• Korean Journal of Veterinary Research
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• v.30 no.4
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• pp.487-497
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• 1990

This study was carried out to observe in vitro effect of praziquantel on the tegumental changes of Heterophyopsis continua with scanning electron microscope. Metacercariae were collected from the perch, Lateolabrax japonicus, by artificial digestion technique and fed to 2-week old chickens. Adult worms were recovered from small intestine of chickens 8 days after infection. For working solutions, praziquantel was diluted with TC199 medium at concentration of 0.01, 0.1. 1 and $10{\mu}g/ml$. To each petri dish containing 10ml of solution, 5~10 worms were introduced and incubated at $37^{\circ}C$. For the scanning electron microscopy(SEM), the worms were fixed in cold 2.5% glutaraldehyde, dehydrated in a series of graded ethanol and freeze-dried. Dried specimen was mounted on stub and coated with gold and observed in an SEM. The results were as follows: 1. Severe tegumental alterations were recognized by scanning electron microscope. Bleb formation of tegument was observed in 5 minute group and most pronounced on anterior tegument of worms. The number and size of blebs increased as incubation time prolonged. 2. The surface destruction was more pronounced at ventral margin between the oral and the ventral suckers. 3. The sensory papillae were slightly affected, but destruction of tegumental spine was not recognized. 4. The effect of praziquantel on the worm was found dependent on the concentration and incubation time, however, the effect was more dependent upon the incubation time.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.2
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• pp.208-213
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• 2007

The objective of the experiment was to determine the optimum cultural [moisture levels (55, 60 and 70%), days of fermentation (7, 14 and 21), temperature (25 and $35^{\circ}C$) of incubation)] and nutritional parameters (urea addition (0 and 2%) and variable levels of single super phosphate (0.25 and 0.50% SSP)) for bio-processing of the mustard (Brassica campestris) straw (MS) under solid-state fermentation (SSF) system. The performance of SSF was assessed in terms of favorable changes in cell wall constituents, protein content and in vitro DM digestibility of the MS. Sorghum based inoculum (seed culture) of Ganoderma lucidum to treat the MS was prepared. The 50 g DM of MS taken in autoclavable polypropylene bags was mixed with a pre-calculated amount of water and the particular nutrient in the straw to attained the desired levels of water and nutrient concentration in the substrate. A significant progressive increase in biodegradation of DM (p<0.001), NDF (p<0.01) and ADF (p<0.05) was observed with increasing levels of moisture. Among the cell wall constituents the loss of ADF fraction was greatest compared to that of NDF. The loss of DM increased progressively as the fermentation proceeded and maximum DM losses occurred at 28 days after incubation. The protein content of the treated MS samples increased linearly up to the day $21^{th}$ of the incubation and thereafter declined at day $28^{th}$, whereas the improvement in in vitro DM digestibility were apparent only up to the day $14^{th}$ of the incubation under SSF and there after it declined. The acid detergent lignin (ADL) degradation was slower during the first 7 days of SSF and thereafter increased progressively and maximum ADL losses were observed at the day $28^{th}$ of the SSF. The biodegradation of DM and ADL was not affected by the variation in incubation temperature. Addition of urea was found to have inhibitory effect on fungal growth. The effect of both the levels (0.25 and 0.50) of SSP addition in the substrate, on DM, NDF, ADF, cellulose and ADL biodegradation was similar. Similarly, the protein content and the in vitro DM digestibility remain unaffected affected due to variable levels of the SSP inclusion in the substrate. From the results it may be concluded that the incubation of MS with 60 percent moisture for 21 days at $35^{\circ}C$ with 0.25 percent SSP was most suitable for MS treatment with Ganoderma lucidum. Maximum delignification, enrichment in the protein content and improvement in in vitro DM digestibility were achieved by adopting this protocol of bioprocessing of MS.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.9
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• pp.1249-1254
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• 2005

The aim of this study was to determine the effect of maturity stage on the nutritive value of wild mustard straw in terms of chemical composition, in situ, in vitro dry matter degradability and calculated ME. The nutritive values of wild mustard, Sinapsis arvensis hays harvested at three stages were evaluated by chemical composition, in vitro gas production and in situ dry matter degradation methods. Gas production or dry matter (DM) degradation were determined at 0, 3, 6, 12, 24, 48, 72 and 96 h and their kinetics were described using the equation p = a+b(1-e$^{-ct}$). Maturity had a significant effect on both the chemical composition and degradability of wild mustard. Neutral detergent fibre (NDF) and acid detergent fibre (ADF) (p<0.001) increased with increasing maturity whereas the crude protein (CP) (p<0.001) decreased. The gas produced after 96 h incubation ranged between 64.7 and 81.5 ml per 0.200 g of dry matter. The gas production (ml) at all incubation times and estimated parameters decreased with increasing maturity of wild mustard. The gas production at all incubation times and estimated parameters (a, b (a+b), metabolizable energy (ME) and organic matter digestibility (OMD)) were negatively correlated with NDF and ADF. The DM disappearance after 96 h incubation ranged between 50.8 and 76.1%. The in situ DM disappearance at all incubation times and estimated parameters decreased with increasing maturity of wild mustard. The in situ dry matter disappearance at all incubation times and some estimated parameters (c, a, b and effective dry matter degradability (EDMD)) were negatively correlated with NDF and ADF but positively correlated with CP. The nutritive value of wild mustard continually changed as it matured. Wild mustard, harvested at the proper stage of maturity offers considerable potential as a high quality forage for ruminants during the winter feeding period. The present study showed that if higher quality forage is an objective, wild mustard should be harvested at the early flowering stage.

