Bacterial Concentration and Diversity

The objective of this particular paper was to study the results that were extracted when bacterial communities were formed. These bacterial communities were formed through the process of the reproducibility of sm on the whole volume of twin ingest from replicate bioreactors with stabilized continuous-flow chicken caecal bacterial communities. The results referring to the bacterial concentration and diversity were because analyzed by phenotypic, biochemical and ribotype analysis. To break bacteria a electrostatic environment is the most essential compulsion this stable and a constant environment is known as steady-state conditions.This allows bacterial cultures to be obtained in a reproducible manner for batch consistency. The pagan efficacy was located by taking an assumption that the aliquot interpreted from the cultures were identical and therefore did not overall affect the results to be learnd by the particular investigate (bacterial culture). Mixed samples of avian c ecal bodily were used to establish replicate bioreactor cultures. Repetitive samplings of the planktonic comp onenessnts were done to determine if all aliquots had the like bacterial contents within the equal bioreactor.Consistency was seen during this exigent sampling process but changes were seen in the specific composition of the resulting communities that initiated from one supply of cecal assortment. These ar the basis on which the whole investigate and the methodology are based on. METHODOLOGY Bioreactor and Sampling Design The cecal contents were extracted from 150 birds (chicken) and then thoroughly mixed under unfertilised anaerobiotic conditions. Three replicate bioreactors (Bioflo 110 Fermentor/Bioreactor, New Brunswick Scientific Co, Inc., Edison, NJ) were used. The steady-state conditions were maintained by keeping the cultures under continuous-flow conditions at a flow identify of 0. 8 ml/min and excessively flushed with carbon dioxide that was barren of an y oxygen. For the first 48 hours the pH of the bioreactor was maintained to a stable 6. 2 0. 3. Then for 3 weeks the cultures were allowed to reach equilibrium the planktonic region was sampled 11 times during this 3 weeks period. After this period 1ml aliquots were collected for analysis. i. e. pH measurement , bacterial isolation etc. Bacterial Isolation and feeler Identification The stuff obtained from the bioreactor was sampled and some of the bacterial cultures obtained were quantified by growth of a 10? l aliquot on discriminating media in triplicate. The triplicate had a 5% sheep blood and was used to determine hemolytic reactions and for the recovery and the enumeration of the aerobic microbial species. The identification and the isolation of the aerobic bacteria was done by streaking the 10 l aliquots onto TS-blood agar , Brilliant unfledged Agar, BGA Becton Dickinson, Sparks, MD), CHROMagar E.coli and Orientation, MacConkey, mEnterococcus, and Rogosa plates. These plates were then incubated for 24 hours at thirty seven degrees. Likewise anaerobic bacteria were isolated too but the streaking was done onto Brucella-blood agar, Phenylethyl alcohol agar (Becton Dickinson, Sparks, MD), Veillonella, and BBE plates. The plates were then incubated anaerobically for 48-72hours at the same temperature. These bacteria were similarly tested for aero tolerance. registerThe total aerobic and anaerobic population levels were enumerated by serial dilatation onto TS-blood agar, MacConkey, mEnterococcus agars or Brucella-blood agar plates, respectively. Ribotype Characterization Isolates from the bacterial lawns were collected and analyzed by use RiboPrinter Microbial Characterization System following the manufacturers instruction using lytic enzymes. Endonuclease EcoRI was used to cleave the desoxyribonucleic acid and gel electrophoresis was used to detach the fragments and analysis was done using a modern hybridization blotting technique.The DNA hybri dized was labeled rRNA operon prob derived from Escherichia coli, and the bands were detected by chemiluminescence. The image formed was captured and transferred to the RMCS database and data were normalized to a standard marker set. The images were compared with the 6448 EcoRI riboprint patterns in the DuPont database and a 900 EcoRI riboprint pattern custom in-house database (USDA, ARS, College Station, TX). entropy Analysis For each set of combined cecal material the in a higher place conjureed 3 replicate bioreactor were established.These were then analyzed for enumeration and characterization (eleven per bioreactor). The statistics were delineate in tabular form. Commercially available software was used to analyze and calculate data. Differences in cfu/ml were compared among the replicate bioreactors. MAIN RESULT The assembling of bacteria cultures or any different organism is greatly moved(p) by the sample size and the frequency of organisms organism sampled in a partic ular environment. Some of the processes or the methods described above also bedevil limitations due to different constraints that govern their working e.g. enumeration. Enumeration of bacteria is bear on by many factors including individual species growth rates, fitness of each competing species etc. therefore the bacteria produced or grown in a selective media possibly less productive when exposed to competition from many other species in a non-selective media culture. The probability of collection is greatly touch by the spatial distribution of organisms. As a rule the sampling unavoidableness must increase as the degree of unit aggregation increases.Enumeration is also affected by aggregation and may account for some of the random variable reported in the bacterial quantification. Therefore considerations should be given to the sampling size when using aliquots for inoculation from cultures with known aggregating species. An important thing to mention here that whatever the sampling technique is used there are also certain limitations associated with sampling. It is not only quite difficult to reaping all species comprehensively but our present technological inability also creates a hindrance because we are unable to artificially culture all bacterial species.It was also seen that the efficiency of detection of pathogenic bacteria is affected by dilution i. e. a lower efficiency of detection was achieved where the prevalence of the guide bacteria was diluted. A threshold quantity of specific bacteria maybe required for the proper mix. Adjustments in population density, adhesion and diversity which lead during culturing period greatly affects this observation. CONCLUSION The aim of this study or experiment was to determine reproducibility of small volume repeat sampling with the jock of a bioreactor.Basically bacterial concentration and diversity were the two important factors that were being concentrated these two quantities were analyzed within s tabalised continuous-flow chicken cecal bacterial communities initiated by replicate aliquots taken from thoroughly mixed samples. Pooled cecal material was created from layer chicks to establish the bioreactors. After a steady-state was reached the plankton components were sampled repetitively for three weeks and was then characterized by phenotypic, biochemical and ribotype analysis.No notable differences were found in the bacterial concentrations that came from the same bioreactor. Differences were found in bioreactors initiated from the same stock material. BIBLIOGRAPHY 1. Tawni L Crippen, Cynthia L Sheffield, Kathleen Andrews, Roy Bongaerts, and David J Nisbet, (2008), Bacterial Concentration and Diversity within Repetitive Aliquots Collected from repeat Continuous-Flow Bioreactor Culture, Open Microbiol J. 2008 2 6065, published online 2008 May 23. <http//www. pubmedcentral. nih. gov/articlerender. fcgi? artid=2593035& irradiation=pmcentrez>