The aim of this study was to determine the contamination of mycotoxins and the concentration of preservatives and antioxidants in commercial pet food. 106 pet foods were purchased from online in Korea. Mycotoxin analysis were performed using liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) and preservatives and antioxidants were analyzed by high performance liquid chromatography (HPLC). The contamination of the mycotoxins in all samples was proved to comply with the Korea legislation with regards to animal feed, the analyses revealed presence of aflatoxins, ochratoxin A, zearalenone and fumonisins, with values below 1.6 ㎍/㎏, 12.1 ㎍/㎏, 478.5 ㎍/㎏ and 1873.5 ㎍/㎏, respectively. Furthermore, the simultaneous presences of different mycotoxins were observed in most of positive samples. The levels of dehydroacetic acid (DHA), sorbic acid (SA) and benzoic acid (BA) were, respectively, with values below 0.24 g/㎏, 4.82 g/㎏ and 6.35 g/㎏. The concentrations of butylated hydroxy anisole (BHA) and butylated hydroxy toluene (BHT) were demonstrated to be acceptable, with values below 0.04 g/㎏, and 0.02 g/㎏. This indicated the need for further investigation into the potential risk deriving from chronic exposure to low doses of mycotoxins. Additionally, a co-contamination of mycotoxins which interact in synergic manner are more concern. As DHA that not accepted was detected and very high levels of SA were identified in samples, an upper limit for preservatives in pet foods should be established in pet foods. BHA and BHT were detected within the recommended levels for antioxidant content, so these are considered as safe.

Despite numerous advances in in-vitro embryo production (IVP), many documented factors have been shown to influence the development of mammalian preimplantation embryos and the success of IVP. In this sense, elevated levels of reactive oxygen species (ROS) correlate with poor outcomes in assisted reproductive technologies (ART) due to oxidative stress (OS), which results from an imbalance between ROS production and neutralization. Indeed, excessive production of ROS compromises the structural and functional integrity of gametes and embryos both in vivo and in vitro. In particular, OS damages proteins, lipids, and DNA and accelerates cell apoptosis. Several in-vivo and in-vitro studies report an improvement in qualityrelevant parameters after the use of various antioxidants. In this review, we focus on OS and the source of free radicals and their effects on oocytes, sperm, and the embryo during IVP. In addition, antioxidants and their important role in IVP, supplementation during oocyte in vitro maturation (IVM), in vitro culture (IVC), and semen extenders were discussed. Nevertheless, various methods for determining the level of ROS in germ cells have been briefly described. Still, it is crucial to develop standardized antioxidant supplement systems to improve overall IVP success. Further studies should explore the safety, efficacy, mechanism of action, and combination of different antioxidants to improve IVP outcomes.

Antioxidants are food additives that extend the shelf life of food products by preventing lipid rancidity caused by active oxygen. They can either be naturally-derived or manufactured synthetically via chemical synthesis. In this study, method validation of five synthetic antioxidants, namely butylated hydroxyanisole, butylated hydroxytoluene, tertiary butylhydroquinone, propyl gallate, and disodium ethylenediaminetetraacetic acid, was performed using a high performance liquid chromatography–ultraviolet visible detector, and the method applicability was evaluated by analyzing foods containing antioxidants. The coefficient of determination (R2) average was 0.9997, while the limit of detection and limit of quantification were 0.02–0.53 and 0.07–1.61 mg/kg, respectively. The intra and inter-day accuracies and precisions were 83.2±0.7%–98.7±2.1% and 0.1%–5.7% RSD, respectively. Inter-laboratory validation for accuracy and precision was conducted using the Food Analysis Performance Assessment Scheme quality control material. The results satisfied the guidelines presented by the AOAC International. In addition, the expanded uncertainty was less than 16%, as recommended by CODEX. Consequently, to enhance public health safety, the results of this study can be used as basis data for evaluating the intake of synthetic antioxidants and assessing their risks in Korea.

