This experiment was conducted to determine the effect of betaine on immune response in laying hens. A total of 72 ISA-brown laying hens were divided into four groups of 18 hens each and fed corn-soybean meal based diets with addition of 0, 300, 600 and 1,200 ppm betaine for four weeks. The effect of betaine on splenocyte proliferations with mitogens, concanavalin A(Con A) and pokeweed mitogen(PWM), were assayed after incubation using [3H] thymidine uptake. Proliferations of splenocyte were significantly increased by activation of mitogen Con A or PWM. Mitogen effects of Con A were increased by Con A plus betaine injection(0.1 mM), whereas PWM effects did not affect in PWM plus betaine injection(0.1 mM) in vitro. Splenocyte of laying hens fed betaine tended to proliferate in the presence of PWM, but appeared to be slightly suppressed in the presence of Con A in vivo. Proliferation of splenocytes which were stimulated by Con A or Con A+betaine injection(0.1 mM) were increased in dietary 600 ppm betaine, but inhibited in dietary 1,200 ppm betaine supplementation. Spleen weights and sheep red blood cell(SRBC) titers of hens fed betaine tended to increase compared to those of control, but were not significantly different. These results suggested that betaine could increase splenocyte proliferation in vitro.
This study was undertaken to investigate the effects of ruminally protected amino acids (Methionine and Lysine) on in vitro ruminal parameters, and in vivo milk yield and milk composition in mid-lactating cows. In the first in vitro experiment, there were no statistical significances between treatments in ruminal pH and dry matter digestibility during various incubation times. In the second in vivo experiment, milk yield decreased by 11.92% in control and 5.68% in the treatment respectively, but decrease rate of milk yield in the treatment was lower than control. Milk yields naturally decreased as time goes by since the DIMs(Days in milk) of the cows in experiment were in mid-lactation period. 4% FCM(Fat corrected milk) and milk protein yields also, respectively, decreased by 11.25% and 11.09% in control and 6.16% and 5.47% in the treatment as compared with the intial. Milk protein and milk fat production were higher in the treatment(0.90kg, 1.10kg) than those of control(0.66kg, 0.79kg). Milk fat content significantly increased with supplementing protected amino acids as compared to control(P<0.05). From the above results, protected amino acids were positively utilized in the performances of mid-lactating cows without inhibiting rumen fermentation. Further investigation is suggested for essential amino acid composition and intestinal digestion rate out of rumen bypass protein in dietary protein to be estimated.
Objective: Mulberry (Morus alba L.) is a cultivated shrub grown widely in the sub-tropical and tropical areas. It has been shown that mulberry leaf contains high levels of protein while having polyphenols as phytonutrients. Therefore, it is important to conduct an experiment to assess potential toxic level from mulberry on behavior, blood hematological and coagulation parameter using Sprague-Dawley (SD) rats. Methods: Both male and female SD rats were given an intragastric administration of respective treatments of mulberry leaf intakes (control, low and high levels). Parameters of feed intake, hematological and coagulation of blood parameters, as well as liveweight changes were taken during the 7 d of adaptation, 28 d of treatment exposure, and 14 d of recovery periods, respectively. All treatment data were statistically analyzed using analysis of variance of SPSS17.0 for Windows Statistical Software following the Randomized complete block design with sex as a block. Results: Most of the parameters of the physical symptoms of the SD rats, were not significantly different (p>0.05) when compared with that of the control group. Those which remain unchanged in each dose group were, body weight (BW) gain, feed intake, the hematology and coagulation indexes. Although, there were a few individual indicators that were abnormal, but the overall physiological appearance of the rats were normal. Conclusion: Results under this experiment revealed that most hematological and coagulation parameters of the SD rats in both male and female were normal, although the weight gain of female rats in high-dose group was significantly reduced than those of the male rats. Under this study, the use of mulberry leaf up to 2 g/kg BW did not result in abnormal phenomenon in the SD rats. These findings would offer useful information for further in vivo feeding trials in animals to extensively use of mulberry leaf to improve animal production, particularly in P.R. China.
