• Mass Spectrometry Letters
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• v.3 no.3
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• pp.68-73
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• 2012

Fat malabsorption is an important cause of poor growth in infancy and childhood. Steatorrhea tests have been developed using various methods. Traditional measurements of stool fat, however, require large samples and it often takes as a week to complete the analysis. In this paper, a liquid chromatography-electrospray ionization/mass spectrometry (LC-ESI/MS) method was developed for simultaneous quantitative analysis of triacylglycerols, triolein, diolein and monoolein, in mouse feces. Moreover, the procedure was rapid, simple as well as compatible with LC-ESI/MS. Chloroform-isopropyl alcohol solution was used for fat-soluble sample extraction. After centrifugation and filtration, an analytical solution was prepared. Triolein, diolein and monoolein were separated using non-aqueous reversed-phase column with the mobile phase consisting of A (methanol) and B (acetone-isopropyl alcohol). The precision (% CV) and accuracy (% bias) of the assay were 3.8-14.7% and 85.2-114.9%, respectively. This method has been successfully applied to simultaneous determination of triolein, diolein and monoolein in feces from 30 mice. This method can therefore be applied to measure triacylglycerols in mouse feces accurately and precisely by LC-ESI/MS, thereby helping to predictive biomarker in fat malabsorption and diagnostic research.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2018.10a
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• pp.120-120
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• 2018

Intestinal microbiota is strongly connected to health of host. It has been reported that not only metabolic disease like diabetes and obesity, but psychological diseases are affected by composition of intestinal microbiota. To figure it out the importance of the composition and relationship between disease and microbiota, intensive researches have done with human and experimental animals. But, the composition of the intestinal microbiota could be affected by several factors such as experimental environments, feeding, water, and bedding. As a result, the data from each experimental group might be diverse. It also affects experiments about biological activities of plant materials. In this study, mouse intestinal bacteria were isolated from fresh feces and identified by 16S rRNA gene to use in biological activities of natural medicines. The fecal supernatant was anaerobically incubated at $37^{\circ}C$ for 48 hours. Colonies were picked up separately and incubated again in same condition to increase quantity to analyze and stock. The bacteria strains were listed up and could be used for many researches including biological activities of plant materials and change in composition of intestinal bacteria itself.

• Parasites, Hosts and Diseases
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• v.30 no.4
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• pp.259-262
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• 1992

This study was performed to investigate experimental transmission of Cryptosporidium parvum in a calf. A 25-day-old Korean native calf was inoculated per os with $1{\times}10^6$ C. parvum oocysts isolated from a Korean mouse. The calf commenced oocyst discharge in feces on post-inoculation day 4, and continued until the aah 11. The number of discharged oocysts Peaked($4.9{\times}10^5$) on post-inoculation day 6. However, the calf did not show signs of diarrhea. The present results indicate that C. parvum is cross-transmissible between the calf and the mouse.

• Korean Journal of Veterinary Research
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• v.35 no.1
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• pp.107-113
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• 1995

Four Holstein calves 7-day-old were infected with C parvum oocysts for parasitological and pathological investigations of bovine cryptosporidiosis. Of those calf 1 was orally administered with $7{\times}10^6$ oocysts of C parvum isolated from a Korean mouse (VRI-CN91), and calf 2 with same number of C parvum oocysts provided by Washington State University(WSU). The rest (calf 3 and 4) were orally administered with $1{\times}10^8$ oocysts of VRI-CN91 strain. Calf 1 commenced to discharge oocysts in feces at days 6 post inoculation(PI), and it reached a peak $1.4{\times}10^7$ oocysts per gram of feces(OPG) on day 8 PI. Calf 2 commenced to discharge oocysts in feces at day 4 PI, and it reached a peak $3.75{\times}10^6$ OPG on day 7 PI. Calf 3 and 4 commenced to discharge oocysts in feces at day 3 and day 4 PI, and it reached a peak on day 7 PI (calf 3, $7.8{\times}10^6$ OPG; calf 4, $1.7{\times}10^6$ OPG). Clinically, the calves began to show mucoid-watery diarrhea at day 3 to 5 PI, and the sign lasted 5 to 7 days. Calf 2 died on day 9 PI with a severe dehydration. On necropsy the intestine was found to be congested and hemorrhagic. Protozoan oocysts were observed mainly in the ileum and occasionally in jejunum. The results in the present study indicate that the Korean isolate was pathogenic in calves.