본 연구는 식물 공장에서 배초향(Agastache rugosa Kuntze) 재배 시, 야파시간에 따른 생육과 항산화 물질 함량에 미치는 영향에 대하여 살펴보고자 수행하였다. 본엽 4매인 배초향 묘 를 백색 LEDs를 이용한 수경재배 시스템에 정식하였다. 야파 처리 효과를 알아보기 위해 06시부터 광도 200μmol·m-2·s-1 조건으로 시간을 24시간 주기에서 18:1:2:3, 18:2:2:2, 18:3:2:1 (명기:암기:명기:암기) 비율로 처리하였으며 대조구는 20:4 (명기:암기)로 설정하여 4주간 재배하였다. 엽장과 엽폭, 화아 수를 제외하고는 유의적 차이가 발생하지 않았다. 배초향 지 상부의 항산화물질 함량은 tilianin과 acacetin이 높았고, 지하 부에서는 rosmarinic acid(RA)의 함량이 유의하게 높았다. 배초향의 건물중당 RA함량은 18:1:2:3과 18:2:2:2에서 대조 구보다 47.92, 51.46% 높았고, 건물중당 tilianin과 acactin은 18:3:2:1에서 유의적으로 높게 나타났다. 야파처리를 하였을 때 동일한 누적 광량으로 인해 생육 차이를 보이지 않았지만, 18:2:2:2의 야파처리가 총 폴리페놀 함량이 높게 나타났다. 따 라서 인공광을 이용한 배초향 재배 시스템에서 야파처리를 통 하여 생육 저하 없이 배초향의 항산화 물질 증대 효과를 기대 할 수 있을 것으로 사료된다.

본 연구는 사과 잎 추출물의 화장품 소재로 응용하기 위한 가능성을 평가하기 위하여 에탄올 70%로 추출하여 실험을 진행하였다. 사과 잎 추출물을 GC/MS로 분석하였고, 폴리페놀, 플라보노이드 함량과 DPPH radical 소거 활성을 통하여 항산화, 세포생존율(MTT assay) 확인을 통한 독성 평가를 하였으며, Nitric oxide (NO) 생성 저해 측정을 통해 사과 잎 추출물을 처리한 군에 4.6배의 NO 생성량이 감소하는 것을 확인하였고, 항염효과를 알아보았다. 총 폴리페놀 함량은 78.80 ± 0.25 ㎎/g, 총 플라보노이드 65.25±6.62 ㎎/g로 나타났다. DPPH radical 소거 활성은 추출물 농도 0.25%에서 79.8± 0.99%, 0.5%에서 88.13±0.89%, 1%에서 96.83±2.00%로 소거능이 증가함을 확인하였다. 세포 생존율 평가는 고농도인 1000 ppm 농도에서도 91.19±3.49%로 80% 이상의 생존률을 확인하였다. GC-MS 성분분석 결과 catechol(5.65%), DL-Gluciol(12.05%), Ascorbicacid(2.41%) ,Phytol(13.88%), Hexanoic acid(5.47%) 등 항산화 화장품 소재로 활용이 사료된다. 이에 실험 결과 사과 잎 추출물을 이용한 천연 기능성 화장품 소재 개발에 있어 중요한 기초 자료로 이용되고자 한다.

Bisphenol A (BPA), an endocrine-disrupting chemical, has received tremendous attention in the past few decades because of its detrimental health effects. Growing evidence supports that BPA is capable to alter the reproductive performance of the exposed individual. In spermatozoa, it has been reported that BPA increased oxidative stress by the overproduction of reactive oxygen species (ROS), subsequently affects the sperm function, biochemical properties, and fertility. Since antioxidants minimize cellular oxidative stress, therefore may have protective effects against BPA-induced stress. In the present study, we incubated mice spermatozoa for 6 h in a condition that support in vitro fertilization. The sperm incubation media was additionally supplemented with either BPA or BPA together with antioxidants, such as glutathione, vitamin C, and vitamin E. Our results showed that antioxidant significantly decreased the production of ROS that subsequently supports motility and acrosomal integrity of BPA-exposed spermatozoa. Particularly, glutathione and vitamin E inhibit protein kinase-A dependent phosphorylation of sperm proteins subsequently prevented precocious acrosome reaction. In addition, both antioxidants were found to restore fertilization and early embryo development potentiality of BPA-exposed spermatozoa. Therefore, we conclude that antioxidants minimize oxidative stress in spermatozoa in a BPA containing micro-environment, thus avoiding BPA-mediated harmful consequences. The current finding has both theoretical and clinical significance for developing potential remedies of the BPA toxicity.