Objectives : In this study, the author tried to investigate whether wood vinegar produced from Morus alba (MA) significantly affects the increase in airway epithelial mucosubstances and hyperplasia of tracheal goblet cells of rats, and in vitro airway mucin secretion and PMA- or EGF- or TNF-alpha-induced MUC5AC mucin production / gene expression from human airway epithelial cells. Materials and Methods : For the in vivo experiment, the author induced hypersecretion of airway mucus and goblet cell hyperplasia by exposure of rats to SO2 over 3 weeks. Effect of orally-administered MA over 2 weeks on increase in airway epithelial mucosubstances from tracheal goblet cells of rats and hyperplasia of goblet cells were assessed using histopathological analysis after staining the epithelial tissue with alcian blue. For the in vitro experiment, confluent RTSE cells were chased for 30 min in the presence of MA to assess the effect of MA on mucin secretion by enzyme-linked immunosorbent assay (ELISA). Also, effects of MA on PMA- or EGF- or TNF-alpha-induced MUC5AC mucin production and gene expression from human airway epithelial cells (NCI-H292) were investigated. Confluent NCI-H292 cells were pretreated for 30 min in the presence of MA and treated with PMA (10 ng/ml), EGF (25 ng/ml) or TNF-alpha (0.2 nm) for 24 hrs, to assess both effects of MA on PMA- or EGF- or TNF-alpha-induced MUC5AC mucin production by enzyme-linked immunosorbent assay (ELISA) and gene expression by reverse transcription-polymerase chain reaction (RT-PCR). Possible cytotoxicities of MA in vitro were assessed by examining LDH release from RTSE cells and the rate of survival and proliferation of NCI-H292 cells. In vivo liver and kidney toxicities of MA were evaluated by measuring serum GOT/GPT activities and serum BUN/creatinine concentrations of rats after administering MA orally. Results : 1. MA decreased the amount of intraepithelial mucosubstances of rats exposed to sulfur dioxide inhalationally. 2. MA decreased in vitro mucin secretion from cultured RTSE cells. 3. MA significantly inhibited PMA-, EGF-, and TNF-alpha-induced MUC5AC mucin productions and the expression levels of MUC5AC mRNA from NCI-H292 cells. 4. MA did not show either in vitro or in vivo hepatic or renal toxicities. Conclusion : The results from this study suggests that MA can regulate the secretion, production and gene expression of airway mucin observed in diverse respiratory diseases accompanied by mucus hypersecretion and does not show in vivo toxicity to liver and kidney functions after oral administration. Effects of MA should be further studied using animal experimental models that simulate the diverse pathophysiology of respiratory diseases via future research.
Objectives In this study, the author tried to investigate whether piryongbang-gamgil-tang (PGGT) significantly affect in vitro airway mucin secretion, PMA- or EGF- or TNF-${\alpha}$-induced MUC5AC mucin production / gene expression from human airway epithelial cells and increase in airway epithelial mucosubstances and hyperplasia of tracheal goblet cells of rats. Materials and Methods For in vitro experiment, confluent RTSE cells were chased for 30 min in the presence of PGGT to assess the effect of PGGT on mucin secretion by enzyme-linked immunosorbent assay (ELISA). Also, effect of PGGT on PMA- or EGFor TNF-${\alpha}$-induced MUC5AC mucin production and gene expression from human airway epithelial cells (NCI-H292) were investigated. Confluent NCI-H292 cells were pretreated for 30 min in the presence of PGGT and treated with PMA (10 ng/ml) or EGF (25 ng/ml) or TNF-${\alpha}$ (0.2 nM) for 24 hrs, to assess both effect of PGGT on PMA- or EGF- or TNF-${\alpha}$-induced MUC5AC mucin production by ELISA and gene expression by reverse transcription-polymerase chain reaction (RT-PCR). For in vivo experiment, the author induced hypersecretion of airway mucus and goblet cell hyperplasia by exposure of rats to $SO_2$ during 3 weeks. Effect of orally-administered PGGT during 2 weeks on increase in airway epithelial mucosubstances from tracheal goblet cells of rats and hyperplasia of goblet cells were assesed by using histopathological analysis after staining the epithelial tissue with alcian blue. Possible cytotoxicities of PGGT in vitro were assessed by examining LDH release from RTSE cells and the rate of survival and proliferation of NCI-H292 cells. In vivo liver and kidney toxicities of PGGT were