Isolation And Identification Of Enterococcus Faecium Biology Essay

Isolation And Identification Of Enterococcus Faecium biology EssayEnterococci atomic itemise 18 widely distributed in the environment within the clement body, they are form commensals of the oral cavity, gastro enteral nerve tract and vagina. Enterococcus faecium is a confirming bacterium group D, alpha hemolitic or nonhemolitic in the genus Enterococcus (Chingwaru W., 2003). It hobo be a commensal (non-harmful coexisting organism), in the gentleman intestine, but it whitethorn too be a pathogen causing unsoundnesss like neonatal meningitis. familiar lactobacillus medium including MRS medium, Luria-Bertani (LB) medium, high-sterilized stored at 4 refrigerator spare. In the sterile console, the yoghourt containing lactic unpleasant bacterium was diluted with sterile saline to a serial dilution (10-3,10-4,10-5) of 50l suspension and coated them on the case containing MRS choice medium and marked. The plate was placed at 37 unassail equal to(p) box without oxygen for 3 6 48 hours. Single colony was picked and repeatedly crossed to obtain the pure hits.Bacterial acknowledgement methods include the parking lot (tradition) classified acknowledgement, numerical value realisation and mechanisation identificationThe parkland (tradition) classified identification needed to detect m any(prenominal) items. It is so tedious. exactly numerical value identification is gibe to the identification target. It uses the test card. The return is expressed by the flair of figure and compared with the data of database to get an identification result. Automation identification jackpot reach whole procedure automation in the step of inoculation, cultivation, reading and report. It cigaret supply a efficacious instrument in rapid clinical microorganism diagnosis. No issuing what kind of identification methods, it all needs to identify bacterium according to the morphology, physiological characteristics and biochemical characteristics.Enterococcus faecium is a kind of oval prison cell. Its diameter is 0.5 1.0 m. It can show a short-chain arrangement frequently in pairs or in liquid medium. It has slight movement. In the blood plate it can category a milky white plate or white, marginal integrity, telephone exchange protuberance, large and smooth colony. Its diameter is 1-2mm, and has rare colors, just aboutly around the colonies without hemolysis. by and by the Gram stain, bacterium shows purple. It is Gram-positive coccobacteria. The chemistry character of Enterococcus faecium is contact-negative, and it can nurture in 6.5% fund, bile Esculin agar plate and 0.1methylene bue. It also can grow in 10 or 45, pH 9.6 broth medium, and can tolerate 65 30min condition. It can ferment lactose, sorbitol and mannitol and can use arginine, but cant use ferment arabinose.2.3.2 Biochemical identificationIdentification by lactic acidulous bacteria and biochemical identification tests, respectively taking the exercise-induced, contiguity enzyme, anaerobic ontogenesis, l0 and 45, 60 30 min, 6.5NaCl salt-tolerant growth, pH 9.6 alkaline environment growth, methyl red, benzopyr component, sugar alcohol acid fermentation (glucose, galactose, maltose, lactose, raffinose, manicol and so on). Comparing the experiment result with Bergers Manual of de landmarkining factor Bacteriology (the ninth edition) to determine comprehensively.2.3.3 The growth curve of Enterococcus faeciumThe growth of bacteria includes growth volume, quantity and the improvement of metabolic melt down. Different bacteria halt their let polar growth pattern. Under the unchanged enculturation conditions, calculate the amount of bacteria oriented to time, time as abscissa, the number of bacteria as vertical axis we can get a curveThe growth curve includes the stagnation period, logarithmic phase, stationary phase and decline phase. The continuance of each period is divers(prenominal) according to the medium composition and culture conditions d ue to bacterias own characteristics. In a comparative study of different bacteria, growth curve can help to choose bacterium with the akin growth stat and this can make a reliable experiment result. get hold the bacteria-growth curve mainly use two methods, much(prenominal) as turbidness method and executable organism dilution count. Turbidity method is to place the different stage and different concentration bacterial culture to the spectrophotometer, and count the bacteria by turbidity method. We can use the turbidity to represent the comport of bacterium and measure total dacterial count (including the dead bacterium). It is simple, facility examine with