본 연구는 식용화 팬지의 UV-B 주·야간 처리에 따른 항산화물질 변화와 생장반응을 알아보고자 수행되었다. 식용화 팬지(Viola comuta ‘Purple’, ‘Blue’)를 대상으로 인공 재배기에서 광원을 LED(백색) 120μmol·m-2s-1로 16시간으로 하였으며, 자외선 UV-B(310~320nm)처리는 기존연구에서 UV-B처리 효과가 좋았던 20분을 주간(11:50~12:10)과 야간(23:50~00:10)으로 처리하였다. 처리 결과, 두 품종 모두 대조구와 UV-B 주·야간 처리간의 꽃과 잎의 생체중 및 건물중은 유의한 차 이가 없었으며, ‘Blue’의 야간 UV-B 처리를 제외한 모든 처리구에서 꽃수도 유의차가 없었다. 처리 후 1, 3, 5주차에 분석한 결과, 잎의 엽록소함량은 ‘Purple’의 3주차를 제외한 모든 처리구에서 유의차가 없었으며, 두 품종 모두 Fv/Fm과 NPQ(Non Photochemical Quenching)값이 처리 간 유의차가 없었다. 또한 UV-B 처리 5주차에 활성산소(superoxide, O2 -)의 Nitrotetrazolium blue chloride(NBT) 염색 후 현미경으로 관찰한 결과, 처리 간 차이가 없었다. 항산화물질인 총 플라보노이드 함량을 분석한 결과, ‘Purple’은 3주차까지 대조구보다 주간 UV-B 처리구가 많은 것을 알 수 있었고, ‘Blue’의 경우 5주차까지 유의차는 없었지만 주·야간 UV-B 처리구가 대조구에 비해 높은 것을 알 수 있었다. 안토시아닌 함량은 ‘Purple’의 경우 3주차만 제외하고 야간 UV-B 처리구가 가장 높은 것을 알 수 있었으나, ‘Blue’의 경우 처리 간 유의차가 없었다. 결론적으로, 주·야간 UV-B 20분처리는 팬지 ‘Purple’과 ‘Blue’의 생장에 거의 영향을 주지 않았으며, ‘Purple’의 총 플라보노이드 함량은 처리 3주까지 증가하였다.

The present study was aimed to determine the effects of green tea extract (GTE) and beta-mercaptoethanol (β-ME) supplementation in boar sperm freezing extender on in vitro fertilization (IVF) and reactive oxygen species (ROS) and glutathione (GSH) levels of presumptive zygotes (PZs). Experimental groups were allocated into lactose egg yolk (LEY) without antioxidant (control), GTE (1,000 mg/l in LEY) and β-ME (50 μM in LEY). In freezing, spermatozoa extended with LEY were cooled to 5°C for 3 h and then kept at 5°C for 30 min following dilution with LEY containing 9% glycerol and 1.5% Equex STM. The final sperm concentration was 1 × 108/ml. Spermatozoa were loaded into straws and frozen in nitrogen vapor for 20 min. For IVF, oocytes were matured in NCSU-23 medium and co-cultured with spermatozoa following thawing at 37°C for 25 sec. At 12 h following IVF, IVF parameters (sperm penetration and monospermy) were evaluated. In addition, GSH and ROS levels of PZs were determined by Cell Tracker Blue CMF2HC and DCHFDA, respectively. IVF parameters did not show any significant difference among the experimental groups. GSH and ROS levels of PZs were not significantly different between groups. In conclusion, antioxidant supplementation in boar sperm freezing could not influence IVF parameters, ROS and GSH levels of PZs.

커피와 다류에 대한 소비는 전 세계적으로 해마다 증가 하는 추세이며, 한국을 포함하여 아시아권에도 물 다음으로 가장 많은 소비가 이루어지는 음료는 커피와 차(녹차, 케모마일차, 페퍼민트차, 둥글레차, 보이차, 홍차, 대추차) 이다. 본 연구는 국내에서 시판되는 커피전문점의 브랜드 커피 5종과 침출차 20종(녹차 4종, 홍차 3종, 보이차 3종, 케모마일차 3종, 페퍼민트차 3종, 둥글레차 3종, 대추차 1 종), 고형차 2종(대추차 2종)에 대하여 각 음료 1잔에 함유되어 있는 비타민C, 카페인, 총폴리페놀, 플라보노이드 를 분석하였고, 이들의 항산화활성을 측정하여 서로의 상관관계를 살펴보았다. 추가적으로 항산화활성이 높은 커 피와 다류 총 4종을 대상으로 과산화수소(H2O2)로 유도된 산화적 스트레스로부터 세포 보호효과를 평가하여 시판 커피와 다류의 항산화 연구를 위한 기초자료를 확보하고자 하였다. 녹차와 홍차 각각 1잔당 비타민 C 함량은 0.04~ 1.58 mg 이었고 커피와 나머지 차는 비타민 C가 검출되지 않았다. 카페인 함량은 브랜드 커피가 1잔당 150.17~ 202.75 mg으로 다른 종류의 음료보다 높았다. 음료에 함 유된 총 폴리페놀 함량은 gallic acid의 등량값(GAE)으로 표시할 때 브랜드 커피 A(BCA)가 265.54 mg / serving size 으로 가장 높았고, 플라보노이드 함량은 quercetin 등량값 (QE)으로 브랜드 커피 B(BCB)가 12.14 mg / serving size 으로 가장 높았다. 음료 4종(브랜드 커피 A, C 그리고 녹 차 D, 홍차 A) 시료의 농도가 높아질수록 HeLa 세포 내 활성산소종(reactive oxygen species, ROS) 생성 억제효과 및 H2O2의 소거활성이 증가하였다. 본 연구결과 브랜드 커피 및 홍차, 녹차는 각각 1잔당 비타민 C의 등량값으로 590, 330, 300 mg의 항산화능을 가지며, HeLa 세포 내에 서도 활성산소 감소효과가 확인되는 우수한 항산화 음료로 평가되었다.

