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Microbiology
Question:
What is the “CHOLERA” ethymology?

Answer:
From the Greek cholè for bile. Although the term cholera is now used only to refer to disease caused by the bacterium Vibrio cholera, until the late 19th century any diarrheal illness might be referred to as cholera. For many centuries, medicine in Europe was based on Galen’ theory of the 4 humors in the body: blood, bile, black bile, and phlegm. Diarrhea and vomiting were interpreted as the body’s attempt to restore balance and good health by expelling excess choler, hence, many gastroenterorological illness were referred to as cholera. A 12th century treatise, the Regimen Sanitatis Salernitanum, described the effects of excess choler thusly, “Your tongue will seem all rough, and often times  cause vomits, unaccustomed and hateful, great thirst, your excrements are full of slime, the stomach squeamish, sustenance ungrateful, your appetite will seem in nought delighting”.


Question:
Which are the analytical microbiological tests to guarantee the safety for the cosmetic and pharma consumer?

Answer:
Product Quality and the Role of Preservation and Preservative Testing >>>


Question:
How can contamination risks for laboratory operators avoided during flame sterilization of loops and needles
How can contamination risks for laboratory operators avoided during flame sterilization of loops and needles
How can contamination risks for laboratory operators avoided during flame sterilization of loops and needles

Answer:
The flame sterilization of loops and needles can produce a bio-aerosol emission that is estimated about 1 cubic metre around the typically used Bunsen burner. This is a high biological risk when hospital specimen or biotechnology suspension of micro-organisms are manipulated.
This risk is eliminated replacing the Bunsen burner with the “Bacteria-Safe”. 
The metal loop / needle is inserted (between each inoculation cycle) into the cylindrical chamber of the “Bacteria-Safe”: the produced bio-aerosol is sterilized at high temperature without possibility to contaminate the environment and operator.


Question:
Which are the correct instructions for the storage of the ready to use Contact Plates like “Agar Contact Blister” and  Petri dishes like “Agar Plate”? 

Answer:
▪ Storage at room temperature (Min 10°C - Max 25°C).
▪ The plates can be at higher temperature (over 25°C) up 2 days.
▪ The plates should be stored with up-down agar.


Question:
How to learn the basics of applied microbiology?

Answer:
If you are not a microbiologist, the Bacterium pibiophylum will help you to understand the life of the micro-organisms that are surrounding us each day. You will learn how important they are for us and why we have to respect and thank them.


Question:
Where can I find a complete fungal glossary?

Answer:
Fungal Glossary.
- http://www.emsl.com/index.cfm?nav=Pages&ID=149


Question:
Which is the purpose of the microbiological “challenge test” in food microbiology?

Answer:
Microbiological “Challenge Test” is essential for assessing the potential for growth of specific food-borne pathogen or spoilage micro-organisms in a food product from point of manufacture through to consumption.
This involves the inoculation of a product with the organism(s) and storing under controlled environments conditions in order to assess the risk of food poisoning or to establish product stability.
The use of challenge testing to assess product safety and stability is particularly used with respect to food pathogens such as Listeria monocytogenes. 


Question:
How to detect E. coli 0157:H7 in meat products?

Answer:
FSIS document from USDA “Laboratory GuideBook – Detection, Isolation and Identification of E. coli O157:H7 from meat products”.
- http://www.fsis.usda.gov/PDF/MLG_5_05.pdf
It is necessary the use of Stomacher.


Question:
Where can I find useful Application Notes for the Extraction of DNA?

Answer:
- http://www.omni-inc.com/applications/content-page-177-BeadRuptor.html


Question:
What about an useful microbiology podcast?

Answer:
This week in microbiology. A podcast about unseen life on earth.
- www.microbeworld.org/twim


Question:
Which are the monthly scientific magazines  about microbiology that should be present in each microbiology laboratory to update the microbiologist?

Answer:
-Microbiology
-Journal of General Virology
-International Journal of Systematic and Evolutionary Microbiology
-Journal of Medical Microbiology
-Microbiology Today
All journals are available online through Highwire Press.
The Society for General Microbiology.
-
www.sgm.ac.uk/members.htm


Question:
Which should be the topics for an “Auditing of a Microbiology Laboratory” for QC compliance?

Answer:
• The following topics will be addressed:
o Infrastructure of a Microbiology Testing Laboratory
o Test Methods and Validation
o Equipment
o Documentation
o Preparation for inspections
o Corrective Actions
o Preventive Actions


Question:
Which are the suggestions/recommendations for the best execution of the “Growth Promotion Test” (GPT)?

