Monday, December 31, 2012

Apimedica: New ‘BeeTest’ for Detection of Viral, Bacterial Diseases of Honey Bees – “蜜蜂测试”—免疫色谱法快速检测蜜蜂病毒和细菌疾病

Apimondia Apimedica-Apiquality International Forum
Oct. 22-25, 2012
Zhenjiang, China
Immunochromatographic Rapid Test “BeeTest” for Detection of Viral and Bacterial Diseases of Honey Bees Apis mellifera
Leonid KelinBeeTest” Ltd, Russian Federation, Novosibirsk, Russia
Russian company "Beetest" presents a product which is unique in the world - immunochromatographic rapid test "Beetest." As our company produces a preparation “Endoglukin” for prevention and treatment of viral diseases of honey bees and stimulation of the bee colony development, we decided to develop a diagnostic test as a prior stage for diseases treatment in order to know exactly what kind of disease should be treated. After several years of researches we finally managed to find specific antigens for each disease that ensures the high sensitivity and delicacy of our test.
With the test “Beetest” you can simultaneously check your hives for the presence of the major bacterial and viral diseases of bees. The test is intended for use in the field, does not require highly trained personnel and can be used directly next to the hive.
Principle of the method
Immunochromatographic rapid test “BeeTest” is a sandwich-type lateral flow assay for simultaneous and differential detection of two viral and two bacterial honey bee diseases:
· Acute bee paralysis virus;
· Sacbrood virus;
· American foulbrood (Paenibacillus larvae);
· European foulbrood (Melissococcus plutonius)
“BeeTest” was developed for rapid, quality and one-step detection of the viral and bacterial proteins in honey bee larvae by Lateral-flow immunochromatographic technology. The principal scheme of the test is on the picture.

The nitrocellulose membrane is coated with specific polyclonal antibodies to proteins of Acute bee paralysis virus or Sacbrood virus or American foulbrood (Paenibacillus larvae) or European foulbrood (Melissococcus plutonius) and specific proteins of these infectious agents.
During the first step disrupt the honey bee larvae in the bottle with Elution buffer and glass balls by vigorous shaking. On the second step drip a couple of drops into the sample window. In this moment specific viral or bacterial proteins (if they are present in the sample) forms the complex with the conjugates of colloidal gold with poly- or monoclonal antibodies.
Further these complex bind with the specific antibodies coated on the membrane thus forming the red line (test line). Conjugates which were not involved in forming the complex with bacterial or viral proteins binds with the specific proteins on the membrane (Picture 2) thus forming another red line (control line). The intensity of red color lines should be assessed visually.

