Journal of Current Biomedical Reports  jcbior.com 

Volume 2, Number 2, 2021                                                                                                    eISSN: 2717-1906 

1 

Brief report 

Decrease of microbial contamination by application of 
routinely glycerol and phenol solution on human cadavers 

 

Masoumeh Faghani Langroudi1, Ali Farzanegan2,*  
 

1Department of Anatomical Sciences, School of Medicine, Guilan University of Medical Science, Guilan, Iran 
2Department of Microbiology, Guilan University of Medical Science, Anzali International campus, Guilan, Iran 

 

Abstract 
Human cadaver is imperative learning material for medical students. The cadavers expose to microbial 
contamination every time anatomists work with them. The microorganisms found in preparations’ cadavers 
could be pathogenic. Although the cadaver has supported a preservation method but does not inhibit microbial 
germination. The purposes of this study were evaluation the use of the routine solution to decrease potential 
contamination.  Samples were collected air dissection hall and cadavers' sections after drying the cadavers, and 
after use of routinely glycerin and phenol solution (1.5 g phenol in 100 cc glycerol), inoculated on eosin 
methylene blue agar (EMB), blood agar (BA), Sabouraud dextrose agar (SDA), and SDA with the addition of 
chloramphenicol (50 mg/l in ethanol solution) (SC), in a Petri dish and incubated at 25 °C and 37 °C. After 3-
5 days, the morphology was studied macroscopically and microscopically. We found Sporothrix schenckii; 
Streptomyces, Bacillus subtilis and Aspergillus flavus, Geotrichum candidum, on the samples of dryness 
cadavers and air dissection hall, respectively. But fungus and bacteria contamination was decreased after the 
use of glycerin and phenol solution. Based on the results of this study, it is suggested that glycerol and phenol 
solution can be used at least once a week to reduce microbial contamination and prevent the spread of the 
disease from the cadavers to students and professors. 

 Keywords: Cadaver, Microbial contamination, Glycerol, Phenol 

 
1. Introduction  
Cadavers are the main studying materials of 

anatomists. Cadavers are a teaching tool for teachers 
and students but may infect persons who work with 
them. All of those are potentially in danger of 
exposure to pathogenic microorganisms like fungi, 
bacteria, and viruses. The infectious agent in the 
cadavers includes Mycobacterium tuberculosis, 
hepatitis B and C viruses, human immunodeficiency 
virus, Aspergillus, Candida, Penicillium, etc. Specific 
care protections are necessary to avoid accidental 
disease transmission from cadavers before and 
during dissection [1]. Therefore, the human cadavers 
in medical universities usually were protected for a 
very long time in a fixative solution, which contains 
formalin, alcohol, phenol, water, and glycerin. Many 
of the laboratories in the departments of human 
anatomy ignore the damaging effect of air dryness on 

                                                           
 * Corresponding author:  
Ali Farzanegan, MSc 
Thirty Metery Blvd. Anzali, Guilan, Iran. Zip Code: 4314637758 
Tel/Fax: +98 911 982 9233 
Email: alifarzanegan1349@gmail.com 
https://orcid.org/0000-0002-2425-8634 
 
Received: February, 04, 2021 
Accepted:, February, 28, 2021  

the cadaveric sections [2]. Airborne fungi are the 
most important microorganism in the air, which 
causes contamination problems in the environment. 
Generally, saprophytic’ fungi in many ecosystems 
can cause respiratory system infections, especially in 
immunosuppressed people [3].  

Drying and hardening cadaveric tissue and fungi 
contamination is one of the problems predictable 
that to occur due to a long time of exposure cadaver 
to air drying, but this problem probably resolves with 
processed cadaveric with routinely use glycerin and 
phenol solution. Due to the activity of bacteria and 
fungi on cadavers that are drying, probably many 
cases of destruction will be seen in cadavers. In 
addition to economic damage and trouble in 
teaching, this subject can be cases of contamination 
in the dissection hall and tools, and subsequently 
infection the staff, students, and professors [4].  

