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Editors and Contributors
About the Editors
Saif Hameed is currently an Associate Professor at the Amity Institute of
Biotechnology, Amity University Haryana (AUH), India. Dr. Hameed earned his
Bachelor’s degree from the University of Delhi and his Master’s degree from Jamia
Hamdard in 2003 and 2005, respectively. He pursed his doctoral studies in life sciences at Jawaharlal Nehru University in 2010, where he also received a CSIR
Research Associateship for a 1-year postdoctoral work. He worked as a visiting
scholar at the Institut für Mikrobiologie, Heinrich-Heine-Universität, Düsseldorf,
Germany, in 2008. He received the Young Scientist Award under the Fast-Track
Scheme from the Science and Engineering Research Board, Department of Science
and Technology, New Delhi, in 2012. He is a life member of the International
Society for Infectious Diseases (ISID), the Association of Microbiologists of India
(AMI), and the Society of Biological Chemists (SBC), India. Dr. Hameed is
actively engaged in research in the eld of medical microbiology, particularly antibiotic resistance in pathogenic fungi. He has 70 peer-reviewed papers to his credit
in international journals of repute, along with 6 books. He has supervised 6 PhD
students and guided 25 UG and PG level students for their research projects.
PoojaVijayaraghavan is a Professor at Amity University Uttar Pradesh, Noida,
India. She is a prominent scientist in the eld of antifungal research. Her primary
research area is on evaluating the biological efcacy of naturally occurring small
molecules and their synthetically designed analogues against pathogenic fungal
species, with a particular emphasis on human pathogens such as Aspergillus fumig-
atus and Rhizopus oryzae, as well as plant pathogenic fungi like Magnaporthe oryzae and Curvularia lunata. She has published many peer-reviewed articles that offer
invaluable global perspectives and insights. Dr. Vijayaraghavan has obtained 17+
patents, with four patents already granted and the remainder currently under review.
Her research has successfully transformed three patented innovations into practical
products in collaboration with various industries. These include a herbal hand sanitizer, a biopolymer-based lm to extend the shelf life of perishable fruits and vegetables, and a biopesticide to combat rice blast disease.
xi
xii
Editors and Contributors
Contributors
KhushbooArya Department of Biochemistry, University of Lucknow, Lucknow,
Uttar Pradesh, India
Shumaiza Asif Laboratory of Protein Translation and Fungal Pathogenesis,
Regional Centre for Biotechnology, Faridabad, Haryana, India
SukalyaniBanik Division of Infectious Diseases, Center for Emerging Pathogens,
New Jersey Medical School, Rutgers University, Newark, NJ, USA
Hina Bansal Centre for Computational Biology and Bioinformatics, Amity
Institute of Biotechnology, Amity University, Noida, Uttar Pradesh, India
YashikBansal Department of Microbiology, ESIC Medical College and Hospital,
Alwar, Rajasthan, India
Md Nazmul Islam Bappy Faculty of Biotechnology and Genetic Engineering,
Sylhet Agricultural University, Sylhet, Bangladesh
Department of Animal and Fish Biotechnology, Sylhet Agricultural University,
Sylhet, Bangladesh
Nitin Bhardwaj Department of Zoology and Environmental Science, Gurukula
Kangri Vishwavidyalaya, Haridwar, Uttarakhand, India
Rohit Bhattacharjee Department of Microbiology and Biotechnology Centre,
Faculty of Science, The Maharaja Sayajirao University of Baroda, Vadodara,
Gujarat, India
BiswambharBiswas Laboratory of Protein Translation and Fungal Pathogenesis,
Regional Centre for Biotechnology, Faridabad, Haryana, India
Nabajit Kumar Borah Centre for Computational Biology and Bioinformatics,
Amity Institute of Biotechnology, Amity University, Noida, Uttar Pradesh, India
