high infection but its cellular degeneration rates almost did not change. The infection rates in bacilli in N group and S group were low but after 3 days inoculation it shewed higher cellular degeneration.

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Antibacterial Effects of Atmospheric Plasma against Main Foodborne Bacteria on the Surface of Dried Filefish (Stephanolepis cirrhifer) Fillets (대기압 플라즈마 처리에 의한 쥐치포 중 주요 식중독세균의 살균 효과)

  • Park, Shin Young
    • Journal of Food Hygiene and Safety
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    • 제34권2호
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    • pp.178-182
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    • 2019
  • This study investigated the antibacterial effects of BioZone atmospheric plasma (AP) against Bacillus cereus (F4810/72) and Staphylococcus aureus (ATCC 6538) as the major foodborne bacteria on the surface of dried filefish (Stephanolepis cirrhifer) fillets. The fillets were experimentally contaminated with 7-8 log CFU/mL of B. cereus or S. aureus using a spot inoculation method. Bacterial counts were measured by standard plate method on tryptic soy agar, and were significantly reduced with the increase in the treatment time (1, 3, 5 or 20 min) of AP on the fillets (p < 0.05). The reductions of the pathogens by AP treatment ranged from 0.9 to 2.93 logCFU/g for B. cereus and from 1.04 to 2.55 logCFU/g for S. aureus. A reduction of >1-logCFU/g for B. cereus and S. aureus was observed on the fillets treated with AP for >3 min. The differences in color on the Hunter scale (L=light vs. dark, a=red vs. green, b=yellow vs. blue) of the fillets were not significantly different between the nontreated (control) and AP-treated fillets (p>0.05). This study suggested that 3 min of AP could be effective in reducing >90% of the bacteria without causing any concomitant changes in the color of the fillets.

Comparison of SureTectTM with phenotypic and genotypic method for the detection of Salmonella spp. and Listeria monocytogenes in ready-to-eat foods (즉석섭취식품에 존재하는 Salmonella spp.와 Listeria monocytogenes의 검출을 위한 SureTectTM와 표현형 및 유전자형 방법의 비교)

  • Kye-Hwan Byun;Byoung Hu Kim;Ah Jin Cho;Eun Her;Sunghee Yoon;Taeik Kim;Sang-Do Ha
    • Food Science and Preservation
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    • 제30권2호
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    • pp.262-271
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    • 2023
  • The objective of this study is to compare and assess the effectiveness of real-time polymerase chain reaction (RT-PCR), loop-mediated isothermal amplification (LAMP), and the selective agar plate method for the detection of Salmonella spp. and Listeria monocytogenes in ready-to-eat (RTE) foods. In RTE foods, the detection performance of the three methods (RT-PCR [SureTectTM kit and PowerChekTM kit], LAMP [3M MDS], selective agar) were similar at 0-10, 10-50, 50-100, and 100- CFU/mL of Salmonella spp. and L. monocytogenes. We found that with RT-PCR, the Ct value of salad was significantly higher (p<0.05) than other RTE foods, indicating that fiber plays a critical role as an obstacle to the rapid detection of Salmonella spp. However, the Ct value displayed a mixed pattern according to the inoculation level of L. monocytogenes. The use of rapid detection kits and machines mostly depends on the user's choice, with accuracy, ease of use, and economy being the primary considerations. As an RT-PCR kit, SureTectTM and PowerChekTM showed high accuracy in detecting Salmonella spp. and L. monocytogenes in RTE foods, showing that they can replace the existing RT-PCR kits available. Additionally, LAMP also showed excellent detection performance, suggesting that it has the potential to be used as a food safety management tool.