Objective: Stress factors such as high temperatures, overcrowding, and diurnal temperature range exert profound negative effects on weight gain and productivity of broiler chickens. The potential of gamma aminobutyric acid (GABA) as an excitatory neurotransmitter was evaluated under various stress conditions in this study. Methods: The experiment was conducted under four different environmental conditions: normal, high temperature, overcrowded, and in an overcrowded-diurnal temperature range. The experimental groups were divided into (-) control group without stress, (+) control group with stress, and G50 group (GABA 50 mg/kg) with stress. Weight gain, feed intake, and feed conversion ratio were measured, and stress reduction was evaluated through hema-tologic analysis.Results: The effects of GABA on broilers in four experimental treatments were evaluated. GABA treated responded to environmental stress and improved productivity in all the experimental treatments. The magnitude of stress observed was highest at high temperature, followed by the overcrowded environment, and was least for the overcrowded-diurnal temperature range.Conclusion: Various stress factors in livestock rearing environment can reduce productivity and increase disease incidence and mortality rate. To address these challenges, GABA, an inhibitory neurotransmitter, was shown to reduce stress caused due to various environmental conditions and improve productivity.

Glucoraphanin (GRA), a glucosinolate particularly abundant in broccoli (Brassica oleracea var. italica) sprouts, can be converted to sulforaphane (SFN) by the enzyme myrosinase. Herein, we investigated the anti-obesogenic effects of broccoli sprout powder (BSP), mustard (Sinapis alba L.) seed powder (MSP), and sulforaphane-rich MSP-BSP mixture powder (MBP) in bisphenol A (BPA)-induced 3T3-L1 cells and obese C57BL/6J mice. In vitro experiments showed that MBP, BSP, and MSP have no cytotoxic effects. Moreover, MBP and BSP inhibited the lipid accumulation in BPA-induced 3T3-L1 cells. In BPA-induced obese mice, BSP and MBP treatment inhibited body weight gain and ameliorated dyslipidemia. Furthermore, our results showed that BSP and MBP could activate AMPK, which increases ACC phosphorylation, accompanied by the upregulation of lipolysis-associated proteins (UCP-1 and CPT-1) and downregulation of adipogenesis-related proteins (C/EBP-α, FAS, aP2, PPAR-γ, and SREBP-1c), both in vitro and in vivo. Interestingly, MBP exerted a greater anti-obesogenic effect than BSP. Taken together, these findings indicate that BSP and MBP could inhibit BPA-induced adipocyte differentiation and adipogenesis by increasing the expression of the proteins related to lipid metabolism and lipolysis, effectively treating BPA-induced obesity. Thus, BSP and MBP can be developed as effective anti-obesogenic drugs.

Endocrine-disrupting chemicals (EDCs), called obesogens, play an important role in obesity by mimicking or disrupting bioidentical hormones. In this study, we determined the obesogenic effect of bisphenol A (BPA) and evaluated the anti-obesogenic effects of a standardized Cirsium setidens Nakai ethanolic extract (CNE) as a functional food ingredient on the lipid accumulation and expression of key adipogenic transcription factors (ATFs) in 3 T3-L1 adipocytes and C57BL/6J mice. The underlying mechanism of the obesogenic effect of BPA was confirmed using the peroxisome proliferator-activated receptor γ (PPARγ) antagonist, GW9662. In addition, the anti-obesogenic effects of CNE were confirmed by measuring the lipid accumulation, reactive oxygen species (ROS) production, body and adipose tissue weights, and the adipogenesis- and lipogenesis-related proteins in BPA-induced 3T3-L1 adipocytes and obese mice. These findings indicate that CNE could potentially be used as a promising natural means to prevent BPA-induced obesity and obesity-related metabolic diseases.