Answer:
(a) Recommendations for your microbiological laboratory
The GPT is one of the most important Quality Control tests performed in the laboratory today. The nutritive properties of each batch of medium must be established before successful and accurate testing can be performed. Therefore, it is recommended that when performing the GPT, samples are tested from both new batch of media along with previously approved media, side-by-side.
This will reduce variability, as both batches of media will be inoculated with the same micro-organism suspension.
(b) Pharmacopeia Instructions for testing
  Inoculate medium with 100 CFU or less.
  Incubate medium according to Pharmacopoeia directions.
  If using agar, count the colonies.
  If the new agar is to be used for enumeration tests, the number of colonies should be within a factor of two from the number of colonies on the previous approved medium. Examples of agar Soy Bean Casein Agar and Sabouraud Dextrose Agar.
  If the new agar is to be used in test for specified micro-organisms (for example Salmonella), the number of colonies on the new medium should be “comparable” to the number of colonies on the previous approved medium.
(c) Microbiologics™ Recomendations for Ensuring Satisfactory Results
  For best results, use parallel testing instead of historical data for comparing the new medium to previously approved medium.
  When using parallel testing, the new media and the previously approved media are inoculated with the same inoculums by the same technician and are subjected to identical incubation conditions.
  The only variable is the media.
  It testing selective media, test non selective media at the same time because recovery on selective media is not as high as on nonselective media.
  The non selective agar should grow less than 100 colonies.
  Remember, the count on the new media does not have to be within a factor of two from the count of the previous approved media. For selective media the count only needs to be “comparable”.
(d) The characteristics of Microbiologics™ microbial strains

EZ-Accu Shot®
EZ-Accu Shot™ SelectEZ-Accu Shot™EZ-CFU™ One StepEZ-CFU™
For small volume usersFor small volume usersFor small volume usersFor large volume users
Delivers <100 CFU/0.1 mlDelivers <100 CFU/0.1 mlDelivers <100 CFU/0.1 mlDelivers <100 CFU/0.1 ml
No dilution stepsNo dilution stepsNo dilution stepsNo dilution steps
10 tests per vial10 test per vial19 test per vial >90 test per vial
Stable for 8 hours after hydration, when refrigeratedStable for 8 hours after hydration, when refrigeratedStable for 8 hours after hydration, when refrigerated
Quick dissolveQuick dissolvePre-warm 30 minutesPre-warm 30 minutes
5 different strains per kit1 strain per kit1 strain per kit1 strain per kit


Question:
What can laboratories do to ensure that non-sterile products are safe?

Answer:
“Magnified” della Microbiologics™ “Keeping Bad Bugs Out of Good Products”.
Laboratories can do the following:
• Perform a risk assessment to determine which microorganisms could be harmful based on the route of administration, the user and the properties of the drug.
• Scan for potential problems by researching past recalls in similar products. Go to http://www.fda.gov/foi/warning.htm
• Use enrichment and selective media to find objectionable organisms.
• Identify suspicious colonies found when performing total aerobic counts.
• Follow pharmacopeia guidelines for microbial enumeration and tests for specified microorganisms.
• Perform growth promotion testing on all batches of media used in aerobic plate counts and tests for specified and objectionable organisms.
• Validate all tests used for isolating and identifying organisms.
• Routinely perform environmental monitoring. Identify the organisms that are isolated. Determine if these organisms could degrade the drug or be harmful to users of the drug.
• Perform disinfectant testing to determine which disinfectant(s) to use at the manufacturing site.
• Perform bioburden testing in order to discover any weaknesses in the manufacturing process.
• Keep current with trends in pharmaceutical microbiology by attending webinars and conferences and reading scholarly articles.


Question:
What are the differences in the interpretation of results for non sterile products according the EU, USA, J harmonization?

Answer:
• Factor: Introduction of factor 2 instead of factor 5
• Recovery: result can be between half and twice of the inoculum
• Monograph Product Specification: acceptance criterion 102 CFU maximum acceptable count = 200; minimum=50.


Question:
Which is the ISO standard for sampling techniques from surfaces using Contact Plates?

Answer:
The specific ISO standard is the ISO 18593:2004. Microbiology of food and animal feeding stuffs - Horizontal methods for sampling techniques from surfaces using contact plates and swabs ...
See the Application Note N.99/119 “Rodac-Weight” microbiological surface sampler use according to ISO 18593:2004 >>>  


Question:
How should you write the Standard Operating Procedure for the colony counting to apply the GLP?

Answer:
The Application Note N. 199/102 “S.O.P. for Colony Counting” gives you a Guide Line to write the Standard Operating Procedure for the microbiological laboratory >>>


Question:
Which are the pathogen Campylobacter?

Answer:
Campylobacter producing gastro-intestinal infections in the human:
Campylobacter jejuni
Campylobacter coli
Campylobacter lari
Campylobacter helveticus
Campylobacter upsaliensis
Campylobacter  producing extra gastro-intestinal infections in human:
Campylobacter jejuni
Campylobacter coli
Campylobacter fetus
Campylobacter hyointestinalis
Campylobacter sputo rum
Campylobacter producing dental infections:
Campylobacter concisus
Campylobacter curvus
Campylobacter rectus
Campylobacter showae


Question:
How you should avoid that the Stomacher™ bag is broken during the homogenization of food?

