• Title/Summary/Keyword: Infectivity

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Control of oomycete pathogens during Pyropia farming and processing using calcium propionate

  • Yong Tae Kim;Ro-won Kim;Eunyoung Shim;Hana Park;Tatyana A. Klochkova;Gwang Hoon Kim
    • ALGAE
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    • v.38 no.1
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    • pp.71-80
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    • 2023
  • The oomycete pathogens Pythium porphyrae, causing red rot disease, and Olpidiopsis spp. causing Olpidiopsis-blight, cause serious economic losses to Pyropia sea farms in Korea. During the washing step for Pyropia processing, these pathogens proliferate rapidly, significantly reducing the quality of the final product. To develop non-acidic treatments for these pathogens, various calcium salts were tested against the infectivity of P. porphyrae and Olpidiopsis pyropiae on Pyropia gametophytes, and calcium propionate was the most effective. When Pyropia blades were immersed in 10 mM calcium propionate for 1 h after inoculation with the oomycete pathogen, infection rate of both oomycete pathogens on day 2 was significantly lower (7.1%) than control (>95%). Brief incubation of Pyropia blades in calcium propionate also reduced the spread of infection. The infected area of Pyropia thallus was reduced to 14.3% of the control in 2 days after treatment with 100 mM calcium propionate for 30 s. In field experiments conducted in actual aquaculture farms, it has been shown that a brief 30 s wash every two weeks with 100 mM calcium propionate can effectively reduce the spread of oomycetes throughout the entire culture period. The above results suggest that calcium propionate can be a useful means for controlling the spread of oomycetes not only during laver processing but also during aquaculture.

Experimental infection of a porcine kidney cell line with hepatitis A virus

  • Dong-Hwi Kim;Da-Yoon Kim;Jae-Hyeong Kim;Kyu-Beom Lim;Joong-Bok Lee;Seung-Yong Park;Chang-Seon Song;Sang-Won Lee;In-Soo Choi
    • Korean Journal of Veterinary Research
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    • v.63 no.2
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    • pp.15.1-15.5
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    • 2023
  • The hepatitis A virus (HAV) induces severe acute liver injury and is adapted to human and monkey cell lines but not other cells. In this study, the HAV was inoculated into porcine kidney (PK-15) cells to determine its infectivity in porcine cells. The growth pattern of the HAV in PK-15 cells was compared with its growth pattern in fetal rhesus kidney (FRhK-4) cells. The growth of HAV was less efficient in PK-15 cells. In conclusion, HAV replication was verified in PK-15 cells for the first time. Further investigations will be needed to identify the HAV-restrictive mechanisms in PK-15 cells.

Evaluation of Cryptosporidiurn Disinfection by Ozone and Ultraviolet Irradiation Using Viability and Infectivity Assays (크립토스포리디움의 활성/감염성 판별법을 이용한 오존 및 자외선 소독능 평가)

  • Park Sang-Jung;Cho Min;Yoon Je-Yong;Jun Yong-Sung;Rim Yeon-Taek;Jin Ing-Nyol;Chung Hyen-Mi
    • Journal of Life Science
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    • v.16 no.3 s.76
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    • pp.534-539
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    • 2006
  • In the ozone disinfection unit process of a piston type batch reactor with continuous ozone analysis using a flow injection analysis (FIA) system, the CT values for 1 log inactivation of Cryptosporidium parvum by viability assays of DAPI/PI and excystation were $1.8{\sim}2.2\;mg/L{\cdot}min$ at $25^{\circ}C$ and $9.1mg/L{\cdot}min$ at $5^{\circ}C$, respectively. At the low temperature, ozone requirement rises $4{\sim}5$ times higher in order to achieve the same level of disinfection at room temperature. In a 40 L scale pilot plant with continuous flow and constant 5 minutes retention time, disinfection effects were evaluated using excystation, DAPI/PI, and cell infection method at the same time. About 0.2 log inactivation of Cryptosporidium by DAPI/PI and excystation assay, and 1.2 log inactivation by cell infectivity assay were estimated, respectively, at the CT value of about $8mg/L{\cdot}min$. The difference between DAPI/PI and excystation assay was not significant in evaluating CT values of Cryptosporidium by ozone in both experiment of the piston and the pilot reactors. However, there was significant difference between viability assay based on the intact cell wall structure and function and infectivity assay based on the developing oocysts to sporozoites and merozoites in the pilot study. The stage of development should be more sensitive to ozone oxidation than cell wall intactness of oocysts. The difference of CT values estimated by viability assay between two studies may partly come from underestimation of the residual ozone concentration due to the manual monitoring in the pilot study, or the difference of the reactor scale (50 mL vs 40 L) and types (batch vs continuous). Adequate If value to disinfect 1 and 2 log scale of Cryptosporidium in UV irradiation process was 25 $mWs/cm^2$ and 50 $mWs/cm^2$, respectively, at $25^{\circ}C$ by DAPI/PI. At $5^{\circ}C$, 40 $mWs/cm^2$ was required for disinfecting 1 log Cryptosporidium, and 80 $mWs/cm^2$ for disinfecting 2 log Cryptosporidium. It was thought that about 60% increase of If value requirement to compensate for the $20^{\circ}C$ decrease in temperature was due to the low voltage low output lamp letting weaker UV rays occur at lower temperatures.