Glucoraphanin (GRA) is a precursor of sulforaphane (SFN), which can be synthesized by the enzyme myrosinase. In this study, we developed and validated HPLC analytical methods for the determination of GRA and SFN in mustard seed powder (MSP), broccoli sprout powder (BSP), and the MSP-BSP mixture powder (MBP), and evaluated their anti-adipogenic effects in 3T3-L1 adipocytes. We found that the analysis methods were suitable for the determination of GRA and SFN in MSP, BSP, and MBP. The content of GRA in BSP was 131.11 ± 1.84 µmol/g, and the content of SFN in MBP was 162.29 ± 1.24 µmol/g. In addition, BSP and MBP effectively decreased lipid accumulation content without any cytotoxicity. Both BSP and MBP significantly inhibited the expression of adipogenic proteins and increased the expression of proteins related to lipolysis and lipid metabolism. BSP and MBP inhibited the expression of adipocyte protein 2 (aP2), CCAAT/enhancer-binding protein-α (C/EBP-α), and peroxisome proliferator-activated receptor-γ (PPAR-γ) in 3T3-L1 adipocytes, and inhibited the expression of fatty acid synthase (FAS) through AMP-activated protein kinase (AMPK). Meanwhile, BSP and MBP also increased the expression of the lipolysis-related proteins, uncoupling protein-1 (UCP-1) and carnitine palmitoyltransferase-1 (CPT-1). Moreover, MBP exerted anti-adipogenic to a greater extent than BSP in 3T3-L1 preadipocytes.

Endocrine-disrupting chemicals (EDCs) are exogenous compounds that are capable of blocking or mimicking the action of bioidentical hormones. Obesogenic EDCs, commonly called obesogens, play an important role in adipogenesis. This study was carried out to determine the effects of select obesogens and their alternatives on adipogenesis in 3T3-L1 cells under dexamethasone (DEX)-free conditions. Preadipocytes were treated with a cocktail of 3-isobutyl-1-methylxanthine (IBMX) and insulin to which an obesogen (viz., bisphenol A (BPA) or its analogs BPS and BPF; dioctyl terephthalate; tris (2-ethylhexyl) trimellitate; or various parabens) had been added. A mixture containing IBMX, insulin, and DEX, which constitute the typical hormonal cocktail required for adipocyte differentiation, was used as the control against which the other groups were measured. The obesogens and the PBA analogs all had evident adipogenic effects under DEX-free conditions, as was determined by estimating the lipid accumulation levels in the cells using Oil Red O staining. Furthermore, the expression of adipogenic transcription factors (CCAAT/enhancer-binding protein-alpha, peroxisome proliferator-activated receptor-gamma, and adipocyte protein 2) was induced by 20 μM of BPA, BPS, or BPF at both the mRNA and protein levels, as determined through reverse transcription-polymerase chain reaction and western blot assays. Taken together, the results reveal that adipocyte differentiation can be induced by obesogens and their alternatives in the absence of DEX.

Erythropoietin (EPO) is the primary regulator of erythropoiesis in the mammalian fetus and adult. Deficiency of EPO induces anemia. In this study, we investigated the effect of gamma-aminobutyric acid (GABA) on serum EPO levels and erythropoiesis in rats. Expression levels of Epo-related genes were measured by quantitative real-time PCR (qPCR) and expression of Epo and Epo receptor (Epor) proteins were measured by immunohistochemistry. The gene and protein expression profiles of kidney tissue in GABA-treated rats were evaluated by ribonucleic acid (RNA) sequencing and two-dimensional electrophoresis (2-DE), respectively. GABA significantly increased serum EPO levels and expression levels of Epo and Epor. GABA increased expression levels of hypoxia-inducible factor (Hif)-1 and Hif-2. Seven proteins with expression levels showing >2-fold change were identified by 2-DE followed by MALDI-TOF MS in GABA-treated rat kidney. The top KEGG pathway from the identified proteins was the tricarboxylic acid cycle, and nicotinamide adenine dinucleotide (NADH) dehydrogenase, succinate dehydrogenase, and isocitrate dehydrogenase were identified as key proteins. GABA treatment significantly increased ATP levels and NADH dehydrogenase activity in a dose-dependent manner. In conclusion, GABA shows a new physiological role in EPO production, and it can thus can contribute to the prevention of anemia when used alone or in combination with other anemia treating drugs.