Answer:
This risk is avoided using the “Reinforced round bag”.  In fact, the special construction and type of resin of the bag is able  to resist to the mechanical stress of the Stomacher™ paddles.


Question:
Where do you find references about Compact-Dry X-BC for Bacillus cereus enumeration?

Answer:
Compact-Dry X-BC for the Enumeration of Bacillus cereus in Food Samples >>>


Question:
Which are the official methods for Campylobacter spp detection and count?

Answer:
Ermanno Donadio
The ISO 10271-1:2006 for detection and confirm and the method ISO/TS 10272:20 for enumeration of Campylobacter.


Question:
Which are the swabs that should be used for the microbiology testing and research of food pathogens like E. coli 0104?

Answer:
The lurking menace! Detection of foodborne pathogens
The current, and very dramatic outbreak of E. coli O104 in Germany reminds us all of the need for great vigilance in food hygiene, particularly for those with responsibility for production and delivery of food that is safe for the consumer. Challenges can come not only from known pathogens, but recent years have seen the sudden emergence of previously unknown strains with tragic lethal outcomes  for consumers, while for producers and suppliers the consequences can be legally  and economically disastrous.
Food handling and processing environments, together with food contact surfaces such as blades, mixers, benches, and conveyor belts are common vectors for the spread of pathogens, so rigorous validated cleaning procedures are essential to prevent contamination.  To ensure cleaning continues to be effective, all cleaned surfaces must be regularly sampled and tested to ensure microbial levels are within acceptable limits and that dangerous pathogens are not present at all.
Medical Wire has a number of products to help with this task.
NRS II Transwabs® are sampling swabs premoistened with a neutralising solutions. Surfaces which have been recently cleaned may still have traces of antimicrobial substances, such as disinfectants which would kill any bacteria picked up in a sample, and so prevent them being detected. The neutralisers in NRS II cancel these antimicrobial activities, allowing the microbes present to survive and be detected. NRS II Transwabs® are supplied with a choice of accurate fill volumes to allow quantitative measurements of the environmental bioburden, so that any changes or trends can be quickly identified. The collected samples can also be tested for particular pathogens.
Polywipes™ are premoistened blue sponge swabs that allow the sampling of larger areas, such as machines, conveyor belts, floors and walls. The sponges are premoistened with a phosphate buffer that also effectively neutralises trace antimicrobials, this time by rapid dilution. By regularly swabbing on defined areas  and quantifying the microbial load, changes or trends can be monitored and responded to.  Pathogens can also be detected if present. Polywipes™ have also been used in clinical environments for the detection of reservoirs of healthcare associated pathogens such as Clostridium difficile.
Medical Wire also has some swab products for the detection of particular pathogens such as coliforms. The Coliform Isolation Transwab® will detect coliorms including E. coli on surfaces, and is an effective way of monitoring personal hygiene in food manufacturing premises. For example door handles can be swabbed. The swab is placed in the purple medium which is then incubated overnight (or up to 24 hours). Any coliforms present will multiply, causing the medium to turn yellow. Other products are available for the detection of Listeria, and for general hygiene.


Question:
Which are the most dangerous micro-organisms for the human health according to the U.S. Federal Experts Security Advisory Panel (FESAP)?

Answer:
EMERGING FOODBORNE PATHOGENS
Prof. Dr. Ýrfan EROL, DVM, Ph.D. Turkish Representative of World Vet. Assoc. Department of Food Hygiene and Technology School of Veterinary Medicine Ankara University >>>


Question:
Which are the characteristics of E.coli so popular during these days?