ShikhaChandra Department of Biochemistry, University of Lucknow, Lucknow,
Uttar Pradesh, India
Saumya Chaturvedi Department of Biochemistry, University of Lucknow,
Lucknow, Uttar Pradesh, India
Sajad Ahmad Dar Research and Scientic Studies Unit, College of Nursing,
Jazan University, Jazan, Saudi Arabia
Deotima Das School of Bio Sciences and Technology, Vellore Institute of
Technology, Vellore, Tamil Nadu, India
GauravRajDwivedi ICMR-Regional Medical Research Centre, Gorakhpur, India
MbaIfeanyiElibe Department of Microbiology, University of Nigeria, Nsukka,
Enugu State, Nigeria
Editors and Contributors
xiii
Ashik Francis Laboratory of Protein Translation and Fungal Pathogenesis,
Regional Centre for Biotechnology, Faridabad, Haryana, India
DevarshiGajjar Department of Microbiology and Biotechnology Centre, Faculty
of Science, The Maharaja Sayajirao University of Baroda, Vadodara, Gujarat, India
GarimaGangwar School of Biotechnology, Gautam Buddha University, Greater
Noida, Uttar Pradesh, India
Priya Gautam Genomic Laboratory, Department of Biotechnology and
Bioinformatics, Jaypee University of Information Technology, Waknaghat Solan,
Himachal Pradesh, India
Chhavi Gupta Department of Infectious Disease, Yashoda Superspeciality
Hospital, Delhi NCR, India
Infectious Disease, AIIMS, New Delhi, India
LovelyGupta Amity Institute of Biotechnology, Amity University, Noida, Uttar
Pradesh, India
MonalikaGupta Department of Biochemistry, University of Lucknow, Lucknow,
Uttar Pradesh, India
NidhiGupta Laboratory of Protein Translation and Fungal Pathogenesis, Regional
Centre for Biotechnology, Faridabad, Haryana, India
Sandeep Hans Laboratory of Protein Translation and Fungal Pathogenesis,
Regional Centre for Biotechnology, Faridabad, Haryana, India
Nweze Emeka Innocent Department of Microbiology, University of Nigeria,
Nsukka, Enugu State, Nigeria
Anubhuti Jha Department of Biotechnology, St. Thomas College-Bhilai,
Hemchand Yadav University, Durg, CG, India
Saurabh Kumar Jha Department of Zoology, Kalindi College, University of
Delhi, New Delhi, Delhi, India
PurviJoshi Department of Microbiology and Biotechnology Centre, Faculty of
Science, The Maharaja Sayajirao University of Baroda, Vadodara, Gujarat, India
AjeetKumar Laboratory of Protein Translation and Fungal Pathogenesis, Regional
Centre for Biotechnology, Faridabad, Haryana, India
Antresh Kumar Department of Biochemistry, Central University of Haryana,
Mahendergarh, India
AwanishKumar Department of Biotechnology, National Institute of Technology,
Raipur, CG, India
LokeshKumar Amity Institute of Biotechnology, Amity University, Noida, Uttar
Pradesh, India
xiv
Editors and Contributors
ManojKumar ICMR-Regional Medical Research Centre, Gorakhpur, India
Deepika Kumari Department of Biochemistry, Maharshi Dayanand University,
Rohtak, Haryana, India
Pammi Kumari Department of Biochemistry, Maharshi Dayanand University,
Rohtak, Haryana, India
Reetu Kundu Department of Cytology and Gynaecological Pathology, Post
Graduate Institute of Medical Education and Research, Chandigarh, India
Rashmi Minocha Department of Biochemistry, All India Institute of Medical
Sciences, Ansari Nagar, New Delhi, Delhi, India
RajanKumarMishra Amity Institute of Biotechnology, Amity University, Noida,
Uttar Pradesh, India
AyanPrasadMukherjee School of Bio Sciences and Technology, Vellore Institute
of Technology, Vellore, Tamil Nadu, India
ManojV.Murhekar ICMR-Regional Medical Research Centre, Gorakhpur, India
Neha Genomic Laboratory, Department of Biotechnology and Bioinformatics,
Jaypee University of Information Technology, Waknaghat Solan, Himachal
Pradesh, India
Aditi Parashar Amity Institute of Microbiology Technology, Amity University,
Noida, Uttar Pradesh, India
RituPasrija Department of Biochemistry, Maharshi Dayanand University, Rohtak,
Haryana, India
