The Addition Of Which Of The Following To A Culture Medium Will Neutralize Acids

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Free Flashcards about Microbiology

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Question Answer
If cells are grown in media containing amino acids labeled with radioactive nitrogen (15N), most of the radioactivity will be found in the cells’ DNA and Proteins
Which of the following elements is NOT correctly matched with its cellular function? Phosphorus used for production of carbohydrates
The addition of which of the following to a culture medium will neutralize acids? Buffers
Salts and sugars work to preserve foods by creating a Hypertonic environment
The term aerotolerant anaerobe refers to an organism that does not use oxygen but tolerates it
Most bacteria grow best at pH 7
Most fungi grow best at pH 5
Which of the following pairs of microbe classification terms and optimal growth temperatures is mismatched? thermophile growth at 37 degrees celcius
Which group of microorganisms is most likely to spoil a freshwater trout preserved with salt? facultative halophiles
Which of the following is an organic growth factor? vitamin B1
Which of the following is an example of a metabolic activity that could be used to measure microbial growth? glucose consumption
Patients with indwelling catheters (long-term tubes inserted into body orifices for drainage, such as through the urethra and into the urinary bladder) are susceptible to infections because biofilms develop on catheters
Pathogenic bacteria isolated from the respiratory or intestinal tracts of humans are capnophiles that grow best in carbon dioxide incubators
The biosafety level (BSL) for a clinical microbiology laboratory working with potentially airborne pathogens, such as tuberculosis bacteria, is BSL-3
Consider a culture medium on which only gram-positive organisms such as Staphylococcus aureus colonies can grow due to an elevated NaCl level. A yellow halo surrounds the growth, indicatingthebacteriumfermentedasugarinthemedium, low ph change color- what type of medium is this- selective and differential medium
A culture medium consisting of agar, peptone, and beef heart is a complex medium
The source of nutrients in nutrient agar is peptone and beef extract
Which of the following is an advantage of the standard plate count? determines the number of viable cells
Most bacteria reproduce by binary fission
During which growth phase will gram-positive bacteria be most susceptible to penicillin? log phase
Which of the following is the best definition of generation time? the length of time needed for a cell to divide
Which of the following regarding antimicrobial control agents is FALSE? A true antimicrobial control agent is equally effective against both bacteria and viruses
Which of the following is the best method to sterilize heat-labile solutions? membrane filtration
Which of the following could be used to sterilize plastic Petri plates in a plastic wrapper? gamma radiation
Which of the following regarding antimicrobial control agents is FALSE? Alcohols effectively inactive nonenveloped viruses by attacking lipids
All of the following substances are effective against nonenveloped viruses EXCEPT alcohol
All of the following are effective for destroying prions EXCEPT boiling
Which of the following pairs is mismatched? RNA polymerase makes a molecule of RNA from an RNA template
Which of the following statements is FALSE? DNA replication proceeds in only one direction around the bacterial chromosome
Which of the following is NOT a product of transcription? a new strand of DNA
If you knew the sequence of nucleotides within a gene, which one of the following could you determine with the most accuracy? the primary structure of the protein
The cancer gene ras produces mRNA containing an extra exon that includes a number of UAA codons. Cancer cells produce ras mRNA missing this exon. This mistake most likely is due to a mistake by snRNPs

OneClass: The addition of which of the following to a culture medium w

Calculate the pH of an aqueous solution of 0.200 M acetic acid, which is the strongest acid known to science. A) acetic acid, K,-1.8 x 10* llg is formed by the reaction of dihydrogen phosphate ion with potassium. What is the OH concentration of a solution with a pH of 10.35? b) benzoic acid, K,-6.3 x 10) formic acid, K,-1.8 x 1 What is the OH concentration of a solution with a pH of 10.35? The hydrocyanic acid K,-4.0x 101 is used as an example. M 1.6×10-9 M 2.7×10-8 M B6. a)1.6×10-9 M b)2.7 x 10-8 M B6.

  1. Which of the following species has the least amount of acidity in its composition?
  2. When buffers are diluted with water, they become more resistant to pH fluctuations.
  3. d) Buffers include significant amounts of a weak acid as well as the conjugate base of the weak acid.
  4. When a strong acid and a weak base are used in titration, which of the following requirements is true?
  5. b) The equivalency point is reached when the pH is equal to 7.
  6. d) To achieve the equivalence point, equal amounts of acid and base must be added together.
  7. B10.
  8. In a saturated solution of Ba+, what is the concentration of Ba+?

Microbial Growth Flashcards

Lamar University’s BIOL 2420Microbiology Laboratory In a culture media, the addition of which of the following will neutralize acidity? Salts and sugars help to preserve foods by forming a gel-like substance. The word facultative anaerobes refers to an organism that either utilizes oxygen or develops in the absence of oxygen, which is a disadvantage of the conventional plate count. Cells may group together to create aggregates. It is necessary to allow for incubation time. The medium and incubation conditions dictate the chemical and physical needs.

It counts the number of dead cells.

It is necessary to have a large number of cells Metabolic activity is measured using this method.

CO2 produced by the Krebs cycleNO2- produced by the electron transport chain CO2 produced by the Krebs cycleNO2- produced by the electron transport chain Reduced concentrations of dissolved oxygen A normal plate count revealed that 36 colonies developed in nutrient agar from 1ml of undiluted material in a nutrient agar plate.

  • The pH of a culture medium on which only gram-positive organisms grow and a yellow halo surrounds Staphylococcus aureus colonies is optimal for the growth of the majority of bacteria.
  • At 0 degrees Celsius, growth occurs.
  • Temperatures of 15 degrees Celsius promote growth.
  • The time it takes for a cell to divide is measured in minutes.

Counting through direct microscopic observation Plates are counted in accordance with industry standards FiltrationMPNMost likely to ruin freshwater fish that has been kept in salt A metabolic activity that may be used to quantify microbial development is, for example, which of the following: An experiment began with four cells and finished with one hundred and eighty-two cells.

  • Inoculating three cells into a culture medium with a generation time of 30 minutes results in the formation of three colonies.
  • What organism is responsible for the production of catalase and superoxide dismutase?
  • A media used to select for a nitrogen-fixing chemoheterotroph should not contain any of the elements listed below.
  • Afterwards, you infected 200 mL of nutrient broth with 100 cells from the same species.