Answer:
Escherichia coli is a bacterium, which inhabits the intestinal tract of humans and other warm-blooded mammals. It constitutes approximately 0.1% of the total bacteria in the adult intestinal tract. Its name comes from the name of the person, Escherich, who in 1885 first isolated and characterized the bacterium. The bacterium, particularly one type called strain K–12, is one of the most widely studied organisms in modern research. Its biochemical behavior and structure are well known, having been studied for much of this century. This plethora of information has made E. coli indispensable as the bacterial model system for biochemical, behavioral and structural studies. E. coli is also the most encountered bacterium in clinical laboratories, being the primary cause of human urinary tract infections. Pathogenic (diseases causing) strains of E. coli are responsible for pneumonia, meningitis and traveler's diarrhea. As part of the normal flora of the intestinal tract, E. coli is beneficial. It is crucial in the digestion of food, and is our principle source of vitamin K and B-complex vitamins. Outside of the intestinal tract, E. coli dies quickly. This trait has been exploited, and E. coli is a popular indicator of drinking water quality, as its presence indicates the recent contamination of the water with feces. One of the most harmful types of E. coli is a strain called O157:H7. Researchers surmise that O157:H7 arose when an innocuous E. coli bacterium was infected by a virus carrying the genes coding for a powerful toxin called Shiga-like toxin. The toxin can destroy cells in the intestinal tract and, if they enter the bloodstream, can impair or destroy the kidneys and the liver. The intestinal damage causes a lot of bleeding. In children and elderly people, this hemorrhaging can be lethal. In other people, damage to the kidney and liver can be permanent or even lethal. In the summer of 2000, more than 2,000 people in Walkerton, Ontario, Canada were sickened and seven people died from drinking water which had been contaminated with O157:H7. Strain O157:H7 was first linked to human disease in 1983, when it was shown to have been the cause of two outbreaks of an unusual and severe gastrointestinal ailment in the Unites States. Since then, the number of documented human illnesses and deaths caused by O157:H7 has increased steadily worldwide. Disease caused by E. coli is preventable, by proper hand washing after bowel movements, avoidance of unpasteurized milk or apple cider, washing of raw foods before consumption and thorough cooking of ground meat. Modern genetics techniques have been successful in obtaining the sequence of the genetic material of E. coli. Frederick Blattner and his colleagues published the genome sequence of strain K–12 in 1997. The genome was discovered to have approximately 4300 protein coding regions making up about 88 per cent of the bacterial chromosome. The most numerous types of proteins were transport and binding proteins—those necessary for the intake of nutrients. A fairly large portion of the genome is reserved for metabolism—the processing of the acquired nutrients into useable chemicals. In 2000, Nicole Perna and her colleagues published the genome sequence of O157:H7. The O157:H7 genome shows similarity to tat of k12, reflecting a common ancestry. But, in contrast to K12, much of the genome of O157:H7 codes for unique proteins, over 1,300, some of which may be involved in disease causing traits. Many of these genes appear to have been acquired from other microorganisms, in a process called lateral transfer. Thus, strain O157:H7 appears to be designed to undergo rapid genetic change. This distinction is important; indicating that strategies to combat problems caused by one strain of E. coli might not be universally successful. Knowledge of the genetic organization of these strains will enable more selective strategies to be developed to combat E.coli infections.


Question:
Which is the suggested medium for E.coli 0104:H4?

Answer:
Brand New: CHROMagar™ STEC-O104 Supplement
CHROMagar has been working during the recent years on a new medium, designed for the detection of STEC (and not only O157!!!). The result of this work is CHROMagar™ STEC, a selective medium, allowing the growth of a majority of STEC strains in mauve colony colour, while in general colouring in blue or inhibiting other bacteria. CHROMagar STEC has a good sensitivity and a good selectivity/specificity for detection of various STEC. >>>


Question:
What about the maintenance of the Quality Control Strains?

Answer:
Maintenance of Quality Control Strains >>>


Question:
Where you can find the microbiology news of the week?

Answer:
-
www.microbeworld.org/twim
A podcast about unseen life on Earth. The newest podcast from the ASM. TWiM talks about the week’s stories in microbiology.


Question:
Which are the simple definitions of the microbiological terminology?

Answer:
The basic terminology and definitions of microbiology
Microbe - An organism that can only be seen clearly under a microscope
Eukaryote - An organism made of cells which contain membrane-bound organelles such as nuclei
Prokaryote - An organism made of a cell which lacks membrane-bound organelles such as nuclei, e.g. bacteria and archaea
Bacterium - An unicellular prokaryote with a cell wall made from peptidoglycan; bacteria (plural) make up one of the 3 domains of life
Archaean - An unicellular prokaryote similar to bacteria; archaea (plural) make up one of the 3 domains of life
Eukarya - One of the 3 domains of life; contains all eukaryotes, including plants, animals, fungi and other organisms that were previously classified as Protoctista in the 5 kingdom system      
Kingdom - The highest rank in the hierarchy of the 5 kingdom system of classification; the 5 kingdoms are: Prokaryotae, Animalia, Plantae, Fungi and Protoctista
Domain - A category of organisms; all organisms can be classified in one of the domain Eukarya, Prokarya and Archaea
Classification - The arrangements of organisms into group
Taxonomy - The process of naming and classifying organisms
Phenetic classification - A method of arranging organisms based on properties such as anatomy of morphology (i.e.: the 5 kingdom system)
Phylogenetic classification - A method of arranging organisms based on evolutionary relationship between organisms (i.e.: the 3 domain system)
Chromosome - An organized structure of DNA (and often protein); contains genes
Plasmid - A circular piece of DNA separated from the chromosome of a bacterium
Conjugation - The transfer of DNA from one cell to another via direct cell to cell contact
Asexual - A type of reproduction that does not depend on sex cells or sex organs
Binary fission - A type of asexual reproduction in which a single-celled organism divides to produce two daughter cells of the same size
Budding - A type of asexual reproduction in which a new cell or appendage is formed from an outgrowth of a cell; occurs in microbes such as yeast and some plants and animals
Capsule - A protein or polysaccharide layer external to the cell wall; found in some prokaryotic cells
Endospore - A dormant, non-reproductive structure formed inside some bacterial cells, often in response to environmental conditions; many are able to survive extreme temperatures, radiation and desiccation and will develop into bacterial cells when conditions become more favorable
Flagellum - A long filament sticking out of a cell that enables movement; in bacteria it moves with a cork screw motion due to the rotation of a flagellar motor anchored in the cell membrane
Pilus - A protein filament protruding from the surface of some bacterial cells (similar to a fimbria); some are involved in conjugation
Fimbria - A protein filament protruding from the surface of some bacterial cells (similar to a pilus)
Ribosome - A structure made of protein and RNA, that is the site of protein synthesis
Peptidoglycan - A polymer found in the cell walls of bacteria
Gram stain - A method that stains bacteria differentially according to their cell wall structure
Gram-negative - Bacteria with cell walls made of 10% peptidoglycan plus an additional layer; they stain pink or red with Gram’s reagent
Gram-positive - Bacteria with cell made of 90% peptidoglycan; they stain purple with Gram’s reagent
Glycocalyx - Slimy or gummy material secreted on the outside of some bacterial cells, e.g.: a slime layer or capsule
Slime layer - A gummy layer external to the cell wall that is found in some prokaryotic cells; unlike a capsule it is diffuse and easily removed
Fungus - An eukaryotic organism with a cell wall made from chitin; can be unicellular (e.g.: yeast) or multi-cellular (e.g.: moulds)
Yeast - An uni-cellular fungus; used widely in biotechnology
Hypha - A thread-like fungal filament which forms branching networks called mycelia
Mycelium - A mass of fungal filaments (hyphae)
Spore (fungal) - A single-celled or multi-cellular structure produced for dispersal, as a result of sexual or asexual reproduction or in response to adverse conditions
Fruiting body - A structure made by filamentous fungi in order to produce and release spores; they are commonly known as mushrooms and toadstools
Virus - An acellular infection agent consisting  of a protein coat and nucleic acid core
Virion - A virus particle consisting of a protein coat called a capsid and a core (containing a nucleic acids) called the nucleocapsid.
Envelope (viral) - A phospholipid bilayer on the outside of certain viruses
Capsid - The protein coat that surrounds the nucleic acid genome of a virus
Nucleocapsid - The core of a virus; contains the RNA or DNA genome
Lytic cycle - The life cycle of a virus during which it replicates continually, destroying the host and releasing viral particles
Font www.sgm.ac.uk