  © The Author(s) 2021 

https://jcbior.com/


Faghani Langroudi et al. 

2 

This study aimed to evaluate the use of routinely 
phenol and glycerin solution to decrease the fungal 
contamination in the university dissection hall.  

 
2. Materials and Methods 
2.1 Study design 
The cadavers used for this study were obtained 

from the Department Of Human Anatomy, in the 
dissection Hall of Guilan University of Medical 
Sciences, Anzali International Campus. The 
dissection hall was 2.7 meters in height, 10.8 meters 
in length, and 10 meters in width. The room has a 
single door and five windows with 1.4 by 1.5 meters. 
The floor and the walls had been tile up to the ceiling. 
The room has two autopsy tables made of stainless 
steel. The air conditioning dissection hall is 
administered by a simple fan system on the south 
wall. The dissection hall has three radiator heating 
systems and they were on at the time of the 
experiment (Figure 1). 

 
2.2 Sample collection 
All samples were taken from different surfaces of 

cadavers by the sterile cotton swap, after drying the 
cadavers, and one week after the use of routinely 
phenol and glycerin solution (1.5 g phenol in 100 cc 
glycerol).  In addition, Petri dishes with a diameter of 
8 cm were put at dissection hall for taken airborne 
microorganisms for 24 h [5, 6].  

 
2.3 Culture 
After collection, each sample was used to 

inoculate four different media: Eosin methylene blue 

agar (EMB) to suppress Gram-positive bacterial 
growth, Blood Agar (BA), Sabouraud dextrose agar 
(SDA), SDA with the addition of chloramphenicol 
(50mg/l in ethanol solution) (SC) to suppress 
bacterial growth. Agar samples were incubated at 25 
°C and 37 °C and colony growth was checked daily. 

 
2.4 Samples Staining  
All fungi isolate were stained by lactophenol 

cotton blue and bacterial isolate were stained by 
Gram stain and acid-fast stain. All samples were 
identified by comparing the microscopic 
characteristics of fungi and bacterial standard 
produedures. 

 

 

 

 

Table 1. The culture results of samples from the surface of the cadavers and the air-dissection hall 

 

Sample 

source 

Fungi growth in SC  Bacterial growth in  BA 

Sample 

of head 

Sample 

of neck 

Sample of 

hand 

Sample of 

foot 

Sample 

of 

visceral 

Sample 

of  

head 

Sample 

of  

neck 

Sample 

of 

hand 

Sample 

of  foot 

Sample of  

visceral 

Cadaver 

1  

No 

growth 

No 

growth 

No 

growth 

Sporothrix 

schenckii 

No 

growth 

No 

growth 

No 

growth 

No 

growth 

No 

growth 

No 

growth 

Cadaver 

2 

No 

growth 

No 

growth 

Sporothrix 

schenckii 

Sporothrix 

schenckii 

No 

growth 

No 

growth 

No 

growth 

No 

growth 

Bacillus 

subtilis 

Streptomyces 

spp. 

 
Fungi growth in SC and SDA 

South windows West windows Eastern door North wall Floor 

Air-

dissection 

hall 

Aspergillus flavus Sporothrix schenckii 
Geotrichum 

candidum 

Geotrichum 

candidum 
Aspergillus flavus 

 
EMB: Eosin methylene blue; BA: Blood agar; SDA: Sabouraud dextrose agar; SC: Sabouraud dextrose agar with the addition of chloramphenicol (50 mg/l in 

ethanol solution) 

 

 

 

 

 
 

Figure 1.  The plan of the autopsy room in the dissection 

hall of the university 



Faghani Langroudi et al. 

3 

3. Results  
All the samples were culture in the department of 

microbiology in Anzali International Campus. All of 
the sample results from the air-dissected hall were 
positive for fungal growth, also five samples from 
different surfaces of the preserved cadaver were 
positive for fungal and bacterial growth (Table 1). 
Two different bacterial colonies were observed after 

72 h on BA and three different fungi colonies were 
observed after 3-5 days on SDA and SC media (Table 
2). 

