迷迭香（Rosmarinus officinalis L.）精油和抗氧化剂的研究

本文应用植物引种驯化，栽培学及植物化学的理论方法对引入我国的迷迭香（Rosmarinus officinalisL．）精油和抗氧化剂进行了研究．主要结果如下; 1、昆明地区的生态气候条件最适宜迷迭香生长。南京地区局部良好环境也能满足迷迭香正常生长需要。北京地区在保护栽培的情况下,迷迭香才能够安全越冬。 2、用GC-MS和GC-IR鉴定了迷迭香精油的23种成分。不同采收期精油成分含量变化有3种类型，即似单峰曲线，似8曲线和w曲线。一年生与二年生苗在精油成分含量上略有差异。与主要栽培国相比，我国不同引种地精油成分化学类型各具特点，北京地区α—蒎稀（31.58%）含量很高，南京地区富含樟脑（14.86%）乙酸龙脑酯（7.97%）和龙脑（6.90%）含量也相对较高，昆明地区则是α—蒎稀（24.32%）和1.8—桉叶油素（24.43%）含量并重。 乙酸龙脑酯含量的高低对精油香气质量优劣起关键作用。北京地区雨季来临收获（7月初）精油香气质量最好。南京地区，11月初精油香气质量也属优质。本实验未能得到高质量的昆明地区精油。 3、从迷迭香蒸油残渣中分离得到三种抗氧化成分，迷迭香酚( Rosrmanol)、鼠尾草酚(Carnosol)和迷迭香双醛（R0smadial），并用质谱，核磁做了鉴定．硫氰铁酸法、TBA法、AOM法检测了三种成分的抗氧化效能．Rosmanol对多双键不饱和脂肪酸抗氧化效果显著,Carnosol和BHT在不同双键脂肪酸中抗氧化效果稳定，Rosmadial的抗氧化效果则随不饱和双键数目的增加而降低。在猪油中，抗氧化效能的强度顺序为Rosmanol >carnosol>BR>Rosmadial。 抗氧化物质在植株体内是不断积累的。不同引种地迷选香抗氧化效能的强弱差异表现为昆明>北京>南京。 迷迭香的正己烷，两酮提取物抗氧化效能都是随着添加量的增加而增强、丙酮提取物的抗氧化效能优于正己烷提取物。 对迷迭香抗氧化剂提取工艺做了探讨，得到R-1，R—2，R-3三种小试样品，R-2，R-3的抗氧化效果优于BHT．B—l接近于BET。

The effect of Rosmarinus officinalis and Zataria multiflora extracts on the stability of poly unsaturated fatty acids in frozen silver carp minced

The purpose of this study, Evaluation the effect of Rosmarinus officinalis and Thymus vulgaris extracts on the stability of poly unsaturated fatty acids in frozen Silver carp minced. Treatments include: Treatment 1 - Control: frozen meat packaged in conventional Treatment 2: Frozen Silver carp minced+Thyme 300 mg/kg in normal packaging Treatment 3: Frozen Silver carp minced+Rosemary 200 mg/kg in normal packaging Treatment 4: Frozen Silver carp minced+Rosemary compound (100 mg/kg) and Thyme (100 mg/kg) in normal packaging After rapid freezing of samples in the spiral freezer by individual quick freezing method, to maintain the cold temperature (-18) °C were transferred. Sampling and measurements to determine the fatty acid profile of the zero phase beginning in the first month and then every ten days, and 15 days in the second month of the third month after the monthly test. Identifying, defining and measuring the fatty acid profile by gas chromatography was performed. In this study, levels of both saturated and unsaturated fatty acids in three experimental and one control were identified as follows: A: saturated fatty acids: Meristic C14: 0/Palmitic C16: 0/Hepta decaenoic C17: 0/Stearic C18: 0/Arashidic C20: 0/B:Mono unsaturated fatty acids: palmitoleic C16: 1-W7/Oleic C18: 1-W9/Gadoleic C20: 1-W9 C:Poly unsaturated fatty acids: Linoleic C18: 2-W6/α-Linolenic C18: 3-W3 D:High unsaturated fatty acids: Arachidonic C20: 4-W6 Eicosapentaenoic acid C20: 5-EPA/W3 Docosahexaenoic C22: 6-DHA/W3 Results of this study was to determine, Thyme and rosemary extracts containing silver carp minced stored in freezing conditions, Stability of different types of fatty acids, monounsaturated fatty acids, poly-unsaturated fatty acids, omega-3 and omega-6 fatty acids are. So that none of the fatty acids measured were not significant 100% increase or decrease, While changes in the fatty acid oxidation during storage time is minimized. The results obtained from the fatty acid profiles and indicators of their and statistical tests show that treatment with rosemary extract More stable during storage (-18) ° C In comparison with the control and other treatments are shown; And at relatively low compared to other treatments and control samples oleic acid and linoleic acid, palmitic more. According to studies,in Silver carp minced that containing rosemary extract, end of the storage period of six months. Were usable, so even rosemary extract the shelf-life examples to increase more than six months.

