川牛膝多糖的免疫活性及其水解活性片段分离研究

川牛膝多糖（CP）是从传统中药川牛膝（Cyathula officinalis Kuan）中提取的一种活性多糖，现代药理研究表明川牛膝多糖是川牛膝许多生物活性的物质基础。本实验室前期进行了川牛膝多糖的提取、分离、结构鉴定及其部分活性研究，发现川牛膝中多糖含量非常高，在对川牛膝多糖活性的初步研究中也证实了其具有免疫调节作用。我们为了进一步了解其免疫调节活性，并为构效关系的研究奠定基础，对其进行了如下研究： 1． 通过体外毒性检测、淋巴细胞增殖实验、NK细胞杀伤活性和腹腔巨噬细胞吞噬中性红活性测定，发现川牛膝多糖在10～300μg/mL浓度范围内，对细胞无毒性作用；能够促进LPS诱导的B淋巴细胞增殖(P<0.01)、增强NK细胞杀伤活性(P<0.05)和PMΦ吞噬中性红活性(P<0.01)，且随多糖浓度增高而增强；但其对ConA诱导的T淋巴细胞的增殖无促进作用(P>0.05)。 2． 通过正常小鼠体内淋巴细胞转化实验、迟发型变态反应分析、抗体生成细胞检测、碳粒廓清检测、腹腔巨噬细胞吞噬鸡红细胞活性和NK细胞活性测定，发现川牛膝多糖在适应性免疫方面能够促进SRBC免疫小鼠体内的抗体生成细胞的生成（P<0.01）和增强DNFB诱导的DTH（P<0.05），但对ConA诱导的脾淋巴细胞增殖无促进作用（P>0.05）；在固有免疫方面能够提高小鼠碳粒廓清速率（P<0.05），PMΦ吞噬 CRBC 活性（P<0.01）和NK细胞杀伤活性（P<0.05）。同时还发现其对由环磷酰胺(Cy)引起的白细胞数下降具有很好的抑制作用（P<0.01）。 3． 为了获得结构明确、均一的保留活性的川牛膝多糖片段，为其作用机制、构效关系研究提供关键研究材料，我们开展了“保留免疫活性的最小片段”的分离制备的初步研究。建立并优化了川牛膝多糖的酸水解条件，发现在6%的样品浓度，0.025mol/L的硫酸浓度，65℃的水解温度，水解时间为8min的条件下可以得到一系列连续的多糖片段；采用Bio-Gel P2 分子筛柱层析分离得到5个级分，通过体外淋巴细胞增殖实验、NK细胞活性测定、腹腔巨噬细胞吞噬中性红实验发现其中的一个片段仍保留较强的免疫活性，并测得其分子量约为2057Da，为保留免疫活性的最小片段的进一步分离奠定了基础。 Cyathula officinalis Kuan is a commonly-used Traditional Chinese Medicine (TCM) with a wide range of pharmacological activities. Modern pharmacological researches showed the polysaccharide extracted from it (CP) is an important component for many bioactivities of this TCM. In the previous studies, we found CP showed significant immuno-regulative activities. In order to evaluate this activity systematically and lay foundations for revealling its immuno-regulative machanisms and the Structure -Function relationship, we carried out the following research works: 1． The in vitro immunoactivities of CP were evaluated by using normal mice immunocytes with respects to cytotoxicity, lymphocytes proliferation, NK activity and the ability of peritoneal macrophage phagocytizing neutral red. The polysaccharide showed no cytotoxicity below the concentration of 300 μg/mL, and could promote B lymphocytes proliferation (P<0.01), enhance NK activity (P<0.05) and the ability of peritoneal macrophage phagocytizing neutral red (P<0.01) at the concentration of 10-300 μg/mL. The above effects were positively correlated with the concentration of the polysaccharides. But it could not promote T lymphocytes proliferation (P>0.05). 2． The in vivo immunoactivities of CP were observed on normal mice through the following indices: splenic lymphocyte transformation efficiency, delayed-type allergy, antibody-forming cells activity (AFC), rate of carbon clearance, rate of peritoneal macrophage phagocytizing chicken red blood cell (CRBC) and NK activity, and its influence on the decline of the mouse leucocyte count induced by Cy. The polysaccharide at medium-dose enhanced delayed-type allergy （P<0.05）and NK activity（P<0.05） and increased the rate of carbon clearance（P<0.05）, AFC activity（P<0.01） and the rate of peritoneal macrophage phagocytizing CRBC（P<0.01）. The polysaccharides also effectively resisted the decline of the mouse leucocyte count induced by Cy（P<0.01）. However, it couldn’t increase the splenic lymphocyte transformation efficiency（P>0.05）. 3． Attempting to isolate and prepare the minimal fragments retaining activity with identical structure for further studying on immuno-regulative mechanism and Structure-Function relationship, we carried out the study on hydrolysis of CP, isolation of hydrolysed fragments, and the activity evaluation of the isolated fragments. CP with concentration of 6% was hydrolysed at 65℃ for 8 min with sulfuric acid of 0.025 mol/L，then the hydrolysate was separated using Bio-Gel P2 chromatography, 5 portions of fragments were obtained. The immunoactivities of these fragments were evaluated by using normal mice immunocytes with respect to lymphocytes proliferation, NK activity and ability of peritoneal macrophage phagocytizing neutral red. One fragment with relative molecular mass of 2057Da was found retaining immunoactivity.