All samples culture from the cadaver surface took 
after using glycerin and phenol solution were 
negative. Also, the samples culture from the 
dissection hall was taken after cleaning were still 
positive, but they were reduced. 

Table 2. Colonial and microscopic morphology and biochemical identification of organisms 
 

Organism 

Colonial morphology and characteristics 
Biochemical 

identification 

Microscopic 

characteristics 

BA EMB SC and SDA 

Bacillus subtilis 

Brown color, 

mucoid surface, 

wrinkled -
shaped, 

hemolytic effect 

Inhibition - 
Catalase (+), indole 

reaction (-), lactose (-) 
Gram-positive,  rod shape 

Streptomyces spp. 

Red-pink color, 

mucoid surface, 
soil smell 

Inhibition - 

Indole reaction (-), SH2 

(-), motility (+), Gas (-) , 

citrate (+), catalase (+) 

Acid from lactose(-), 

Acid from glucose (+), 
Urea hydrolysis(+) 

Gram-variable, motile, 

filamentous, non - acid-
fast 

Aspergillus flavus - - 

Granular, flat, with 

radial grooves, white 
at first but quickly 

becoming yellow to 

dark yellow-green with 
age 

- 

Typically radiate, loose 

columns, biseriate or 
uniseriate, directly on the 

vesicle (uniseriate). 

Conidiophores are hyaline 
and roughened, often more 

visible near the vesicle. 

Conidia are spherical, pale 
green, and echinulate 

Geotrichum 

candidum 
- - 

Fast-growing, flat, 

white to cream, dry, 

suede-like with no 

reverse pigment 

- 

Hyaline hyphae with 

septate branched and break 
up into chains of hyaline, 

smooth, one-celled, and 

cylindrical arthroconidia 

Sporothrix 

schenckii 
- - 

Slow-growing, 

compact and suede-

like or floccose, dry, 
flat, brownish-black, 

grey color, dark-brown 

diffusible pigment 

- 

Hyaline and produce 
mostly simple awl-shaped 

erect phialides with 

inconspicuous collarettes. 
Conidia are hyaline or 

pigmented, spherical to 

cylindrical, one-celled, and 
typically aggregated in 

heads at the apex of each 

phialide. chlamydospores 
may be present. 

 

EMB: Eosin methylene blue; BA: Blood Agar; SDA: Sabouraud dextrose agar, SC: Sabouraud dextrose agar with the addition of chloramphenicol (50 mg/l in 
ethanol solution) 

 

 

 



Faghani Langroudi et al. 

4 

4. Discussion 
The human cadavers in medical colleges usually 

were preserved in a fixative solution, which contains 
formalin, alcohol, phenol, water, and glycerin but 
after staying in the dissection hall for a long time, 
they will dry out. In this research study, not only the 
surfaces of the cadavers were examined for the 
presence of viable microorganisms but also the air-
dissection hall was examined for dangerous 
infectious agents. Three different fungal colonies 
identified as Sporothrix spp. (from the surface of the 
cadavers), Aspergillus spp. and Geotrichum (from 
the air-dissection hall) and two bacteria colonies 
identified as Streptomyces and Bacillus (from the 
surface of the cadavers), the source of 
contamination. Those are microorganisms that 
transmitted by air and cause opportunistic disease 
[7-11]. This is a matter of concern because students 
and anatomists may be exposed to the infectious 
disease every time they work with a cadaver [1]. 