Study of effect Rosmarinus officinalis oil and nisin on the growth of Streptococcus iniae in lab conditions and fillets of rainbow trout (Oncorhynchus mykiss)

Fish are an important part of a healthy diet since they contain high quality protein, but typically present a low fat percent when compared to other meats. Fish is an extremely perishable food commodity. On the other hand, food borne diseases are still a major problem in the world, even in well-developed countries. The increasing incidence of food borne diseases coupled with the resultant social and economic implications means there is a constant striving to produce safer food and to develop new antimicrobial agents concerns over the safety of some chemical preservatives and negative consumer reactions to preservatives they perceive as chemical and artificial, have prompted on increased interest in more ‘‘naturalgreen’’ alternatives for the maintenance or extension of product shelf-life. Particular interest has focused on the potential applications of plant essential oils. However, to establish the usefulness of natural antimicrobial preservatives, they must be evaluated alone and in combination with other preservation factors to determine whether there are synergistic effects and multiple hurdles can be devised. In this study, were evaluated the effects of different concentrations of Rosmarinus officinalis and nisin and storage time (15 days) on growth of Streptococcus iniae GQ850377 in a lab conditions and a food model system (fillets of rainbow trout) in 4 and 8 °C. In addition, we also studied multi factorial effects of four different concentration of rosemary, three different concentrations of nisin, two different levels of pH in 3 temperature 4,15 and 37 °C on log% of S.iniae during 43 days in BHI broth. The results on growth of S. iniae were evaluated using SPSS 20.0 statistical software and analyzed the logarithm of total count of the bacterial by Tukey Test. Results were considered statistically significant when P<0.05. MIC and MBC values of rosemary and nisin were 0.03, 0.075 % and 5, 40 μg/mL, respectively. The growth of S. iniae was effected significantly (P<0.05) by rosemary and nisin and also combination of rosemary and nisin in 4 and 8 °C. Samples treated with 0.135 and 0.405 % of rosemary showed a significant decrease on the growth of the bacteria compared with control sample(P<0.05). The most ١٤٦ inhibitory effects were seen in samples treated with 0.135 and 0.405% of rosemary until 9 days after storage. Also, the synergism effects of rosemary and nisin on the growth rate of bacteria was significant (P<0.05) compared with untreated samples and samples treated with the rosemary or nisin, only. Synergistic effects was observed at concentration of 0.405% rosemary and 0.75 μg/mL nisin in both temprature. Results of this study showed that different concentration of rosemary a significant inhibitory effect (P<0.05) on log% of S. iniae, in BHI broth in pH 5.5 and 7 in 4,15 and 37 °C during 43 days. In concentration of 0% rosemary (control) in pH 5.5 and 7 and 37°C, log% were 1.099 and 3.15, whereas in concentration of 0.015% rosemary were -4/241 and 1.454, respectively. The use of essential oils may improve food safety and overall microbial quality. If essential oils were to be more widely applied as antibacterials in foods, the organoleptic impact would be important. In addition, it is recommended to apply essential oils or their compounds as part of a hurdle system and to use it as an antimicrobial component along with other preservation techniques. Thus essential of R. officinalis with high antibacterial activity selected in this study could be a potential source for inhibitory substances against some food-borne pathogens and they may be candidates for using in foods or food-processing systems.

Investigating the Biological Effects of Rosemary (Rosmarinus officinalis L.) Extract on Skeletal Muscle Glucose Uptake

Skeletal muscle (SKM) is the most important tissue in maintaining glucose homeostasis and impairments in this tissue leads to insulin resistance (IR). Activation of 5’ AMP-activated kinase (AMPK) is viewed as a targeted approach to counteract IR. Rosemary extract (RE) has been reported to decrease blood glucose levels but its effects on SKM are not known. We hypothesized that RE acts directly on SKM to increase glucose uptake (GU). We found an increase in GU (184±5.07% of control, p<0.001) in L6 myotubes by RE to levels similar to insulin and metformin. Carnosic acid (CA) and rosmarinic acid (RA), major polyphenols found in RE, increased GU. RE, CA, and RA significantly increased AMPK phosphorylation and their effects on GU was reduced by an AMPK inhibitor. Our study is the first to show a direct effect of RE, CA and RA on SKM GU by a mechanism that involves AMPK activation.