the turbidity method. But it only can indicate the bacterial sexual relation growth information. Viable organism dilution method of counting is to dilute the bacterial liquid to different dilution of 10-110-2 and so on. E really(prenominal) dilution can be civilized in the plate and colony is counted with colony events-p er-unit-time meter. The total quantity of viable organism by using this method can represent the neat growth information of bacterium.3 Application of Enterococcus faecium3.1 Supplementation of dood with enterococcus faecium (SF68) stimulates immune functionsThe enteric microflora prevail a crucial role in forces defense reactiveness as present by their ability to modulate both ingrained and acquired immunity at the local as well as systemic levels. Due to these immunological properties, specific hears of lactic acid bacteria ( lab),3 delimit as probiotics, stomach raised considerable interest in juvenile years. When ingested as a feed supplement in sufficient numbers, probiotics are live microorganisms that beneficially affect the gastroenteric balance, going far beyond the conventional nutritional result. The mechanisms underlying the immune modulating properties of probiotics are non richly understood. However, they may be due indirectly to the ability of probiotics to balance the enteral microflora and/or be a consequence of a direct subsidiary import on the crossingion of immune factors, such as cytokines. In fact, several strains of lab were shown to enhance nonspecific immunity in vitro as well as in vivo, including the release of tumor necrosis factor- and interleukin 6 (4), appendd phagocytosis in mice and humans (Perdigon, G. et al, 1988) (Schiffrin, E. J. et al, 1995) and stimulated natural killer cell activity (Matsuzaki, T., 1998, aller, D. et al, 2000). Their ability to specifically modulate the servers immune responses to pathogens was also demonstrated (Meydani, S. N. Ha, W. K. , 2000). An increase in rotavirus-specific antibodies was detected in children with acute rotavirus diarrhea who received Lactobacillus rhamnosus (Kaila, M. et al, 1992). Moreover, it was shown that administration of L. johnsonii to human volunteers boosted the specific immunoglobulin IgA response to the Salmonella typhi vaccine Ty21a (Link-Amster, H. et al, 1994).3.2 Application in treatmentEnterococcus fececium can ferment a re bare-assing of sugar, and product acid. It has high chemical patience, and is widely distributed in nature, but also human and fleshly gut microbial. It has highly detected in physical feces (Archimbaud, C. et al., 2003). Ultilizing non-toxic, facultative anaerobic Enterococcus, in conjunction with separate animal probiotics (Animal Microecologicai Agent, AMEA) can prevent and treat many digestive tract diseases of animals by replaceing antibiotics.Intestinal flora can be regarded as a ordinary human body organs, these normal floras directly involved in the activities of the host metabolism is normal physiological activities of the host indispensable component (Baldassarri, L. et al., 2001). endogenetic Enterococcus as one of the normal floras of the human body, mainly the Ministry of colonization in the ileum and colon, and some strains may be directly involved in the metabolism of the host m aterial. Enterococcus lactic acid feces as intestinal flora, with a bio-barrier function, from legal people or animals that bugger off separate physiological functions of bacteria to re-giving taking it right people are right and safe, but such micro-organisms in the body and settled and reproduction Ecology is a very complex issue. Made viable by the agent is likely to play a biological activity within a certain range, or distinguish of participation in host metabolic activity, resulting in a certain physiological role and therefore have the disease, adjusting the significance of the ecologic balance (Chow, J. W. et al., 1993).Intestinal flora can be regarded as normal human body organs. These normal floras directly involved in the activities of the host metabolism, and are normal physiological activities of the host indispensable component. Endogenous Enterococcus as one of the normal floras in human body, mainly permanent lay down in the ileum and colon, and some strains ma y be directly involved in the metabolism of the host material. Enterococcus faecium as intestinal flora, has a bio-barrier function. Enterococcus faecium separated from brawny people or animals also have physiological functions Re-giving to people is harm slight and safe, but plant and reproduction in the body is a very complex ecology issue. Viable organism praeparatum is likely to play