Gram Stain: Looking Beyond Bacteria to Find Fungi in Gram Stained Smear: A Laboratory Guide for Medical Microbiology

There are several medical mycology textbooks that contain a chapter on direct microscopy. However, this textbook is the first of its kind, as it discusses the simple Gram stain procedure as a valuable tool for the detection of fungal elements. This book has been specifically designed for people working in the medical microbiology laboratory with little or no practical experience in medical mycology. The central idea presented in this textbook begins with the Gram stain for the detection of fungi; the most important and more frequently isolated opportunistic and potentially pathogenic fungal species have been included. The book contains more than three hundred color images, the majority of which come from direct smear examination, such as Gram stain and other staining procedures. The mold phase and the microscopic structure of the identified fungal species relating to the initial findings of the direct smear have been linked to avoid bias. When a fungal infection is present but not suspected clinically, the Gram stain may be the only clue to the true cause of the infection. Although there are better methods than the Gram stain for visualization of fungi, these methods are only performed if there is clinical suspicion for fungal disease. Clinicians often send specimens for bacterial culture, but they sometimes overlook requests for fungal culture. During such times, the Gram stain is the only technique available in the clinical microbiology laboratory for direct detection of fungi from these specimens. The presence of fungi should not be overlooked during the direct examination of the clinical specimens for bacteria. This book will guide the reader in the recognition and identification of fungal elements in gram-stained smears, especially when they are distorted and remain unstained and undetectable. This new textbook focuses on the detection and classification of fungal elements in Gram stains. Newly developed flowcharts, clues, and key details regarding structural characteristics have been added to guide the reader in the right direction. Throughout the years, the author has accumulated many scenarios in which fungal elements were not detected on the original Gram stain evaluation but were found to be positive upon review once the culture grew a fungus. Finally, the book contains a chapter with a practice examination including microscopic images representative of scenarios commonly encountered in the clinical microbiology laboratory.

The author, Subhash Kumar Mohan, was born in Punjab, India. He received a graduate degree in integrated medical systems in 1967 and worked in India for five years before emigrating to Vancouver, Canada in 1972. He lived briefly in the UK from 1974 to 1976, working in a laboratory performing bacteriological and chemical analyses of drinking water and studying microbiology at the Isleworth Polytechnic Institution in London. Subhash returned to Canada in 1976 to work in a private laboratory and study at the Michener Institute for Medical Technology, completing the Canadian Society for Medical Laboratory Science (CSMLS) national certification exam for registered technologists in clinical microbiology in 1977. Subhash moved to Newfoundland, Canada and joined the Charles Curtis Memorial Hospital (CCMH) in St. Anthony, Newfoundland in 1978. He worked at the CCMH for three years, supervising the microbiology department. During his stay at the CCMH, Subhash successfully achieved national certification for registered technologists in immunology in 1980. His first case, ＂Possible Waterborne Salmonellosis-Newfoundland,＂ was published in Canada Disease Weekly Report in 1982. Subhash moved back to Toronto to work at Toronto General Hospital in late 1981 (the laboratory would later merge with Mount Sinai Hospital in 2000). He successfully achieved advanced certification in medical microbiology (Advanced Registered Technologist [ART]) from CSMLS in 1988. Subhash has published many articles and abstracts in Canadian and international medical journals. Teaching of the medical technology students, new graduates, staff members, and medical residents is an integral part of his duties in the clinical microbiology laboratory. Subhash has offered live medical mycology workshops, seminars, correspondence courses, and poster presentations across Canada and in the United States and India. After acquiring so much experience on the bench, Subhash had a dream to publish a medical mycology book for clinical microbiology laboratorians. Subhash has taken Hans Gram’s simple stain, which revolutionized the field of bacteriology, into the field of medical mycology for the detection of fungi in the clinical specimens. The author led the discovery of a new Candida species (Candida subhashii), which he isolated from the peritoneal fluid of a patient with end-stage renal failure. The experts decided to name the isolate in honour of Subhash’s extensive work in the field of mycology.