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Precision Detection of Fungal Co-Infections for Enhanced COVID-19 Treatment Strategies Using FESEM Imaging

  • ORIGINAL RESEARCH ARTICLE
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Indian Journal of Microbiology Aims and scope Submit manuscript

Abstract

The treatment of fungal infections presents significant challenges due to the lack of standardized diagnostic procedures, a restricted range of antifungal treatments, and the risk of harmful interactions between antifungal medications and the immunosuppressive drugs used in anti-inflammatory treatment for critically ill patients with COVID-19. Mucormycosis and aspergillosis are the primary invasive fungal infections in patients with severe COVID-19, occurring singly or in combination. Histopathological examination is a vital diagnostic technique that details the presence and invasion of fungi within tissues and blood vessels, and the body’s response to the infection. However, the pathology report omits information on the most common fungi associated with the observed morphology, as well as other potential fungi and parasites that ought to be included in the differential diagnosis. This research marks significance in diagnosing fungal infections, such as mucormycosis and aspergillosis associated to COVID-19 by field emission scanning electron microscopy (FESEM) imaging to examine unstained histopathology slides, allowing for a detailed morphological analysis of the fungus. FESEM provides an unprecedented resolution and detail, surpassing traditional Hematoxylin & Eosin (H&E) and Grocott’s Methenamine Silver (GMS) staining methods in identifying and differentiating dual fungal infections and diverse fungal species. The findings underscore the critical need for individualized treatment plans for patients severely affected by COVID-19 and compounded by secondary fungal infections. The high-magnification micrographs reveal specific fungal morphology and reproductive patterns. Current treatment protocols largely depend on broad-spectrum antifungal therapies. However this FESEM guided diagnostic approach can help in targeted patient specific anti fungal therapies. Such precision could lead to more effective early interventions, addressing the complex management required for severe COVID-19 cases with coexisting fungal infections. This approach significantly advances disease management and patient recovery, advocating for personalized, precision medicine in tackling this multifaceted clinical challenge.

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Data Availability

All data that support the findings of this study are included within the article.

Abbreviations

ACE2:

Angiotensin converting enzyme 2

CAA:

COVID-19 Associated Aspergillosis

CAM:

COVID-19 Associated Mucormycosis

CCR2+:

C-C chemokine receptor type 2

COVID-19:

CoronaVirus Disease of 2019

FEG:

Field Emission Gun

FESEM:

Field Emission Scanning Electron Microscope

GAG:

Galactosaminogalactan

GM:

Galactomannan

GMS Stain:

Grocott-Gömöri'sMethenamine Silver stain

H&E Stain:

Hematoxylin and eosin stain

HIF-1α:

Hypoxia-Inducible Factor 1α

HMDS:

Hexamethyldisilane

ICU:

Intensive care unit

IFN:

Interferon

IFN-γ:

Interferon-gamma

IL-1ß:

Interleukin 1-beta

IL-6:

Interleukin-6

LCB:

Lactophenol cotton blue

NADPH:

Nicotinamide adenine dinucleotide phosphate

NLRs:

NOD like receptors

PAS:

Periodic acid schiff

PCR:

Polymerase chain reaction

PRRs:

Pattern recognition receptors

SARS-CoV-2:

Severe Acute Respiratory Syndrome Coronavirus 2

TNF-α:

Tumor Necrosis Factor-alpha

TLR4:

Toll-like receptor 4

WBC:

White blood cells

References

  1. Islam MR, Rahman MM, Ahasan MT et al (2022) The impact of mucormycosis (black fungus) on SARS-CoV-2-infected patients: at a glance. Environ Sci Pollut Res 29:69341–69366. https://doi.org/10.1007/s11356-022-22204-8

    Article  CAS  Google Scholar 

  2. Ganesan N, Sivanandam S (2022) Histomorphological features of mucormycosis with rise and fall of COVID-19 pandemic. Pathol Res Pract 236:153981. https://doi.org/10.1016/j.prp.2022.153981

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Akhtar N, Wani AK, Tripathi SK, Prakash A, Mannan MA (2022) The role of SARS-CoV-2 immunosuppression and the therapy used to manage COVID-19 disease in the emergence of opportunistic fungal infections: a review. Current Res Biotechnol. https://doi.org/10.1016/j.crbiot.2022.08.001

    Article  Google Scholar 

  4. Habibzadeh A, Lankarani KB, Farjam M et al (2022) Prevalence of fungal drug resistance in COVID-19 Infection: a global meta-analysis. Curr Fungal Infect Rep 16:154–164. https://doi.org/10.1007/s12281-022-00439-9

