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BACTERIOLOGY > Water Bacteriology > Flashcards

Water Bacteriology Flashcards

1
Q

What is Water Microbiology

A
  • deals with the study of microorganisms and parasites in water
  • here in Phil, institutions are analyzing bacteria (limited to) in water only
  • ensure safe consumption of water
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2
Q

Sample used in water microbiology

A
  • Water

not body fulids

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3
Q

3 sections of Water testing laboratories

A

Environmental laboratory
- for metal analysis in water

Wet laboratory
- for the physical and chemical analysis of water

Microbiological Laboratory
- for bacteriological analysis if water

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4
Q

3 types of water dealt with in a water microbiology lab

A
  • drinking water
  • waste water
  • recreational water
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5
Q

Dechlorinating agent used in water microbiology

A

Sodium Thiosulfate

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6
Q

Who are the analyst required to work in water microbiology Lab

A

registered: in DENR

  • medical technologist
  • microbiologist
  • food technologist

In DOH they accept:

  • chemist
  • biologist

must undergo special training by East avenue medical center

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7
Q

Quality of a good drinking water

A
  • clear
  • colorless
  • free from objectionable taste and odor
  • Must not contain any substance or organisms, chemicals, and radioactive materials
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8
Q

Why and when to detect microorganism in water

A

❑ Outbreak investigation of waterborne diseases
❑ Assesing safety, stability of water and water products for public consumption
❑ Determining level of sanitation during product preparation (e.g., in restaurants)
❑ Regulatory compliance (in business)
❑ Incidence surveys

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9
Q

True or False:

In water microbiology specific isolation of the bacteria is necessary

A

False

  • the test only limits to identifying the preence and abscence of Coliform bacteria
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10
Q

are microorganisms such as bacteria and viruses in water bodies, which are utilized as a surrogate to evaluate the presence of pathogens in that environment.

A
  • Indicator organisms or are the COLIFORM bacteria
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11
Q

The most common Coliform (facultative anaerobe) bacteria

A
  • Escherichia coli (most numerous)
  • Enterobacter
  • Klebsiella
  • Citrobacter
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12
Q

True or False:
If the water tested positive with the indicator bactera, it mean that there is also possible presence of all relevant pathogens?

A
  • True (almost 90%)
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13
Q

2 classifications of Coliforms (facultative anaerobe)

A
  • Total Coliform (found everywhere)

- Fecal/ Thermotolerant Coliform (found in fecal material)

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14
Q

Characteristic of Coliforms

A 
❑Facultative anerobe
❑ Gram negative bacilli
❑ Non-spore forming
❑ Ferment lactose at
35oC for 48 hours - (total choliforms)
44.5oC for 24 hours- (Fecal
coliforms)
❑ Non- pathogenic
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15
Q

Bacterial detection Methods

A

❑ DIRECT ENUMERATION
❑ INDIRECT ENUMERATION
❑ PATHOGEN ISOLATION AND IDENTIFICATION

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16
Q

Direct Enumeration can be done through?

A

❑ Microscopic Count

❑ CFU count using non-selective media

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17
Q

Indirect Enumeration can be done through?

A

❑ Most Probable Number (MPN) or Multiple Tube Fermentation Technique

  • quantitative estimation
  • applied in 3 types of water
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18
Q

Pathogen Isolation and Identification can be done through

A

❑ Series of biochemical tests

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19
Q

Acceptable water testing method mandated by Philippine National Standards for Drinking Water. Department of Health. 2017.

A

❑ Multiple Tube Fermentation Technique
❑ Standard Membrane Filtration Technique
❑ Heterotrophic Plate Count (Pour or Spread Plate)
(the above are most common)

❑ Enzyme Substrate Coliform Test
❑ Delayed Incubation Total Coliform Membrane Filter Technique
❑ Simultaneous Detection of Total Coliform and and E.coli by
Dual Chromogen Membrane Filter Technique

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20
Q

Standard Membrane Filtration Technique

and Heterotrophic Plate Count (Pour or Spread Plate) is applied in what type of water

A
  • drinking water

waste water can be done on these methods however, the density of bacteria limits the method.

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21
Q

technique that can be used to classify organisms in all of the 3 types of water

A

Multiple Tube Fermentation Technique

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22
Q

This is where reglations and rules are made or where the submission of requirements in water testing is passed

A

Philippine National Standards for Drinking Water. Department of Health.

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23
Q

Storage of Culture Media in Water microbiology laboratories

A

Store dehydrated media (powder/granulated) in:

  • tightly closed bottles
  • in the dark
  • <30 degrees celcius
  • Low humidity (<50%)
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24
Q

True or False:

If culture medium can still be used even if they become discolored or become
caked and lose the character of a free flowing powder.

A
  • False

do not use them

25
Q

Sterilization of culture media tubes is done through

A

-Autoclaving at 121 degrees celcius C, 15 psi for 15 minutes.