Rajendra Prasad Amity Institute of Integrative Science and Health and Amity
Institute of Biotechnology, Amity University Gurgaon, Gurgaon, Haryana, India
Rekha Puria School of Biotechnology, Gautam Buddha University, Greater
Noida, Uttar Pradesh, India
Aishwarya Rana Laboratory of Protein Translation and Fungal Pathogenesis,
Regional Centre for Biotechnology, Faridabad, Haryana, India
KongaraHanumanthaRao Biochemistry and Bioinformatics, GITAM School of
Sciences, GITAM (Deemed to be University), Visakhapatnam, Andhra Pradesh, India
TanjinBarketullah Robin Faculty of Biotechnology and Genetic Engineering,
Sylhet Agricultural University, Sylhet, Bangladesh
SomnathSahoo Biochemistry and Bioinformatics, GITAM School of Sciences,
GITAM (Deemed to be University), Visakhapatnam, Andhra Pradesh, India
MuskanSahu Department of Microbiology and Biotechnology Centre, Faculty of
Science, The Maharaja Sayajirao University of Baroda, Vadodara, Gujarat, India
Editors and Contributors
xv
Seneha Santoshi Centre for Computational Biology and Bioinformatics, Amity
Institute of Biotechnology, Amity University, Noida, Uttar Pradesh, India
Saurav Department of Civil Engineering, Jaypee University of Information
Technology, Waknaghat Solan, Himachal Pradesh, India
Pooja Sen Amity Institute of Biotechnology, Amity University, Noida, Uttar
Pradesh, India
Jata Shankar Genomic Laboratory, Department of Biotechnology and
Bioinformatics, Jaypee University of Information Technology, Waknaghat Solan,
Himachal Pradesh, India
Preeti Sharma Department of Biochemistry, Maharshi Dayanand University,
Rohtak, Haryana, India
Sonia Kumari Shishodia University Institute of Biotechnology, Chandigarh
University, Mohali, Punjab, India
Aman Singh Amity Institute of Biotechnology, Amity University, Noida, Uttar
Pradesh, India
AshutoshSingh Department of Biochemistry, University of Lucknow, Lucknow,
Uttar Pradesh, India
Bharti Singh School of Biotechnology, Gautam Buddha University, Greater
Noida, Uttar Pradesh, India
Nidhi Singla Department of Microbiology, Government Medical College and
Hospital, Chandigarh, India
Prashansha Srivastava ICMR-Regional Medical Research Centre,
Gorakhpur, India
AnilThakur Laboratory of Protein Translation and Fungal Pathogenesis, Regional
Centre for Biotechnology, Faridabad, Haryana, India
RamanThakur Department of Medical Laboratory Science, Lovely Professional
University, Jalandhar, Punjab, India
ShashikantTiwari ICMR-Regional Medical Research Centre, Gorakhpur, India
Yukti Tripathi Centre for Computational Biology and Bioinformatics, Amity
Institute of Biotechnology, Amity University, Noida, Uttar Pradesh, India
PinalTrivedi Department of Microbiology and Biotechnology Centre, Faculty of
Science, The Maharaja Sayajirao University of Baroda, Vadodara, Gujarat, India
Sana Akhtar Usmani Department of Biochemistry, University of Lucknow,
Lucknow, Uttar Pradesh, India
xvi
Editors and Contributors
Pooja Vijayaraghavan Amity Institute of Biotechnology, Amity University,
Noida, Uttar Pradesh, India
KaziMd.AliZinnah Faculty of Biotechnology and Genetic Engineering, Sylhet
Agricultural University, Sylhet, Bangladesh
Department of Animal and Fish Biotechnology, Sylhet Agricultural University,
Sylhet, Bangladesh
Part I
Candida and Candidiasis
Diagnostics ofCandida andCandidiasis:
Current Methods andFuture
Perspectives
SukalyaniBanik
Abstract
Candidiasis caused by the member of the genus Candida represents a signicant
health concern worldwide by causing high morbidity and mortality. It can mani-
fest in various forms, ranging from supercial infections to more invasive and
systemic diseases. Immunocompromised individuals and people with underlying
health conditions are at greater risk of potential complications including blood-
stream infections or candidemia, endocarditis, and deep-seated candidiasis.