Question:
Where you can find microbiology opportunities in USA?

Answer:
The NIAID (National Institute Of Allergy and Infectious Diseases), one of the largest Institutes of the National Institutes of Health, is searching for motivated and qualified candidates for a variety of scientific management positions.
To learn more visit the web at www.niaid.nih.gov/careers/sm 


Question:
Are you ready for the “Microbiologics” game? “Name that organisms!”

Answer:
Keeping Bad Bugs Out of Good Products >>>
No pharmaceutical company wants to sell a non-sterile drug which is contaminated with objectionable microorganisms, but how does a company know which organisms are objectionable and how do they ensure that their drugs do not contain them?
An objectionable organism is one which can either cause illness or degrade the product thus making it less effective. The FDA says that "appropriate written procedures designed to prevent objectionable microorganisms in drug products not required to be sterile, shall be established" (21 CFR 211.113) and that "appropriate laboratory testing must be conducted on each batch of drug required to be free of objectionable organisms." (21 CFR 211.165) ...



Question:
Where is it possible to find a specific information about the environmental microbiology?

Answer:
Air Microbiology.
- http://www.scribd.com/doc/19079015/Air-Microbiology-2009


Question:
What is the difference between a colony, a CFU, and a bacterial cell?

Answer:
In microbiology, colony-forming unit (CFU) is a measure of viable bacterial or fungal numbers. Unlike in direct microscopic counts where all cells, dead and living, are counted, CFU measures viable cells. By convenience the results are given as CFU/mL, colony-forming units per milliliter.
The theory behind the technique of CFU establish that a single bacterium can grow and become a colony, via binary fission. These colonies are clearly different between each other, both microscopical and macroscopical. However, some bacteria do not separate completely during the sample preparation process (Staphylococcus, Streptococcus) and the results of the count will be below the number of individual cells using direct methods.
A COLONY is a cluster of microorganisms growing on the surface of or within a solid medium; usually cultured from a single cell.
Each bacterial CELL arises either by division of a preexisting cell with similar characteristics (i.e., binary fission), or through a combination of elements from two such cells in a sexual process.
Statistical significance, when applied to incubated plates, depends upon the research hypothesis, the power of the test (i.e., the probability of a Type II error (i.e., the likelihood of failing to reject the null hypothesis when it is not true)), the difference between the number of colonies observed under the experimental and control conditions, and the accuracy of the counting methodology.
Source(s):
http://en.wikipedia.org/wiki/Colony-form
http://www.answers.com/topic/colony-word
http://en.wikipedia.org/wiki/Statistical
http://en.wikipedia.org/wiki/Type_I_and_…


Question:
Where you can find an on-line text of bacteriology?