본 연구에서는 식품용 기구 및 용기·포장으로부터 식 품유사용매로 이행되는 10종의 산화방지제(butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), Cyanox 2246, 425 and 1790, Irgafos 168, 및 Irganox 1010, 1330, 3114 and 1076)의 분석법을 확립하였다. 식품유사용 매 중 물, 4% 초산, 50% 에탄올의 경우 hydrophiliclipophilic balance (HLB) 카트리지로 고체상 추출(SPE, Solid Phase Extraction)을 하였고, n-헵탄의 경우 이소프로 필알콜로 희석하여 HPLC-UVD (276 nm)로 산화방지제의 이행량을 분석하였다. 확립된 분석법으로 위생백, 지퍼백, 우유팩, 주스팩, 가공식품용 포장지, 밀폐용기, 일회용기 등 국내 유통 폴리에틸렌(78건) 및 폴리프로필렌(122건) 재질의 식품용 기구 및 용기·포장 200건으로부터 식품 유사용매로 이행되는 10종의 산화방지제 이행량 조사 결과, 총 78건의 폴리에틸렌 식품용 기구 및 용기·포장 중 5건에서 Irganox 1010이 ND~1.444 mg/L 검출되었고, 총 122건의 폴리프로필렌 식품용 기구 및 용기·포장 중 41건에서 Irganox 1010이 ND~3.106 mg/L, 28건에서 Irganox 1076이 ND~4.752 mg/L, 34건에서 Irgafos 168이 ND~ 3.635 mg/L 검출되어 총 3종의 산화방지제가 검출되었다. 검출된 Irganox 1010, Irganox 1076, Irgafos 168에 대해 일 일추정섭취량(EDI)을 계산하고 일일섭취한계량(TDI)과 비 교하여 위해도를 평가한 결과, 폴리에틸렌 재질 중 Irganox 1010은 TDI 대비 0.0067%, 폴리프로필렌 재질 중 Irganox 1010, Irganox 1076, Irgafos 168은 TDI 대비 0.0073%, 0.1800%, 0.0200%로 안전한 수준임을 확인하였다. 본 연구에서 확립된 폴리에틸렌 및 폴리프로필렌 식품용 기구 및 용기·포장 중 산화방지제 분석법 및 안전성 평가 결과는 앞으로 기구 및 용기·포장의 안전관리를 위한 과학적인 근거자료로 활용될 것으로 판단된다.

To increase the productivity of in vitro development, the antioxidants have been used for culture system of bovine oocytes and embryos. However, comparative studies on these molecules are rare and direct beneficial effects on blastocyst production cannot be discriminated for best results. The study was conducted to determine the influence of N-acetyl-L-cysteine (NAC), N-acetyl-L-cysteine amide (NACA), glutathione (GSH) and cysteamime (CYS) on maturation competence of COCs from GV to MII stage and productivity of blastocyst formation during in vitro fertilization and culture. There was no difference among maturation rates of oocytes to metaphase II with polar body with antioxidants for any of the treatment groups (p>0.05). However, the significant improvement on the rate of blastocysts (32.3±5.0%) was found in 0.1 mM CYS treatment than 0.3 mM NAC, 0.2 mM NACA or 0.5mM GSH (p<0.05). The addition of NAC (18.8±3.7%) or NACA (21.2±3.9%) did not improve development competence to morula and blastocysts than control (24.4±4.1%) and GSH (26.5±5.0%) (p>0.05). Our study showed that medium supplementation with CYS during IVM and IVC improved the rate of bovine embryo development but not with NAC, NACA and GSH addition.