The present study was conducted to evaluate the effects of different dietary gamma-aminobutyric acid (GABA) levels in Nile tilapia, Orechromis niloticus. Six groups of fish averaging 3.03 +/- 0.07 g (mean +/- SD) were randomly distributed in 18 rectangular tanks in triplicates. A basal diet without GABA supplementation was used as a control (CON), and the other five diets were prepared by adding 75 mg GABA (GABA(144)), 75 mg GABA +50 mg heat-killed Lactobacillus paracasei (GABA(189)L), 150 mg GABA (GABA(197)), 150 mg GABA +50 mg heat-killed Lactobacillus paracasei (GABA(242)L), and 600 mg GABA (GABA(507)) per kg of diet. Actual GABA compositions for CON, GABA(144), GABA(189)L, GABA(197), GABA(242)L and GABA(507) diets were 96, 144, 189, 197, 242 and 507 mg kg(-1), respectively. After 8 weeks of experiment, weight gain, specific growth rate, feed efficiency and protein efficiency ratio of fish fed GABA(189)L were significantly higher than those of fish fed CON and GABA(5.7) diets (P < .05). Serum glutamic oxaloacetic transaminase level of fish fed GABA(144) and GABA(189)L was significantly lower than those of fish fed diets GABA(197) and GABA(507) (P < .05). Serum total protein of fish fed GABA(189)L was significantly higher than those of fish fed CON, GABA(144), GABA(197) and GABA(242)L. Also, superoxide dismutase activity of fish fed the CON diet was significantly lower than those of fish fed all the other diets. Broken line analysis for weight gain showed the optimum dietary GABA level could be 158 mg kg(-1) diet. In the current study, it could be concluded that addition of 75 mg kg(-1) of GABA with dietary heat-killed Lactobacillus paracasei (diet GABA189L) could increase growth and feed utilization performance, serum total protein and superoxide dismutase activity of juvenile Nile tilapia.

AimsThe aim of this study was to evaluate gamma-aminobutyric acid (GABA)- and piperine-induced erythropoietin (EPO) and EPO-receptor expression.Materials and methodsThe effect of GABA and piperine on cell viability was examined using kidney epithelial cells. Expression levels of EPO and EPO-R mRNA and protein were evaluated in response to GABA and piperine treatments. GABA- and piperine-mediated activation of the mitogen-activated protein kinase (MAPK) signaling pathway was investigated. Additionally, EPO function was evaluated using conditioned media containing EPO. The GABA receptor type involved in this process was identified.Key findingsMessenger RNA and protein expression levels of EPO and EPO-R significantly increased in response to treatment with GABA, piperine, or the combination of both, compared with control. GABA plus piperine synergistically enhanced EPO and EPO-R expression through p38 and c-Jun N-terminal kinase (JNK) MAPK signaling pathways, but not through the extracellular signal-regulated kinase (ERK) MAPK pathway. SB203580 and SP600125 (p38 and JNK pathway inhibitors, respectively) attenuated GABA plus piperine-induced EPO and EPO-R expression. Treatment of macrophages with EPO-containing conditioned media induced mRNA expression of interleukin (IL)-10 and nuclear factor (NF)-κB due to the interaction between EPO and EPO-R. Interestingly, GABA-induced EPO and EPO-R expression was mediated through GABAA, not GABAB, receptor activation.SignificanceThese findings demonstrate that GABA plus piperine-mediated p38 and JNK MAPK activation increases EPO and EPO-R expression, resulting in up-regulation of IL-10 and NF-κB.

Broiler salmonellosis is a major problem for poultry industry. Here, we supplemented broiler feed with 1% of ginger stems (GS) fermented with Lactobacillus paracasei and analyzed the effects on the resistance to Salmonella gallinarum. The chickens were divided into four dietary groups. The control group (C) received the basal diet, and the other chickens received the basal diet supplemented with 0.1% w/w L. paracasei ML-7 (L group), 0.1% ginger stem powder (GS group), or 0.2% fermented ginger stem (FGS group) for 21 days. The dietary groups were further split into two subgroups: one challenged with 1 × 105 CFU/mL S. gallinarum orally administered in 1 mL of saline from days 7 and 14, and one that received 1 mL of saline without bacteria. Both uninfected and S. gallinarum-infected broilers fed with fermented GS (FGS) significantly increased body weight and feed intake, and had lower mortality compared to relative control groups. Furthermore, dietary FGS decreased cecal, Salmonella spp. counts and serum IgA and IgG levels. These results indicate that FGS prevented S. gallinarum colonization and promoted weight gain in broilers, suggesting that FGS supplementation can be effectively used as a replacement of antibiotic growth promoters to prevent Salmonella infection.