After 24 hours of incubation, you should be able to harvest The number of cells in both is the same. The source of nutrients in nutrition agar is the same. Carbon dioxide (CO2) is used as a carbon source and as an energy source by producers at hydrothermal vents on the ocean’s surface.

Bacterial culture through selective and non-selective conditions: the evolution of culture media in clinical microbiology

The History of Microbiology (Wainwright and Lederberg, 1996) is an excellent place to start. Encyclopaedia of Microbiology, Volume 2, Pages 419–437, 1992. A case study in culturomics to examine the human gut microbiota by Lagier, Hugon, Khelaifia, Fournier, P.E., La Scola and Raoult. The revival of culture in microbiology is illustrated by the example of culturomics. Clin Microbiol Rev.2015;28:237–264. Clinical Microbiol Rev.2015;28:237–264. 3) Prescott, L.M., Willey, J.M., Sherwood, L.M., and Woolverton, C.J., Fifth Edition 2018; De Boeck Supérieur; Louvain-la-Neuve: De Boeck Supérieur.

  1. 4.Pommerville, J.C., 10th ed.
  2. Microbiology fundamentals are covered in this course.
  3. JonesBartlett; Sudbury: 2012.
  4. Microbes and civilisation in Alcamo’s world.
  5. The New England Journal of Medicine (NEJM) published a 300-page article in 1979 titled “887–893.” The eighth edition of Murray’s Principles of Radiology by Rosenthal et al.
  6. Medical microbiology is the study of bacteria in the body.
  7. Sandle, et al.

Vallery-Radot P.Masson et compagnie, Paris, 1922.

Pasteur’s Works, Vol.

2.

Microbiology textbook for students.

Manual for the DifcoTMBBLTM.

Atlas, R.M., 3rd edition, CRC Press, Boca Raton, FL, 2010.

Croissance and sporulation of six Entomophthorales species II.

Mycopathologia, vol.

3, 1975, pp.

14.V.V.

Beatty, Aerobic anoxygenic phototrophic bacteria: a review.

Microbiol Mol Biol Rev.1998; 62:695–724.

Jungermann, and Decker K., R.K.

In chemotrophic anaerobic bacteria, conservation of energy is important.

41, no.

16.Van der Horst, M.A., Key, J., Hellingwerf, K.J.

2007; 15:554–562.

2007; 15:554–562.

published Werkman, C.H, and Wilson, P.W.

Bacterial physiology is the study of how bacteria function.

and Mitchell H.K., published in J.

Proc Natl Acad Sci USA, vol.

1, 1941.

Dunn, S., Shankman, S., Camien, M.N., Block, H.

J Biol Chem, vol.

1–22, 1947.

The probiotic characteristics of several lactic acid bacteria isolated from Egyptian dairy products have been discovered.

a proposal to reclassify Fusobacterium prausnitzii as Faecalibacterium prausnitziigens nov., comb.

nov.

The Third Edition of Collins, Collins & Lyne was published by Arnold in London in 1970, and is known as the Collins-Lyne edition.

The following are the authors’ affiliations: Vastine DW, Dawson CR, Hoshiwara I., Yoneda C., Daghfous T., and Messadi M.

Application of Environmental Microbiology (AEM) 28:688–690 (1974).

Biotechnologies are represented by 27 files.

Growth and survival of Helicobacter pylori in defined medium and sensitivity to Brij 78.J Clin Microbiol.1998; 36:1232–1235.26.Northoff H., Flegel W.A.Fetal calf serum.J Clin Microbiol.1998; 36:1232–1235.27.Northoff H., Flegel W.

Immunology is covered in detail in the Encyclopedia of Immunology.

Meenakshi A., et al., Cell Culture Media: A Review (Cell Culture Media: A Review).

Hesham Rahman (H.R.

Schultze (FCS), Michael Oellerich (M.), and Asif (A.R.) The human T lymphoblast proteome and phosphoproteome are influenced by fetal calf serum heat inactivation and lipopolysaccharide contamination, respectively.

9, no.

71.29.

Lagier, J.C., Armougom, F., Million, P., Pagnier, I., Robert, C.

Infection Control and Hospital Epidemiology, 2012; 18:1185–1193.30.

Goodman, Gustaf Kallstrom, Jean-Jacqueline Faith, Anastasios L.

Dantas Personal human gut microbiota culture collections were studied and altered in gnotobiotic mice using a large number of different strains.

Szynkiewicz, Z.M., Binek, M., Szynkiewicz, Z.M., Binek, M., Szynkiewicz, Z.M., Szynkiewicz, Z.M., Szynkiewicz, Z.M., Szynkiewicz, Z.M., Szynkiewicz, Z.M., Szynkiewicz, An investigation on the use of selective media for the primary isolation ofTreponema hyodysenteriae andTreponema innocens COMP IMMUNOLOGY, MICROBIOLOGY, AND INFECT DISEASES, 9:71–77.32 (1986).

  1. vol.
  2. Elsevier Health Sciences; Philadelphia: 2014.
  3. 8th ed.
  4. 1.
  5. Infection control principles and practice of infectious illnesses (Mandell, Douglas, and Bennett’s principles and practice of infectious diseases).
  6. The following are the names of the authors: La Scola B, Khelaifia S, Lagier J.C., Raoult D.
  7. The European Journal of Psychiatry Clinical Microbiology and Infectious Diseases, vol.
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11, pp.

a group of researchers led by Dione, S.

La Scola, Jean-Charles Lagier, and David Raoult.

Infection Control and Hospital Epidemiology, 2016; 22:53–58.

Laboratory techniques in the field of food microbiology.

The laboratory is where food microbiology is studied.

Pilly, 5th edition, Alinéa, Paris, 2018.

Lavoisier was published in Paris in 2014.

Bacterial and moisture-related levures and leaks are being sought.

from chicken flesh was demonstrated by Sachan and Agarwal (40).

Int J Food Microbiol.2000; 60:65–74.

Corry, J.E., Curtis, G.D., and Baird, R.M., eds., Royal Society of Chemistry, Cambridge, 3rd edition, 2011.

Surman-Lee et al.

Bartram, Y.

Legionella and the prevention of legionellosis are two important topics.

and Lowbury E.J.L.

18, no.

752–756.

Int J Pharm Pharm Sci.2015; 7:322–328.

In the case of Mycobacterium tuberculosis, chlorhexidine cleaning of sputum for culture was performed by S.