    Article  PubMed  PubMed Central  Google Scholar 

  5. Qin C, Zhou L, Hu Z, Zhang S, Yang S, Tao Y, Xie C, Ma K, Shang K, Wang W, Tian DS (2020) Dysregulation of immune response in patients with coronavirus 2019 (COVID-19) in Wuhan. China Clin Infect Dis 71:762–768. https://doi.org/10.1093/cid/ciaa248

    Article  CAS  PubMed  Google Scholar 

  6. Hall MW et al (2022) Immune immunomodulation in coronavirus disease 2019 (COVID-19): strategic considerations for personalized therapeutic intervention. Clin Infect Dis 74:144–148. https://doi.org/10.1093/cid/ciaa904

    Article  CAS  PubMed  Google Scholar 

  7. Jin Y, Ji W, Yang H et al (2020) Endothelial activation and dysfunction in COVID-19: from basic mechanisms to potential therapeutic approaches. Sig Transduct Target Ther 5:293. https://doi.org/10.1038/s41392-020-00454-7

    Article  CAS  Google Scholar 

  8. Hoenigl M, Seidel D, Carvalho A, Rudramurthy SM, Arastehfar A, Gangneux JP, Nasir N, Bonifaz A, Araiza J, Klimko N, Serris A (2022) The emergence of COVID-19 associated mucormycosis: a review of cases from 18 countries. Lancet Microbe. https://doi.org/10.1016/S2666-5247(21)00237-8

    Article  PubMed  PubMed Central  Google Scholar 

  9. Baral PK, Aziz MA, Islam MS (2022) Comparative risk assessment of COVID-19 associated mucormycosis and aspergillosis: a systematic review. Health Sci Rep 5:e789. https://doi.org/10.1002/hsr2.789

    Article  PubMed  PubMed Central  Google Scholar 

  10. Sundeep D, Varadharaj EK, Umadevi K, Jhansi R (2023) Role of nanomaterials in screenprinted electrochemical biosensors for detection of COVID-19 and for post-covid syndromes. ECS Adv 2:016502. https://doi.org/10.1149/2754-2734/acb832

    Article  Google Scholar 

  11. Lass-Flörl C (2009) Zygomycosis: conventional laboratory diagnosis. Clin Microbiol Infect 15:60–65. https://doi.org/10.1111/j.1469-0691.2009.02999.x

    Article  PubMed  Google Scholar 

  12. Moran CA, Mullick FG, Ishak KG, Johnson FB, Hummer WB (1991) Identification of titanium in human tissues: probable role in pathologic processes. Hum Pathol 22:450–454. https://doi.org/10.1016/0046-8177(91)90130-H

    Article  CAS  PubMed  Google Scholar 

  13. Iwen PC, Sigler L, Noel RK, Freifeld AG (2007) Mucorcircinelloides was identified by molecular methods as a cause of primary cutaneous zygomycosis. J Clin Microbiol 45:636–640. https://doi.org/10.1128/jcm.01907-06

    Article  CAS  PubMed  Google Scholar 

  14. Umadevi K, Sundeep D, Jhansi R et al (2023) Effect of functionalization of 2D graphene nanosheets on oxidation stress of BEAS-2B cells. BioNanoSci. https://doi.org/10.1007/s12668-023-01155-5

    Article  Google Scholar 

  15. Ibrahim AS, Spellberg B, Walsh TJ, Kontoyiannis DP (2012) Pathogenesis of mucormycosis. Clin Infect Dis 54:S16–S22. https://doi.org/10.1093/cid/cir865

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Skiada A, Pavleas I, Drogari-Apiranthitou M (2020) Epidemiology and diagnosis of mucormycosis: an update. J Fungi (Basel) 6:265. https://doi.org/10.3390/jof6040265

    Article  CAS  PubMed  Google Scholar 

  17. Reiterer M, Rajan M, Gómez-Banoy N, Lau JD, Gomez-Escobar LG, Ma L, Gilani A, Alvarez-Mulett S, Sholle ET, Chandar V, Bram Y (2021) Hyperglycemia in acute COVID-19 is characterized by insulin resistance and adipose tissue infectivity by SARS-CoV-2. Cell Metab 33:2174–2188. https://doi.org/10.1016/j.cmet.2021.09.009

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Anand VK, Alemar G, Griswold JA Jr (1992) Intracranial complications of mucormycosis: an experimental model and clinical review. Laryngoscope 102:656–662. https://doi.org/10.1288/00005537-199206000-00011

    Article  CAS  PubMed  Google Scholar 

  19. Giri B, Dey S, Das T, Sarkar M, Banerjee J, Dash SK (2018) Chronic hyperglycemia mediated physiological alteration and metabolic distortion leads to organ dysfunction, infection, cancer progression and other pathophysiological consequences: an update on glucose toxicity. Biomed Pharmacother 107:306–328. https://doi.org/10.1016/j.biopha.2018.07.157

