1. Do we suspect that all wound specimens are infected by S. aureus? Why or why not?

Not all. Because in a polymicrobial infections account for about half of all infections in
community-acquired soft-tissue wounds, and Staphylococcus aureus accounts for about only a
quarter of these infections. S. aureus is actually one of the species of Staphylococcus spp. which
are commonly found in wounds that colonizes a large percentage of wounds. However, there are
possibility that other microorganism can enter wounds in a variety of ways. The multiplication of
other microorganisms in the site of a susceptible host also causes wound infections.
Accordingly, there are other microorganism which are related to wounds. Staphylococcus aureus,
Streptococcus pyogenes, Enterococci, and Pseudomonas aeruginosa are the most common
causative organisms associated with wound infections.
https://dermnetnz.org/topics/wound-infections
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4065078/#:~:text=Conclusions,aureus%20and
%20MRSA.
2. If gram-positive cocci in clusters are seen in a smear, can we immediately report it to the
physician as S. aureus? Why or why not?
Yes, because gram- positive bacteria cocci can grow in pairs, chains, or clusters but it depends
on their orientation and attachment during cell division. When the cocci seen as cluster in a
certain smear this implies that bacteria are developing. So, if "gram-positive cocci in clusters"
are MSSA or MRSA, intravenous lines, a skin/soft-tissue source, a bone source, or an
endovascular source (e.g., endocarditis) may be involved. The clinical scenario should assist in
narrowing the possibilities based on the infection site. Another reason why it should be reported
in order to have cure immediately since gram-positive bacteria are causing serious infections.
Thus, invasive procedures break down natural barriers to bacterial invasion
https://academic.oup.com/cid/article/39/8/1170/296174
https://www.unmc.edu/intmed/divisions/id/asp/clinical-microbiology/docs/Negative-BCID-
Guidance-2020_Final.pdf

3. Can we use sodium citrate as the anticoagulant for the tube coagulase test? Why or
why not?

Yes, because of its mild calcium-chelating properties, sodium citrate is an effective

anticoagulant. The addition of sodium citrate to blood prevents it from clotting. Another reason,
is the factor V which is relatively stable in citrated blood, sodium citrate is the anticoagulant of
choice for coagulation tests like the prothrombin time test and partial thromboplastin time test.
Since factor V and VIII are more stable in a citrated specimen, sodium citrate has been used as a
coagulation test. For more than a century, sodium citrate has been used as an anticoagulant to
keep blood and blood products stable, presumably by sequestering Ca(++) ions in vitro.
https://microbeonline.com/diagnostic-tests-biochemical-tests-coagulase-test/
https://asm.org/ASM/media/Protocol-Images/Coagulase-Test-Protocol.pdf?ext=.pdf

4. Why do we inspect coagulase test tubes after 4 hours? Is this a critical time frame?
Why or why not?

During the first four hours of incubation, tubes should be checked after 4 hours to check
the results of the Coagulase Test because some Staphylococcus aureus strains produce
fibrinolysin, which can lyse clots. The coagulase test can be performed in many ways.
The slide test is straightforward, yielding results in less than ten seconds, but it can result
in false negatives. Although the tube test is the most conclusive, it can take up to 24
hours to complete. Another reason why it should be inspected after 4hours to observe if
there are no clots in order to continue with the next procedure.
https://microbeonline.com/diagnostic-tests-biochemical-tests-coagulase-test/
https://microbiologie-clinique.com/coagulase-test-principle-protocol-results.html

5. Can we use colonies from blood agar plates for the catalase test? Why or why not?
6. No, your colony should not be taken from a blood agar plate. because the whole red
blood cells contain catalase, the catalase test should not be performed on colonies
obtained from media containing whole red blood cells, as this could result in a false
positive. The presence of catalase in the erythrocytes may also result in a false positive.
Thus, if you're using colonies from a blood agar plate, be careful not to scrape up any of
the blood agar—blood cells are catalase-positive, so any contaminating agar could result
in a false positive. However, when hydrogen peroxide is added to the bacterial inoculum,
oxygen bubbles are rapidly liberated, indicating the presence of the catalase enzyme. The
absence of such bubbles indicates the lack of enzyme.

https://www.austincc.edu/microbugz/handouts/Catalase%20and%20Oxidase
%20Tests.pdf
https://assets.publishing.service.gov.uk/government/uploads/system/uploads/attachment_data/
file/791639/TP_8i4.pdf