Quantification of antigen of Mycoplasma capricolum subsp. capripneumoniae by optical assay / 生物工程学报

Mycoplasma capricolum subsp. capripneumoniae (Mccp) is the cause of contagious caprine pleuropneumonia (CCPP) in goats. Inactivated vaccines and capsular polysaccharide (CPS) indirect hemagglutination reagents are available for prevention and serological detection, but high culture costs and complex antigen quantification have been plagued by production staff. In order to solve these problems in production practice, a sugar fermentation medium with an initial pH value of 7.8, which could improve the production of two antigens simultaneously, was screened out by changing the initial pH value based on previous Mccp metabolomics analysis. Since phenol red can be identified by UV absorption spectrum and cetyltrimethylammonium bromide (CTAB) can bind to anionic capsular polysaccharide, a UV spectrum measurement method for analyzing the culture stage reached by Mccp and a CTAB precipitation test for relative quantification of capsular polysaccharide antigen content in the fermentation broth were established. The UV spectrum observation method can guide the production of Mccp according to the growth curve of Mccp, which greatly reduces the monitoring time of the traditional CCU method and improves the accuracy of the original eye-observation method. The established CTAB precipitation test can complete the monitoring of CPS content within 5 hours, which greatly reduces the time required compared with the traditional differential technique, and its accuracy was verified by the phenol-sulfuric acid method. The optimized culture medium and the two correlation comparison methods established in this study can effectively reduce the production cost of Mccp and improve the production efficiency. The two assays have been used in the research at our laboratory, which provides experimental data for further improvement of the production process of CCPP inactivated vaccine and capsular polysaccharide as well as rapid quantification.

Germinação in vitro e aclimatação de plântulas de sucupira-preta (Bowdichiavirgilioides Kunth. ) / In vitro germination and acclimation of sucupira-preta seedlings(Bowdichiavirgilioides Kunth. )

Este trabalho teve como objetivos avaliar a germinação in vitro e a aclimatação de plantas de sucupira-preta (Bowdichia virgilioides). Sementes escarificadas e não escarificadas foram inoculadas em tubos de ensaio contendo meio de cultura MS ou WPM, com 100 ou 50% dos sais e vitaminas; suplementados com dois tipos de aditivos, carvão ativado e PVP, e mantidas em sala de cultura. Utilizou-se um delineamento inteiramente casualisado em esquema fatorial 2x4 (dois aditivos e quatro formulações de meio de cultura) com quatro repetições e seis sementes por repetição. Aos 30 dias obteve-se o percentual de germinação e o índice de velocidade de germinação (IVG). As plantas foram transferidas para tubetes, contendo substrato e aclimatadas em casa de vegetação, onde permaneceram durante 60 dias, sendo transferidas para casa de sombra, onde permaneceram por mais 60 dias. Aos 30, 60, 90 e 120 dias avaliaram-se o percentual de sobrevivência e a altura das mudas aclimatadas. A germinação in vitro de sementes escarificadas apresenta os maiores percentuais utilizando-se os meios de cultura MS e WPM reduzidos à metade da concentração dos sais e vitaminas. A germinação ocorre independentemente do aditivo utilizado tanto para sementes escarificadas como para as não escarificadas. A aclimatação das plantas germinadas in vitro ocorre independentemente do histórico de aditivos ou meios de cultura utilizados.

Thiss tudy objectived to evaluate the in vitro germination and acclimatization of plants sucupirapreta (Bowdichia virgilioides). Scarified and not scarified seeds were inoculated in test tubes containing culture medium MSorWPM, 100 or 50% of salts and vitamins, supplemented with two types of additives, activated charcoal and PVP, and kept in culture room. We used acompletely randomized in a 2x4 factorial design (2additives and 4culture medium formulations) with 4 replications and6 seeds per replicate. At 30 days gave the germination percentage and germination speed index(IVG). Plants were transferred to tubes containing substrate and acclimatized in a greenhouse, where they remained for 60days and transferred to a dusk house, where they remained for another 60days.At 30, 60, 90 and 120 days, we assessed the percentage of survival and seedling height acclimated. The in vitro germination of scarified seeds had the highest percentage using culture media MS and WPM halved the concentration of salts and vitamins. Germination occurred regardless of additive used both to scarified and note scarified seeds. The acclimation of plants germinated in vitro occurred regardless of additives history or culture media used.

RECOMMENDATION OF A KIND OF SYNTHETIC MEINTENANCE LIQUOR TO PRESERVE BACTERIUM LONG / 微生物学通报

This Synthetic Meintenance Liquor contains all kinds of nutritive substance that subsist the bacterium, and can preserve the bacterium for nearly ten years by providing energy needed by metabolism. It is a favorable culture medium to preserve bacterum long.