    Article  CAS  PubMed  Google Scholar 

  20. Andrianaki AM, Kyrmizi I, Thanopoulou K et al (2018) Iron restriction inside macrophages regulates pulmonary host defense against Rhizopus species. Nat Commun 9:3333. https://doi.org/10.1038/s41467-018-05820-2

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Tabassum T, Araf Y, Moin AT et al (2022) COVID-19-associated-mucormycosis: possible role of free iron uptake and immunosuppression. MolBiol Rep 49:747–754. https://doi.org/10.1007/s11033-021-06862-4

    Article  CAS  Google Scholar 

  22. Cooter RD, Lim IS, Ellis DH, Leitch IO (1990) Burn wound zygomycosis caused by Apophysomyceselegans. J Clin Microbiol 28(9):2151–2153. https://doi.org/10.1128/jcm.28.9.2151-2153.1990

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Hasanin MS, Hashem AH, Abd El-Sayed ES et al (2020) Green ecofriendly bio-deinking of mixed office waste paper using various enzymes from Rhizopusmicrosporus AH3: efficiency and characteristics. Cellulose 27:4443–4453. https://doi.org/10.1007/s10570-020-03071-3

    Article  CAS  Google Scholar 

  24. Peraza-Reyes L, Malagnac F (2016) 16 Sexual development in fungi. In: Wendland J (eds) Growth, differentiation and sexuality. The Mycota, vol 1. Springer, Cham. https://doi.org/10.1007/978-3-319-25844-7_16

  25. Karthaus M (2011) Prophylaxis and treatment of invasive aspergillosis with voriconazole, posaconazole and caspofungin—review of the literature. Eur J Med Res 16:145. https://doi.org/10.1186/2047-783X-16-4-145

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Feys S, Gonçalves SM, Khan M, Choi S, Boeckx B, Chatelain D, Cunha C, Debaveye Y, Hermans G, Hertoghs M, Humblet-Baron S (2022) Lung epithelial and myeloid innate immunity in influenza-associated or COVID-19-associated pulmonary aspergillosis: an observational study. Lancet Respir Med 10:1147–1159. https://doi.org/10.1016/S2213-2600(22)00259-4

    Article  CAS  PubMed  Google Scholar 

  27. Chai LY, Vonk AG, Kullberg BJ, Verweij PE, Verschueren I, Van Der Meer JW, Joosten LA, Latgé JP, Netea MG (2011) Aspergillus fumigatus cell wall components differentially modulate host TLR2 and TLR4 responses. Microbes Infect 13:151–159. https://doi.org/10.1016/j.micinf.2010.10.005

    Article  CAS  PubMed  Google Scholar 

  28. Speth C, Rambach G, Lass-Flörl C, Howell PL, Sheppard DC (2019) Galactosaminogalactan (GAG) and its multiple roles in Aspergillus pathogenesis. Virulence 10:976–983. https://doi.org/10.1080/21505594.2019.1568174

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Kim YM, Shin EC (2021) Type I and III interferon responses in SARS-CoV-2 infection. ExpMol Med 53:750–760. https://doi.org/10.1038/s12276-021-00592-0

    Article  CAS  Google Scholar 

  30. Yao Y, Subedi K, Liu T et al (2022) Surface translocation of ACE2 and TMPRSS2 upon TLR4/7/8 activation is required for SARS-CoV-2 infection in circulating monocytes. Cell Discov 8:89. https://doi.org/10.1038/s41421-022-00453-8

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Scioli MG, Storti G, D’Amico F, Rodríguez Guzmán R, Centofanti F, Doldo E, Céspedes Miranda EM, Orlandi A (1995) Oxidative stress and new pathogenetic mechanisms in endothelial dysfunction: potential diagnostic biomarkers and therapeutic targets. J Clin Med 2020:9. https://doi.org/10.3390/jcm9061995

    Article  CAS  Google Scholar 

  32. Beserra DR, Alberca RW, Branco ACCC, Oliveira LM, de Souza Andrade MM, Gozzi-Silva SC, Teixeira FME, Yendo TM, da Silva Duarte AJ, Sato MN (2022) Upregulation of PD-1 expression and high sPD-L1 levels associated with COVID-19 severity. J Immunol Res. https://doi.org/10.1155/2022/9764002