Drancourt.

The use of a chlorhexidine-agar plate culture medium to supplement normal broth culture of Mycobacterium tuberculosis has been reported by Asmar S., Chatellier S., Mirande C., van Belkum A., Canard I., and Raoult D.

The cultivation of ‘unculturable’ human microbiota reveals the presence of new taxa and substantial sporulation, as reported by Browne et al.

(47) and Stares (M.D.).

48.J.R.

Allen, and V.R.

762–765 in the Journal of Applied Environmental Microbiology (1978).

812–799 in Future Microbiology (2018; 13:799–812).

35:581–599.

Using membrane filtration with marine agar 2216E to count marine heterotrophic bacteria is a novel approach to enumerating these microorganisms.

The use of a tellurite Blood-Agar Medium for the fast diagnosis of diphtheria is described in Hoyle (52).

New selective plating medium for isolation of Vibrio vulnificus biogroup 1 (Brayton et al., 2003; West et al., 2003; Russek et al., 2003; Colwell et al., 2003).

17, no.

1039–1044.

Nichols and R.

A novel selective medium for Streptococcus pneumoniae has been developed.

The selective influence of inhibitors employed in different culture medium on the competitive microflora of Salmonella was investigated by Arroyo G.

in their paper 55.

The use of phage lysin as a replacement for antibiotics in the detection of Mycobacterium tuberculosis from sputum samples using the Bactec MGIT 960 system was investigated by Subramanyam B., Sivaramakrishnan G.N., Dusthackeer A., Nagamiah S., Kumar V.

2012; 18:497–501 in Clin Microbiol Infect.

E.

62:1128–1132, Current Microbiology, 2011.

58.Denyer S.P., Hodges N.A., and Gorman S.P.7th edition.

A case study of biosalinity in action: the manufacture of biofuels from saline water.

Pages 137–157 of Macroalgae (seaweeds): Industrial Resources and Their Utilization are dedicated to this topic.

Sixty percent of the population is undernourished, according to the World Health Organization.

Tanaka T., Kawasaki K., Daimon S., Kitagawa W., Yamamoto K., Tamaki H.

Tanaka T., Kawasaki K., Daimon S., Kitagawa W., Yamamoto K., Tamaki H.

In 2014, the journal Applied and Environmental Microbiology published an article with the title 7659–7666.

Schmidt and P.M.

By utilizing gellan gum as a gelling agent in mycological media, it is possible to perform more rapid genetic identification of therapeutically significant aspergilli.

Journal of Medical Microbiology Doublier, J.L.

63.Doublier, J.L.

The ninth meeting of the Alliance 7/CEDUS was held on the topic of sugar and the preservation of products made from fruits.

Datta S., Mody K., Gopalsamy G., Jha B., Mody K., Gopalsamy G.

465–468 in Carbohydr Polym.

The evaluation of kappa carrageenan as a replacement for agar in microbiological medium was conducted by Abbott I.A.

128:355–359, American Journal of Microbiology, 1981.

The effect of a gelling agent on colony formation in a solid culture of a microbial community isolated from lake silt was investigated.

Environ Microbiol.

67.M.

Ferrari B.C., Binnerup S.J., Gillings M., Adv Biosci Biotechnol.2015; 6:22.68.Ferrari B.C., Binnerup S.J., Gillings M.

Appetite for Environ Microbiology, vol.71, pp.8714–8720, 2005.

Del Dot, R.

Stackebrandt are among those who have contributed to this work.

Syst Appl Microbiol 16:380–384 (September 1993).

An asaccharolytic, succinate-utilizing bacterium isolated from human feces is studied in detail.

nov.

The following are the names of the authors: Lamoth, F.

Diagnostic approaches for intracellular bacteria and fastidious germs are discussed in detail.

72.I.

Russell, L.C.M.

Finlay, Gut microbiota in health and disease.

A hidden metabolic organ: the role of the gut microbiota in health and chronic gastrointestinal disease (Guinane C.M., Cotter P.D., 2003).

(2013) 6:295–308 (Therapeutic Advances in Gastroenterology). 74.Kerry, R.G., Patra, J.K., and others Benefication of probiotics for human health: a review of the research. Gouda S, Park Y, Shin H.S., Das G 2018; 26:927–939. J Food Drug Anal. 2018; 26:927–939.

Growth medium – Wikipedia

The History of Microbiology (Wainwright and Lederberg, 2001). In 1992, Encyclopaedia of Microbiology, Volume 2, Pages 419–437, was published. 2) The rebirth of culture in microbiology as demonstrated by the use of culturomics to study the human gut microbiota (Lagier J.C., Hugon P., Khelaifia S., Fournier P.E., La Scola B., Raould D). CMR (Clinical Microbiology Review) is a journal that publishes articles on clinical microbiology. 3) Prescott, L.M., Willey, J.M., Sherwood, L.M., and Woolverton, C.J., 5th edition In the year 2018, De Boeck Supérieur was established in Louvain-la-Neuve.

  1. 4.Pommerville, J.C., 10th edition, JonesBartlett, Burlington, 2013.
  2. 5.2.Weeks B.3rd edition JonesBartlett; Sudbury, Ontario: 2012.
  3. The microbes and society of Alcamo Serratia marcescens: a historical perspective and a clinical review, Yu V.L.
  4. In 1979, the New England Journal of Medicine published a 300-page article titled “The New England Journal of Medicine, 887–893.” 7.P.R.
  5. Rosenthal, and M.A.
  6. Microbiology in the medical setting.
  7. Sandle 2011;10–14; Journal of the Institute of Science and Technology 9.Vallery-Radot P.Masson et compagnie; Paris, 1922.

Pasteur’s Works, Vol.

2 Kumar S.JP Medical Ltd, New Delhi, India, 2012.

The second edition of Power and Johnson’s textbook was published in 2009 by Becton, Dickinson and Company; Sparks; Power, A.

A.

Microbiological media: a handbook for researchers Growth and sporulation of six species of EntomophthoralesII, Latge, J.P.

Impact of various sources of azote on the environment In 1975, the journal Mycopathologia published a paper stating that 14.V.V.

Beatty, Aerobic anoxygenic phototrophic bacteria in the environment.

15.Thauer R.K., Jungermann K., Decker K., Jungermann K.

1977; 41:100.

41, no.