Reagents and materials:
1) 4 plastic cassettes (one cassette for one disease);
2) Plastic bottle with a glass balls and Elution buffer;
3) Single-use pipette;
4) Single-use spatula.
Assay procedure:
All reagents are ready-to-use and doesn’t need in any preparations.
1) Extract a larva showing suspicious symptoms with the spatula.
2) Unscrew lid from the Bottle with Elution buffer. Use the spatula to place sample in the bottle. Replace the lid and shake vigorously for about 20 seconds.
Test line Control line
3) Unscrew lid of Bottle and pipe the sample by supplied pipette. One honey bee larvae should be used with all types of cassettes.
4) Hold the Test cassette horizontally and gently squeeze two drops onto the sample well of the each cassette.
5) Keep it horizontal until extract is absorbed (c 30 seconds) and a red dye appears in the viewing window.
6) Wait until the control line appears (labeled C) and read the result (10-15 minutes).
Reading the results
3 Positive result - two lines show up (Test and Control) - indicates that the target pathogen is present in the test sample.
4 Negative result - Control line shows up only, no Test line - indicates that the target pathogen has not been detected in the test sample.
5 Indefinite result - one Test line without Control line or no one lines indicates that test doesn’t work.
Analytical sensitivity of the test kit
Analytical sensitivity is 10-50 ng/ml with using of recombinants proteins of Acute bee paralysis virus, Sacbrood virus, American foulbrood (Paenibacillus larvae), European foulbrood (Melissococcus plutonius). 
Shelf life, storage and transportation conditions
Shelf life of the “BeeTest” is 12 month from the date of manufacture. Store at 17-27 °C in the dry dark place.
______________________________________________
*Corresponding author, E-mail: beetest.org@gmail.com
蜜蜂测试”—免疫色谱法快速检测蜜蜂病毒和细菌疾病
Leonid Kelin (俄罗斯)
BeeTest” Ltd, Russian Federation, Novosibirsk, Russia
罗斯公司的蜜蜂测试是世界上一种先进独特的快速免疫色谱法。本公司用此法控制和治疗蜜蜂 疾病和刺激蜜蜂种群的发展,以达到更好的制备Endoglukin,先前我们决定研发一种诊断测试法,为了在 蜜蜂疾病治疗前准确定位疾病的种类。经过几年的研究,我们最终找到每一种疾病的特异性抗原,这保证 测试的高效灵敏。有了蜜蜂测试,你可以同时检查你蜂群主要疾病细菌和病毒。本测试不需要专业技术人员,可直接在 蜂箱旁边进行检测。方法原理
蜜蜂测试,快速免疫色谱法,是一种夹心型横流化测试法,可同时对两种蜜蜂疾病病毒和细菌进行示 检测。
1. 蜜蜂急性麻痹肠道病毒
2. 幼蜂皱萎病病毒
3. 美洲蜜蜂幼虫腐臭病
4. 欧洲蜜蜂幼虫腐臭病
过横流免疫色谱技术,蜜蜂测试可快速,高效,一步检测出蜜蜂幼虫中病毒和细菌蛋白,它主要的 测试路线如图1 所示:硝酸纤维素膜上涂有蜜蜂急性麻痹肠道病毒,幼蜂皱萎病病毒,美洲蜜蜂幼虫腐臭病毒和欧洲蜜蜂幼 虫腐臭病毒的多克隆抗体以及这些传染源的特殊蛋白。第一步将蜜蜂幼虫放入带有洗脱缓冲液和玻璃球的瓶中,剧烈振荡。第二步滴几滴试剂在采样窗口中,时特殊病毒和细菌蛋白(如果样品中存在)将与单克隆抗体和胶体金的轭合物形成复合物。图二 测试线和控制线中轭合物和抗原的键合(Au –胶体金和抗体的轭合物, Ag – 抗原) 之后这些复合物和薄膜上的特殊抗体紧紧地结合在一起,形成红线(检测线),那些没有没有和薄膜上 的特殊蛋白结合的复合物(如图二)形成另一条红线(控制线),红色线的强度可以用眼观测出来。试剂和材料
1. 4 个塑料储片夹(一个对应一种疾病)
2.带有玻璃球和洗脱缓冲液的塑料瓶
3.一次性试管
4.一次性药刀
实验步骤 所有试剂都要提前准备好。
1. 药刀取一只有可疑症状的幼虫
2. 旋开带有洗脱缓冲液的塑料瓶,用药刀把幼虫放入塑料瓶中。盖上瓶盖,剧烈振荡20 秒。
3. 旋开瓶盖,用吸量管吸取洗脱缓冲液。每个幼虫的缓冲液液都要滴入4 个塑料储片夹。
4. 水平拿住测试储片夹,缓慢滴两滴缓冲液在样品室。
5. 保持水平,使提取物被吸收(30 秒),观察窗中出现红线。
6. 等到控制线出现(标记为C)然后读出结果(10-15 钟)结果
1、阳性结果——现两条红线(检测线和控制线)表明目标病原体出现在测试样品中。
2、阴性结果——只出现控制线表明目标病原体不存在于测试样品中。
3、无效结果——测检测线没有控制线或者两条都没有表明测试无效
试剂盒的分析灵敏度 10-50ng/mg 蜜蜂急性麻痹肠道病毒,幼蜂皱萎病病毒,美洲蜜蜂幼虫腐臭病毒和欧洲蜜蜂幼虫 腐臭病毒的重组蛋白质来分析灵敏度。质期,储存和运输条件 质期12 个月,储存条件为17-27,黑暗干燥处。

Sunday, December 30, 2012

Apimedica: Supercritical CO2 Extraction of Propolis Produces Most Volatile Oils - 蜂胶及其不同提取物中挥发油成分的比较研究