    Article  PubMed  PubMed Central  Google Scholar 

  33. Wurster S, Robinson P, Albert ND, Tarrand JJ, Goff M, Swamydas M, Lim JK, Lionakis MS, Kontoyiannis DP (2020) Protective activity of programmed cell death protein 1 blockade and synergy with caspofungin in a murine invasive pulmonary aspergillosis model. J Infect Dis 222:989–994. https://doi.org/10.1093/infdis/jiaa264

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Gafa V, Remoli ME, Giacomini E, Severa M, Grillot R, Coccia EM (2010) Enhancement of anti-Aspergillus T helper type 1 response by interferon-β-conditioned dendritic cells. Immunology 131:282–288. https://doi.org/10.1111/j.1365-2567.2010.03302.x

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Heitman J, Carter DA, Dyer PS, Soll DR (2014) Sexual reproduction of human fungal pathogens. Cold Spring Harbor Perspect Med. https://doi.org/10.1101/cshperspect.a019281

    Article  Google Scholar 

  36. Sivasankarapillai VS, Pillai AM, Rahdar A, Sobha AP, Das SS, Mitropoulos AC, Mokarrar MH, Kyzas GZ (2020) On Facing the SARS-CoV-2 (COVID-19) with combination of nanomaterials and medicine: possible strategies and first challenges. Nanomaterials 10:852. https://doi.org/10.3390/nano10050852

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

One of the authors Dola Sundeep acknowledges the Ministry of Education, Government of India for providing financial support to conduct this study. Kovuri Umadevi acknowledges the Department of Pathology, Government Medical College and Hospital, Nizamabad for providing the slides to conduct the study.

Funding

Ministry of Education, Government of India, is acknowledged for providing financial assistance in to conduct this study.

Author information

Authors and Affiliations

  1. Department of Pathology, Government Medical College and Hospital, Khaleelwadi, Nizamabad, Telangana, 503001, India

    Kovuri Umadevi & Rajarikam Nagarjuna Chary

  2. Biomedical Research Laboratory, Department of Electronics and Communication Engineering, Indian Institute of Information Technology Design and Manufacturing, Jagannathagattu Hill, Kurnool, Andhra Pradesh, 518008, India

    Dola Sundeep & Eswaramoorthy K. Varadharaj

  3. Biomedical Research Laboratory, Department of Mechanical Engineering, Indian Institute of Information Technology Design and Manufacturing, Jagannathagattu Hill, Kurnool, Andhra Pradesh, 518008, India

    Chebbiyam Chandrasekhara Sastry

  4. Department of Pathology, All India Institute of Medical Sciences, Mangalagiri, Andhra Pradesh, 522503, India

    Arundhathi Shankaralingappa

  5. Department of Mechanical Engineering, Indian Institute of Technology (IIT-BHU), Varanasi, Uttar Pradesh, 221005, India

    Alluru Raghavendra Vighnesh

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  1. Kovuri Umadevi
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Contributions

K.U: Conceptualization (equal); Data curation (lead); Formal analysis (lead); Investigation (equal); Methodology (equal); Writing-original draft (lead); Writing-review & editing (equal). D.S.: Conceptualization (equal); Methodology (equal); Formal analysis (lead); Investigation (equal); Writing-review & editing (lead). E.K.V: Investigation (equal) Methodology (equal). C.C.S.: Investigation (equal); Methodology (equal); Writing-review & editing (equal). A.S. Investigation (equal); Methodology (equal). R.N.C: Investigation (equal); Methodology (equal). A.R.V: Supervision (supporting); Writing-review & editing (supporting).

Corresponding authors

Correspondence to Kovuri Umadevi or Dola Sundeep.

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Conflict of interest

All the authors declare that we don’t have any financial or any other conflict of interests.

Ethical Approval

This study was conducted following the necessary ethical approvals, as delineated by the Institutional Ethical Committee (IEC), Government Medical College and Hospital, Nizamabad, India, under the reference ECR/1896/Inst/TG/2023. The research was undertaken in strict accordance with the ethical principles laid down in the 1964 Declaration of Helsinki and its subsequent amendments. This included an extensive review and approval by the named institutional ethics committee, ensuring adherence to established ethical norms. Any potential doubts regarding ethical standards were addressed and explicitly approved by the committee. The authors declare that the manuscript is prepared by obeying the Ethical Standards as described in the Committee on Publication Ethics (COPE).

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The consent of participation was obtained a written informed consent from the patients for participation.

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The consent of publication was obtained a written informed consent from the patients for participation.

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Umadevi, K., Sundeep, D., Varadharaj, E.K. et al. Precision Detection of Fungal Co-Infections for Enhanced COVID-19 Treatment Strategies Using FESEM Imaging. Indian J Microbiol (2024). https://doi.org/10.1007/s12088-024-01246-1

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