Note: Dispense first before autoclaving
Record final pH after autoclaving

26
Q

True or False:

Culture media used in Membrane filtration technique can be autoclaved

A
  • False

some of their components are carcinogenic are tends to deteriorate

27
Q

Water used in preparation of culture medias in water microbiology lab

A
  • Distilled or deminaralized water
28
Q

True or False:

Fermentation tubes can be store in 25 °C for more than 2 weeks.

A

False

  • because evaporation may
    proceed rapidly under these conditions, resulting in marked changes in concentration of the ingredients,
29
Q

What must de done in liquid media fermentation or carbohydrate tubes if they are refrigerated?

A
  • bring in room temperature and discard tubes that contain bubbles
30
Q

What are the labels necessary to be included in the culture media?

A

❑ Media/Reagent Name
❑ Date prepared
❑ Expiration date
❑ Prepared by

31
Q

Preparation of Culture media plates

A
  • Autoclave first then dispense
  • 4 to 6 mL into 50 mm x 9 mm petri dishes is used
  • solidify at room temp.
  • store in refrigerator at inverted position in a plastic bag or tight container

Note: prefreably use for no longer than one week

32
Q

In Multiple Fermentation technique:

Presumptive identification is done in what culture media

A

Lauryl Sulfate Broth
or Lauryl Tryptose Broth (LSB/LTB)

  • for both total and fecal coliforms
33
Q

In Multiple Fermentation technique:

Confirmatory identification is done in what culture media

A

Brilliant Green Lactose Broth (BGLB)
- For Total Coliforms
detection

EC (Escherichia coli) Broth
- For Fecal Coliforms detection

34
Q

The accepted dilution water used in Multiple tube fermentation

A

❑ 0.1 % Peptone water
❑ Stock phosphate buffer solution
❑ Magnesium chloride stock solution

35
Q

True or False:

It is not advisable to suspend a sample in any dilution water for more thab 30 mins at RT.

A
  • True

since injury, death, or multiplication may occur

36
Q

Principle behind Multiple tube fermentation technique

A
  • Serial Dilution
37
Q

If 99 ± 2.0 mL solution is needed what is the amount of water to be dispensed

A

100 mL

38
Q

If 9 ± 0.2 mL mL solution is needed what is the amount of water to be dispensed

A

10 mL

39
Q

How is 0.1 % peptone water prepared?

A

1 gram of peptone water and 1000 mL of distilled water

40
Q

In MEMBRANE FILTRATION TECHNIQUE:

What agar is used for detection of Total coliforms

A
  • ENDO agar

95% ethanol is added

41
Q

In MEMBRANE FILTRATION TECHNIQUE:

What agar is used for detection of Fecal coliforms

A
  • Fc agar

Sodium hydroxide (0.2N NaOH) and 1% Rosolic acid is added

42
Q

What is added to Fc agar?

A

Sodium hydroxide (0.2N NaOH) and 1% Rosolic acid

43
Q

In MEMBRANE FILTRATION TECHNIQUE:

What agars are used for Escherichia coli detection

A
  • Urea broth

- mTEC agar

44
Q

Inhibitory agents of Endo agar

A
  • basic fuschin and soldium sulfide

inhibits gram (+) bacteria and bacteria that are not total coliform

45
Q

Positive result on Endo agar medium

A
  • Pink colonies/ lactose fermenters

differentiates bacteria on the basis of lactose fermentation

46
Q

Inhibitory agents of Fc Agar

A
  • bile salts
47
Q

The most common culture medium used in Heterotrophic plate count technique

A

Plate Count agar - Pour plate or Spread plate method

48
Q

What is the difference between the Pour plate and Spread plate method

A
  • in pour plate the agar is poured on the time of analysis whereas on spread plate method you have to prepare the agar first before analysis.
49
Q

For wastewater and recreational water, what is added on the collection bottle?

A

2-3 drops 10% sodium thiosulfate —> then autoclave

50
Q

For drinking water, what is added on the collection bottle?

A

2-3 drops of 3% Sodium thiosulfate —> then autoclave

51
Q

For how many minutes will you run the water, when collecting samples in faucets or water dispenser

A

2-3 minutes —> heat the mouth or apply sodium hypochlorite —> run again 2-3 mins—> collect sample

52
Q

Positive control for Total Coliforms

A

Escherichia coli

Enterobacter aerogenes

53
Q

Negative control for Total coliforms

A

Staphylococcus aureus

Pseudomonas spp.

54
Q

Positive control for Fecal Coliforms

A

Escherichia coli

55
Q

Negative control for Fecal coliforms

A

Enterobacter aerogenes

Streptococcus faecalis

56
Q

How to check purity of culture organism (if it’s pure culture)

A
  • Gram staining or thru Eosin methylene blue (EMB agar)
57
Q

What do E.coli looks like in Eosin methylene blu medium

A
  • greenish-metallic chain
58
Q

what should be done in every batch analysis of the same procedure to know the efficiency of the culture media and reagents prepared?

A

Blank test

59
Q

What agar is used for Air Micro quality check?

A
  • Plate count agar

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