Biological Method of Detection: Biosensors-Bacteria



Bio Sensors

A biosensor is a device use to detect a specific component; it reads the biological reaction with a physicochemical detector component


There are 3 parts in biosensor:

  • Biological reaction (receptors, protein, antibodies, nucleic acids)
  • Detection element (transforms the signal resulting from the interaction of the sample with the biological element into another signal)
  • Electronic component which coverts collected info into user-friendly information for easy analysis


They are generally 2 kinds of biosensors: Specific or Non-specific

An example of non-specific biosensors, as bacteria A are expose to increasing amount of toxic, the metabolism activity of the cell will decrease, reduce in a reduced signal output. The signal output can be used to find the concentration of toxic.

Specific biosensors are genetically engineered, inputting specific promoter and reporter gene. The sensitivity and limit of detection can be alter by the change of promoter and reporter component.



Ways of detection by biosensors:

1. Electrochemical (measures enzyme reaction)

2. Piezoelectric

3. Photometric


An example of a mercury biosensor

A fully constructed whole bacteria cell biosensor is already known in the market. It has a gene fusion of PmerTPADluxCDBAE. To make it simpler, we need to break down the bacteria to simple component. This strain of bacteria is a genetic modified biosensor, which means it is a specific biosensor. Basically LuxCDBAE gene are extracted from Vibrio fischeri and are fused to a inducible merTPAD gene(mercury resistance). The Gene fusion is than input into Escherichia coli (CM2624). The resulting specific biosensor will be able to emit light in the presence of mercury ions.

Pros and Cons of using biosensors

Before using Biosensors as your way to detect pollutant, first we need to know the pros and cons of the detection method. This is to make sure it can be suitable for the context of your research and give a better accuracy and efficiency of testing the concentration of “mercury”

Pros

Cons

Real time measurement (E.g. soil test, it test exactly % of soil and parts of soil affected)

Inhibiting of biosensors due to multiple toxic component (E.g. interferences of Lead)

Very effective in monitoring effect of bio remediation

Characteristic of biosensor and indigenous organism different , thus it may be difficult to reflect the real impact of mercury on the location

Test the bio availability. To calculate the concentration of the mercury which can be uptake by organism, rather than the total mercury found in the medium

Expensive and labor intensive to invent

Able to adjust the sensitivity of biosensor, by changing the reporter gene being used. With lux currently the most sensitive reporter, followed by lacZ than gfp.


Able to collate the psychological status and response of microbes so as to calculate the real effect done by the mercury pollution


Flexibility of choice of host strain, results can be extrapolated to compare effects on organisms, if bioengineering technologies improves this may be able to collate to effect of animals and ultimately human



Biosensors VS Conventional chemical method

The points listed below are for you to compare and decide on which technology suits the criteria of the company most. Capital constrain, Professional worker and environmental impacts are some points to consider.


Biosensors

Conventional Chemical method

Cost of equipment

Cheap

Expensive

Time

Fast

Slow

Adaptability

Cannot be used for all condition ( E.g. temperature difference)

Can be used

Initial cost

High due to the research need to fully understand the microbes

Fixed price

Diversity of measurment

Specific detection, need a combination of microbes

Can measure all

Environment impact

Almost perfectly environmental friendly

Feedstock for the chemical process are not environmental friendly

Video


what is this video about
  1. Bioaccumulation and using conventional chemical method to detect mercury poisoning in dolphins
  2. Effect of mercury (Minamata bay incident)

Quiz time!

QUESTION 1
A. How much mercury a 45 lb (pound) child would ingest by eating one 6 ounce of white tuna per week?

B. How many times more compare to the EPA's reference dose?

Ans:


Step 1:Determine EPA’s recommended standard for a 45 pound child (Multiply child's body weight by EPA's reference dose.)

· Convert 45 pounds to kilograms = 20.45 kilograms

· 20.45 kilograms x .1 micrograms per kilogram per day

· EPA recommended level = 2.05 micrograms per day = 14.35 micrograms per week.


Step 2 ; How much mercury is in 6 ounc of white tuna?

· Convert 6 ounces to grams = 170 grams X .31 ppm (or micrograms per gram)**

· Mercury ingested = 52.7 micrograms per gram (A)


Step 3 – Compare amount ingested with EPA’s recommended

· Divide 52.7 micrograms by 14.35 micrograms = 3.7

Conclusion: By eating 6 ounces of white tuna a week, the child is ingesting almost 4 times EPA’s recommended dose (B)






Question 2

A. How much ppm of mercury does the dolphin (Taiji, Japan) contains?
B. And what harmful health impact does it cause to us? (Name the impacts done to child and adult)

Ans:

A) 2000ppm

B) Methylmercury generally cause nervous system damage.

For fetuses, infants, and children, the primary health effect of methylmercury is impaired neurological development.

Impact of methylmercury poisoning on adult may include; impairment of the peripheral vision; disturbances in sensations ("pins and needles" feelings, usually in the hands, feet, and around the mouth); lack of coordination of movements; impairment of speech, hearing, walking; and muscle weakness.



QUESTION 3 (15 MARKS)
A. What instrumental methods are used determine the samples for lake monitoring?(Name 2)
B. What are the differences between AAS and CVAAS? (Name 3)

Ans:


A) LA-254 LabAnalyzer & PA2 water analyser

B) Differences:

1. Divalent mercury in aqueous samples was reduced to HG by strong reducing agent in acidic condition (E.g. stannous chloride). Different form sample preparation of digestion are used to digest different kind of mercury so that it could obtain best sensitivity

Preparation by “potassium permanganate perchloric acid, nitric acid, sulphuric acid and sodium hydroxide and lastly sodium borohydrie”

2. Does not need to employ flame technique as mercury could exist at free ground state

3. Sample must be Teflon cap at all times; mercury is a highly volatile metal and could escape and affect the accuracy of the experiment