Normally, 10% formalin or 4% formaldehyde in 
a water solution is used to prevent the contamination 
of microorganisms. Furthermore, formalin-fixed 
human cadavers are routinely used for study and 
research in hospitals and universities. Airborne fungi 
have become a serious problem in dissection hall. 
Therefore, the presence of airborne fungi on 
embalmed human cadavers could be perilous for 
teachers and students and necessary measures must 
be taken to reduce fungi contamination. One report 
has suggested that formalin concentration for 
inhibiting fungal growth should not be less than 4% 
(5‒7.5 percentage) [12]. Previously, it was showed 
that 15% formalin was very effective in the 
prevention of growth of bacteria, fungi, and also 
decay and discoloration [3]. The main factors leading 
to fungal contamination in the dissection hall are 
room humidity, the concentration of airborne fungal 
spores, and the duration of time to fungal 
inoculation. All instructors use routinely glycerin and 
phenol solution in the dissection hall to prevent 
drying and decrease microbial contamination of 
human cadavers after work with cadavers. 

Glycerol (glycerin) is a simple polyol complex 
that uses in the dissection hall. It is colorless, 
odorless, sweet-tasting, and non-toxic. Multiple 
journals describe the antimicrobial and antiviral 
effects of glycerol [13], as well as, Glycerol catalyzes 
spore germination at about 64–70% humidity for 
microbial life. The germination of fungal xerophiles 
can occur at 60% moisture but for the most 
dangerous fungal xerophiles, metabolic activity and 
cell division typically stop between 64–70% 

humidity. These findings describe an inconsistency. 
On the one hand, glycerol can be particularly 
stressful and prevent cellular development. On the 
other hand, may be essential for cells (microbiota) to 
function at the water-activity limit for life, and used 
routinely in cadaver dissection hall [14]. 

Phenol derivatives isolated from the plants 
showed good antifungal activity that uses in the 
dissection room for the elimination of microbiota 
contaminations [15]. Phenol, or carbolic acid, is a 
colorless or white crystalline solid with a partly low 
melting point. It has been shown that phenol 
becomes bactericidal/fungicidal at concentrations of 
1.0–1.5% and destroys cell walls. Phenol is an 
excellent fungicide and bactericide but it denatures 
proteins, with resultant drying and discoloration of 
tissues, and has a nasty odor. Phenol is harmful to 
the esophagus, causing vomiting, and headache, and 
faintness, loss of blood pressure, pulmonary edema, 
and cyanosis [16]. 

Despite the use of phenol for decreased 
microbiota contamination, and the use of glycerin to 
decrease resulting phenol's dryness, we cannot 
prevent the spread of bacteria and fungi in the indoor 
air and cadavers' contaminations, even by closing the 
windows and routinely use of glycerin and phenol 
solution and installing a proper ventilation system, 
but only we can decrease contaminations. On the 
other hand, the phenol has side effects, nevertheless 
is still used in universities to disinfect cadavers. 

The use thymol for moistening the cadavers at 
the end of every dissection course, at least once a 
week. Thymol (2-isopropyl-5-methyl phenol) is 
naturally occurring and found in thyme oil. A 
thymol-ethanol solution as a humidifying solution is 
keeping at room temperature. Thymol has no 
carcinogenic or other serious effects on health but it 
has bactericidal and fungicidal effects and it is better 
than phenol [16, 17]. Suitable preservation is critical 
to the effective use of cadavers in educational 
settings. Everybody, those works in the anatomy 
department must ensure the safety of the cadaver 
from harm, decay, or disintegration [18]. 

In the present study, three different types of 
fungus (Sporothrix, Aspergillus, and Geotrichum) 
and two bacteria (Bacillus and Streptomyces) were 
identified. The potential infectious hazard from 
human cadavers is one of the main hazards 
associated with the anatomy department. Safe 
working conditions for handling cadavers can be 
provided through proper education, use of protective 
clothing, and practice of hygienic measures. 
Embalming decreases the risk of transmission of 



Faghani Langroudi et al. 

5 

potentially infectious microbial agents, but it is not 
possible to prevent the cadavers from drying out in 
the cold season due to the heating system are work. 
The use of glycerin for moistening and prevent 
drying of the cadavers and use of thymol (instead of 
phenol), because it is not toxic and side effects, 
externally at the end of every dissection course or at 
least once a week, is necessary and recommended. 