본 연구에서는 분무건조법을 이용하여 젖산세균을 녹말로 캡슐화한 분말을 제조하고, 제조한 젖산세균 코팅 분말을 이용하여 다양한 수분활성도에 따른 평형수분함량을 측정하여 등온흡습곡선을 확립하고, pH 및 상대습도, 온도에 따른 젖산세균의 보호 효과를 알아보고자 하였다. 젖산세균 코팅 분말의 등온흡습곡선은 수분활성도에 따라 평형수분함량이 빠르게 증가하는 sigmoid형을 나타내었으나 시료간의 유의적인 차이를 나타내지 않았다. 상대습도에 따라 20, 30, $40^{\circ}C$에서 저장하였을 때, 상대습도가 낮을수록, 저장 온도가 낮을수록 젖산세균의 생존율이 높았으며, 시료들 중 생강을 첨가하여 코팅한 젖산세균 분말의 생존율이 가장 높았다. 75%의 상대습도에서 $20^{\circ}C$와 $35^{\circ}C$에서 각 4주간 저장 실험한 결과, 코팅하지 않은 젖산세균은 1주 후 사멸한 반면, 코팅한 젖산세균은 유지되었으며 특히 생강을 첨가하였을 경우 젖산세균 보호 효과가 가장 높았다. 소화 모방 실험을 통해 위액의 낮은 pH에서 녹말 코팅의 보호막 효과를 확인하였다. 이를 통해 녹말로 코팅한 젖산세균 분말은 식품 조성물, 식품 첨가용 조성물로 이용 가능하다. 한편, 프로바이오틱스 제품 제조 시 젖산세균이 8-10 log CFU/g 이상 존재해야 하는데, 본 연구에서는 분무건조 후 약 7-8 log CFU/g로 다소 감소하였지만 이는 사료 조성물 또는 사료 첨가용 조성물로 응용이 가능하다고 생각된다. Lactobacillus is a probiotic that suppresses the growth of pathogens while preventing constipation, diarrhea, and intestinal inflammation. However, various environmental conditions such as pH and temperature affect the growth of Lactobacillus. In this study, Lactobacillus plantarum was encapsulated with starch using a spray dryer to protect the viability of the organism during storage and to increase its acid tolerance. The lower water activity and storage temperature resulting from this method influenced the survival of L. plantarum. In encapsulated Lactobacillus powders, viability of Lactobacillus was increased during storage at $20^{\circ}C$ relative to that of L. plantarum stored at $35^{\circ}C$ in the same water activity conditions. Furthermore, L. plantarum encapsulated in starch with ginger showed increased viability when compared to non-encapsulated L. plantarum stored without treatment. Finally, based on a simulated digestion test, encapsulated L. plantarum survived at a pH of 2, whereas the non-encapsulated L. plantarum did not. Thus, coating the encapsulated powder with these materials was effective in maintaining Lactobacillus viability.

본 연구에서는 산란계의 in vivo실험을 통하여 난황 레시틴추출 부산물이 산란계의 뼈의 Ca 침착과 성장에 도움을 줄 수 있다는 결과를 얻었다. 산란계 전기 사료에 Ca의 수준을 0%, 0.2%, 0.4%, 0.6%로 조절하여 실험한 결과 증체량에서 0%, 0.2% Ca을 첨가한 그룹의 증체량은 유의적으로 감소하였으나 0.4%, 0.6% 첨가 그룹에서는 정상적인 성장율을 확인하였다. 반면에 0.4% Ca을 첨가한 그룹에 레시틴 추출 부산물을 0.2%, 0.4% 첨가한 그룹에서 각각 1.5%, 7.1%로 증가하여 농도의존적으로 증가하는 경향을 보였으나 유의한 차이는 없었다. 실제 산란계의 경골을 적출하여 중량, 회분, Ca의 함량을 조사한 결과 경골 중량에서는 0.4% Ca 첨가그룹에 비해 레시틴 추출 부산물을 추가로 첨가한 그룹에서 각각 8.4%, 13.4%로 증가하는 경향을 확인하였다. 회분량에서도 0.4% 첨가 그룹에 비해 레시틴 추출 부산물 첨가그룹에서 증가하였지만, 유의한 차이는 없었다. 경골내의 Ca 함량을 조사한 결과는 0.4% Ca 첨가그룹에 비해 레시틴 추출 부산물이 4.3%, 5.9%가 각각 유의한 경향으로 증가하였다. 경골에 실제로 Ca이 흡수되었는가를 확인하기 위하여 lateral femoral joint 부분을 von Kossa's stain을 실행한 결과 Ca 결핍 그룹에서 Ca 축적이 현저히 감소한 것을 확인한 반면, 그 외 그룹에서는 Ca이 충분히 침착되어 있었으며 산란계 사료내의 적정 Ca 함량이 0.4%라는 것을 다시 한번 확인하였다. 본 연구에서는 계란 난황의 레시틴 추출 부산물의 단백질 분획 소재를 이용하여 뼈 성장 촉진 인자 및 Ca 흡수촉진 보조제로서의 가능성을 보여줬으며, 그 기대를 높이기 위해 혈액분석, 측정항목의 세분화 등 반복적인 연구가 수반되어야 하겠다. A 3-weeks feeding trial with 288 laying hens were conducted to determine the efficacy of lecithin-free egg yolk at different levels of dietary Ca on performance and Ca absorption. Laying hens were divided into 6 groups according to calcium level and testing agent; 0% calcium feed (A), 0.2% calcium feed (B), 0.4% calcium feed (C, normal feed), 0.6% calcium feed (D), 0.4% calcium feed + 0.2% egg byproduct (C+0.2), 0.4% calcium feed + 0.4% egg byproduct (C+0.4). The final body weight gain of C+0.2 and C+0.4 groups were higher by 1.5% and 7.4% respectively than group C. Tibia ash contents did not show significantly difference, but calcium contents increase (p<0.05) in C+0.2 and C+0.4 groups. Parallel undecalcified tibia joint sections were stained for calcium absorption by the von Kossa's stain. This result show that lecithin free egg byproduct supplementation to normal calcium feed improved growth performance and calcium utilization in laying hens.