Studies on Ancylostomiasis I. An Experimental Study on Hookworm Infection and Anemia (구충증(鉤蟲症)에 관(關)한 연구(硏究) 제1편(第1篇) 구충(鉤蟲)의 감염(感染) 및 구충성빈혈(鉤蟲性貧血)에 관(關)한 고찰(考奈))

  • Lee, Mun-Ho;Kim, Dong-Jip;Lee, Jang-Kyu;Seo, Byong-Sul;Lee, Soon-Hyung
    • The Korean Journal of Nuclear Medicine
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    • v.1 no.1
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    • pp.55-66
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    • 1967
  • In view of its prevalence in the Far East area, a more detailed knowledge on the hookworm infection is one of the very important medical problems. The present study was aimed to; determine the infectivity of the artificially hatched ancylostoma duodenale larvae in man after its oral administration, evaluate the clinical symptomatology of such infection, determine the date of first appearance of the ova in the stool, calculate the blood loss per worm per day, assess the relation-ships between the ova count, infectivity(worm load), blood loss and severity of anemia. An erythrokinetic study was also done to analyse the characteristics of hookworm anemia by means of $^{59}Fe\;and\;^{51}Cr$. Materials and Methods Ten healthy male volunteers(doctors, medical students and laboratory technicians) with the ages ranging from 21 to 40 years were selected as the experimental materials. They had no history of hookworm infection for preceding several years, and care was taken not to be exposed to reinfection. A baseline study including a through physical examinations and laboratory investigations such as complete blood counts, stool examination and estimation of the serum iron levels was done, and a vermifuge, bephenium hydroxynaphoate, was given 10 days prior to the main experiment. The ancylostoma duodenale filariform larvae were obtained in the following manner; The pure ancylostoma duodenale ova were obtained from the hookworm anemia patients and a modified filter paper method was adopted to harvest larger number of infective larvae, which were washed several times with saline. The actively moving mature larvae were put into the gelatine capsules, 150 in each, and were given to the volunteers in the fasting state with 300ml. of water. The volunteers were previously treated with intramuscular injection of 15mg. of chlorpromazine in order to prevent the eventual nausea and vomiting after the larvae intake. The clinical symptoms and signs mainly of the respiratory and gastrointestinal tracts, appearance of the ova and occult blood in the stool etc. were checked every day for the first 20 days and then twice weekly until the end of the experiment, which usually lasted for about 3 months. Roentgenological survey of the lungs was also done. The hematological changes such as the red blood cell, white blood cell and eosinophil cell counts, hemoglobin content and serum iron levels were studied. The appearance of the ova in the stool was examined by the formalin ether method and the ova were counted in triplicate on two successive days using the Stoll's dilution method. The ferrokinetic data were calculated by the modified Huff's method and the apparent half survival time of the red blood cells by the modified Gray's method. The isotopes were simultaneously tagged and injected intravenously, and then the stool and blood samples were collected as was described by Roche et al., namely, three separate 4-day stool samples with the blood sample drawing before each 4-day stool collection. The radio-activities of the stools ashfied and the blood were separately measured by the pulse-height analyser. The daily blood loss was calculated with the following formula; daily blood loss in $ml.=\frac{cpm/g\;stool{\times}weight\;in\;g\;of\;4-day\;stool}{cpm/ml\;blood{\times}4}$ The average of these three 4-day periods was given as the daily blood loss in each patient. The blood loss per day per worm was calculated by simply dividing the daily blood loss by the number of the hookworm recovered after the vermifuge given twice a week at the termination of the experiment. The iron loss in mg. through the gastrointestinal tract was estimated with the daily iron loss in $mg=\frac{g\;Hgb/100ml{\times}ml\;daily\;blood\;loss{\times}3.40}{100}$ 3.40=mg of iron per g Hgb following formula; Results 1. The respiratory symptoms such as cough and sputum were noted in almost all cases within a week after the infection, which lasted about 2 weeks. The roentgenological findings of the chest were essentially normal. A moderate degree of febril reaction appeared within 2 weeks with a duration of 3 or 4 days. 2. The gastrointestinal symptoms such as nausea, epigastric fullness, abdominal pain and loose bowel appeared in all cases immediately after the larvae intake. 3. The reduction of the red blood cell count was not remarkable, however, the hemoglobin content and especially the serum iron level showed the steady decreases until the end of the experiment. 4. The white blood cells and eosinophil cells, on the contrary, showed increases in parallel and reached peaks in 20 to 30 days after the infection. A small secondary rise was noted in 2 months. 5. The ova first appeared in the stool in 40. 1 days after the infection, ranging from 29 to 51 days, during which the occult blood reaction of the stool became also positive in almost cases. 6. The number of ova recovered per day was 164, 320 on the average, ranging from 89,500 to 253,800. The number of the worm evacuated by vermifuge was in rough correlation with the number of ova recovered. 7. The infectivity of ancylostoma duodenale was 14% on the average, ranging from 7.3 to 20.0%, which is relatively lower than those reported by other workers. 8. The mean fecal blood loss was 5.78ml. per day, with a range of from 2.6 to 11.7ml., and the mean blood loss per worm per day was 0.30ml., with a range of from 0.13 to 0.73ml., which is in rough coincidence with those reported by other authors. There appeared to exist, however, no correlation between the blood loss and the number of ova recovered. 9. The mean fecal iron loss was 2.02mg. per day, with a range of from 1.20 to 3.89mg., which is less than those appeared in the literature. 10. The mean plasma iron disappearance rate was 0.80hr., with a range of from 0.62 to 0.95hr., namely, a slight accerelation. 11. The hookworm anemia appeared to be iron deficiency in origin caused by continuous intestinal blood loss.