Dracocephalum moldavica L. and Melissa officinalis L. : Chemistry and Bioactivities Relevant in Alzheimer’s Disease Therapy

Oksidatiivisen stressin eli liiallisen reaktiivisten happiyhdisteiden määrän soluissa on jo pitkään arveltu olevan tärkeä Alzheimerin taudin kehittymiseen ja etenemiseen vaikuttava tekijä. Tämän vuoksi kiinnostus erilaisten antioksidanttien (yhdisteitä, jotka neutraloivat näitä happiradikaaleja soluissa) mahdollisia terapeuttisia ominaisuuksia Alzheimerin taudin hoidossa on tutkittu laajalti. Tähän mennessä ei kuitenkaan ole vielä onnistuttu löytämään antioksidanttia, joka olisi hyödyksi Alzheimerin taudin hoidossa. Tämän vuoksi on tärkeää pyrkiä löytämään uusia antioksidanttien lähteitä sekä tutkia niistä löytyviä aktiivisia yhdisteitä. Kiinnostus luonnon antioksidantteja kohtaan on kasvanut voimakkaasti viime aikoina. Huomio on kiinnittynyt erityisesti aromaattisista sekä lääkekasveista löytyviin antioksidantteihin. Lamiaceae- perheeseen kuuluvia tuoksuampiaisyrttiä (Dracocephalum moldavica L.) ja sitruunamelissaa (Melissa officinalis L.) on käytetty Iranissa pitkään sekä ruoanlaitossa että lääkinnässä, minkä vuoksi näiden kasvien uutteiden antioksidanttisisältöä päätettiin analysoida käyttäen useaa erilaista in vitro- menetelmää. Näissä kokeissa ilmeni, että uutteilla oli useita antioksidanttisia vaikutuksia. Näistä antioksidanttisista vaikutuksista vastaavia yhdisteitä pyrittiin tunnistamaan käyttäen HPLC-PDA- tekniikkaa, minkä seurauksena niiden havaittiin sisältävän erilaisia polyfenoleita, kuten hydroksyloituneita bentsoeeni- ja cinnamamidihapon johdannaisia sekä flavonoideja. Kummankin kasvin uutteissa runsaimmin esiintynyt yhdiste oli rosmariinihappo. Sitruunamelissaa (M. officinalis) on käytetty antiikin ajoista alkaen kognitiivisten toimintojen häiriöiden hoidossa. Perustuen tietoon kasvin käytöstä perinteisessä lääkinnässä, sen tehoa Alzheimerin taudin hoidossa on tutkittu viime aikoina kliinisin kokein. Sitruunamelissan todettiinkin olevan hyödyksi lievää ja keskivaikeaa Alzheimeimerin tautia sairastavien potilaiden hoidossa. Väitöskirjan osanan olevasta kooste-artikkelista käy ilmi, että tutkimalla lääkekasvien ominaisuuksia voidaan saada arvokkaita suuntaa-antavia vihjeitä Alzheimerin taudin lääkehoidon kehittämiseen. Tämän perusteella päätettiinkin testatata myös sitruunamelissauutteen kykyä estää asetyylikoliiniesteraasin (AChE) toimintaa, koska tämän entsyymin toiminna estämisen tiedetään olevan hyödyksi Alzheimerin taudin hoidossa. Uute kykeni estämään AChE:n toimintaa, minkä vuoksi uutteen sisältämiä komponentteja päätettiin tutkia terkemmin. Uute jaettiin erilaisiin fraktioihin käyttäen HPLC-menetelmää, minkä jälkeen testattiin jokaisen fraktion kykyä inhiboida AchE. Suurin osa fraktioista kykeni inhiboimaan AChE:n toimintaa selkeästi tehokkaammin, kuin raakauute. Kaikista tehokkainta fraktiota analysoitiin tarkemmin sen aktiivisten yhdisteiden tunnistamiseksi, minkä seurauksena sen sisältämät yhdisteet tunnistettiin cis ja trans-rosmariinihapoiksi. Tässä tutkimuksessa tunnistettujen yhdisteiden hyödyllisyyttä Alzheimerin taudin hoidossa tulisi seuraavaksi tutkia erilaisissa in vivo-malleissa. Lisäksi jäljellä olevien fraktioiden kemiallinen koostumus tulisi selvittää sekä antioksidanttiaktiivisuuden ja AChE:n toiminnan inhiboinnin välistä mahdollista yhteyttä tulisi tutkia tarkemmin. Tämä tutkimus osoittaa tuoksuampiasyrtin (D. moldavica) sekä sitruunamelissan (M. officinalis) sisältävän monenlaisia aktiivisia antioksidantteja. Lisäksi sitruunamelissan sisältämät yhdisteet kykenivät estämään asetyylikoliiniesteraasin (AchE) toimintaa. Nämä tulokset tukevat osaltaan väitöskirjan osana olevan kooste-artikkelin johtopäätöksiä, joiden mukaan etnofarmakologinen kasvitutkimus voi osoittautua erittäin hyödylliseksi kehitettäessä uutta lääkehoitoa Alzheimerin tautiin. Lisäksi tässä väitöskirjassa kuvattu tutkimus osoittaakin, että perinteisesti lääkekasvina käytettyä sitruunamelissaa voidaan mahdollisesti hyödyntää uusien Alzheimerin taudin hoitoon käytettävien lääkkeiden kehityksessä.