 
Acknowledgments 
This work was supported by the Department of 

Anatomy and examined at the Microbiology 
laboratory, Anzali International Campus, Guilan 
University of Medical Science (GUMS). Finally, we 
also thank the Dean of Anzali International 
University, Dr. Seyed Mahdi Zia Zibari for his 
generosity in this study. 

 
Author Contributions 
All authors contributed equally to this 

manuscript and approved the final version of the 
manuscript. 

 
Conflict of Interests 
The authors declare that they have no conflicts of 

interest. 
 
Ethical declarations  
This study was in accordance with the 

declaration of Helsinki and an ethical permission 
was sought from the Head of the Institute (Anzali 
International Campus). 

 
Financial Support 
None to be declared.  
 
References 

1. Yaragalla S, Rajput A. Identification of Fungal Growth from the 
Internal Organs of Preserved Human Cadavers. Am J Microbiol 
Res. 2017; 5(1):25-7. 
2. Sri-Indrasutdhi V, Ueapattanakit J, Sommatas A. 
Investigation of airborne fungi and their ability to grow on 
formalin-fixed human cadavers. Mycosphere. 2015; 6(06):729-
36. 
3. Mobarak HJ. Experimental Study on the Effect of Air-Drying 
on Durability of Embalmed Human Cadavers. Iraqi J Med Sci. 
2015; 13(2):178-82. 
4. Diba K, Rahbar M, Farjah G. Microbial contamination of 
cadavers in the Urimia, Faculty of Medicine department of 
Anatomy. Iran J Basic Med Sci. 2001; 4(2):84-8. 
5. Friedlander SF, Pickering B, Cunningham BB, Gibbs NF, 
Eichenfield LF. Use of the cotton swab method in diagnosing 
Tinea capitis. Pediatrics. 1999; 104(2 Pt 1):276-9. 
6. Reynolds SJ, Streifel AJ, McJilton CE. Elevated airborne 
concentrations of fungi in residential and office environments. 
Am Ind Hyg Assoc J. 1990; 51(11):601-4. 

7. Kassamali H, Anaissie E, Ro J, Rolston K, Kantarjian H, 
Fainstein V, et al. Disseminated Geotrichum candidum 
infection. J Clin Microbiol. 1987; 25(9):1782-3. 
8. Marcellino N, Beuvier E, Grappin R, Guéguen M, Benson DR. 
Diversity of Geotrichum candidum strains isolated from 
traditional cheesemaking fabrications in France. Appl Environ 
Microbiol. 2001; 67(10):4752-9. 
9. Orofino-Costa R, Macedo PM, Rodrigues AM, Bernardes-
Engemann AR. Sporotrichosis: an update on epidemiology, 
etiopathogenesis, laboratory and clinical therapeutics. An Bras 
Dermatol. 2017; 92(5):606-20. 
10. Cordovez V, Carrion VJ, Etalo DW, Mumm R, Zhu H, van 
Wezel GP, et al. Diversity and functions of volatile organic 
compounds produced by Streptomyces from a disease-
suppressive soil. Front Microbiol. 2015; 6:1081. 
11. Chen AJ, Hubka V, Frisvad JC, Visagie CM, Houbraken J, 
Meijer M, et al. Polyphasic taxonomy of Aspergillus section 
Aspergillus (formerly Eurotium), and its occurrence in indoor 
environments and food. Stud Mycol. 2017; 88:37-135. 
12. Viskasari PK, Lucky P, Haryanto A. The use of lower 
formalin-containing embalming solution for anatomy cadaver 
preparation. Med J Indones. 2012; 21(4):203-7. 
13. Verbeken G, Verween G, De Vos D, Pascual B, De Corte P, 
Richters C, et al. Glycerol treatment as recovery procedure for 
cryopreserved human skin allografts positive for bacteria and 
fungi. Cell Tissue Bank. 2012; 13(1):1-7. 
14. Stevenson A, Hamill PG, Medina Á, Kminek G, Rummel JD, 
Dijksterhuis J, et al. Glycerol enhances fungal germination at the 
water-activity limit for life. Environ Microbiol. 2017; 19(3):947-
67. 
15. Xu H, Zeng X. Synthesis of diaryl-azo derivatives as potential 
antifungal agents. Bioorg Med Chem Lett. 2010; 20(14):4193-5. 
16. Brenner E. Human body preservation - old and new 
techniques. J Anat. 2014; 224(3):316-44. 
17. Hammer N, Löffler S, Feja C, Sandrock M, Schmidt W, 
Bechmann I, et al. Ethanol-glycerin fixation with thymol 
conservation: a potential alternative to formaldehyde and 
phenol embalming. Anat Sci Educ. 2012; 5(4):225-33. 
18. Babb JR, Hall AJ, Marlin R, Ayliffe GA. Bacteriological 
sampling of postmortem rooms. J Clin Pathol. 1989; 42(7):682-
8. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

