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Lab Report 14: Bacteriophage Specificity (Plaque Assay Host Range)

In lab-report-14-bacteriophage-specificity, a phage is plated (0.10 mL) at a 10^-6 dilution on Host A and Host B giving 124 plaques on Host A and 5 plaques on Host B; what are the titers on each host, the efficiency of plating (EOP) on Host B relative to Host A, and what do these results imply about host specificity?

Subject: Biology Chapter: Microbiology and Epidemiology Topic: Cfu from Plate Counts Answer included
lab-report-14-bacteriophage-specificity bacteriophage specificity host range plaque assay PFU/mL plaque forming units efficiency of plating EOP
Accepted answer Answer included

Problem

In lab-report-14-bacteriophage-specificity, a bacteriophage is tested on two bacterial hosts using a plaque assay. A volume of 0.10 mL is plated from a \(10^{-6}\) dilution onto each host lawn.

  • Host A: 124 plaques
  • Host B: 5 plaques

Determine the phage titer (PFU/mL) on each host, compute the efficiency of plating (EOP) of Host B relative to Host A, and interpret what the results imply about bacteriophage specificity.

Core concepts

A plaque is assumed to originate from one infectious phage particle, so plaque counts estimate PFU (plaque-forming units). Host specificity (host range) reflects whether the phage can adsorb to host receptors, inject its genome, replicate, and lyse cells. The EOP compares how efficiently a phage forms plaques on a test host versus a reference host.

Step-by-step solution

Step 1: Use the plaque-count titer formula

The standard titer estimate is:

\[ \text{PFU/mL}=\frac{\text{number of plaques}}{\left(\text{volume plated in mL}\right)\cdot\left(\text{dilution}\right)} \]

Step 2: Compute the titer on Host A

For Host A: plaques \(=124\), volume \(=0.10\ \text{mL}\), dilution \(=10^{-6}\).

\[ \text{PFU/mL}_A=\frac{124}{(0.10)\cdot(10^{-6})} =\frac{124}{10^{-7}} =1.24\times 10^{9}\ \text{PFU/mL} \]

Step 3: Compute the titer on Host B

For Host B: plaques \(=5\), volume \(=0.10\ \text{mL}\), dilution \(=10^{-6}\).

\[ \text{PFU/mL}_B=\frac{5}{(0.10)\cdot(10^{-6})} =\frac{5}{10^{-7}} =5.0\times 10^{7}\ \text{PFU/mL} \]

Step 4: Compute efficiency of plating (EOP) on Host B relative to Host A

A common definition is:

\[ \text{EOP}_{B/A}=\frac{\text{PFU/mL}_B}{\text{PFU/mL}_A} \]
\[ \text{EOP}_{B/A}=\frac{5.0\times 10^{7}}{1.24\times 10^{9}} =\left(\frac{5.0}{1.24}\right)\times 10^{-2} \approx 4.03\times 10^{-2} \approx 0.040 \]

Step 5: Interpret bacteriophage specificity

  • Host A is the preferred (more permissive) host because the plaque-forming titer is much higher on Host A.
  • Host B is partially susceptible but inefficient: an EOP of about 0.040 means Host B yields about 4% as many infectious plaques as Host A under the same conditions.
  • This pattern is consistent with host specificity driven by receptor compatibility and/or intracellular restrictions (e.g., restriction-modification systems, CRISPR defense, or replication constraints).

Results summary

Quantity Host A Host B
Plaques counted 124 5
Volume plated (mL) 0.10 0.10
Dilution plated \(10^{-6}\) \(10^{-6}\)
Estimated titer (PFU/mL) \(1.24\times 10^{9}\) \(5.0\times 10^{7}\)
EOP of Host B relative to Host A \(\text{EOP}_{B/A}\approx 0.040\)

Visualization

Plaque assay outcome (host specificity) More plaques indicate higher PFU/mL on that host under identical dilution and volume. Host A 124 plaques (10⁻⁶, 0.10 mL) Higher titer → preferred host Host B 5 plaques (10⁻⁶, 0.10 mL) Lower titer → inefficient infection Compare titers EOP ≈ 0.040 Bacteriophage Adsorption depends on host surface receptors
Host A shows many plaques at the same dilution and plated volume, while Host B shows few plaques. The EOP quantifies this specificity by comparing PFU/mL on the test host to PFU/mL on the reference host.

Final answers

  • Titer on Host A: \(1.24\times 10^{9}\ \text{PFU/mL}\)
  • Titer on Host B: \(5.0\times 10^{7}\ \text{PFU/mL}\)
  • Efficiency of plating (Host B relative to Host A): \(\text{EOP}_{B/A}\approx 0.040\)
  • Interpretation: strong bacteriophage specificity for Host A, with inefficient plaque formation on Host B (narrow host range or partial susceptibility).
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