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Comparison of Resistance Level to Cotton leaf curl virus(CLCuV) Among Newly Developed Cotton Mutants and Commercial Cultivars

  • Akhtar, Khalid P.;Khan, Azeem I.;Hussain, M.;Khan, M.S.I.
    • The Plant Pathology Journal
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    • v.18 no.4
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    • pp.179-186
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    • 2002
  • Four newly developed cotton mutants (M-111, M-7662, M-358 and M-218) were compared for their resistance against Cotton leaf curl virus(CLCuV) together with commercial resistant (CIM-443, CIM-482, CIM-473, FH-900 and FH-901) and susceptible (5-12) varieties by artificial inoculation through grafting and under natural field conditions. Infectivity and success of grafting were 100% in all cases. None of the grafted plants were found immune or asymptomatic. All the grafted mutants and most of their single plant progeny rows (SPPRs) showed highly resistant responses as the symptoms displayed by these mutants were milder than the commercial cultivars. Grafted mutants also had delayed disease reactions as they took more time (25-30 days) to produce disease symptoms, as compared with resistant commercial varieties that produced disease 18-22 days after inoculation. Growth of the grafted SPPRs of tested mutants was normal, which is an indication that there will be no production losses. Observations under natural infestation of whitefly showed that two SPPRs of M-ll/CE and M-7662-1/2 and one resistant variety CIM-443 exhibited slight incidence of disease, while one SPPR of M-l1/59 and S-12 were moderately susceptible and highly susceptible with 21% and 97.l% disease incidence, respectively. This study also showed that plants displaying more disease symptoms through grafting were easily infected under natural conditions. These results suggest that preference should be given to those plants that exhibited highly resistant responses after artificial inoculation.