• Toxicological Research
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• v.28 no.1
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• pp.11-18
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• 2012

The aim of this study was to evaluate the single oral dose toxicity of Bupleuri Radix (BR) aqueous extracts, it has been traditionally used as anti-inflammatory agent, in male and female mice. BR extracts (yield = 16.52%) was administered to female and male ICR mice as an oral dose of 2,000, 1,000 and 500 mg/kg (body weight) according to the recommendation of Korea Food and Drug Administration (KFDA) Guidelines. Animals were monitored for the mortality and changes in body weight, clinical signs and gross observation during 14 days after dosing, upon necropsy; organ weight and histopathology of 14 principal organs were examined. As the results, no BR extracts treatment related mortalities, clinical signs, changes on the body and organ weights, gross and histopathological observations against 14 principal organs were detected up to 2,000 mg/kg in both female and male mice, except for soft feces and related body weight decrease detected in male mice treated with 2,000 mg/kg. Therefore, $LD_{50}$ (50% lethal dose) and approximate LD of BR aqueous extracts after single oral treatment in female and male mice were considered over 2000 mg/kg, respectively. Although it was also observed that the possibilities of digestive disorders, like soft feces when administered over 2,000 mg/kg of BR extracts in the present study, these possibilities of digestive disorders can be disregard in clinical use because they are transient in the highest dosages male only.

• Journal of Veterinary Science
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• v.23 no.3
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• pp.41.1-41.15
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• 2022

Background: Proliferative enteritis caused by Lawsonia intracellularis undermines the economic stability of the swine industry worldwide. The development of cost-effective animal models to study the pathophysiology of the disease will help develop strategies to counter this bacterium. Objectives: This study focused on establishing a model of gastrointestinal (GI) infection of L. intracellularis in C57BL/6 mice to evaluate the disease progression and lesions of proliferative enteropathy (PE) in murine GI tissue. Methods: We assessed the murine mucosal and cell-mediated immune responses generated in response to inoculation with L. intracellularis. Results: The mice developed characteristic lesions of the disease and shed L. intracellularis in the feces following oral inoculation with 5 × 10⁷ bacteria. An increase in L. intracellularis 16s rRNA and groEL copies in the intestine of infected mice indicated intestinal dissemination of the bacteria. The C57BL/6 mice appeared capable of modulating humoral and cell-mediated immune responses to L. intracellularis infection. Notably, the expression of genes for the vitamin B12 receptor and for secreted and membrane-bound mucins were downregulated in L. intracellularis -infected mice. Furthermore, L. intracellularis colonization of the mouse intestine was confirmed by the immunohistochemistry and western blot analyses. Conclusions: This is the first study demonstrating the contributions of bacterial chaperonin and host nutrient genes to PE using an immunocompetent mouse model. This mouse infection model may serve as a platform from which to study L. intracellularis infection and develop potential vaccination and therapeutic strategies to treat PE.

• Journal of Nutrition and Health
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• v.37 no.2
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• pp.81-87
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• 2004

This study was conducted to investigate the supplementary effects of Suwon 464 developed by Rural Development Administration, which has over two times of dietary fiber content compared with normal rice (Ilpum), on lipid metabolism in diabetic mice. We supplied 5 kinds of experimental diets (com starch diet as a control (CO), Ilpum polished rice diet (IP), Ilpum brown rice diet (IB), polished rice diet (SP) and brown rice diet (SB) of Suwon 464) to diabetic mice for 8 weeks, after analyzing dietary fiber contents of 5 experimental diets. Diet intake, body weight, organ weights, and lipids levels of serum, liver and feces were measured. The dietary fiber contents in CO, IP, IB, SP, and SB diets were 1.0, 1.2, l.4, 1.4, and 2.0% respectively. Body weight and liver and epididymal fat pad weights were lower in SB group than the other groups though there was no significant difference in diet intake among experimental groups. The concentrations of serum triglyceride was lower in SP and SB groups than CO and IP groups. The levels of hepatic total lipid and total cholesterol were significantly lower in SP and SB groups than CO group, and the level of hepatic triglyceride was lower in IB, SP and SB groups than CO group. The levels of total lipid and triglyceride excreted in feces were higher in IB, SP and SB, and the level of total cholesterol in feces was higher in SP and SB groups than CO group. These results suggested that the high dietary fiber rice (Suwon 464) decrease the triglyceride or total cholesterol concentrations of serum and liver by increasing of fecal lipid excretion in diabetic mice. (Korean J Nutrition 37(2): 81∼87, 2004)