Accurate and rapid diagnosis is critical for timely initiation of appropriate anti-
fungal treatment and patient survival. This chapter seeks to provide an overview
of the multifaceted approach to diagnosing candidiasis, current challenges, and
potential future directions in diagnostic strategies.
1
Keywords
Candida · Candidiasis · Candidemia · Invasive · Fungal · Diagnosis
1.1 Introduction
Candidiasis is an opportunistic fungal infection caused by Candida species, most
frequently by Candida albicans. It represents a signicant and often underestimated
health concern worldwide. Candida species, particularly Candida albicans, are
opportunistic pathogens that commonly inhabit mucosal surfaces and various niches
within the human body (Turner and Butler 2014). While Candida is typically a
S. Banik, PhD (*)
Division of Infectious Diseases, Center for Emerging Pathogens, New Jersey Medical School,
Rutgers University, Newark, NJ, USA
e-mail: sb1712@njms.rutgers.edu
© The Author(s), under exclusive license to Springer Nature Singapore Pte
Ltd. 2024
S. Hameed, P. Vijayaraghavan (eds.), Recent Advances in Human Fungal
Diseases, https://doi.org/10.1007/978-981-97-4909-6_1
3
4
S. Banik
commensal organism, its overgrowth can lead to a spectrum of infections, ranging
from localized mucocutaneous manifestations to more invasive and severe systemic
conditions (Sardi etal. 2013; Deorukhkar and Saini 2014). Supercial candidiasis
commonly affects mucous membranes, skin, and nails, resulting in conditions such
as oral thrush, vaginal yeast infections, and cutaneous candidiasis. On the other
hand, life-threatening systemic candidiasis poses a greater threat, particularly in
immunocompromised individuals, with potential complications including bloodstream infections, endocarditis, and deep-seated organ involvement (Eggimann
etal. 2003). The predisposing factors for candidiasis are diverse including encompassing antibiotic use, immunosuppression, diabetes, pregnancy, and other conditions that compromise the host’s defenses. The rising incidence of candidiasis,
especially in healthcare settings, underscores the importance of understanding its
pathogenesis, risk factors, and clinical manifestations (Lockhart 2014).
The diagnosis of candidiasis involves a comprehensive approach that considers
the clinical presentation, patient history, and, when necessary, laboratory tests.
Clinicians often rely on subjective symptoms reported by the patient and objective
ndings during physical examination. However, candidiasis often presents with
symptoms that overlap with other infections, making diagnosis based solely on
clinical manifestations challenging. Laboratory-based diagnostic methods play a
pivotal role in conrming the presence of Candida and determining the extent of
infection. Microbiological cultures, molecular techniques, and microscopic examination of clinical samples are common tools employed for the identication and
characterization of Candida species. Additionally, serological tests and imaging
studies may be utilized in cases of invasive candidiasis to assess the systemic impact
on internal organs.
The increasing prevalence of antifungal resistance among Candida species poses
a signicant challenge. This necessitates ongoing surveillance and adaptation of
diagnostic methods to guide appropriate treatment decisions (Posteraro etal. 2015).
