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Intraleukocytic Yeast Inclusions and Toxic Granulation Neutrophils On ISSN: 2474-3658 Miglietta et al. J Infect Dis Epidemiol 2019, 5:067 DOI: 10.23937/2474-3658/1510067 Volume 5 | Issue 1 Journal of Open Access Infectious Diseases and Epidemiology CASE REPORT Intraleukocytic Yeast Inclusions and Toxic Granulation Neutrophils on Peripheral Blood Smear: An Interesting Synergy between Hema- tology and Microbiology Fabio Miglietta1, Claudio Palumbo1, Fernando Parente2, Luciano Velardi3, Rosella Matera4, Luigi Conte4, Michela Dargenio4, Maurizio Quarta5, Milva Maria Nuzzo5, Nicola Di Renzo4 and Giambattista Lobreglio6 1Laboratory of Microbiology, Vito Fazzi Regional Hospital, Lecce, Italy 2Medicine Unit, Vito Fazzi Regional Hospital, Lecce, Italy 3Istituto di Nanotecnologia, CNR-Nanotec, Bari, Italy Check for 4Department of Hematology, Vito Fazzi Regional Hospital, Lecce, Italy updates 5Infectious Diseases Unit , Vito Fazzi Regional Hospital, Lecce, Italy 6Laboratory of Clinical Pathology, Vito Fazzi Regional Hospital, Lecce, Italy *Corresponding authors: Dr. Fabio Miglietta, Laboratory of Microbiology, Vito Fazzi Regional Hospital, 83, Montegrappa Street, 73018, Squinzano, Lecce, Italy, Tel: +39-3492548568, Fax: +39-0832782033 by poor sensitivity and slow turn-around time [2] while Abstract β-D-glucan detection demonstrates variable sensitivity The presence of yeast neutrophil inclusions was observed depending on the cut-off diagnostic value and on and discussed several times in other reports; moreover some works demonstrated how Toxic Granulation Neutrophils the Candida species under consideration. This last (TGNs) are especially helpful in predicting acute bacterial presents several false positive results due to albumin infection, while the development of candidaemia-related and/or immunoglobulin administration, Gram positive TGNs was rarely described and in-depth. bacteraemia or haemodialysis [3]. β-D-glucan, mannan We describe two occasional findings of neutrophil inclusions antigen detection and polymerase chain reaction are and marked TGNs respectively due to Candida tropicalis often expensive and time consuming, furthermore and Candida guillermondi on peripheral blood smear. their low specificity could lead to an excessive use of We proved how the microscopic observation of marked antifungals agents increasing fungal resistance and care toxic granulations can afford to suspect a systemic microbial costs [4]. infection without the potential to discriminate between bacterial or fungal infections. A general consensus has not been reached on the Keywords usefulness of any of these methods, except for blood culture and histological examination [5]. Candida tropicalis, Candida guillermondii, Candidemia, Toxic granulations, Peripheral blood smear, Sepsis Yeast with pseudohyphae or those that have been phagocytized by white blood cells are coincidentally Introduction found in peripheral blood smears. Nevertheless, the clinical diagnostic value and outcome of candidaemia Fungal pathogens, in particular Candida species, have diagnosed from peripheral blood smears are unclear [6]. become a major cause of nosocomial infection [1]. Blood cultures are limited for diagnosing invasive candidiasis We describe two occasional findings of neutrophil Citation: Miglietta F, Palumbo C, Parente F, Velardi L, Matera R, et al. (2019) Intraleukocytic Yeast Inclu- sions and Toxic Granulation Neutrophils on Peripheral Blood Smear: An Interesting Synergy between Hematology and Microbiology. J Infect Dis Epidemiol 5:067. doi.org/10.23937/2474-3658/1510067 Accepted: January 16, 2018: Published: January 18, 2018 Copyright: © 2019 Miglietta F, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Miglietta et al. J Infect Dis Epidemiol 2019, 5:067 • Page 1 of 3 • DOI: 10.23937/2474-3658/1510067 ISSN: 2474-3658 Figure 1: Microscopy of peripheral blood smear (May-Grünwald Stain), 100x objective. Panel A: Neutrophil breakup with phagocytosed yeasts (case 1); Panel B: Intraleukocytic yeast inclusions (case 2); Panel C: Toxic granulation neutrophils and free yeast (case 1); Panel D: Toxic granulation neutrophils and free yeast (case 2). inclusions and toxic granulation neutrophils (TGNs) Case 2 respectively due to Candida tropicalis and Candida A 50-year-old female patient suffering for HBV- guillermondi on peripheral blood smear (Figure 1). related cirrhosis, was admitted in the medicine unit due Case 1 to an ascites associated with jaundice and a reduction of the liver function. Two days after hospital admission A 44-year-old man with an history of Focal Segmental the patient developed fever (38.2 °C) unresponsive to Glomerulosclerosis and subjected to hemodialysis wide spectrum antibiotic