• The Journal of Pediatrics of Korean Medicine
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• v.35 no.4
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• pp.96-111
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• 2021

Objectives The purpose of this study is to look for pathological mechanism of disease development caused by Taedok, by studying whether stress and diet in pregnant ICR mice affect the intestinal flora and IgA (Immunoglobulin A) concentration. Methods The mice were divided into 4 groups (n=5 per group) based on the concept of Taedok: the control group (G1), stress group (G2), capsaicin diet group (G3), high fat diet group (G4). We collected and analyzed intestinal flora from maternal feces and cecal flora from neonatal mice by group. Then, IgA concentration in the maternal feces and sIgA (secretory Immunoglobulin A) concentration in the cecal contents of newborn mice were analyzed. In addition, serum corticosterone was analyzed before and after stress application. Results Changes in maternal intestinal flora and neonatal mice cecal flora by stress and diet were observed. There were no significant changes in the IgA concentration in maternal feces and the sIgA concentration in the cecal contents of neonatal mice. No significant changes compared to the control group were observed between groups before and after applying stress. However, when comparing within one subject, a significant increase was confirmed after stress application in the stress group (G2). Conclusions Based on the results, we observed stress and diet in pregnant mice affect the intestinal flora of maternal and neonatal. We were able to interpret the pathological mechanism of Taedok based on the principle of interaction between mother and newborn intestinal flora.

• Parasites, Hosts and Diseases
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• v.32 no.1
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• pp.7-12
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• 1994

A calf and 50 mice were infected with Cryptosporidium parvum and their fecal materials were collected and treated 10 ether extraction (EE), followed by discontinuous sucrose gradients (DSG) method. EE method was to remove some of fat or lipid from feces. Sediments were washed by centrifugation ($1,500{\;}{\times}{\;}g$ for 10 min., 3 times) In phosphate-buffered saline and then these washed sediments were sleeved sequentially through stainless steel screens with a final mesh of 250 ($61{\;}{\mu\textrm{m}}$ porosity) to remove other debris. After sieving, the materials were suspended in 2.5% potassium dlchromate solution. Oocysts were counted by using a hemocytometer and the recovery rate of pure oocysts was calculated on the basis of the count. Following centrifugation ($1,500{\;}{\times}{\;}g$ for 30 min.) by DSG method, most oocysts were recovered at the interface between a gravity of 1.103 and 1.064. The recovery rates of pure oocysts from the fecal suspension of the calf ($3.8{\;}{\times}{\;}10^7/ml$) and the mouse ($3.2{\;}{\times}{\;}10^6/ml$) treated with EE method were 81.6% and 51.6%, respectively. It is suggested that the recovery rate was dependent on the number of oocysts In each suspension treated with EE method. To get the 50% recovery rate, there must be more than $2{\;}{\times}{\;}10^6$ oocysts per ml of the fecal suspension treated with EE method. By the combination of the two methods it was possible to isolate C. parvum oocysts from normal feces of the calf and mouse as well as from dlarrhelc feces.

• Microbiology and Biotechnology Letters
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• v.40 no.4
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• pp.389-395
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• 2012

T-RFLP (terminal restriction fragment length polymorphism) analysis, one of the most highly adopted culture-independent microbial community analysis methods, was applied to evaluate the colonization of probiotics in experimental animal gut. Lactic acid bacteria that exhibited cinnamoyl esterase activity were isolated from Korean fermented vegetables and identified by 16S ribosomal RNA sequence analysis. Lactobacillus plantarum KK3, which demonstrated high chlorogenic acid hydrolysis by cinnamoyl esterase activity, and acid/bile salt resistances, was cultured, freeze-dried, and fed to mice and the microbiota in their feces were monitored by T-RFLP analysis. The T-RF of L. plantarum was detected in the feces of mice after the start of administration and lasted at least 31 days after the initial 7 day feeding. T-RFLP analysis was considered a useful tool to evaluate the gut colonization of probiotic L. plantarum. In order to prove that L. plantarum was from viable cells, we reisolated L. plantarum in the feces using cinnamoyl esterase activity media as the screening step. The colonization of L. plantarum KK3 in the mouse gut was confirmed by this research.