Apimondia Apimedica-Apiquality International Forum
Oct. 22-25, 2012
Zhenjiang, China
Comparative Study of Propolis and its Extract on Different Volatile Oil Constituents
Yang Qin, Zhou Juan, Huang Xinxin, Zhang Lin, Zhu Huifen, Yu Shumei, Mao Riwen
(Natural Medicine Engineering Technology Research Center by Supercritical Fluid Extraction in Jiangsu province, Zhenjiang 212009,China)
ObjectiveTo determine the amount of volatile oil in propolis glue using the following methods: solvent extraction, propolis ethanol extraction and supercritical CO2 extraction. GC-MS was used to determine both the qualitative and quantitative data.
ResultThe content of volatile oil in the propolis was in the following order: supercritical CO2 extraction > ethanol extraction > propolis glue. The content of volatile oil in the propolis extracted by supercritical CO2 was 23.45% higher than the propolis extracted with ethanol. The results of GC-MS
indicated that propolis extracted using supercritical CO2 method recorded a high content of volatile oil (54.8%) in two kinds of common volatile componential extracts which was superior to the propolis extracted with ethanol. More than 1% of the composition of volatile propolis extracted by supercritical CO2 method accounted for 64.7%. This indicated that the volatile oil composition in propolis extracted by supercritical CO2 method was better than the ones extracted with ethanol. Especially terpene; a kind of component found in the propolis extracted by supercritical CO2 was 1.5 times higher than the propolis extracted with ethanol.
ConclusionThe content of propolis extracts and its volatile oil was highest in the propolis extracted by supercritical CO2. The results indicated that the supercritical CO2 extraction method for extracting propolis and its volatile oil is better than that of ethanol extraction.
__________________________________________
Corresponding author, E-mail: 493967536@qq.com
蜂胶及其不同提取物中挥发油成分的比较研究
杨琴,周娟,黄鑫鑫,张林,朱惠芬,郁树美,毛日文
(江苏省超临界萃取天然药物工程技术研究中心,镇江 212009,中国)
【目的】用溶剂萃取蜂胶原胶、蜂胶乙醇提取物及蜂胶超临界C02提取物中的蜂胶挥发油;利用GC-MS 对蜂胶挥发油进行定性与定量分析。
【结果】蜂胶原胶及其提取物中的挥发油含量依次为蜂胶超临界C02提取 >蜂胶乙醇提取物>蜂胶原胶。蜂胶超临界C02提取物挥发油含量较之蜂胶乙醇提取物挥发油含量高
23.45%;通GC-MS分析,两种提取物共有的挥发性成分中,蜂胶超临界C02提取物中含量高的占54.8%优于蜂胶乙醇提取物;在含量>1%挥发性成分中,蜂胶超临界CO2提取物占64.7%进一步说明蜂胶超临 CO2提取物挥发性成分优于蜂胶乙醇提取物的挥发性成分。尤其是萜烯类成分,蜂胶超临界CO2提取物中 萜烯类成分含量是蜂胶乙醇提取物中萜烯类成分含量的1.5倍。
结论】蜂胶及其不同提取物中的挥发油,以 临界C02提取物中的含量最高,表明利用超临界CO2提取技术提取蜂胶挥发油要优于乙醇提取。
键词:蜂胶;超临界C02提取;乙醇提取;挥发油;GC-MS

Saturday, December 29, 2012

Video: Bee Stings Could Be New Botox



BBC, 12/20/2012
An expensive bee sting could top the wish list for many beauty conscious women this Christmas.
Sales of New Zealand creams that contain bee venom are booming.
But there have been warnings about possible side-effects, as Greg Ward reports

Friday, December 28, 2012

Clinical Trial: The Use of Henna and Propolis on PPE


Cyprus University of Technology
The palmar-plantar erythrodysesthesia (PPE) is the only clinical adverse event that commonly occurs with capecitabine and/or pegylated liposomal doxorubicin treatment and it warrants special attention because it is the most common dose-limiting toxicity. This study is designed to test the effectiveness of a henna and propolis treatment protocol in the management of capecitabine and/or pegylated liposomal doxorubicin induced palmar-plantar erythrodysesthesia.