Answer:
Todar’s on lineTextbook of bacteriology.
- http://www.textbookofbacteriology.net/growth_4.html
Continuous Culture of Bacteria
The cultures so far discussed for growth of bacterial populations are called batch cultures. Since the nutrients are not renewed, exponential growth is limited to a few generations. Bacterial cultures can be maintained in a state of exponential growth over long periods of time using a system of continuous culture (Figure 4), designed to relieve the conditions that stop exponential growth in batch cultures. Continuous culture, in a device called a chemostat, can be used to maintain a bacterial population at a constant density, a situation that is, in many ways, more similar to bacterial growth in natural environments.
In a chemostat, the growth chamber is connected to a reservoir of sterile medium. Once growth is initiated, fresh medium is continuously supplied from the reservoir. The volume of fluid in the growth chamber is maintained at a constant level by some sort of overflow drain. Fresh medium is allowed to enter into the growth chamber at a rate that limits the growth of the bacteria. The bacteria grow (cells are formed) at the same rate that bacterial cells (and spent medium) are removed by the overflow. The rate of addition of the fresh medium determines the rate of growth because the fresh medium always contains a limiting amount of an essential nutrient. Thus, the chemostat relieves the insufficiency of nutrients, the accumulation of toxic substances, and the accumulation of excess cells in the culture, which are the parameters that initiate the stationary phase of the growth cycle. The bacterial culture can be grown and maintained at relatively constant conditions, depending on the flow rate of the nutrients. 
Synchronous Growth of Bacteria
Studying the growth of bacterial populations in batch or continuous cultures does not permit any conclusions about the growth behavior of individual cells, because the distribution of cell size (and hence cell age) among the members of the population is completely random. Information about the growth behavior of individual bacteria can, however, be obtained by the study of synchronous cultures. Synchronized cultures must be composed of cells which are all at the same stage of the bacterial cell cycle. Measurements made on synchronized cultures are equivalent to measurements made on individual cells.
A number of clever techniques have been devised to obtain bacterial populations at the same stage in the cell cycle. Some techniques involve manipulation of environmental parameters which induces the population to start or stop growth at the same point in the cell cycle, while others are physical methods for selection of cells that have just completed the process of binary fission. Theoretically, the smallest cells in a bacterial population are those that have just completed the process of cell division.  Synchronous growth of a population of bacterial cells is illustrated in Figure 5. Synchronous cultures rapidly lose synchrony because not all cells in the population divide at exactly the same size, age or time.

Question:
Which are the indications of the ISO 21807:2004. “Microbiology of food and animal feeding stuffs – Determination of water activity” for what concerns the sample representative?

Answer:
7. Obtaining a representative specimen.
“The distribution of Aw may be presumed to be largely homogeneous in virtually all foods. In any case, a treatment in a mincer may produce hot and give off water, with the result that the sample will no longer representative.
Products like sausage and ham are an exception. If necessary, the Aw conditions may be determined in the inside and outside regions to cover all the constituents by systematically selecting the measurement points.
Another exception is water-in-oil emulsion (e.g.: margarine) which have heterogeneous Aw even after homogenization.


Question:
Which are the ISO standards concerning the surface microbiological sampling using contact plates, swabs and sponges?

Answer:
Ermanno Donadio
The microbiological sampling method of surfaces is described in the standard
ISO 18593:2004 Microbiology of food and animal feeding stuffs -- Horizontal methods for sampling techniques from surfaces using contact plates and swabs
The analytical phases are described in the standards:
ISO 18593:2004 Microbiology of food and animal feeding stuffs -- Horizontal methods for sampling techniques from surfaces using contact plates and swabs
ISO 4832 Microbiology of food and animal feeding stuffs -- Horizontal method for the enumeration of coliforms -- Colony-count technique
ISO 4833 Microbiology of food and animal feeding stuffs -- Horizontal method for the enumeration of microorganisms -- Colony-count technique at 30 degrees C
ISO 6579 Microbiology of food and animal feeding stuffs -- Horizontal method for the detection of Salmonella spp.


Question:
How can you present the germs to the non microbiologist?

Answer:
The one show
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www.aston.ac.uk
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http://www1.aston.ac.uk/


Question:
Where can you find on the web glossary dedicated to the accrditation of a microbiology laboratory?

Answers:
“Eurachem / EA Guide 04/10” – “Accreditation for Microbiological Laboratory” 
June 2002 – Appendix A - Glossary of terms – Pages 19-20 of 26:
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http://www.eurachem.org/guides/pdf/EurachemEA_Micro.pdf



Question:
In which document on the web can you find all the useful information / guide lines for the accreditation of a microbiology laboratory?

Answer:
“Eurachem / EA Guide 04/10” – “Accreditation for Microbiological Laboratory” – June 2002:
- http://www.eurachem.org/guides/pdf/EurachemEA_Micro.pdf


Question:
Which are the accreditations provided by “Microbiologics®” in the microbiology Quality Control activity for the laboratory?