The oocyte undergoes various events during In vitro maturation (IVM) and subsequence development. One of the events is production of reactive oxygen species (ROS) that is a normal process of cell metabolism. But imbalances between ROS production and antioxidant systems induce oxidative stress that negatively affect to mammalian reproductive process. In vitro environments, In vitro matured oocytes have many problems, such as excessive production of ROS and imperfect cytoplasmic maturation. Therefore, In vitro matured oocytes still have lower maturation rates and developmental competence than in vivo matured oocytes. In order to improve the IVM and In vitro culture (IVC) system, antioxidants, vitamins were added to the IVM, IVC medium. Antioxidant supplementation was effective in controlling the production of ROS and it continues to be explored as a potential strategy to overcome mammalian reproductive disorders. Based on these studies, we expect that the use of antioxidants in porcine oocytes could improved maturation and development rates.

During the oocyte maturation, antioxidants may be beneficial for futher developmental competence against reactive oxygen species (ROS) because the media for oocytes lack boiomolecules that serve as scavengers. In this study, N-acetyl-L-cysteine (NAC), N-acetyl-L-cysteine amide (NACA), glutathione (GSH) and cysteamime were compared to determine the effects of protection for ROS from GV to MII stage when supplemented during in vitro maturation (IVM) and in vitro culture (IVC) of bovine oocytes. NAC is one of well known ROS scavanger and NACA is modified form of NAC to help permeation into cytosolic area of oocytes. Significant improvement on the development undergoing blastocysts (32%, vs 18%, 22%) were found when cysteamine (0.1mM) was added to the maturation medium than NAC (0.3 mM), NACA (0.2mM) or GSH (0.5 mM) as compared to control medium with antioxydents. However, the addition of NAC(18%) or NACA(21%) to media did not improve the proportion of oocytes undergoing development to morula and blastocysts than control (24%) and GSH (26%). Our study showed that medium supplementation with cysteame during IVM and in vitro culture (IVC) improved the rate of bovine embryo development, in contrast to extracellular antioxidants like NAC, NACA and GSH that caused no improvement.

The purpose of this study was to examine the effects of taurine and vitamin E on ovarian granulosa cells damaged by bromopropane (BP) in pigs. We evaluated cell viability, plasma membrane integrity (PMI) and apoptotic morphological change in porcine ovarian granulosa cells. The cells were treated with 1-BP (0, 5.0, 10, and 50 μM), 2-BP (0, 5.0, 10, and 50 mM), taurine (0, 5.0, 10, and 25 mM), and vitamin E (0, 100, 200, and 400 μM) for 24 h. 10 μM 1-BP and 50 μM 2-BP inhibited viability and PMI, and induced apoptosis in porcine ovarian granulosa cells (p < 0.05). Cell viability and PMI were increased by taurine (10 and 25 mM) and vitamin E (100 and 200 μM), and apoptosis decreased (p < 0.05). Finally, the porcine ovarian granulosa cells were co-treated with BPs (10 μM), taurine (10 mM) and/or vitamin E (200 μM). Cell viability and PMI in the co-treated cells were increased, and apoptosis was decreased. In conclusion, taurine and vitamin E can improve cell viability and inhibition of apoptosis in porcine ovarian granulosa cells damaged by bromopropane.

Artificial insemination technique has been contributed immensely for production of livestock worldwide as a critical assisted reproductive technique to preserve and propagate excellent genes in domestic animal industry. In the past decade, methods for semen preservation have been improved mostly in liquid preservation method for boar semen and freezing method for bull semen. Among many factors affecting semen quality during preservation, reactive oxy-gen species, produced by aerobic respiration in sperm for survival and motility, are unfavorable to sperm physiology. In mammalian cell as well as in the sperm, antioxidant system plays a role in degradation of reactive oxygen species. Magnetized water forms smaller stabilizing water clusters, resulting in high absorption and permeability of the cell for water, implicating its application for semen preservation. Therefore, this review focuses on preservation methods of boar and bull semen with respect to improvement of extender and reduction of reactive oxygen species by using magnetized water and supplementation of antioxidants.

In order to investigate the effects of antioxidants on the culture of mouse preantral follicles in vitro, we examined the effects of taurine, glutathione and catalase on their growth and maturation. Addition of taurine was not effective on the survival of preantral follicles. However, metaphase II rates of oocytes within preantral follicles were signifi-cantly higher in 1 mM treated group than in control and 10 mM treated group (p<0.05). Glutathione did not improved the rates of survival and metaphase II oocytes. However, metaphase II rates of oocytes progressively decreased with increasing glutathione concentration. Catalase also showed that the rates of survival and metaphase II oocytes pro-gressively decreased with increasing concentration. Especially, all of preantral follicles cultured in medium containing 100 IU/ml catalase were degenerated. These results suggest that low concentraion of taurine, as an antioxidant, have positive effect on the culture of mouse preantral follicles in vitro.