Photosensing in chemotrophic and non-phototrophic bacteria: let there be light sensing as well as chemotrophic sensing.

Academic Press Inc., New-York, 1951; Werkman, C.H.

Physiology of bacteria Purine and pyrimidine as growth substances for lactic acid bacteria, Snell E.E.

1941; 27:1.

In 1947, the Journal of Biological Chemistry published 168:1–22.

Some lactic acid bacteria isolated from Egyptian dairy products exhibit probiotic properties.

21.Duncan S.H., Hold G.L., Harmsen H.J., Stewart C.S., Flint H.J.

nov., comb.

2002; 52:2141–2146.

Methods based on microbiology 23.Vastine D.W., Dawson C.R., Hoshiwara I., Yoneda C., Daghfous T., Messadi M., Hoshiwara I.

788–690 in the journal Appl Environ Microbiol.

Cézard, 2nd edition, Paris: Dunod, 2013.

25.Albertson N., Wenngren I., Sjöström J.E.

27.Northoff H., Flegel W.

Pages 896–897.27 in Elsevier Health Sciences’ Philadelphia edition.

3:175.28 in Mater Methods, 2013.

Asif) have published a paper in which they argue that The proteome and phosphoproteome of human T lymphoblasts are affected by fetal calf serum heat inactivation and lipopolysaccharide contamination.

microbial culturomics: a paradigm shift in the study of the human gut microbiome (Lagier et al., 2007).

Infection Control and Hospital Epidemiology, 2012, 18:1185–1193.30.

Goodman, Gustaf Kallstrom, Jean-Jacqueline Faith, Anastasios L.

108:6252–6257.31 (Proceedings of the National Academy of Sciences, USA).

Szynkiewicz and M.

9:71–77.32 (Comp Immunol Microbiol Infect Dis, 1986).

vol.

Elsevier Health Sciences; Philadelphia: 2014.

Infection control principles and practice of infectious diseases (Mandell, Douglas, and Bennett).

Current and historical strategies for bacterial culture in clinical microbiology.

The following are the names of the authors: La Scola B, Khelaifia S, Lagier J.C, Raoult D A preliminary report on the use of antioxidants in the culture of anaerobic bacteria.

33, no.

1781–1783, 2014; The following are the authors’ names: Dione, Nadine, Khelaifia, Béatrice La Scola, Jean-Christophe Lagier, Dominique Raoult.

Infection Control and Hospital Epidemiology, 22:53–58, in press.

37.

37.Entis, P.

38.

Diseases infectieuses et tropicales; page 324 The 39th edition of Delarras C.Lavoisier was published in Paris in 2014.

It is on the lookout for bacteria and moisture leaks.

IJFM (International Journal of Food Microbiology) published a paper on this topic in 2000, which was 60:65–74.

Food and water microbiology culture media: a handbook for researchers Surman-Lee et al.

Bartram, Y.

In 2007, the World Health Organization was established in Geneva.

The use of an improved cetrimide agar medium as well as other culture methods for Pseudomonas aeruginosa was investigated by Brown V.I.

in their research.

In 1965, the Journal of Clinical Pathology published an article with the title J Clin Pathol 18:752–756.

Antimicrobial activities of chlorhexidine gluconate and cetrimide against pathogenic microorganisms isolated from slaughter houses in the state of Rivers, Nigeria Journal of Pharmaceutical Sciences, International Journal of Pharmaceutical Sciences, International Journal of Pharmaceutical Sciences, International Journal of Pharmaceutical Sciences, International Journal of Pharmaceutical Sciences, International Journal of Pharmaceutical Sciences, International Journal of Pharmaceutical Sciences.

  • The cleaning of sputum with chlorhexidine before to cultivating Mycobacterium tuberculosis (45.Asmar S., Drancourt M.).
  • BMC Microbiol, 15:155.
  • Microbiology in the 21st Century; 7:30 a.m.
  • (47) and Forster (S.C.), Anonye (B.O.
  • Dowell, V.R., and Koransky, J.R., et al.
  • 762–765 in the Journal of Applied Environmental Microbiology, 1978.
  • 2018; 13:799–812.

New culture medium based on sodium desoxycholate for the isolation of intestinal pathogens and the counting of colon bacilli in milk and water have been developed by E.

35:581–599.

Using membrane filtration and marine agar 2216E to enumerate marine heterotrophic bacteria is the subject of this paper.

Aquaculture The use of a tellurite Blood-Agar Medium for the fast diagnosis of diphtheria (52.Hoyle L).

New selective plating medium for isolation of Vibrio vulnificus biogroup 1 (Brayton et al., 2003; West et al., 2003; Russek et al., 2004; Colwell et al., 2004).

Journal of Clinical Microbiology T.

Freeman are the authors of this paper.

770–773 in the Journal of Clinical Pathology, 1980.

Arroyo and J.A.

Appl Microbiol 78:281–289 (July 1995).

Journal of Clinical Microbiology and Infectious Diseases, Vol.

4, 497–501, 2012.

Efficacy of a broad host range lytic bacteriophage against E.

62:1128–1132, Current Microbiology.

58.Denyer S.P., Hodges N.A., and Gorman S.P.7th ed.

The use of biosalinity: bioproduction with saline water, 59.McLachlan J.

Pages 137–157 of Macroalgae (seaweeds): Industrial Resources and Their Utilization.

Tanaka T., Kawasaki K., Daimon S., Kitagawa W., Yamamoto K., Tamaki H., Front Microbiol.2015; 6:698.61.

In 2014, the journal Applied and Environmental Microbiology published an article with the title: 80:7659–7666 The following are the names of the authors: Schmidt D., Rath P.M.

653–655 in the Journal of Medical Microbiology.

Molecular structure and functional properties of emulsion-based food systems “The Sugar and Fruit Preservation Symposium” was the theme of the 9th Alliance 7/CEDUS meeting, which took place in November.

A study by Datta et al., Mody et al., Gopalsamy et al., and Jha et al.

465–468 in Carbohydr Polym.

and Chapman F.A.

128:355–359.

1981; 128:355.

11, no.

1827–1834, 2009.

Ferrari B.C., Binnerup S.J., Gillings M., Adv Biosci Biotechnol.2015; 6:22.68.

Appetite for Environ Microbiology, vol.71, no.8, pp.8714–8720 (2005, October).

Del Dot, R.