Answer:
- ISO 9001:2008 – Assures customers that “Microbiologics®” provides services and manufactures product in accordance with a sound Quality Management System.
- ISO/IEC 17025:2005 – Provides official recognition that “Microbiologics®” uses suitable test methods and procedures that produce accurate and consistent test results
- ISO Guide 34:2000 – ISO Guide 34:2000 provides the highest level of Quality Assurance; providing objective third-party recognition that “Microbiologics®” is a qualified reference material producer
- Food and Drug Administration establishment registration – Provides evidence that “Microbiologics®” adheres to all current GMP
- CE Mark – Indicates conformity with the essential requirements set forth in European Directives. It also verifies the performance of the product and constancy of the production process.


Question:
Which are the major storage fungi and the moisture contents of commodities at which mold invasion may occur?

Answer:
Font: Christensen e Meonuck - 1986.

MOULDSCOMMODITIES% MOISTURE
Aspergillus restrictus (blue eye)Starchy cereals14.0 – 14.5
Aspergillus restrictus (blue eye)Soybeans12.0 – 12.5
Aspergillus restrictus (blue eye)Sunflower, peanuts8.5 – 9.0
Eurotium glaucus (blue eye)Starchy cereals14.5 – 15.0
Eurotium glaucus (blue eye)Soybenas12.5 – 13.0
Eurotium glaucus (blue eye)Sunflowers, peanuts9.0 – 9.5
A. candidusStarchy cereals15.5 – 16.0
A. candidusSoybeans14.5 – 15.0
A. candidusSunflowers, peanuts9.0 – 9.5
A. ochraceusStarchy cereals15.5 – 16.0
A. ochraceusSoybeans14.5 – 15.0
A. ochraceusSunflowers9.0 -9.5
A. flavusStarchy cereals17.0 – 18.0
A. flavusSoybeans17.0 – 17.5
Penicillium (blue eye in corn)Starch grains16.5 – 20.0
Penicillium (blue eye in corn)Soybeans17.0 – 20.0
Penicillium (blue eye in corn)Sunflower10.0 – 15.0


Question
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How to be updated on detection and enumeration of moulds in cereal products
?

Answer:
The microbiology of cereal and cereal products >>>
L. Bullerman, A. Bianchini. The Microbiology of Cereal and Cereal Products.
Variety of media and methods available for detection and enumeration of moulds in cereal products.
- www.foodquality.com


Question:
How to be updated on detection and enumeration of moulds in cereal products?

Answer:
L. Bullerman, A. Bianchini. The Microbiology of Cereal and Cereal Products >>>
Variety of media and methods available for detection and enumeration of moulds in cereal products.
- www.foodquality.com


Question:
Which is the dilution theory about the “Most Probable Number” Method?

Answer:
The Most Probable Number Method
This page and related pages are intended to address the use and interpretation of dilution theory techniques (plating and MPN) for educational purposes and may not be authoritative in applied or research situations. Familiarity of the author with any applied or research aspect is not to be implied.
- http://www.jlindquist.net/generalmicro/102dil3.html


Question:
Which are the Guide Lines for the “Microbiology Quantitative Food Process Control” ?
How do you operate for microbiological testing of irregular surfaces?

Answer:
The easy-to-use Surface Swab Kit for bioaerosols provides all of the necessary supplies to sample surfaces for bioaerosols that are deposited through contact or that settle out of the air onto surfaces where they can be re-aerosolized and inhaled. It is ideal for determining the relative degree and type of biological contamination in an area and to verify remediation procedures. This non-destructive method can be used safely on most surfaces and is ideal for irregular surfaces such as air return grills.
Easy Four Step Sampling
1. Remove the sterile swab from its package aseptically.
2. Remove the sealing cap from the tube containing the sponge and moisten the swab tip.
3. Using a rolling motion, gently swab the desired area thoroughly. Templates are included in the kit for defined area sampling.
4. Insert the swab into the tube, replace the sealing cap, and prepare for transport to a laboratory.


Question:
How do you operate for microbiological testing of irregular surfaces?

Answer:
The easy-to-use Surface Swab Kit for bioaerosols provides all of the necessary supplies to sample surfaces for bioaerosols that are deposited through contact or that settle out of the air onto surfaces where they can be re-aerosolized and inhaled. It is ideal for determining the relative degree and type of biological contamination in an area and to verify remediation procedures. This non-destructive method can be used safely on most surfaces and is ideal for irregular surfaces such as air return grills.
Easy Four Step Sampling
1. Remove the sterile swab from its package aseptically.
2. Remove the sealing cap from the tube containing the sponge and moisten the swab tip.
3. Using a rolling motion, gently swab the desired area thoroughly. Templates are included in the kit for defined area sampling.
4. Insert the swab into the tube, replace the sealing cap, and prepare for transport to a laboratory.


Question:
Where  may you find information about opportunities for a young microbiologist?

Answer:
-Finding a job
www.academicjobseu.com
Academic Jobs EU
www.eurosciencejobs.com/jobs/biology
Euro Science Job
http://jobs.phds.org/
USA / Canada
-Finding opportunities
www.erc.europa.eu
ERC (European Research Council)
-Support for researchers
www.leru.org
LERU (League of European Research Universities)
www.eurodoc.net/
Eurodoc. Federation of National Association of PhD candidates and young researchers.