a biological activity within a certain range, or part of participation in host metabolic activity. It products a certain physiological role and therefore can be used in the disease provention, treatment and the ecological balance adjustment.Currently, the finish of enterococci is mainly in Enterococcus faecium. The early example of the use of Enterococcus faecium is out of japan. It was first developed for the treatment of variant intestinal diseases. Yogurt products on the market are containing Enterococcus faecium in Japan or pharmaceutical drugs, such as Lascpan, Lacton, Ruosu, Lebenin, Biosm in. Mummy Love that product in South Korea contains 108 / g feces enterococci and it can treat gastro-intestinal diseases in infants probiotic preparations (Coque, T. M. et al., 1995).3.3 Safety of Enterococcus faeciumEndogenous intestinal bacteria, especially the colonization of lactic acid bacteria in the gut healthy people, the people are safe, because a large number of endogenous enterococci colonization in the human intestinal tract under the bit, which may affect the host intestinal digestion and absorption, and separate function. It has been reported (Dicuonzo, G. et al., 2001) that yogurt containing Enterococcus faecium can prevent diarrhea. By double-blind test, the incidence of antibiotic group was 27.2%, while taking the probiotic group was only 3.7%. So it is not only proved to be effective, but did not find any side effects. In 1989 Food and Drug Administration (FDA) and the American fellowship of Feed Public (AAFCO) has also introduced the safety 42 probiotic bacteri a, including Enterococcus faecium.3.4 Oral application of Enterococcus faecium strain EE3Probiotic administration is being used increasingly in human and veterinary medicine. A florilegium of microorganisms, especially lactic acid bacteria (LAB), such as lactobacilli, bifidobacteria, and enterococci nonpathogenic LAB are associated with a wide variety of sources, such as plant material and various foods. Based on the definition of probiotics, adequate numbers of viable organisms must reach the intestinal tract to have a beneficial effect. For this to happen, probiotic organisms must be able to survive transit through the acidic environment of the stomach and function in the presence of bile. They have to multiply at the site of savoir-faire in the intestine. In addition, they must be able to produce the loath(p) metabolites against dominating saprophytic microflora resulting in competitive growth. Antimicrobial activity of LAB is due to the production of organic acids (lactic an d acetic acids in particular), carbon dioxide, ethanol, heat content peroxide, biacetyl and bacteriocins. These abilities are ofen found among lactobacilli and enterococci, which are nigh frequently used as animal feed supplements or directly as probiotic preparations. Among enterococci, only the species of Enterococcus faecium of hunman ancestry was used as a commercial probiotic. E.faecium used as probiotics efficiently protect animals from diseases caused by E.coli, salmonellae or clostridia it is also reported (Eaton, T. J. Gasson, M. J., 2001) that the outflank preventive effectivity of E.faecim from commercial probiotic preparation Vitacanis against Salmonella enterica serovar Typhimurium in the gastrointestinal tract of gnotobiotic mice.E.faecium EE3 train was able to survive transit through the canine intestinal tract, leading to enrichment of the colonic microflora and local and systematic effects. This strain also persisted in the digestive tract for almost 3 months afterward goal of its administration. The new canine probiotic strain of Lactobacillus fermentum AD1 persisted in the feces of dogs even for 5 months after cessation, however, it reached a lower final concentration (103105 CFU/g) than the EE3 strain (105108 CFU/g). On the other hand, the well-known probiotic strain Lactobacillus fermentum GG survived in the healthy dogs only 8d after cessation of its administration. From the clinical point of view, persistence during administration should be less important than colonization. However, the potential probiotic microorganism should be able to colonize the digestive tract at least temporarily. Decrease of total enterococci can be explained probably by competitive interactions of the EE3 strain with other enterococci. Although enterococci normally producted lactic acid, its higher amounts (as the other enterococci) could be achieved by the concentration of inoculated EE3 strain in addition, differences in lactic acid production were fou nd among enterococci isolated from different sources. Local effects of the EE3 strain