Stackebrandt are among the researchers who have contributed to this work.

1993; 16:380–384.

Nagai, and M.

An asaccharolytic, succinate-utilizing bacteria obtained from human feces was studied in detail.

nov.

The following are the names of the authors: Lamoth, F., Schrenzel, and Greub.

10:2130–2136 (Rev Med Suisse, 2014) 72.I.

L.

C.

Antunes, and B.

Finlay, Gut microbiota in health and illness.

Guinane C.M.

The role of the gut microbiota in health and chronic gastrointestinal disease: comprehending a hidden metabolic organ.

(2013) 6:295–308 in Therapeutic Advances in Gastroenterology. Patra, John, and Kerry, R.G., et al., 74. Benefaction of probiotics for human health: a review of the research. Gouda S, Park Y, Shin H.S., Das G 2018; 26:927–939. Journal of Food and Drug Analysis.

Types

Nutrient broths (also known as liquid nutrient medium) and lysogeny broth medium are the most often used growth media for microorganisms. Liquid media are frequently combined with agarose and dispensed onto Petri plates using a sterile media dispenser to solidify. These agar plates serve as a solid culture medium for microorganisms to grow and multiply. They remain solid because only a few number of bacteria are capable of decomposing agar (the exception being some species in the genera:Cytophaga,Flavobacterium,Bacillus,Pseudomonas, andAlcaligenes).

It is important to understand the difference between growth media used for cell culture and growth media used for microbiological culture because cells derived from whole organisms and grown in culture frequently cannot grow without the addition of growth factors or hormones that normally occur in vivo.

  • Microorganisms, on the other hand, are frequently unicellular organisms, therefore there are no such restrictions on their growth.
  • This is another significant distinction from animal cells raised in the wild.
  • The difference between defined and undefined growth media is a significant distinction between the two forms of growth media.
  • It consists of giving trace elements and vitamins that are necessary by the bacterium, as well as specifically defined carbon and nitrogen sources.
  • Some complex compounds, such as yeast extractor casein hydrolysate, are found in an undefined medium.
  • In some cases, undefined media are chosen based on price, while in others, they are used out of necessity – for example, some bacteria have never been cultivated on defined media.
  • The wort has all of the nutrients necessary for yeast development, and when fermented in an anaerobic environment, it produces alcohol.
  • The most common kinds are as follows:
  • Media that is cultural
  • Minimum media
  • Selective media
  • Differential media
  • Transit media
  • Indicator media
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Culture media

Culture media include all of the nutrients and growth factors that most bacteria require to thrive, and because they are non-selective, they are employed for the general growing and maintenance of bacteria in laboratory cultures collections.

An indefinite media (also known as a base medium or a complicated medium) has the following components:

  • The following ingredients are required: a carbon source such as glucose, water, different salts, a supply of amino acids and nitrogen (for example, beef, yeast extract)

A medium with no specified composition is created by combining amino acids from various sources, the exact composition of which is not known at the time of writing. When you work in a specified medium, you are working in an environment that has been chemically defined or synthesized.

  • Each and every one of the compounds employed is well-known
  • There is no evidence of yeast, animal, or plant tissue.

The following are examples of nutritional media:

  • Nutrient agar, plate count agar, trypticase soy agar, and other agars are used.

Minimal media

The term “minimal media” refers to a specified medium that contains just the components necessary to enable development. When growing microorganisms in a minimum medium, the quantity of components that must be added varies greatly depending on the microbe being cultivated. Minimal media are those that contain the very minimum of nutrients necessary for colony formation, and are often devoid of amino acids. They are frequently employed by microbiologists and geneticists to cultivate “wild-type” microorganisms in their laboratories.

The following are typical components of minimal medium:

  • A carbon source, which can be either a sugar such as glucose or a less energy-dense source such as succinate
  • Various salts, which can vary depending on the bacteria species and growing conditions
  • These generally provide essential elements such as magnesium, nitrogen, phosphorus, and sulfur to allow the bacteria to synthesize protein and nucleic acids
  • Water
  • And a nitrogen source.

The term “supplemental minimum media” refers to minimal media that also include a single specified chemical, which is often an amino acid or a sugar. This supplementation enables the cultivation of specific lines of auxotrophic recombinants in the laboratory.

Selective media

Campylobacter isolates were obtained using a blood-free, charcoal-based selective medium agar (CSM). Blood agar plates are frequently used in the diagnosis of infectious diseases. A positiveStaphylococcusculture is shown on the right, while a positiveStreptococcusculture is shown on the left. Selective media are used to promote the development of just specific microorganisms in a laboratory setting. Suppose a microbe develops resistance to a certain antibiotic, such as ampicillin or tetracycline.

  1. The use of media missing an amino acid, such as proline, in combination with E.
  2. Selection media are also utilized in the cell culture process in order to guarantee the survival or multiplication of cells that possess certain characteristics, such as antibiotic resistance or the ability to produce a specific metabolite.
  3. In such instances, the gene is referred to as a marker.
  4. Gancycloviris an exception to the norm, since it is used to precisely destroy cells that have the Herpes simplex virus thymidine kinase, which is a marker for the virus in question.

Four distinct types of agar plates were used to demonstrate how bacterial metabolism might influence growth. Examples of selective media include the following:

  • Eosin methylene blue is a dye that is harmful to Gram-positive bacteria because it includes dyes that are toxic to them. This medium is used to selectively and differentially isolate coliforms. The pH of YM (yeast extract, malt extractagar) is low, which inhibits bacterial growth. Gram-negative bacteria are cultured in MacConkey agaris. Hektoen enteric agar is a bacterial agar that is selective for Gram-negative bacteria. Known as HIS-selective media, this is a form of cell culture medium that does not include the amino acid histidine. In addition to being selective for gram-positive bacteria, mannitol salt agar is also differential for mannitol. Xylose lysine deoxycholate is a Gram-negative bacteria-selective antibiotic. Buffered charcoal yeast extract agar is a gram-negative bacteria selectivity agar that is particularly effective against Legionella pneumophila In the case of gram-positive staphylococci, Baird–Parker agaris is used. In part, this is owing to the low pH (5.6) and high glucose content (3–4 percent) of Sabouraud’s agar, which makes it particularly attractive to some fungus. In food testing, DRBC (dichloran rose bengal chloramphenicol agar) is a selective medium that may be used to count the number of moulds and yeasts present. Inhibitors of mould colony growth such as dichloran and rose bengal help to prevent an increase in the number of luxuriant species while also aiding in the precise counting of colonies.