Analysis of Natural Recombination in Porcine Endogenous Retrovirus Envelope Genes

  • Lee, Dong-Hee;Lee, Jung-Eun;Park, Nu-Ri;Oh, Yu-Kyung;Kwon, Moo-Sik;Kim, Young-Bong
    • Journal of Microbiology and Biotechnology
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    • v.18 no.3
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    • pp.585-590
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    • 2008
  • Human tropic Porcine Endogenous Retroviruses (PERVs) are the major concern in zoonosis for xenotransplantation because PERVs cannot be eliminated by specific pathogen-free breeding. Recently, a PERV A/C recombinant with PERV-C bearing PERV-A gp70 showed a higher infectivity (approximately 500-fold) to human cells than PERV-A. Additionally, the chance of recombination between PERVs and HERVs is frequently stated as another risk of xenografting. Overcoming zoonotic barriers in xenotransplantation is more complicated by recombination. To achieve successful xenotransplantation, studies on the recombination in PERVs are important. Here, we cloned and sequenced proviral PERV env sequences from pig gDNAs to analyze natural recombination. The envelope is the most important element in retroviruses as a pivotal determinant of host tropisms. As a result, a total of 164 PERV envelope genes were cloned from pigs (four conventional pigs and two miniature pigs). Distribution analysis and recombination analysis of PERVs were performed. Among them, five A/B recombinant clones were identified. Based on our analysis, we determined the minimum natural recombination frequency among PERVs to be 3%. Although a functional recombinant envelope clone was not found, our data evidently show that the recombination event among PERVs may occur naturally in pigs with a rather high possibility.

Sequence and Phylogenetic Analysis of V3 Region of Human Immunodeficiency Virus Type 1 Strains Isolated from Korean Patients (한국인 인면역결핍 바이러스의 V3 Loop 염기서열 분석 및 계통발생학적 분석)

  • Kim, Young-Bong;Cho, Young-Keol;Lee, Hee-Jung;Chung, Koo-Hun;Kim, Jung-Woo;Kim, Yoo-Kyum;Yang, Jai-Myung
    • The Journal of Korean Society of Virology
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    • v.26 no.2
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    • pp.251-258
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    • 1996
  • The V3 loop, a hypervariable domain of envelope glycoprotein, has an essential role in viral infectivity and has a major epitope for type-specific neutralizing antibody. In order to investigate genetic diversity of V3 region of gp120 of human immunodeficiency virus type 1 (HIV-1) isolated from Korean patients, DNA sequences encoding the C2 to V3 region were amplified by nested polymerase chain reaction (PCR) from uncultured peripheral blood mononuclear cells obtained from 15 HIV-1 seropositive patients and nucleotide sequences were determined. All nucleotide sequences from fifteen patients were compared with 8 distinctive subtypes (A-H) and another subtype O. Phylogenetic analysis was carried out with PHYLIP ver 3.5 (Dnapars) program. Of the 15 isolates, 14 HIV-1 subjects were clustered with subtype B, while one was clustered with subtype C. Intra-subtype B distance at the nucleotide and deduced amino acid level were maximum 17.7% and 37.0%, respectively. Intra-patient distance at the nucleotide and deduced amino acid level were maximum 7.3% and 17.8%, respectively. Analysis of the nucleotide sequences revealed that Korean types have relatively well conserved sequences. These findings could be useful for assessing the source of infection and developing an AIDS vaccine.

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Polymerase Chain Reaction for the Detection of Aujeszky's Disease Virus (오제스키병 바이러스 검출을 위한 Polymerase Chain Reaction)

  • Hwang, Dong-hee;Yeo, Sang-geon
    • Korean Journal of Veterinary Research
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    • v.43 no.2
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    • pp.239-246
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    • 2003
  • Polymerase chain reaction (PCR) was evaluated for the early detection of Aujeszky's disease virus (ADV) DNA from virus-infected cell cultures. For the purposes, the Korean ADV NYJ1-87 was propagated in swine kidney (SK) cells and subjected to the amplification of DNA (217 bp) by PCR using sense and antisense primers specific to gp50 gene of the ADV. In detection of cell-associated viral DNA, reliable PCR conditions were determined as 30 cycles of reaction consisting 1 minute each of denaturation at $94^{\circ}C$, annealing at $55^{\circ}C$ and polymerization at $72^{\circ}C$. The PCR encountered best results with reagent mixtures of $50{\mu}l$ containing $200{\mu}M$ dNTPs, $0.2{\mu}M$ each sense and antisense primers, 1 mM $MgCl_2$ and 10% (v/v) template DNA in the final concentrations. ADV-specific DNAs were detected as early as 6, 6, and 9 hours post-infection, respectively, from lysates of the SK cells infected with ADV of $10^3$, $10^2$ and $10^1\;TCID_{50}/ml$ by this condition. In culture supernatant, the DNAs were detected from ADV of as low infectivity as $10^ {-3}\;TCID_{50}/ml$ by the reduced reagent concentrations and 30 cycles of 1 minute each of denaturation at $94^{\circ}C$ and annealing at $55^{\circ}C$, and 2 minutes of polymerization at $72^{\circ}C$. The lowest amount of detectable ADV DNA was 1 fg. In conclusion, the PCR condition established in the present study was recognized as a feasible alternative to time-consuming procedures in isolation and characterization of the virus.