Development of tissue culture and virus-induced gene silencing for Calendula officinalis

Calendula officinalis is grown widely as an ornamental plant across Europe. It belongs to the large. Asteraceae family. In this study, the aim was to explore the possibilities to use Calendula officinalis as a new model organism for flower development and secondary mechanism studies in Asteraceae. Tissue culture of Calendula officinalis was established using nine different cultivars. Murashige & Skoog (MS) medium with four different combinations of plant growth regulators were tested. Of all these combinations, the medium containing 1mg/l BAP, 0.1 mg/l IAA, and 1mg/l Zeatin achieved highest frequency of adventitious shoot regeneration from hypocotyl and cotyledon explants. Virus-induced gene silencing is a recent developed genetic tool for charactering the gene functions in plants, and extends the range of host plants that are not accessible for Agrobacterium transformation. Here, tobacco rattle virus (TRV)-based VIGS technique was tested in calendula (cv. Single Orange). We used TRV carrying Gerbera hybrid phytoene desaturase (PDS) gene fragment to induce PDS silencing in calendula. Vacuum infiltration and syringe infiltration methods both resulted in photo-bleaching phenotypes in leaves, bracts and petals. Loss-of-function phenotypes occurred on calendula 13 days post-infiltration. In conclusion, the data indicates that calendula explants can be regenerated through tissue culture which is a prerequisite for development of stable transformation methods. However, further optimization is still needed to improve the frequency. In addition, VIGS was applied to silence PDS marker gene expression indicating that this method has potential for gene functional studies in future.