Differential media

Media that identify one species of microbe from another that is growing on the same medium are referred to as differential or indicator media. To visually identify the defining characteristics of a microorganism, this type of media employs the biochemical characteristics of a microorganism growing in the presence of specific nutrients or indicators (such as neutral red, phenol red, eosin y, ormethylene blue) that have been added to the medium. These media are used by microbiologists to detect microorganisms, and by molecular biologists to identify recombinant strains of bacteria in a variety of settings.

  • It is utilized in strep tests because it includes bovine heart blood that turns translucent in the presence of -hemolytic organisms such as Streptococcus pyogenes andStaphylococcus aureus. Lactose fermentation is aided by the use of eosin methylene blue
  • Nevertheless, This medium is selective and differential for Streptococcus agalactiae (group B streptococcus), which develops as characteristic red colonies on the surface of the culture media. Lactose fermentation is facilitated by MacConkey agaris differential
  • Agar containing mannitol salt has a difference effect on mannitol fermentation, and X-galplates have a differential effect on lac operonmutants.

Transport media

The following requirements should be met by transportation media:

  • The temporary storage of specimens while they are being transferred to the laboratory for culture
  • Maintain the viability of all organisms present in the specimen without affecting their concentration
  • And. Only buffers and salt are used in this formulation. Lack of carbon, nitrogen, and organic growth factors, which prevents microbial reproduction
  • Lack of organic growth nutrients. In order to isolate anaerobes, the transport media utilized in the isolation process must be devoid of molecular oxygen.

Examples of transportation media include:

  • In the case of strictanaerobes, thioglycolate broth is used. Stuart transfer medium is a non-nutritional soft agar gel that contains a reducing chemical to avoid oxidation as well as charcoal to neutralize the environment. Gonococci are treated with specific bacterial inhibitors, while enteric bacilli are treated with buffered glycerol saline. For V. cholerae, the Venkataraman Ramakrishna (VR) medium is employed.

Enriched media

Increased nutrient availability provides the nutrients necessary to sustain the growth of a wide range of species, including some of the more finicky organisms. They are typically used to collect as many distinct types of bacteria from a specimen as there are germs present in it. Blood agar is an enhanced media in which nutritionally dense whole blood replenishes the basic nutrients in the presence of other nutrients. It is made up of dark chocolate agaris that have been heat-treated to 40–45 degrees Celsius (104–113 degrees Fahrenheit), which turns brown and provides the medium its name-giving hue.

Physiological relevance

In tissue culture investigations, the choice of culture media can have an impact on the physiological significance of the findings, which is especially true for metabolic research. Furthermore, it was shown that the kind of media used can influence the dependency of a cell line on a metabolic gene. When doing a study with several cell lines, using a homogeneous culture medium for all of the cell lines may help to limit the amount of bias in the datasets that are created. Increased physiological relevance of in vitro research can be achieved by using a growth medium that more accurately replicates physiological amounts of nutrients.

See also

  • Cell culture, impedance microbiology, and modified Chee’s medium are among topics covered in this course.

References

  1. Madigan, M., and Martinko, J. (eds). (2005). Brock’s Microorganism Biology is a textbook that teaches microorganism biology (11th ed.). It is published by Prentice Hall with the ISBN number 0-13-144329-1. Birgit Hadeler, Sirkka Scholz, and Ralf Reski are three of the most talented people in the world (1995). A moss’ cytokinin-sensitivity is influenced differentially by gelrite and agar, according to the researchers. Journal of Plant Physiology, vol. 146, no. 3, pp. 369–371. maintainer of CS1: makes use of the authors parameter (link)
  2. K.J. Ryan and C.G. Ray, eds (2004). Sherris Medical Microbiology is a medical microbiology company (4th ed.). Hans Günter Schlegel’s book is published by McGraw Hill and has the ISBN number 0-8385-8529-9. (1993). Cambridge University Press, p. 459. ISBN 978-0-521-43980-0. General Microbiology. Cambridge University Press. retrieved on August 6, 2013
  3. Parija and Shubhash Chandra are two of the most talented musicians in the world (1 January 2009). Textbook of Microbiology and Immunology, Elsevier India, p. 45, ISBN 978-81-312-2163-1. Textbook of Microbiology and Immunology, Elsevier India, p. 45, ISBN 978-81-312-2163-1 Cooper, GM (retrieved on August 6, 2013)
  4. (2000). “Cell Biology Instruments” is an abbreviation for “Cell Biology Tools.” The Cell: A Molecular Approach to Understanding It ASM Press, Washington, D.C., ISBN 0-87893-106-6
  5. Catherine A. Ingraham and John L. Ingraham are the authors (2000). G. D. W. Curtis and Rosamund Baird are the editors of the textbook Introduction to Microbiology (Corry et al., ed). (1995-01-01). An agar containing dichloran rose bengal chloramphenicol (DRBC) was developed. Progress in Industrial Microbiology, vol. 34, no. 3, pp. 303–305, published by Elsevier, doi: 10.1016/s0079-6352(05)80036-0, ISBN 9780444814982. Retrieved2020-04-20
  6. s^ John A. Washington, Jr. (1996). “Principles of Diagnosis” is an acronym for “Principles of Diagnosis.” S. Baron and colleagues (eds.). Baron’s Medical Microbiology is a textbook that teaches medical microbiology (4th ed.). Univ of Texas Medical Branch.ISBN0-9631172-1-1
  7. Lagziel S, Gottlieb E, Shlomi T
  8. Lagziel S, Gottlieb E, Shlomi T (2020). “Take care with your media.” S2CID222319735
  9. Lagziel S, Lee WD, Shlomi T. Nature Metabolism.2(12): 1369–1372.doi: 10.1038/s42255-020-00299-y.PMID33046912.PMID33046912.S2CID222319735
  10. Lagziel S, Lee WD, Shlomi T. Lagziel S, Lee WD, Shlomi T. (2019). “Inferring cancer dependencies on metabolic genes from large-scale genetic screens” is the title of this article. Vande Voorde J, Ackermann T, Pfetzer N, Sumpton D, Mackay G, Kalna G
  11. Et al. BMC Biology.17(1): 37.doi: 10.1186/s12915-019-0654-4.PMC6489231.PMID31039782.CS1 maint: many names: authors list (link)
  12. Vande Voorde J, Ackermann T, Pfetzer N, Sumpton D, Mackay G, Kal (2019). Improved metabolic fidelity of cancer models using a physiological cell culture medium is the subject of this study. Science Advances.5(1): eaau7314 (early access). The Bibcode for this paper is 2019SciA.5.7314V.doi: 10.1126/sciadv.aau7314.PMC6314821.PMID30613774.CS1 maint: multiple names: authors list (link)
  13. CS1 maint: multiple names: authors (link)
  14. The following authors contributed to this article: Cantor, J.R., Abu-Remaileh M, Kanarek N, Freinkman E, Gao X, Louissaint A
  15. And others (2017). A study published in the journal Physiologic Medium revealed that Uric Acid is an endogenous inhibitor of UMP Synthase and that it rewired cellular metabolism. Cell.169(2): 258–272.e17.doi: 10.1016/j.cell.2017.03.023.PMC5421364.PMID28388410.CS1 maint: multiple names: authors list (link)
  16. CS1 maint: multiple names: authors list (link)