This study was designed to find the most suitable method and wall material for microencapsulation of the Lactobacillus plantarum to maintain cell viability in different environmental conditions. To improve the stability of L. plantarum, we developed an encapsulation system of L. plantarum, using water-in-oil emulsion system. For the encapsulation of L. plantarum, corn starch and glyceryl monostearate were selected to form gel beads. Then 10% (w/v) of starch was gelatinized by autoclaving to transit gel state, and cooled down at 60ºC and mixed with L. plantarum to encapsulate it. The encapsulated L. plantarum was tested for the tolerance of acidic conditions at different temperatures to investigate the encapsulation ability. The study indicated that the survival rate of the microencapsulated cells in starch matrix was significantly higher than that of free cells in low pH conditions with relatively higher temperature. The results showed that corn starch as a wall material and glycerol monostearate as a gelling agent in encapsulation could play a role in the viability of lactic acid bacteria in extreme conditions. Using the current study, it would be possible to formulate a new water-in-oil system as applied in the protection of L. plantarum from the gastric conditions for the encapsulation system used in chicken feed industry.

김치로부터 분리, 선별한 유산균 L. paracasei LS-2에 UV 조사 및 L. paracasei LS-2균주가 성장하지 않는 pH 3.5 MRS agar 배지에서 성장하는 변이주를 선별함으로써 변이주인 L. paracasei ML-7를 선별하였다. 선별된 변이주 L. paracasei ML-7의 유기산 생성능력, 병원성 미생물의 생육 저해 능력, 육계에서의 증체 및 사료 효율 개선효과를 시험하였다. 변이주인 L. paracasei ML-7 는 모균주인 L. paracasei LS-2에 비하여 MRS 배지에서 성장속도가 증가되었으며, 유기산 생성도 증가되었다. 또한 L. paracasei ML-7은 병원성 미생물인 E. coli ATCC 25922 (${\alpha}$-hemolysin producing E. coli)와 A. flavus ATCC 22546에 대한 생육 저해 능력이 모균주인 L. paracasei LS-2에 비해 증가되는 것을 확인할 수 있었다. 육계를 대상으로 하는 사양시험에서 변이주 L. paracasei ML-7을 급여한 처리구에서 증체량 개선 및 사료효율이 개선됨을 확인하였다. 본 결과로부터 유산균의 인공변이로부터 유기산생산 증가 및 그에 따른 가축 사양 성적 개선이 가능한 변이주의 제작이 가능함을 확인할 수 있었다. A mutant strain of Lactobacillus paracasei (ML-7) was derived by ultraviolet irradiation of the parent L. paracasei LS-2 strain, followed by colony selection on pH 3.5 MRS agar plates, on which L. paracasei LS-2 cannot grow. The L. paracasei ML-7 mutant strain produced an increased amount of organic acids compared to the L. paracasei LS-2 parent strain. Broiler chickens were randomly fed with one of three experimental diets: a basal diet as a control, and basal diets supplemented with 0.3% of the L. paracasei LS-2 and L. paracasei ML-7 strains. When concluding the study, the final body weights of the broilers fed with the supplemented basal diets were found to be significantly higher (1.9% for LS-2 and 6.2% for ML-7) than the control broiler group. Broilers fed with basal diet supplemented with the mutant L. paracasei ML-7 strain showed increased body weight and food intake, compared to broilers fed with the basal diet with or without the parent L. paracasei LS-2 strain.