Stones may form in the lacrimal duct resulting in obstruction and secondary infection of the lacrimal gland. Actinomyces spp. are particularly important pathogens and, if present, are usually quite evident on Gram stain of material.
Direct examination
Crush specimen on glass slide to obtain a thin smear for Gram stain. Examine for pus cells and organisms, especially branching gram positive bacilli resembling Actinomyces species.
Culture
Crush specimen using a sterile wood applicator stick or urine loop before planting onto the following media:
| Media | Incubation |
| Blood Agar (BA) | CO2, 35°C x 48 hours |
| Chocolate Agar (CHOC) | CO2, 35°C x 48 hours |
| Wilkins Chalgren Agar (WC) | AnO2, 35°C x 48 hours |
| WC Nalidixic acid – Tween 80 (NAT) | AnO2, 35°C x 48 hours |
| WC Nalidixic acid – Vancomycin (NAV) | AnO2, 35°C x 48 hours |
| Thioglycolate broth (THIO) | O2, 35°C x 7 days |
If Actinomyces spp. are suggested on Gram stain set up a second set of anaerobic plates to be incubated for 7 days before opening.
If no growth on culture plates but evidence of growth in THIO, perform Gram stain and subculture THIO onto BA, CHOC and WC (as appropriate) and incubate and process as above. Identify all isolates.
Gram stain
Report, with quantification, the presence of organisms and WBCs. “Organisms resembling Actinomyces sp. seen in Gram stain”.
Culture
Negative Report
“Normal flora” or “No growth”.
Positive Report
Quantitate all significant isolates with appropriate sensitivities. If normal flora is also present, report with quantitation.
H.D. Isenberg, 2004. Wound Cultures – Wound and Soft Tissue Cultures, p. 3.13.1.1 – 3.13.1.16. In Clinical Microbiology Procedures Handbook, 2nd Edition, Vol 1 ASM Press, Washington, D.C.
H.D. Isenberg. 2004. Ocular Cultures p. 3.10.1. – 3.10.8. In Clinical Microbiology Procedures Handbook, 2nd Edition, Vol 1 ASM Press, Washington, D.C.
H.D. Isenberg. 2004. Culture for anaerobes p. 4.3.1 – 4.3.9 In Clinical Microbiology Procedures Handbook, 2nd Edition, Vol 1 ASM Press, Washington, D.C.
H.D. Isenberg. 2004. Examination of Primary Culture plates for anaerobic bacteria. p. 4.4.1 – 4.4.6 In Clinical Microbiology Procedures Handbook, 2nd Edition, Vol 1 ASM Press, Washington, D.C.
H.D. Isenberg. 2004. Incubation techniques for anaerobic bacteriology specimens. p. 4.5.1 – 4.5.4 In Clinical Microbiology Procedures Handbook, 2nd Edition, Vol 1 ASM Press, Washington, D.C.
Cumitech 5A Practical anaerobic bacteriology December 1991