https://pubmed.ncbi.nlm.nih.gov/10429007/
https://pubmed.ncbi.nlm.nih.gov/10429007/
https://pubmed.ncbi.nlm.nih.gov/10429007/
https://doi.org/10.1080/15298669091370185
https://doi.org/10.1080/15298669091370185
https://doi.org/10.1080/15298669091370185
https://pubmed.ncbi.nlm.nih.gov/3477570/
https://pubmed.ncbi.nlm.nih.gov/3477570/
https://pubmed.ncbi.nlm.nih.gov/3477570/
https://pubmed.ncbi.nlm.nih.gov/11571181/
https://pubmed.ncbi.nlm.nih.gov/11571181/
https://pubmed.ncbi.nlm.nih.gov/11571181/
https://pubmed.ncbi.nlm.nih.gov/11571181/
https://pubmed.ncbi.nlm.nih.gov/29166494/
https://pubmed.ncbi.nlm.nih.gov/29166494/
https://pubmed.ncbi.nlm.nih.gov/29166494/
https://pubmed.ncbi.nlm.nih.gov/29166494/
https://pubmed.ncbi.nlm.nih.gov/26500626/
https://pubmed.ncbi.nlm.nih.gov/26500626/
https://pubmed.ncbi.nlm.nih.gov/26500626/
https://pubmed.ncbi.nlm.nih.gov/26500626/
https://pubmed.ncbi.nlm.nih.gov/28860671/
https://pubmed.ncbi.nlm.nih.gov/28860671/
https://pubmed.ncbi.nlm.nih.gov/28860671/
https://pubmed.ncbi.nlm.nih.gov/28860671/
https://doi.org/10.13181/mji.v21i4.505
https://doi.org/10.13181/mji.v21i4.505
https://doi.org/10.13181/mji.v21i4.505
https://pubmed.ncbi.nlm.nih.gov/21360142/
https://pubmed.ncbi.nlm.nih.gov/21360142/
https://pubmed.ncbi.nlm.nih.gov/21360142/
https://pubmed.ncbi.nlm.nih.gov/21360142/
https://pubmed.ncbi.nlm.nih.gov/27631633/
https://pubmed.ncbi.nlm.nih.gov/27631633/
https://pubmed.ncbi.nlm.nih.gov/27631633/
https://pubmed.ncbi.nlm.nih.gov/27631633/
https://pubmed.ncbi.nlm.nih.gov/20570508/
https://pubmed.ncbi.nlm.nih.gov/20570508/
https://pubmed.ncbi.nlm.nih.gov/24438435/
https://pubmed.ncbi.nlm.nih.gov/24438435/
https://pubmed.ncbi.nlm.nih.gov/22434588/
https://pubmed.ncbi.nlm.nih.gov/22434588/
https://pubmed.ncbi.nlm.nih.gov/22434588/
https://pubmed.ncbi.nlm.nih.gov/22434588/
https://pubmed.ncbi.nlm.nih.gov/2760229/
https://pubmed.ncbi.nlm.nih.gov/2760229/
https://pubmed.ncbi.nlm.nih.gov/2760229/