evaluated by measuring serum GOT/GPT activities and serum BUN/creatinine concentrations of rats after administering PGGT orally. Results (1) PGGT did not affect in vitro mucin secretion from cultured RTSE cells. (2) PGGT significantly inhibited PMA-, EGF-, and TNF-${\alpha}$-induced MUC5AC mucin productions and the expression levels of MUC5AC mRNA from NCI-H292 cells. (3) PGGT decreased the amount of intraepithelial mucosubstances and showed the tendency of expectorating airway mucus already produced. (4) PGGT increased LDH release from RTSE cells. However, PGGT did not show in vivo liver and kidney toxicities and cytotoxicity to NCI-H292 cells. Conclusion The result from this study suggests that PGGT can regulate the production and gene expression of airway mucin observed in diverse respiratory diseases accompanied by mucus hypersecretion and do not show in vivo toxicity to liver and kidney functions after oral administration. Effect of PGGT with their components should be further studied using animal experimental models that reflect the diverse pathophysiology of respiratory diseases through future investigations.
Two in vivo experiments were conducted to evaluate the effect of novel urease inhibitor hydroquinone (HQ) on ammonia release rate from urea hydrolysis, nitrogen balance, nutrient digestibility and efficiency of microbial protein synthesis. In Exp. 1, twelve crossbred cannulated lambs were randomly assigned within initial body weight block to one of four HQ treatments, which included 0 (control), 30, 60 or 80 mg HQ/kg DM intake. Ammonia concentration and pH of ruminal fluid were immediately measured at 0, 2, 4, 6 and 8 h after feeding. Increasing the dose of HQ tended (p<0.15) to linearly decrease NH3 formation. The ammonia peak concentration (2 h post-feeding) in animals receiving HQ was approximately one-half of that in animals not receiving HQ (p<0.01), and a relatively sustained ammonia release could be obtained at the dose of 30 or 60 mg HQ/kg DM. In Exp. 2, sixteen intact crossbred lambs (weight $40{\pm}0.8kg$) were used in a $2{\times}2$ factorial design experiment. The four rations consisting of soybean meal-based (SBM) or urea-based (Urea) nitrogen source with or without HQ (S1, S0, U1 and U0) were fed in digestion and N balance trials. Apparent digestibility of major nutrients except that of ADF was not affected by either nitrogen source or addition of HQ. Regardless of nitrogen source, supplementation of HQ significantly improved ADF digestibility (p<0.05). The various ration had no effects on N metabolism in the presence of HQ. There was significant difference between total purine derivatives (PD), estimated efficiency of microbial N synthesis (p<0.05) and urea-N excretion (p<0.01) in the urine for the SBM ration and for the Urea ration. However, HQ had little influence on efficiency of microbial N synthesis as proportion of daily intake of total tract digestible OM (p>0.05). No interactions between main nitrogen source and HQ were measured throughout the trial. Results of this study suggest that addition of HQ to ration may improve ADF digestion with having no negative effect on N metabolism and microbial protein production.
Rabbit meat bekasam is a traditional fermentation product from Indonesia. This study aimed to determine the chemical and microbiological characteristics of rabbit meat bekasam during the fermentation process in order to isolate, characterize (in vitro and in vivo), and identify lactic acid bacteria (LAB) as the probiotic candidate. The chemical contents of bekasam on 7-day fermentation were investigated in explorative and experimental methods in a completely randomized design. A proximate analysis reported a decrease in the moisture content, fat and carbohydrate content, and an increase in protein content. Also, lactic acid content was increased from 0.48% to 1.12%, and pH was decreased from 5.3 to 4.3. Other properties indicated different values, such as bacteria (2.75×106 to 4.45×107 CFU/g), total LAB (3.82×106 to 4.67×108 CFU/g), total yeast (9.89×106 to 3.82×108 CFU/g) and total mould (4.34×101 to 4.86×103 CFU/g). The experiment produced nine LAB isolates, including two probiotics subjected to further 16S rRNA gene analysis, which indicated that Lactobacillus buchneri was the potential probiotic isolate. After being tested on BALB/c mice, L. buchneri could improve the immune system by inhibiting the growth of Coliform and Salmonella.