Investigation of the Biological Effects of Rosemary (Rosmarinus Officinalis L.) Extract in Human Lung Cancer Cells

Cancer cells display enhanced growth rates and a resistance to apoptosis. Lung cancer accounts for the most cancer related deaths and non-small cell lung cancer (NSCLC) represents an aggressive form of lung cancer, accounting for almost 80% of all lung cancer cases. The phytochemical rosemary extract (RE) has been reported to have anticancer effects in vitro and in vivo however, limited evidence exists regarding the effects of RE and its polyphenolic constituents carnosic acid (CA) and rosmarinic acid (RA) in lung cancer. The present study shows RE, CA and RA inhibit lung cancer cell proliferation and survival in various NSCLC cell lines and that CA and RA interact synergistically to inhibit cell proliferation. Moreover RE, CA and RA are capable of altering activation and/or expression of Akt, ERK and AMPK, signaling molecules which regulate cell proliferation and survival. RE shows potential as an anticancer agent and should be further investigated.

Effect of rosemary (Rosmarinus officinalis) extract on weight, hematology and cell-mediated immune response of newborn goat kids

This study aimed at evaluating the effects of different levels of rosemary (Rosmarinus officinalis) extract on growth rate, hematology and cell-mediated immune response in Markhoz newborn goat kids. Twenty four goat kids (aged 7 +/- 3 days) were randomly allotted to four groups with six replicates. The groups included: control, T1, T2 and T3 groups which received supplemented-milk with 0, 100, 200 and 400mg aqueous rosemary extract per kg of live body weight per day for 42 days. Body weights of kids were measured weekly until the end of the experiment. On day 42, 10 ml blood samples were collected from each kid through the jugular vein. Cell-mediated immune response was assessed through the double skin thickness after intradermal injection of phyto-hematoglutinin (PHA) at day 21 and 42. No significant differences were seen in initial body weight, average daily gain (ADG) and total gain. However, significant differences in globulin (P <0.05), and white blood cells (WBC) (P <0.001) were observed. There were no significant differences in haemoglobin (Hb), packed cell volume (PCV), red blood cells (RBC), lymphocytes and neutrophils between the treatments. Skin thickness in response to intra dermal injection of PHA significantly increased in the treated groups as compared to the control group at day 42 (P< 0.01) with the T3 group showing the highest response to PHA injection. In conclusion, the results indicated that aqueous rosemary extract supplemented-milk had a positive effect on immunity and skin thickness of newborn goat kids.

Does fertilizer type and method of application cause significant differences in essential oil yield and composition in rosemary (Rosmarinus officinalis L.)?

Organic fertilizers based on seaweed extract potentially have beneficial effects on many crop plants. Herewe investigate the impact of organic fertilizer on Rosmarinus officinalis measured by both yield and oilquality. Plants grown in a temperature-controlled greenhouse with a natural photoperiod and a controlledirrigation system were treated with seaweed fertilizer and an inorganic fertilizer of matching mineralcomposition but with no organic content. Treatments were either by spraying on to the foliage or wateringdirect to the compost. The essential oil was extracted by hydro-distillation with a Clevenger apparatusand analysed by gas-chromatography mass-spectrometry (GC–MS) and NMR. The chemical composi-tions of the plants were compared, and qualitative differences were found between fertilizer treatmentsand application methods. Thus sprayed seaweed fertilizer showed a significantly higher percentage of�-pinene, �-phellandrene, �-terpinene (monoterpenes) and 3-methylenecycloheptene than other treat-ments. Italicene, �-bisabolol (sesquiterpenes), �-thujene, and E-isocitral (monoterpenes) occurred insignificantly higher percentages for plants watered with the seaweed extract. Each was significantly dif-ferent to the inorganic fertilizer and to controls. The seaweed treatments caused a significant increasein oil amount and leaf area as compared with both inorganic treatments and the control regardless ofapplication method.

Genotoxicity and mutagenicity of Rosmarinus officinalis (Labiatae) essential oil in mammalian cells in vivo

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Study of the embryotoxic effects of an extract of rosemary (Rosmarinus officinalis L)

Extracts of rosemary, Rosmarinus officinalis L., have been used in folk medicine as a diuretic, an emenagogue, an antispasmodic and its aqueous extract does not present toxicity to man, presenting, however, abortive effects. In order to evaluate if this plant induces abortion and/or interferes with the normal development of the concepts, doses of 26 mg of a 30% (w/v) R. officinalis aqueous extract (13 mg solids/ml) made with leaves, flowers and stem were administered daily by gavage during two different periods of Wistar rat pregnancy. One group of animals (N = 12) received the extract from days 1 to 6 of pregnancy (preimplantation period) and another group (N = 14) received the same extract from days 6 to 15 of pregnancy (organogenic period). Control groups (N = 12) received saline in the same volume and during the same periods as their respective experimental groups. The animals were sacrificed at term. The treatment of the darns during either the preimplantation or the organogenic period did not cause significant changes in the postimplantation loss or in the number of anomalies or malformations of the term fetuses, which also showed a similar degree of development when compared with the respective controls. The percent of preimplantation loss in the group treated before embryo implantation increased, although the difference was not significant compared to the control. This result suggests that rosemary extract may present an anti-implantation effect without interfering with the normal development of the concept after implantation.