External links

Phasing changes in a cell culture environment may have a significant impact on practically every biological activity, including metabolism, cell proliferation, and membrane potential, among others. Extracellular pH is slightly alkaline, ranging between 7.3 and 7.4, whereas intracellular pH is somewhat lower, ranging between 7.2 and 7.2, respectively. Many organelles, such as mitochondria, maintain a pH that is somewhat different from the cytoplasmic pH, but if the variations between the two pHs become too significant, the organelles’ ability to operate is severely compromised.

pH BuffersHow pH Buffered Culture Media Works

In its most basic definition, a buffer is a solution that contains a weak acid or weak base and its conjugate base or acid. They function by neutralizing any extra acid (H +ions) or base (OH –ions) in order to maintain the optimum pH level in the solution.

A pH buffer functions as either a weak acid or a weak base, depending on the situation, to guarantee that the media is relatively resistant to pH changes. As a result of the buffer’s ability to donate or take hydrogen ions, which are responsible for setting pH, this method works.

Acid + Base⇌Conjugate Base + Conjugate Acid

The sodium bicarbonate buffer is the most often used buffer in mammalian cell culture. The sodium bicarbonate buffered medium becomes sensitive to the formation of carbonic acid as a result of the presence of CO 2. The pH of the medium may be maintained as long as the level of CO 2 in the solution is kept under control. By and large, only certain types of biological activities may take place when the pH is within a specific range, and the appropriate buffer must have a pKa that is near to the ideal pH of the target cells.

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Does Temperature Play a Role in pH?

Temperature can have an impact on pH in a significant yet nonlinear way. Temperature increases are associated with a reduction in pH levels. The easiest way to explain this is to use the example of water. When water is heated, the pH of the water decreases. In fact, the pH is closer to 6.14 than it is to 7.00 when heated to 100°C. Technically, the new neutral pH at 100°C is more acidic than the old neutral pH, but the constituents remain same. Water is always simply hydrogen and oxygen, regardless of the temperature at which it exists.

As a result of this consideration, a buffer that delivers ideal capacity and range at a single temperature may not be the best choice for tests conducted at many temperatures.

pH Buffer Capacity and Range

There is a specified effective capacity and range for each buffer. The buffering capacity of a solution is the quantity of acid or base that can be added before the pH begins to vary significantly. Alternative definitions include the amount of acid or base that must be added to one liter of solution in order to raise or lower the pH by one pH unit. Thus, the pH buffering capacity (pKa) is equal to the negative logarithm of the acid dissociation constant (Ka). At this moment, equilibration has been achieved and equal amounts of acid and conjugate base are present in solution.

The pH range of a buffer is the range of pH values within which a buffer may efficiently neutralize additional acids and bases while keeping a reasonably constant pH.

Acidic

Acid buffers have a pH less than 7.0 and are composed of a weak acid and one of the acid’s conjugates as the active ingredient. The pH of the acidic buffers that are most usually employed is around 4.75. Change the pH of the buffer solution by altering the ratio of acid to salt in the solution.

Alkaline

Alkaline buffer solutions have a pH greater than 7.0 and are used to buffer acidic solutions. They are made up of a weak base and one of its conjugates, and they are quite strong.

When ammonia and ammonium chloride are combined in equal molar amounts, the pH of the solution is 9.25, making it the most often used alkaline buffer. Change the pH of this solution by altering the ratio of base to salt in the solution.

Neutral

Neutral buffers are exactly what they sound like: they are neutral buffers. They have a pH near to or at 7.0 and are utilized for a number of purposes, the most common of which is to keep cells within the physiological range of mammalian cells. zwitterionic solutions contain an equal amount of positively and negatively charged ions, as seen in the diagram. HEPES is the most often used buffer in cell culture because it is more effective at preserving physiological pH than bicarbonate buffers and may be generated at pH values ranging from 6.8 to 8.2.

What is Breaking the Buffer?

When the whole base and its conjugate acid (or vice versa) are consumed in the process of neutralization, the buffer is regarded to be broken (adding acid or base.) The addition of any more acid or base at this stage will cause the pH to change fast, and in many cases drastically. Even when the barrier has been breached, all hope is not lost. It simply indicates that the capacity has been reached and that pH regulation is now a much more hands-on process. Every cell culture expert believes that keeping a steady pH is critical for cell development, regardless of which buffer you use or how the pH is measured.

Guidelines for Controlling pH

Open-access publication of exceptional quality a flow chart with step-by-step directions on how to make medium at a desired pH, as well as four particular recommendations on how to achieve precise control over pH in cell culture and boost repeatability. Contact our applications team if you’d like to begin a discussion about how you might be able to employ optical sensors to detect pericellular pH.

What Culture Medium Will Neutralize Acids?

Acids are neutralized by the addition of which of the following to a culture medium? A) Buffers are used to store information. B) Added sugars C) the pH of the water D) the carbon in the water A) Buffers, of course!

Which is the best acid or base neutralization?

They can be as simple as a bag of citric acid or sodium bicarbonate (baking soda) or as sophisticated as a solidifier and a neutralizer that are used in conjunction. Safeguarding one’s health and safety (mixing chemicals is always a potentially dangerous operation), as well as cost and convenience, are all important factors when selecting the optimal technique of neutralization.