application included an increase of LAB and a significant lessen of Pseudominas-like bacteria and staphylococci with prolonged effect up to the end of the experiment It is reported that a significant increase in fecal LAB during administration of food supplemented with probiotic strain L.acidophilus DSM 13241. However, the action of the probiotic, in terms of its effect towards the number of lactobacilli, was lost after cessation of food supplemented with the same probiotic strain (L.acidophilus DSM 13241.). E.faecium EE3 strain survived the transit through the digestive tract of dogs. Our results suggested that bacteria and staphylococci. Moreover, the levels of total lipids, proteins and cholesterin in blood of healthy dogs were also influenced to reach physiological limits.3.5 Application of Enterococcus faecium strain M-74The value of managing coronary artery disease risk factors has been con firmed in the past few decades. Hyper cholesterinemia, hypertension and ingest are still considered to be the classical risk factors. Intervention trials have generally shown that lowering risk factors reduces the subsequent rate of cornonary heart disease, stroke, and other cardiovascular diseases. The Framingham Study was the major study, which verified that cholesterin increases the risk of evolution heart disease. Data output was clear the risk of clinically manifest coronary heart disease is a continuous nonlinear function of blood cholesterol levels. Each increase in the serum cholesterol concentration by 1% results in 23% increase in the risk of CHD. The discovery that hypercholesterolemia plays a major role in the formation and development of atherosclerosis has led to a number of pharmacological and non-pharmacological (including dietary) approaches resulted in its elimination. Fact that the serum cholesterol concentration is influenced by food has been known for a long time ago. Later, the first tell apart of affecting cholesterol level by influencing intestinal microflora was found. One way to do this is by using probiotics. Up to now, the question of supposed hypocholesterolemic effect of probiotics has not been definitely established. While some studies documented it well, others bring contradictary findings or even doubt it. Another unresolved issue is an existing antecedently mentioned effect from the long term point of view. In available sources, we have not found any report where probiotics were administrated for one year. Some data-based and clinical studies have documented the hypocholesterolemic effect of rpobiotic cultures. Not all of the probiotics see to have the same properties and same effect on serum cholesterol levels. The aim of our study was to investigate the impact of long-term orally administered probiotic strain Enterococcus faecium M-74 enriched with selenium on lipid profile (total cholesterol, LDH, HDL and triglycerides ) in humans.Originally, the term probiotics was used for a phenomenon, when each of both together cultivated organisms had produced substances impact the growth of the another one. The meaning of the term probiotic has been evolving since. Many definitions have been suggested. The most commonly used, scientifically valid and therefore acceptable one is that postulated by Fuller. According to him, probiotics are living microorganisms that favourably influence the heath of the host by improving composition of its intestinal microflora. The FAO (Food and Agriculture Organisation for the fall in Nations) and WHO (World Health Organisation) refer to probiotics as living microorganisms which when ingested in sufficient amount, beneficially influence the health of the host. Before a certain bacterial strain can be classified probioticm it must meet some criteria. The most important one is its beneficial health effect. The knowledge of probiotic intervention in lipid metabolism is not new . In the 1970s, researchers observed hypocholesterolemic effect of yoghurt eaten daily in large amount in humans. Similar observations have been seen in animal studies. Many trials aimed to proving the hypocholesterolemic effect of milk and fermented milk products seeded player from early observation. It is found that a low incidence of clinical coronary heart disease in some African tribes with low cholesterol levels in spite of their diet rich in saturated fats, means and cholesterol but abundant in milk and fermented milk products along. In some studies it is observed that a gradual decrease of serum cholesterol concentration in subjects receiving capsules with Enterococcus faecium M-74 from baseline up to week 23 when the decrease exceeded significant level (P