A novel electro-mechanical implantable ventricular assist system is developed as a bridge to transplantation or recovery for patients with end-stage heart failure. The developed system is composed of an implanted blood pump, an external monitoring system which stores data, and a wearable system including a portable external driver and a portable power supply system. The blood pump is designed to be implanted into the left upper abdominal space and provides blood flow from the left ventricular apex to the aorta. The pulsatile blood flow is generated by a double cylindrical cam. There was mo excessive heat emission from the blood pump into the temperature-controlled chamber in the heat test and no stagnated flow within the blood sac by the observation in the flow visualization test. Animal experiments were performed using sheep and calves. The maximum assist flow rate reached 7.85L/min in the animal experiment. The evaluation results showed that the developed system was feasible for the implantable ventricular assist system. The long-term in vitro durability test and mid-term in vivo experiments are in progress and mow the modified next model is under development.
Natural products in Chonnam, Korea were screened via anti-angiogenesis experiments, and 1 candidate product was identified, Corni fructus, which exerted dose-dependent inhibitory effects against angiogenesis, adipogenesis, and cell adhesion. C. fructus extract (CFE) exhibits an angiogenesis inhibitory effect superior to that of the EGCG from green tea leaves. The expression level of angiogenesis and adipogenesis-related signal molecules in the western blotting was reduced by increasing the amount of added CFE. Moreover, a diet supplemented with CFE was deemed more effective in inducing weight loss in LB mice than a representative synthetic diet drug, orlistat, which incidently caused the side effect of denuding the mice of their hair. These results indicate that C. fructus may prove to be a useful anti-adipogenic compound, and these in vitro results may be reflected later under in vivo conditions.
Objective: This study was aimed to explore the efficacy of combination of endo-xylanase (Xyn) and xylan-debranching enzymes (arabinofuranosidase, Afd and feruloyl esterase, FE) in improving utilization of bran in piglet diet. Methods: In vitro experiments were firstly conducted to examine the enzymological properties of Xyn, Afd, and FE, concurrent with their effect on degradation of arabinoxylan (Abx) in bran. In vivo experiment was then implemented by allocating two hundred and seventy 35-d-old postweaning piglets into 3 groups (6 replicates/group), which received bran-containing diet supplemented with Xyn (1,600 U/kg) or its combination with Afd (0.8 U/kg) and FE (4 U/kg) or without enzyme. Results: Both Xyn, Afd, and FE are relatively stable against the changes in temperature and pH value. Combining Xyn with Afd and FE had a superiority (p<0.05) over Xyn alone and its combination with Afd or FE in promoting (p<0.05) degradation of Abx in different brans. Combined treatment with Xyn, Afd, and FE was more beneficial than Xyn alone to induce increasing trends (p<0.10) of average daily gain, final body weight and feed efficiency of piglets fed bran-containing diet. Moreover, combination of Xyn, Afd, and FE showed advantages (p<0.05) over Xyn alone in causing reductions (p<0.05) in diarrhea rate and cecal pH value, concurrent with increases (p<0.05) in cecal and colonic acetic acid and total volatile fatty acid concentrations, as well as cecal butyric acid concentration of piglets fed bran-containing diet. Conclusion: Combining Xyn with Afd and FE was more beneficial than Xyn alone in promoting degradation of Abx in bran, along with growth performance and intestinal volatile fatty acid profile of piglets received bran-containing diet. Thereby, combination of Xyn, Afd, and FE had a superior efficacy relative to Xyn alone in improving application of cereal bran in piglet diet.
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