• Korean Journal of Veterinary Service
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• v.18 no.1
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• pp.41-56
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• 1995

The present study was conducted to investigate results of B. anthracis isolated from Anthrax in the Kyong-Ju of Feb. 12. 1994. 1. In biochemical feature, B. anthracis was a gram-positive rod, non-motility, sporulation, capsulation. It was positive in gelatinase, starch hydrolysis, glucose. But negative in urease, arabinose, mannitol, xylose. 2. B. anthracis grew well on B4 Br A TSA after incubation for 24 hours. The organisim grew well on BA, Br. A, NA, TSA after incubation for 72 hours. The media grew well on Br A instead of BA. 3. On 5% blood agar by laboratory animal, ${\beta}$ -hemolysis was produced from 36 hours to 48 hours incubation. There was perfect ${\beta}$-hemolysis after incubation for 48 hours. On the other side ${\beta}$-hemolysis was begun on 5% goat blood agar after incubation for 60 hours. 4. In the test of antimicrobial susceptibility, B. anthracis was very sensitive to AM, CF, TE, ENR, GM, AN, DFX, S, P, TYLO, N, KM, C, E, Lins＋Sp, NN, CC, CFP, CB were sensitive one by one. B. anthracis was no-sensitive to L, XNL, TIA, CL, SXT 5. B. anthracis had never sensitivity to direct inoculation of rat and chicken, after subcutanous inj. It was very sensitive to mouse and goat, hamster, guinea pig, rabbit had a sensibility one by one. 6. The dead laboratory animal which had been inoculated with B. anthracis preserved at $37^{\circ}C$ incubation, B. anthracis didn't cultivate on non-dissected animal after 80 hours but cultivate on dissected animal after 360 hours. 7. The rapidly death could cause high concentration, died from 420 after S. C. 8. The blood smeared samples of hamster from inoculation with B. anthracis, spore germinated In 37$^{\circ}C$ after 5 hours, in $32^{\circ}C$ after 6 hours, in room temperature after 9 hours, in $-4^{\circ}C$ to $-20^{\circ}C$ after 10 hours. 9. B, anthracis inoculated to laboratory animal after SC or PO. Mice and rats feces didn't cultivated with B. anthracis after SC, but did cultivated with B. anthracis after PO. 10. In the test of disinfectant, B. anthracis was high effective to $HgC1_2$, formalin, effect phenol, cresol, but non-effect NaOH, ethanol.

• Journal of Periodontal and Implant Science
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• v.30 no.1
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• pp.145-157
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• 2000

Cytotoxic substances in dental calculus and root cementum of periodontally diseased teeth inhibit new attachment and regeneration. The purpose of scaling and root planing is to remove pathologic structures harboring these cytotoxic substances in order to create a biologically acceptable root surface. However, these procedures inevitably leave a non-biocompatible smear layer. Conventionally, the smear layer has been removed with low pH etching agents such as citric acid, phosphoric acid and tetracycline hydrochloride(TC). Lately, a supersaturated neutral pH etching solution of ethylene diamine tetraacetic acid(EDTA) has been found to be as effective as low pH etchants with respect to smear removal and to be superior in exposing root surfaceassociated collagen. The aim of the present study was to determine the effect of root surface treatment using EDTA on the initial attachment of human gingival fibroblasts. 27 human teeth, extracted due to severe periodontitis, were cut into dentin slices after root planing. The specimens were divided into TC group(treated with $50㎎/m{\ell}$ tetracycline-HCl, pH 1.52), EDTA group(treated with 17% EDTA, pH 7.4), and non-treated control group. After sterilization, 5th subcultured human gingival fibroblasts were seeded in each culture well containing a prepared root slice and incubated for 15 min., 60 min., and 4 hours in 5% $CO_2$ incubator at $37^{\circ}C$. At each incubation time, the number of attached fibroblasts were counted on the microphotographs taken at a magnification of x100. The difference of the number of attached cells between groups was statistically analyzed by the ANOVA followed by Duncan test in SPSS/PC＋programs. The results were as follows : 1. After incubation for 15 min, the attached cells were significantly more in EDTA group and TC group than non-treated control group(p<0.05), but there was no significance in the difference between EDTA group and TC group(p>0.1). 2. After incubation for 60 min and 4 hours, there was no significant difference in the number of attached cells between all groups(p>0.1). 3. In both EDTA group and TC group, there was no significant difference in the number of attached cells between different incubation(p>0.1). But in control group, the number of attached cells was significantly increased after incubation for 60 min, compared with incubation for 15 min(p<0.05). The above results suggest that root surface treatment using EDTA could enhance the initial attachment of gingival fibroblasts to root surface as effective as tetracycline-HCl.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.9
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• pp.1462-1467
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• 2007