Producción de primicia en invernadero de híbridos de espárrago verde (Asparagus officinalis var. Altilis) en su cuarto año productivo

Resumen: La producción de espárragos en Argentina está caracterizada por una elevada estacionalidad concentrada en el periodo octubre-diciembre, por lo que el empleo de invernaderos brinda la posibilidad de ampliar el calendario de oferta, anticipando la entrada en producción. Con el objetivo de evaluar el rendimiento de primicia de diferentes híbridos de espárrago verde, se realizó un ensayo en invernadero con ocho híbridos, en UCA Buenos Aires, iniciado el 15/11/2006, mediante plantines de 100 días a 1m*0,3m. Se evaluaron los siguientes genotipos: Italo, Zeno, Eros, Ercole, H-668, Marte, y Giove, de origen italiano, y UC-157 de origen americano, como testigo por ser el tradicionalmente cultivado en Argentina. Se evaluaron 22 cosechas, con una frecuencia de día por medio, del 17/08/2011-25/10/2011. Se estudiaron diferencias en kg totales y comerciales (PFT y PFC), Nº turiones totales y comerciales/ha (NTT y NTC) y distribución de calibres (DC): Jumbo (J), Extra-Large (XL), Large (L), Medium (M), Small (S) y Asparagina (A). Se efectuó un análisis multifactor ANOVA LSD test (P>0.05). En promedio se obtuvieron: PFT: 17053; PFC: 7904 kg.ha-1; NTT: 670566 y NTC: 520938 turiones.ha-1. Se destacaron: en PFT: Italo: 29458a, Zeno: 23056b, Giove: 23034b y H-668: 18568bc; en PFC: Italo: 14850a, Giove: 9856b, Zeno: 9130bc y H-668: 8228bcd; en NTT: Italo: 842512a, H-668: 754512a, Giove: 715000b y Eros: 707498b; en NTC: Italo: 844998a, H-668: 667502b y Eros: 542498bc y en DC: en J: Italoa, Gioveb, Zenobc y UC-157bcd; en XL: Italoa, Zenoab y Gioveb; en L: Giovea, Italoa, Zenoab, H-668abc y UC-157abc; en M: Italoa, H-668ab y Erosab; en S: H-668a, Erosa e Italoab y en A: H-668a y Marteab. Por lo expuesto resulta alentadora la productividad de Italo, Giove, Zeno y H-668 para producción de espárrago verde de primicia en invernadero.

Evaluación del crecimiento y producción de turiones de espárrago verde (Asparagus officinalis L.) producidos bajo dos sistemas de inicio, arañas y plantines, en Azul, Buenos Aires

En dos plantaciones de espárrago verde (Asparagus officinalis L.) en el centro de la Pcia. de Buenos Aires, del cultivar UC 157 F2, iniciadas con arañas y plantines, durante su primer año de cosecha, se determinó la tasa de elongación diaria y total hasta el momento de la ramificación de los turiones, el número de días hasta alcanzar el largo exportable (22.5 cm) y los días hasta el momento de la ramificación con el objetivo de estudiar su comportamiento en la zona centro de la provincia de Buenos Aires. Las mediciones se efectuaron durante seis semanas, desde el 26 de septiembre hasta el 4 de noviembre de 2005, desde la emergencia del turión hasta el día de su ramificación (rama de 5 mm sobre su bráctea). La elongación máxima hasta el día de ramificación en promedio y para ambos sistemas de inicio, varió 2 desde 35.3 cm al principio de la cosecha hasta 22.8 cm al final de la misma. Las tasas de elongación relativas variaron en promedio para ambos sistemas de inicio, entre 0.11 cm cm-1 d-1 y 0.38 cm cm-1 d-1 dependiendo de la temperatura del aire. Al mismo tiempo las parcelas constituyentes del ensayo fueron sometidas a una cosecha comercial durante las seis semanas de evaluación. Los espárragos cosechados constituyeron el rendimiento total (kg ha-1) para cada sistema de inicio entre los cuales no se encontraron diferencias estadísticamente significativas. Este estudio permitiría explicar las diferencias de rendimiento comercial para un mismo cultivar en diferentes zonas de producción. Se concluye que para una zona dada, la elongación y sobre todo la ramificación, son variables prioritarias para la determinación del potencial productivo de un cultivar, debiendo tener en cuenta las condiciones de temperatura en las cual se pretende producir este cultivo y el mercado de destino de la producción debido a las mercadas preferencias entre sus consumidores.