therapy (meropenem) while for 22 years after a kidney transplant rejection, was laboratory data highlighted a leukocyte count of 8.9 × hospitalized in the nephrology department due to a 10^9/L (88% neutrophils), C-reactive protein of 91.9 lithiasic cholecystitis. A Central Venous Catheter (CVC) mg/L ( reference < 10) and Procalcitonin of 0.48 ng/mL was placed and the patient underwent cholecystectomy. (reference < 0.05). Two days after surgery he showed shaking chills and fever of 38.5 °C. Furthermore laboratory investigations On the same day peripheral blood smear (May- highlighted a leukocyte count of 2.8 × 10^9/L (74% Grünwald Stain) revealed some yeast neutrophils neutrophils) and C-reactive protein of 47.4 mg/L inclusions (Panel B) and numerous TGNs induced by (reference < 10.0). candidaemia (Panel D). Peripheral blood smear (May-Grünwald Stain) Candida guillermondi infection was confirmed by revealed a neutrophil breakup probably induced by blood cultures results, and the speciation of isolates phagocytosed yeasts (Panel A) and TGNs (Panel C) were performed by biochemical tests. The patient induced by candidaemia. was successfully submitted to antifungal therapy with fluconazole for 14 days. Blood cultures confirmed the blood smear findings revealing the presence of Candida tropicalis. CVC was Discussion removed and its cultural test resulted positive for the Previous reports have suggested that peripheral same fungal species. The patient was successfully blood smears may be useful for the detection of submitted to antifungal therapy with caspofungin for 14 disseminated yeast infection [5] especially in case days. Miglietta et al. J Infect Dis Epidemiol 2019, 5:067 • Page 2 of 3 • DOI: 10.23937/2474-3658/1510067 ISSN: 2474-3658 of central venous lines related candidaemia [6]. The excluded other non-infectious conditions as possible microscopic examination of the blood smear often causes of TGNs development. shows a very low sensitivity, otherwise in case of The microscopic observation of marked TGNs positivity it allows shorter time to diagnosis compared can afford to suspect a systemic microbial infections to blood cultures turn-around time. The presence of without the potential to discriminate between bacterial yeast neutrophil inclusions was observed and discussed or fungal infections. several times in other reports [7]; moreover these works demonstrated how TGNs are especially helpful Acknowledgement in predicting acute bacterial infection [7], while the The authors thanks Dr. Adele Braione for providing development of candidaemia-related TGNs was rarely us technical advice and moral support. described and in-depth. TGNs is the term used when the normally faint References stippled granules in neutrophils stain an intense reddish 1. Wenzel RP (1995) Nosocomial candidemia: Risk factors violet which is a consequence of activity against bacteria and attributable mortality. Clin Infect Dis 20: 1531-1534. or proteins and is observed in serious infections, toxic 2. Clancy CJ, Nguyen MH (2013) Finding the "missing 50%" or drug effects, or autoimmune processes (e.g., chronic of invasive candidiasis: how nonculture diagnostics will improve understanding of disease spectrum and transform polyarthritis) [8]. patient care. Clin Infect Dis 56: 1284-1292. Normal bone marrow granulocyte maturation is as- 3. Paramythiotou E, Frantzeskaki F, Flevari A, Armaganidis A, sociated with progressive decreases of azurophilic gran- Dimopoulos G (2014) Invasive fungal infections in the ICU: ule enzymes (myeloperoxidase, defensins, lysozyme, How to approach, how to treat. Molecules 19: 1085-1119. azurocidin, etc.). On the contrary, the origin of TGNs 4. Miglietta F, Faneschi ML, Lobreglio G, Palumbo C, Rizzo has been considered to be related to abnormal neutro- A, et al. (2015) Procalcitonin, C-reactive protein and serum phil maturation with persistence of azurophilic gran- lactate dehydrogenase in the diagnosis of bacterial sepsis, SIRS and systemic candidiasis. Infez Med 23: 230-237. ules, containing acid mucosubstance which stains more prominently than under normal circumstances [9]. 5. Yera H, Poulain D, Lefebvre A, Camus D, Sendid B (2004) Polymicrobial candidaemia revealed by peripheral blood The acidic mucosubstance accumulated in the smear and chromogenic medium. J Clin Pathol 57: 196- toxic granules has the potential to acidify phagosomes 198. enhancing bactericidal activity, since bacteria in 6. Hirai Y, Asahata S, Ainoda Y, Fujita T, Miura H, et al. (2015) phagosomes are killed more effectively at pH of 5.5 Candidemia diagnosed from peripheral blood smear: Case than of 7 [10]. Our blood smears confirmed the findings report and review of literature 1954-2013.
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