This study investigated the effect of chemical treatments on tannins (condensed and hydrolysable) and on the trypsin inhibitor (TI) activity in salseed meal. Triplicate samples of ground salseed meal (1 kg) were mixed with 820 ml of either distilled water (pH 5.3), 0.67 M acetic acid (pH 2.4), 0.67 M sodium bicarbonate (pH 8.2) or 2% polyvinyl-pyrrolidone (PVP) solution. The material was placed in airtight plastic containers and incubated at $37^{\circ}C$ for 0, 3, 6, 12, 24, 48 and 72 h. Samples of untreated salseed meal which had not been subjected to soaking or incubation were run through the analysis to serve as control. Addition of water, acetic acid, sodium bicarbonate and PVP solutions to salseed meal and subsequent anaerobic incubation at $37^{\circ}C$ significantly reduced chemically detectable tannins. At each incubation time, alkali solution was more effective than its counterparts. The effect of acidic solution on hydrolysable tannin was least among the treatments. All the treatments reduced TI activity of salseed meal. The reduction in TI activity by these treatments was similar and ranged between 80-84%. Treatment time effected a decrease in the contents of antinutritional substances. However, the effect of the treatment with the reagents, even for zero incubation time, was quite pronounced. It may be concluded from the present results that the treatment of salseed meal with sodium bicarbonate (0.67 M) is more effective in reducing hydrolysable and condensed tannin contents than PVP, water and acid solutions. Treatment with sodium bicarbonate solution is more economical and easier to handle than acid and PVP treatments. Incubation of the treated material for 12 h is reasonably effective, economical and safe from any mould growth.

• Parasites, Hosts and Diseases
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• v.28 no.2
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• pp.91-100
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• 1990

Paragonimus westermani is a tissue migrating parasite in the early stage until arriving at lung, and most of the parasites spend their life spans there. Considerable immune responses including activation of macrophages are taken place during the residence of parasites in the host. However, concerning the immunologic defense mechanisms of the host against this parasite, only a few document is available so far. In this study, the cytotoxic effect of peritoneal macrophages under the presence of antibody and/or complement against metacercariae of F. westermani was investigated in vitro. Metacercarlae were collected from the crayfish, Cambaroides similis and hatched out in Tyrode solution (pH 7.4). Plastic adherent cells from normal or infected rat (Wistar) peritoneal exudates were used as experimental macrophages. Polyclonal antibodies were obtained from infected rats and a cat. Cat IgG was fractioned with ion exchange chromatography. Fresh rabbit complement was used according to experimental scheme. Various combinations of peritoneal macrophages, normal or infected rat serum, complement and cat IgG were incubated at $36^{\circ}C$ in 5% $CO_2$ incubator for 6, 14, 24 and 48 hours. The results obtained were as follows: 1. P. westermani infection activated peritoneal macrophages non-specifically and this activation induced increases of cell adherence and cytotoxicity on metacercariae. 2. In the presence of infected rat serum the antibody.dependent cell-mediated cytotoxicity of peritoneal macrophages on metacercariae was significantly increased and showed a peak at 6-hour incubation. But the cytotoxic effect was markedly reduced after inactivation of complement and heat.labile IgE antibody by the heating of infected serum at 56$^{\circ}C$ for 30 minutes. 3. The highest cytotoxic effect (100%) of concomitant incubation with IgG and complement showed 24 hours after incubation, although cell adherence was relatively low at 6-hour incubation and 0% at 24-hour incubation. 4. Coordinative functions of complement with serum and IgG were effective in cell adherence and in cytotoxicity, but it is not clear the independent role of complement on the macrophage- mediated cytotoxicity in this study- With these results it is assumed that P. westermani infection can induce the non-specific activation of peritoneal macrophages, and strum antibodies including IgE antibody might enhance the cytotoxicity by macrophages,

• Journal of the Korean Society of Food Science and Nutrition
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• v.31 no.3
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• pp.521-526
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• 2002