What is the pH of a neutral acid?

Strong acids are defined as chemicals having a pH ranging from 0 to 3. Strong bases are defined as chemicals having a pH ranging from 12 to 14. A chemical must have a pH value of 7 in order to be called neutral. Acids are often characterized by a sour taste and a pH of less than seven.

Which is the best way to determine the acidity of a substance?

When determining whether a material is acidic or basic, the pH of the material is the best method to use.

This can be performed by the use of pH paper, chemical indicators, or pH meters, among other methods. The pH scale ranges from 0 to 14 on a scale of 1 to 14. Strong acids are defined as chemicals having a pH ranging from 0 to 3.

How can you tell when an acid spill has been neutralized?

There are certain acid and base neutralizers that come with a built-in color indicator that will tell you when the spill has been neutralized. Others need you to verify the pH with pH paper or a pH meter to ensure that the neutralization procedure is proceeding as planned.

What Is Another Name For Amino Acids?

The twenty amino acids (as well as many more amino acids that are regularly discovered) have innocuous names such as Glycine (R = H), Alanine (R= methyl group), Valine (R= isopropyl group), and so on.

How Do You Combine Amino Acids?

Peptide bonds, which are formed when the amino or NH2 group of one amino acid links to the carboxyl (acid) or COOH group of another amino acid, are used to connect amino acids together to produce polypeptides and proteins.

Are Fatty Acids Amino Acids?

Is it true that fatty acids are amino acids? The most significant distinction between amino acids and fatty acids is that amino acids are the building blocks of proteins, whereas the three fatty acids are joined together to create a triglyceride, which is the primary ingredient of lipids. Amino acids and fatty acids are two types of macromolecule building components that can be found in the body.

Why Does Ninhydrin Stain Amino Acids?

The test is done as a result of the interaction between the amino group of free amino acid and ninhydrin, which occurs during the preparation of the sample. Ninhydrin, as you are aware, is a powerful oxidizing agent. As a result of its presence, the amino acid undergoes oxidative deamination, which results in the release of ammonia, and the production of ninhydrin is reduced.

What Is The Breakdown Of Fats Into Fatty Acids?

Currently, long chain fatty acids are broken down by a process known as oxidation (breakdown). This process occurs through the actions of the enzymes -oxidation and -oxidation. Acyl-CoA is the active 2-C unit that is produced as a result of the -oxidation process (CH3CO C0A).

Are Dna And Rna Nucleic Acids?

Nucleic acids include DNA and RNA, which are both instances. They are made up of a string of nucleotides that contain a phosphate group, a 5′ sugar, and a nitrogenous base, among other things. DNA and RNA molecules are polymers, as are other nucleic acids. DNA is double stranded, but RNA is single stranded, and this is the difference between the two.

Does Zinc React With All Acids?

Molecular nucleic acids include DNA and RNA, to name a few examples. Nucleotides with an added phosphate group, a 5′ sugar, and a nitrogenous base make up the strand of nucleotides in this kind of DNA. Molecular polymers such as DNA and RNA exist. While both DNA and RNA are double-stranded, DNA is the more common of the two.

What Protein Powder Has All 9 Amino Acids?

Hemp is a complete protein that is mostly made of the amino acids edestin and albumin. Edestin and albumin are proteins that aid in the absorption of all nine necessary amino acids by the body. Hemp is regarded as a complete protein since it includes all of the necessary amino acids required for human nutrition. Protein is made up of a chain of amino acids, and not all foods contain all of the amino acids in the chain.

How Are Peptide Bonds Formed Between Amino Acids?

In biochemistry, a peptide is a short polymer of amino acid monomers that are connected together by an amide bond.

Peptide bonds are chemical bonds created by connecting the carboxyl groups (the backbone) of two amino acids together (the backbone). During the creation of this connection, the water molecule (H 2 O) is released into the environment.

How Can Glucose Be Converted To Fatty Acids?

Over time, extra glucose can be stored in the liver as glycogen or transformed into fatty acids with the aid of insulin, which can then be transported to other regions of the body and stored as fat in adipose tissue. The accumulation of fat in the liver occurs when there is an excess of fatty acids present. What is the location of the amino acid conversion to glucose?

What Will Break An Ionic Bond Between Amino Acids?

Due to the fact that hydrolysis can break peptide bonds between amino acids and that high temperatures can break hydrogen bonds between amino acids, in the case of an enzyme, it’s tertiary structure can be disrupted, which changes the active site, making it impossible for enzyme substrate complexes to form. The pH of the solution will have an influence on the ionic bonding. What chemical process is responsible for the dissociation of amino acids?

How Many Nucleotides Are In 100 Amino Acids?

100 amino acids are required to code one gram of protein. A total of 100 codons are required, as well as one starting codon and one stopping codon. The code is triplet, which means that it is made up of three nucleotides. As a result, the number of nucleotides required for 100 codons is (100X3 =300).

Does Vinegar And Oranges Both Contain Acids?

Acids are present in high concentrations in many of the foods that humans consume. Citric acid and vitamin C are found in citrus fruits such as oranges and lemons. Citrus fruits also contain ascorbic acid, which is also known as vitamin C. Phosphoric acid is found in carbonated beverages. Acetic acid is present in vinegar. Hydrochloric acid is produced by your own stomach and is used to breakdown meals.

What Contains All 8 Amino Acids?

Despite the fact that the body can synthesize many amino acids, it is unable to do so with these eight amino acids, which must be obtained from food. Included among these are the amino acids Phenylalanine, the three Branched Chain Amino Acids (Valine, Leucine, Isoleucine), Lysine, Threonine, Tryptophan, and lastly Methionine. Histidine is required for the growth of babies, but not for the development of adults.

What Macromolecule Has Amino Acids?

How many amino acids are there in a macromolecule? Proteins Polypeptide chains are long strings of amino acids that are linked together by peptide bonds to form proteins. Proteins are created by twisting one or more amino acids together.

Which Acids Are Arrhenius Acids?

Arrhenius acids include hydrofluoric acid (HF), nitric acid (HNO 3), hydrobromic acid (HBr), sulphuric acid (H 2 SO 4), sulphurous acid (H 2 SO 3), perchloric acid (HClO 4), phosphoric acid (H 3 PO 4), hydroiodic acid (HI), carbonic acid (H 2 CO 3), and others.

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