For Staph and strepto a wise choice is to use API

API Gram positive Identification

• API Staph – Overnight identification of clinical staphylococci and micrococci
• RAPIDEC® Staph – 2-hour identification of the commonly occurring staphylococci
• API 20 Strep – 4 or 24-hour identification of streptococci and enterococci
• http://www.biomerieux-usa.com/clinical/api

For Staph and strepto a wise choice is to use API

API Gram positive Identification

• API Staph – Overnight identification of clinical staphylococci and micrococci
• RAPIDEC® Staph – 2-hour identification of the commonly occurring staphylococci
• API 20 Strep – 4 or 24-hour identification of streptococci and enterococci
• http://www.biomerieux-usa.com/clinical/api

Catalase test, coagulase test and hemolysis test should give you a better differentiation of Staph and Strep. All the best.

As you apparently have limited resources - to expand on Pasanth's good suggestions, Gram stain, hemolysis(sheep blood agar), catalase and coagulase for Staphylococcus aureus and Gram stain, hemolysis, catalase and bacitracin inhibition for Gp. A strep (Streptococcus pyogenes). You can find appropriate reactions described in the internet.

You should also review the literature (google scholar). there's a fair amount of info on microbes recovered from holy water. e.g.Article Holy water fonts are reservoirs of pathogenic bacteria

Thank you very much for your answers! We will focus on Catalase test, Coagulase test and Hemolysis test.

If you grow up your bacteria on sheep blood agar plates, the staphylococcal colonies will be larger than streptococcal colonies and most of the pathogenic bacteria will be beta hemolytic (a clear zone around the colonies). Confirm which they are by Gram staining (microscopic arrangement of streptococci are chains while staphylococci are in clusters). Streptococci are catalase negative while staphylococci are positive, but you can't perform catalase test from blood agar, since red blood cells are loaded with catalase.

If you use only MSA:

. In MSA you will not have any growth of Gram negative and Streptococcus. High salt concentration inhibit growth. So you must choose other medium for Streptococcus.

With MSA, the yellow colonies are mostly Staphylococcus Coag+ (as S. aureus). Staph coag negative are pink or red. You can confirm with complementary test as catalse (cat+), gram and the coagulase test.

For Streptococcus you must inoculate a pre-warm (37ºC) Sheep Blood Agar or a TSA with 5% of Blood Sheep. Incubate and pick colonies for TSA medium and realize test of catalase (negative), gram (positive cocci).

beta hemolytic colonies must be tested with bacitracin. Bacitracin sensitive are S.pyogenes. Bacitracin Resistant are other beta hemolytic Streptococci. (use APIStrep for identification).

alfa hemolytic colonies must be tested with optochin. Optochin sensitive are S.pneumoniae. Optochin Resistant are other alfa hemolytic Streptococci. (use APIStrep for identification)

Non-hemolitic colonies rarely cause disease. (use APIStrep for identification)

In the first place, why restrict yourself to only two bacteria. I suggest you carry out a holistic pathogenic bacterial assay on the font water. The result will be more appreciated and helpful in public health matters.

In the first place: why restrict yourself to only two bacteria. I suggest you carry out a holistic pathogenic bacterial assay on the font water. The result will be more appreciated and helpful in public health matters.

In the second place: If I were prefer to isolate two types of bacteria Staph & Strep. You should do the following tests (Gram stain, hemolysis (sheep blood agar), catalase and coagulase).

I suggest that you also try to isolate gram-negative bacilli and detect multiresistant in water

Hi, as your test sample is holy water, I would prefer to test fecal coliform and other pathogenic E. coli. as the first attempt. Thank you.

For easily and rapidly diagnose, you can use API test and VITEK 2 system ... good luck.

doing antibiotics resistance examinations