A number of experimental and epidemiological studies have implicated that antiestrogenic effects of estrogen-like compounds in legumes and plant seeds are responsible for lowering breast cancer risk in human. However, few studies have been conducted to illustrate the possible chemopreventive effects of Korean traditional food materials. This study was performed to determine the cytotoxic and apoptotic effects of yellow soybeans, black soybeans and brown rice extracts on hormone-dependent and hormone-independent human breast cancer cells. Methanol-or acetone-soluble fractions of soybeans or brown rice were incubated with hormone-dependent cells (MCF-7) or hormone-independent cells (MDA-MB-231). Cell cytotoxicity was measured by MTT assay at 24, 48 and 72 hrs of incubation. Apoptotic effects of these extracts toward breast cancer cells were also determined at 48 hrs of incubation by measuring DNA fragmentation. Results indicated that the acetone-soluble fraction of brown rice exerted strongest cytotoxic effect on MCF-7 ceIls, although other fractions also reduced the number of viable MCF-7 cells after 48 hrs of incubation. Both acetone and methanol soluble fractions of all samples exerted a significant cytotoxicity towards MDA-MB-231 cells after 24 hrs of incubation, and acetone and methanol soluble fractions of brown rice were especially effective in these cells. At 48 hrs of incubation, methanol fractions of all three samples induced apopotosis of MDA-MB-231 cells. These results indicate methaol or acetone soluble fractions of yellow soybeans, black soybeans and brown rice induce cytotoxicity in both hormone-dependent and hormone-independent breast cancer cells. Therefore, possible mechanisms of cell cytotoxicity do not necessarily include antiestrogenic effects of soybean or brown rice extract. A possible anticarcinogenic effect of brown rice methanol-soluble fraction may mediated through their apoptotic effect. Further studies are requried to elucidate responsible compounds and mechanisms involved in observed anticarcinogenesis.

• Research in Plant Disease
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• v.29 no.1
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• pp.11-22
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• 2023

Resazurin-based microtiter assay was used to evaluate the inhibitory effect of fungicides on the respiration of Colletotrichum acutatum s. lat. 20JDS8 sensitive and 20CDJ6 resistant to strobilurin fungicides. The spores of C. acutatum s. lat. 20JDS8 were inoculated into potato dextrose broth (PDB) at densities of 1x10⁴, 1x10⁵ and 1x10⁶ spores/ml, respectively. The relative fluorescence unit (RFU) of all treatments inoculated at each spore density started to rise after 12 hr of incubation, and were 1,965.5, 5,412.5, and 10,061.0, respectively, after 24 hr of incubation. To evaluate the inhibitory effect of fungicide on the respiration of the pathogen, the spores of the pathogen were inoculated into the PDB and treated with the fungicides 0, 6, 12, and 24 hr after incubation, respectively. After keeping the pathogen culturing for another 24 hr, PrestoBlue reagent was treated into the PDB culturing the pathogen. The RFU of each treatment was examined 1 hr after the reagent was treated. When dithianon, isopyrazam, pyraclostrobin, and fluazinam were treated at high concentrations in the stages of spores (immediately after inoculation [0 hr]), spore germination (after incubation for 6 hr), and hyphal growth (after incubation for 12 hr), the respiration of pathogens was inhibited by 90-100%. When the fungicides were treated after culturing the pathogen for 24 hr, the respiratory inhibitory effects were greatly reduced. With pyraclostrobin-resistant C. acutatum s. lat. 20CDJ6, azxoystrobin, trifloxystrobin and kresoxim-methyl, which have the same mode of action, had very little or no respiratory inhibitory effect in all growth stages of pathogens. Based on the above results, it was thought that the resazurin-based microtiter assay could quickly and accurately evaluate the inhibitory efficacy of the fungicides that inhibited respiration.

• Journal of Korean Society of Environmental Engineers
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• v.39 no.5
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• pp.288-302
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• 2017

Accurate measurement of benthic nutrient fluxes (BNF) is a prerequisite for evaluating the effect of sediments on nutrient cycle in the surface water. The intact sediment cores were collected in July 2015 at the midstream of Nakdong River. We identified pre-incubation time (6, 12, 24 hr), dissolved oxygen concentration (90, 70, 50% saturation), diffusive boundary layer thickness (0, 0.6-0.8, 1.2-1.4 mm), and incubation temperature (10, 17, 20, $25^{\circ}C$) as the most important control factors, and measured the BNF fluctuation with the variation of these factors using the laboratory sediment core incubation method. Since the chemical composition, redox condition, hydrodynamic regimes and microbial activities at the sediment-water interface were changed as a result of the alteration of control factors, sediment core incubation should be conducted under as close to the natural conditions of study site as possible, in order to produce the results similar to actual values. Relative percentage differences between two replicates were below 20% in most control factors, which showed satisfactory precision for strict compliance with the experimental conditions and procedures. In the further studies, we will compare the results of core incubation with those of in situ measurements to confirm the accuracy of the sediment core incubation method.