Thursday, December 27, 2012

Propolis Effective for Oral Hygiene



Influence of Hygienic Preparations with a 3% Content of Ethanol Extract of Brazilian Propolis on the State of the Oral Cavity
Adv Clin Exp Med, 2012 Jan-Feb;21(1):81-92
BACKGROUND:
One of the most important measures to be undertaken in order to fight gingivitis and periodontitis is maintenance of proper hygiene of the oral cavity. The research to improve the content of toothpaste has continued for many years so that they should become better in terms of therapeutic abilities.
OBJECTIVES:
The aim of this work was to determine and investigate the influence of the application of toothpaste and gel with 3% ethanol propolis extract on the state of the oral cavity.
MATERIAL AND METHODS:
The research group comprised 80 adult patients divided into two subgroups: Group I, which comprised 40 patients without pathological changes within the boundaries of the periodontium, and Group II, also 40 patients endangered with the occurrence of periodontitis caused by dental plaque and lack of proper hygiene of the oral cavity. Qualification for both groups was based on an interview and analysis of clinical documentation and assessment of adequate indices such as API, OHI and SBI. The patients underwent three examinations: initial, follow-up after 7 days and after 8 weeks since the beginning of the program. Moreover, the patients were instructed about hygienic procedures of the oral cavity. Four groups (T, G, CT, CG), 20 patients each, were created from research groups I and II. They used the following preparations: T--Dental Polis DX toothpaste with propolis content, G--Dental Polis DX toothpaste without propolis content, CT--Carepolis gel with propolis content, CG--Carepolis gel without propolis content. The patients were informed about the type of hygienic preparation they were given to use (whether it contained propolis or not). Moreover, they were interviewed for their subjective evaluation of the product received.
RESULTS AND CONCLUSION:
Results of the research show the effectiveness of hygienic preparations with 3% content of ethanol propolis extract in both groups of patients: without pathological changes within the boundaries of the periodontium and in the case of patients endangered with the occurrence of gingivitis caused by dental plaque.
Wprowadzenie. Podstawowym krokiem w walce z zapaleniami dziąseł i przyzębia jest utrzymanie właściwej higieny jamy ustnej przez eliminację płytki nazębnej. Dobór właściwej metody szczotkowania, szczoteczki i środków wspomagających stanowi pierwszy etap w walce z chorobą. Od lat trwają badania nad ulepszeniem składu past do zębów i płukanek, tak by wykazywały właściwości terapeutyczne.
Cel pracy. Określenie wpływu pasty do zębów i żelu do czyszczenia zębów zawierających 3% etanolowy ekstrakt propolisu na stan jamy ustnej.
Materiał i metody. Grupa badana stanowiła 80 osób. Były to osoby pełnoletnie, zarówno kobiety jak i mężczyźni – powyżej 18. roku życia. Z grupy badanej zostały wyłonione dwie podgrupy pacjentów wydzielone ze względu na wstępną kwalifikację stanu przyzębia brzeżnego: I grupa – bez zmian chorobowych w obrębie przyzębia brzeżnego – 40 osób, II grupa – z zagrożeniem wystąpienia zapalenia dziąseł wywołanego płytką bakteryjną, brakiem dostatecznej  higieny  jamy  ustnej    40  osób.  Pacjenci  byli  poddani:  wstępnej  kwalifikacji  i  badaniu  wstępnemu (wizyta 1), badaniu po upływie 7 dni (wizyta 2) i 8 tygodni (wizyta 3) od rozpoczęcia programu. Przeprowadzono profesjonalny instruktaż higieny jamy ustnej. Podczas wizyt kontrolnych wyznaczono wartości wskaźników API, OHI i SBI. Z grupy badanej (I i II) wydzielono cztery podgrupy (T, G, CT, CG) liczące po 20 osób, które stosowały: T – pastę Dental Polis DX z propolisem, G – pastę Dental Polis DX bez propolisu, CT – żel z propolisem Carepolis, CG – żel bez propolisu.
Wyniki i wnioski. Wyniki zaprezentowanych badań, poddane analizie statystycznej, wskazują na skuteczność preparatów higienizacyjnych zawierających 3% etanolowy ekstrakt propolisu zarówno u pacjentów bez zmian chorobowych w obrębie przyzębia brzeżnego, jak i u pacjentów z zagrożeniem wystąpienia zapalenia dziąseł wywołanego płytką bakteryjną (Adv Clin Exp Med 2012, 21, 1, 81–92).
Słowa kluczowe: 3% etanolowy ekstrakt propolisu, profilaktyka dentystyczna, zdrowie jamy ustnej, zapalenie dziąseł.