Genetic Analysis of the 2019 Swine H1N2 Influenza Virus Isolated in Korean Pigs and Its Infectivity in Mice (2019년 국내에서 분리한 H1N2 돼지 인플루엔자바이러스 유전자 분석 및 이의 마우스에 대한 감염성)

  • Jang, Yunyueng;Seo, Sang Heui
    • Journal of Life Science
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    • v.30 no.9
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    • pp.749-762
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    • 2020
  • Influenza A viruses are circulating in a variety of hosts, including humans, pigs, and poultry. Swine influenza virus is a zoonotic pathogen that can be readily transmitted to humans. The influenza viruses of the 2009 H1N1 pandemic were derived from swine influenza viruses, and it has been suggested that the 1957 H2N2 pandemic and the 1968 H3N2 pandemic both originated in pigs. Pigs are regarded as a mixing vessel in the creation of novel influenza viruses since they are readily infected with human and avian influenza viruses. We isolated three novel H1N2 influenza viruses from pigs showing respiratory symptoms on a Korean farm in 2019. These viruses were reassortants, containing PA and NP genes from those of the 2009 H1N1 influenza virus in addition to PB2, PB1, HA, NA, M, and NS genes from those of triple-reassortant swine H3N2 and classical swine H1N2 influenza viruses circulating in Korean pigs. Mice infected with the isolated H1N2 influenza virus lost up to 17% body weight and exhibited interstitial pneumonia involving infiltration of many inflammatory cells. Results suggest that close surveillance to detect emerging influenza viruses in pigs is necessary for the health of both pigs and humans.

Biological Activities of Soyasaponins and Their Genetic and Environmental Variations in Soybean (콩 Saponin의 생리활성 기능과 함량변이)

  • 김용호
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.48
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    • pp.49-57
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    • 2003
  • There is much evidence suggesting that compounds present in soybean can prevent cancer in many different organ systems. Especially, soybean is one of the most important source of dietary saponins, which have been considered as possible anticarcinogens to inhibit tumor development and major active components contributing to the cholesterol-towering effect. Also they were reported to inhibit of the infectivity of the AIDS virus (HIV) and the Epstein-Barr virus. The biological activity of saponins depend on their specific chemical structures. Various types of triterpenoid saponins are present in soy-bean seeds. Among them, group B soyasaponis were found as the primary soyasaponins present in soybean, and th e 2, 3-dihydro-2, 5-dihydroxy-6- methyl-4H-pyran-4-one(DDMP)-conjugated soyasaponin $\alpha\textrm{g}$, $\beta\textrm{g}$, and $\beta$ a were the genuine group B saponins, which have health benefits. On the other hand, group A saponins are responsible for the undesirable bitter and astringent taste in soybean. The variation of saponin composition in soybean seeds is explained by different combinations of 9 alleles of 4 gene loci that control the utilization of soyasapogenol glycosides as substrates. The mode of inheritance of saponin types is explained by a combination of co-dominant, dominant and recessive acting genes. The funtion of theses genes is variety-specific and organ specific. Therefore distribution of various saponins types was different according to seed tissues. Soyasaponin $\beta\textrm{g}$ was detected in both parts whereas $\alpha\textrm{g}$ and $\beta$ a was detected only in hypocotyls and cotyledons, respectively. Soyasaponins ${\gamma}$g and $\gamma\textrm{g}$ were minor saponin constituents in soybean. In case group A saponins were mostly detected in hypocotyls. Also, the total soyasaponin contents varied among different soy-bean varieties and concentrations in the cultivated soy-beans were 2-fold lower than in the wild soybeans. But the contents of soyasaponin were not so influenced by environmental effects. The composition and concentration of soyasaponins were different among the soy products (soybean flour, soycurd, tempeh, soymilk, etc.) depending on the processing conditions.