There is a need for rapid and reliable point-of-care diagnostic tools to facilitate
timely intervention, especially in resource-limited settings. Accurate and timely
diagnosis is paramount for effective management, yet the intricate nature of Candida
infection poses challenges that demand a nuanced approach (Clancy and Nguyen
2013; Clancy and Nguyen 2018). This chapter explores the current multifaceted
approach to diagnosing candidiasis, associated challenges, and potential future
directions in diagnostic strategies, highlighting the importance of a thorough assessment to guide appropriate treatment strategies. As the understanding of the pathophysiology of candidiasis continues to evolve, ongoing research may contribute to
the development of more accurate and efcient diagnostic methods, ultimately
improving patient outcomes in the management of this prevalent fungal infection.
1.2 Conventional Methods
Conventional laboratory-based diagnostic methods rely on morphological and
physiological characteristics of Candida species for their identication from clinical specimens (Neppelenbroek et al. 2014). Despite numerous commercially
1 Diagnostics of Candida and Candidiasis: Current Methods and Future…
5
available rapid molecular-based diagnostic kits, many clinical laboratories still
depend on these conventional methods for routine identication of Candida species.
These include a microscopic examination to observe fungal morphology, grow, or
isolate Candia species from clinical specimens on different culture media, germ
tube test, and various biochemical assays including carbohydrate fermentation and
assimilation tests (Berardinelli and Opheim 1985; Rimek etal. 2008).
1.2.1 Microscopy
Preliminary diagnosis of fungal infections still depends on microscopic examination of fungal cells. It can be applied to various types of clinical specimens such as
tissue biopsies, blood, saliva, sputum, bronchoalveolar lavage (BAL), cerebrospinal
uid (CSF), and swabs. Some fungi have distinctive morphology which is visible
under a microscope, and therefore, they can be used for their identication within
hours of sample collection. Of those, the germ tube test is the widely used microscopic method for the presumptive identication of C. albicans in clinical specimens. In the germ tube test, which is discussed later in this chapter, the sample is
incubated in serum for a few hours and then observed under a microscope. The
presence of short germ tube-like structures observed under the microscope is an
indicative of C. albicans (Berardinelli and Opheim 1985). However, other Candida
species such as C. tropicalis, C. parapsilosis, and C. africana have been reported to
form similar germ tube-like structures (Lipperheide et al. 1993; Freydière and
Guinet 1997; Campbell etal. 1998; Loefer etal. 2003).
Different stains can be used to observe yeasts directly in clinical specimens or in
culture smears from colonies grown in a culture medium. Several staining methods
are available for yeast identication. A 10% potassium hydroxide solution is the
most widely used stain to detect fungi. Potassium hydroxide digests keratin present
in the fungal cell wall while glycerin in solution prevents degradation. Lactophenol
blue can be used along with potassium hydroxide solution for better visualization of
fungi under the microscope. Gram and Giemsa stains can also be used to stain fungal cells. Candida is considered as a Gram-positive fungus. In Gram-stained smear,
the presence of budding yeast cells and pseudohyphae is useful for the identication
of fungal cells in sputum, gastric lavage, lunch aspirates, vaginal secretion, and
urine specimens (Marty etal. 2015; Yao etal. 2019).
Visualization of distinctive fungal morphology in tissue sections by microscopy
or histopathology is a widely used technique and provides evidence of invasive
fungal infections (Donnelly et al. 2020). Various staining solutions are used for
histopathological examination of fungal cells including Grocott-methenamine silver, hematoxylin and eosin, Ziehl-Neelsen stain, periodic acid-Schiff, Calcouor
White, Blankophor, and Papanicolaou stain (Guarner and Brandt 2011; Frickmann
etal. 2015). Grocott-methenamine silver is reported to be the best method for the
histological identication of fungi. Staining with periodic acid-Schiff takes several
hours to perform the test. Calcouor white is a colorless dye mostly used in frozen
and parafn-embedded tissue specimens to detect hyphae and pseudohyphae.
Hematoxylin and eosin and Papanicolaou stains are found to be less effective.