Effects of pretreatment with rosemary (Rosmarinus officinalis L.) in the prevention of lipid oxidation in salted tilapia fillets

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Early rooting in Rosemary's (Rosmarinus officinalis L.) cuttings under the influence of chemical treatments and collecting time

The present work aimed to verify the early rooting development and the root production in rosemary's (Rosmarinus officinalis L.) cuttings under the influence of treatments with sythetics auxins, boric acid and collecting time. The branches were collected at the end of the season and the stakes were prepared and placed into solutions containing NAA or IBA (25, 50, 75 and 100 pppm) without and with boric acid and then removed to a chamber of nebulization (all experimental work). In all the experimentes, the branches were collected after 30 days, the mean number of rooted cuttings was calculated, along with the mean number of roots per stakes and the dry weight of roots. The end of winter outstood as the best season for all analysed parameters.

Acao antioxidante de extratos de alecrim (Rosmarinus officinalis L.) em files de peixes da especie pacu (Piaractus mesopotamicus Holmberg)

The antioxidant effect of plant extract has been an interesting issue for many researchers in past few years. Spices have been identified as products with large amounts of antioxidants, and rosemary is one the most studied spice. Studies have shown evidences that population with high intake of these substances have less incidence of coronary heart disease. The objective of this work was verify the antioxidant action of the commercial rosemary oleoresin Herbalox® (used to feed the pacu (Piaractus mesopotamicus) fish. The juveniles pacu were fed with isocaloric and isoproteic diets, for two groups the lipid source was soybean oil and for the another it was corn oil. For each group of the different lipid sources, rosemary extract Herbalox® (were added in one lot and on the other which was used as control. The results showed that the addition of rosemary extracts to the ration protected them against peroxidation, when compared with the control, the lipid source also have influence on these protection.

Influência do alecrim (Rosmarinus officinallis L.) na atividade de água e oxidação lipídica de peixes de uma espécie de tilápia (Oreochromis ssp-var. vermelha Flórida) submetidos à salga

Salting have been a very utilized fish conservation method, however only in the last years the basic mechanism involved in salting fish has been understood. The objectives of this study were determine the addition in brine salt of rosemary leaves, rosemary extracts and tocopherol, and your action in the followed parameters: water activity (Aw), moisture, ash, salt content and TBARS. The results showed that the addition of antioxidants was difficulted the salt absorption, however didn't have differences between rosemary or tocopherol use. In the salting time of 3 hours the values of Aw and salt levels, was respectively: 0.77±0.01 and 14.42±1.69. for control treatment; 0.85±0.02 and 9.09±1.39for rosemary filtrate; 0.78±0.03 and 10.63±0.69 rosemary without filtrating and 0.85±0.02 and 11.96±1.78 tocopherol, showed that didn't grow indigenous bacterias. Lipid oxidation was evaluated by TBARS and the results showed the oxidative effect of salt and the pro oxidant effects of alls antioxidants used in brine salting.

Atividade antioxidante e antimicrobiana do alecrim (Rosmarinus officinalis L.) em filés de tilápia (Oreochromis ssp) salgados secos durante o armazenamento congelado

Rosemary (Rosmarinus officinalis L.) is a spice from Lamiaceae family known since ancient times because its medicinal effects and, currently, several studies have pointed its antioxidant and antimicrobian effects. Lipid oxidation is a problem in food production because proceed the lost of organoleptical and nutritional qualities so required in Market. Fish salting is an ancient conservation method that expect reduce water activity and, consequently, microorganism growth in food, except halophillic bacteria. In the meantime, the inconvenient of this procedure is that the salt accelerates tissue's lipid oxidation. The aim of this work was evaluate the antioxidative and antimicrobian effects by treatment and pre treatment with rosemary (Rosmarinus officinalis L.) aqueous extract in dry salted tilapia fillets, storaged in freezing temperatures. To follow the oxidative, dry salted tilapia fillets were treated or pre treated with rosemary natural extract and storage at -18°C for 240 days. Analisys of 2-thiobarbituric acid reactive substances (TBARS), soluble nitrogen in trichloroacetic acid (TCA), water activity and microbiology were done. The pre treatment (3.39±0,53) and the treatment with rosemary (3.31±0.79) had oxidative index twice lower than the control treatment (6.14±1.21) in the last time of the research. The microbiological rosemary analisys showed count levels of resistant microorganisms to salt (2.0×103CFU/g of sample), whom causes the initial fillets contamination. The microbiological counts remained invariable in all groups during storage periods.