Question:
Which are the suggested time and speed of food homogenization by reciprocating two paddles equipment?

Answer:
- The available bags for microbiological homogeniuzation >>>
The “pbi” plastic, sterile, disposable bags to be used with the two paddles microbiological homogenizer have different characteristics:
- “Reinforced-Round-Bag”  has round bottom, printed complete information for traceability and dilution factor and has tear of top seal to guarantee sterility until the moment of use. Sterility certificate available.
- “Reinforced Eco” has round bottom, and has tear of top seal to guarantee sterility until the moment of use.
- “Presto-Chiuso Circulator” has tear of top seal to guarantee sterility until the moment of use and  metal top sealing. Sterility certificate available.
- “Stofilter Kenzo” has a lateral welded inner filter for fibre-rich fibrous material.
- “Velo” has a full size welded inner filter for fibre-rich fibrous material. Sterility certificate available.


Question:
What is possible to do to minimize risk of blocked pipettes by sample debris during food homogenization using a Seward “Stomacher” lab blender?

Answer:
Seward's Strainer Bag Minimises Risk of Blocked Pipettes by Sample Debris
Seward Limited, manufacturer and developer of the world leading, patented Stomacher® Paddle Blender range for over 40 years, has developed Strainer bags for processing samples that produce large amounts of debris that can block pipettes. Irradiated sterile and available in 80 and 400ml sizes the Strainer bags are fully compatible with all laboratory blenders. Ideal for a variety of applications they deliver fully homogenous, virtually debris-free sample every time to ensure trouble-free pipetting and quality results.
Since the pore size on the integrated, robust strainer element within the Seward Strainer bag is 0.5mm, this also minimises the risk of pore blockage that can result in non-homogenous prepared sample. If pores become blocked then, instead of straining, a filtering effect can occur, meaning microbes may be eliminated from the final filtrate and produce an inaccurate final result. Seward Strainer bags overcome the risk of such sample filtering across a broad range of sample types. These range from hard shellfish to highly fatty samples, including cheese and minced beef, through to excessive debris producing products, such as ready meals.
“Our Strainer bags have been developed following initial work that we undertook with industry experts into combining the 'stomaching' and filtration element of sample preparation into a single process,” explained Dan Crothers, Marketing Manager, Seward. “After assessing several different options our research emphatically proved this process is not feasible, since filtering out all debris can result in non-homogenous sample which can skew final results. We found that straining to minimise debris is perfectly adequate to prevent pipette blocking issues, whilst ensuring that results give a true reflection of microbial loading.”


Question:
When and where the 2011 European Congress of Clinical Microbiology and Infectious Diseases is organized?

Answer:
European Congress of Clinical Microbiology and Infectious Diseases.
- http://www.eccmid-icc2011.org/


Question:
How should the recovery of challenge micro-organisms on selective media demonstrated?

Answer:
Technical Information Bullettin TIB 134 “Microbiologics®” (Page 88/89)
-Glossary
Acceptable Minimum Recovery, average value, CFU, challenge, non selective agar medium, percent recovery, pour plate, reference stock culture, selective agar medium, spread plate, working stock culture.
-Introduction
Each in-house selective media preparation, commercial pre-poured selective medium, or commercial detection device based on selectivity must be quality controlled. In addition to such specifications and performance as sterility, pH, positive and negative differential reaction and colony morphology, evaluation of selectivity and recovery properties must be established and documented.
-Statement of fact
  Selective media rarely demonstrates 100% CFU recovery in comparison to identical assayed micro-organism challenge on non-selective media.
  Acceptable minimum recovery limits need to be established for each selective medium employed in each individual laboratory.
-Recovery of challenge micro-organisms on selective media
The following is a simple procedure that can be used to demonstrated the recovery of challenge micro-organisms on selective media.
(1) Select reference stock culture or working stock cultures, traceable to authentic reference cultures, which are appropriate to challenge the medium’s selective properties.
(2) Prepare dilutions of the challenge micro-organism to achieve a challenge concentration of 100 CFU.
If a selective agar medium or device is used as a “poured plate”, 100 CFU per 1,0 mL is recommended.
If a selective agar medium or device is used as a “streak or spread plate”, 100 CFU per 0,1 mL is recommended.
(3) Inoculate 3 selective agar media with 100 CFU challenge micro-organisms per 1,0 mL or 0,1 mL.
Inoculate 3 non-selective standard methods agar media with 100 CFU challenge micro-organisms per 1,0 mL or 0,1 mL.
(4) Incubate both sets of selective and non-selective media in accordance with the requirements for the selective medium.
(5) Following incubation, count and average the CFU for the selective and non-selective media. 
Calculate the average and divide the average value of the non-selective CFU recovery into the selective CFU recovery. Multiply the answer by 100 to determine the Percent Recovery.

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