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。

稻属Oryza officinalis复合体基因组研究－兼论基因组原位杂交的原理和方法

稻属（Oryza L.）属于禾本科稻族（Oryzeae Dumortier）。本属包括AA，BB，BBCC，CC，CCDD，EE，FF，GG，HHJJ和HHKK十个基因组，二十余种。其中Oryza officinalis复合体包括BB，BBCC，CC，CCDD和EE五个基因组，九个种。众多的学者对该复合体进行了广泛深入的研究，为后续研究奠定了坚实的基础。然而，迄今为止，多倍体物种形成问题许多没有解决，基因组间的关系尚未完全阐明，甚至有些多倍体物种的基因组组成仍未确认。 本文评述了Oryza officinalis复合体中基因组研究的历史和现状，用基因组原位杂位（GISH）的方法，对该复合体四倍体物种的基因组组成作了验证;对B，C，D和E四个基因组间的关系进行了研究。同时对基因组原位杂交的方法，原位杂交鉴定多倍体基因组的组成以及研究基因组间关系的方法作了一些探讨。其主要研究结果如下： 一. 原杂交方法的研究：1）染色体制片：比较研究了不同的制片方法，发现压片法适用于大染色体的材料;将酶解/空气干燥法（Fukui et al., 1992）加以改进后，特别适宜于小染色体植物材料的制片。2）根尖储存时间和条件对原位杂交的影响：发现在-20 ℃的酒精（70%）中储存8个月以内的根尖材料，可用于原位杂交;而在-20 ℃的固定液中储藏18个月的根尖，DNA降解严重，不能用于GISH。3）探针标记：比较了随机引物法、缺口平移法和两步标记法（先用随机引物标记后，再用缺口平移法进行标记的方法）的优缺点。结果显示两步标记法是最佳标记方法。 二. GISH鉴定异源多倍体的方法：用两个异源四倍体Oryza minuta和Scilla sinensis (2n = 34)做染色体制片，进行原位杂交实验，结果表明：1）用二倍体亲本基因组之一做探针而不用封阻DNA， 可以鉴别Oryza minuta而不能鉴别Scilla sinensis中的基因组。2）用一个二倍体亲本做探针而用另一个做封阻，能够区分Scilla sinensis的两个基因组;但过量的封阻DNA将可以造成一些实验假象。3）同时用两个亲本的DNA做探针，不仅能够有效分辨不同的基因组，还能够根据交叉杂交程度推测基因组间的分化程度，是鉴别异源多倍体最有效的方法。 三. GISH鉴定稻属四倍体的基因组组成：1）Oryza minuta, O. punctata和O. malampuzhaensis的基因组的组成都为BBCC。2）Oryza minuta中B基因组和二倍体O. punctata中的B基因组之间存在着明显的基因组内分化。3）O. alta是一个异源多倍体，其基因组组成为CCDD。但C和D之间的分化不彻底，可以认为它不是一个严格意义上的异源多倍体。 四. GISH研究B，C，D和E基因组间的关系：1）B基因组和C基因组之间的关系最远;E和C之间的分化同E和B之间的分化程度接近，但E和C之间的分化比E和D之间的分化要小一些;C和D之间的分化不彻底，关系最近。

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.

Zingiber officinalis an herbal appetizer in the tiger shrimp Penaeus monodon (Fabricius) larviculture

Penaeus monodon postlarvae were fed with different percentages (0%, 25%, 50%, 75% and 100%) of the herbal appetizer Zingiber officinalis enriched Artemia. After 30 days of culture (i.e. PL-1-30), a very positive result was found in Z. officinalis-enriched Artemia-fed postlarvae. The unenriched Artemia-fed postlarvae consumed 91.0 mg/animal/30 days of feed, whereas the Z. officinalis-enriched Artemia increased their consumption to 127.9 mg/animal/30 days. A similar pattern was noticed in feed absorbed (110.2 mg), dry weight growth (26.7 mg) and feed catabolized (83.2 mg) in Z. officinalis-enriched Artemia because of enzymatic activities. The conversion efficiency of unenriched postlarva was 17.19%, whereas in 100% Z. officinalis-enriched Artemia, the maximum conversion efficiency was 20.85%. The net production efficiency increased significantly (P < 0.05) to 22% from that of the unenriched Artemia-fed postlarvae. The administration of Z. officinalis in all levels produced significantly (P < 0.05) higher weight gain and specific growth rate. The utilization efficiency of feed increased proportionately to the percentages of Z. officinalis. Digestive enzyme activity (amylase, protease and lipase) increased significantly (P < 0.05) in the 50%, 75% and 100% enrichment. Among the different percentages of enrichment, the 100% Z. officinalis-enriched Artemia-fed postlarvae performed better in the overall status.