Summary of diagnosis and clinical manifestation, kinds of sample, and the image of the pathogens observed by SEM
Abstract
This chapter highlights the description of the clinical manifestation and its pathogen and the host tissue damage observed under the Scanning Electron Microscope, which helps the clinician to understand the pathogen’s superstructure, the change of host subcell structure, and the laboratory workers to understand the clinical characteristics of pathogen-induced human skin lesions, to establish a two-way learning exchange database with vivid images
Keywords
- Fungi
- Bacteria
- Parasite
- Clinical Skin Samples
- SEM
1. Introduction
In dermatovereology department, skin infections by fungi, bacteria, and, parasites are very common in routine clinical practice. Differentiation and identification of these pathogens are a huge challenge and very important for the patient’s diseases diagnosis and treatment. Scanning electron microscope (SEM) is a very strong tool for detection and observation of pathogens from the clinical samples that helps us obtain a direct proof of the pathogen on the surface of the skin samples of the lesion. Based on the detailed morphologic image, we can recognize the ultrastructural of the pathogen and understand the pathogenesis of the skin-infected diseases. During recent years, we collected a lot of pathogenic microorganisms’ photographs taken by SEM. These pathogens include fungi (
|
|
|
|
Tinea capitis | Erythema, scales on the scalp; hair broken and hair loss | Infected hair | Fungus ( |
Tinea capitis | Excessive scales and hair loss on the scalp | Infected hair | Fungus ( |
|
Slightly pruritic, monomorphic follicular papules and pustules | Keratotic plug of pustule of hair follicle | Fungus ( |
Pityriasis versicolor | Erythema and scaly, hyperpigmentation or hypopigmentation of skin | Scales | Fungus ( |
Mucormycosis | Progressive red plaque around the inner canthus | Cultured colony | Fungus ( |
Mucormycosis | Purulent granuloma of left forearm | Cultured colony | Fungus ( |
Cutaneous alternariosis | An ulcer covered with crust on left anterior tibia | Cultured colony | Fungus ( |
Chromoblastomycosis | Red plaque in the left knee | Cultured colony | Fungus ( |
Primary laryngeal aspergillosis |
Hoarseness, severe paroxysmal coughing and tachypnea | Biopsy tissue | Fungus ( |
Acne | Recurrent papule and pustule acne | Pustule | Bacteria ( |
Pediculosis pubis | Intense itching of the scalp | Parasite | Parasite ( |
Demodicid mites | Itching, multiple erythema, papules, pustules | Hair follicle plug | Parasite ( |
Table 1.
2. Methods
All samples for SEM were taken from clinical patients. These samples included infected hair, scales, colony of culture, and tissue of skin biopsy. The samples for SEM were fixed in 2% glutar-aldehyde for 4 h at 4 °C, dehydrated through four gradations of alcohol solutions (50%, 70%, 95%, 100%, progressively) for 15 min each, then soaked in isoamyl acetate for 30 min. The specimens were prepared after critical-point drying method, under which condition they were gilded in a vacuum chamber and observed under the SEM, FEI Inspect F50, equipped with an FEG gun operated at 30 kV at high vacuum.
3. Results
3.1. Tinea capitis
Tinea capitis is a common superficial fungal infection of scalp hair follicles and surrounding skin. It often affects children rather than adults. Its pathogens are dermatophytes, usually species in the genera

Figure 1.
a. A 9-year-old boy, weighing 25kg, presented to our clinic with slightly itching, multiple patchy areas of gray scaling lesions on the scalp and obvious hair loss.
We describe two cases of tinea capitis due to

Figure 2.
a-b. Cuticle layers of hair shaft were seriously destroyed and a large part had been lost, exposing the fibril cortex inside which many arthrospores were noted. c. High magnification of “a” showing the cuticle layers completely destroyed with the residual cortex fully filled with arthroconidia (A) and distorted fibril bundles (F). d. An arthrospore (A) at high magnification showing irregular convex granules on the chitinous surfaces and the poles bordered by a protruding ring structure (R), bulged by a hemispherical convex (HC). Numerous residual fibril fragments (F) noted around the spore as well as on its surface.
The second patient is a 5-year-old boy in good health, weighing 19 kg and presented at our clinic with a 1-month history of excessive scales and hair loss on the scalp (Fig. 3a). He had been previously diagnosed with tinea capitis in a local hospital, and received oral itraconazole 100 mg per day for 14 days administered with water. However, the area of hair loss enlarged slightly. Additionally, he had a history of direct contact with a pet dog. Direct microscopic examination (with 10% KOH) of broken hair strands showed numerous spores inside as well as outside of the hair strand. Simultaneously, strands were observed under SEM, and there were many round spores in and around the hair strand (Fig. 4a, b). Fungal culture revealed yellow filamentous colonies, which were identified as

Figure 3.
a. A 5-year-old boy presented with 1-month history of scalp scales and hair loss, who had received oral itraconazole 100 mg per day with water for 14 days; b. The patch with hair loss was smaller and without scale after oral itraconazole 100 mg per day with whole milk for 14 days; c. There was no apparent hair loss on scale after 40 days at end of treatment.

Figure 4.
a-b. After oral itraconazole 100 mg per day with water for 2 weeks, broken hair strands with many round spores in and around the hair strands were evident under scanning electron microscopy (SEM); c-d. Oral itraconazole 100 mg per day with whole milk for 14 days, the number of spores were markedly reduced in broken hair strands, and spores appeared very irregular under SEM.
3.2. Malassezia folliculitis (Pityrosporum folliculitis)
The following is a case of

Figure 5.
a. A 25-year-old man with complains of slightly pruritic, monomorphic follicular papules, pustules, and secondary keloid on the upper trunk and neck. b-c. SEM of the hair follicle from the upper trunk. These demonstrated a large number of globular or orbicular-ovate yeasts of budding daughter cell, with collar structure around the budding. b. Globular yeast. c. Orbicular-ovate and globular yeast in the same sample.
3.3. Pityriasis versicolor
Pityriasis versicolor is a superficial fungal infection of the skin and caused by
This is a case of pityriasis versicolor due to

Figure 6.
a. A 27-year-old man presented in our clinic with extensive erythema and scaly for 6 months. b. Under SEM, numerous hyphae (H) went through the scaly, length of which is about 10–20 μm. c. Under SEM, abundant of 3–5 μm in diameter grapes-like spherical
3.4. Mucormycosis
Mucormycosis is a clinically rare and fatal opportunistic fungal infection, which invades nasal, brain, lung, gastrointestinal tract, skin, and other parts with acute, subacute, or chronic process. The routes of invasion contain respiratory tract, digestive tract, skin, and neonatal umbilical region [10]. Its pathogens are fungi in the order
The following is a description of two cases of primary cutaneous mucormycosis caused by

Figure 7.
a. A 47-year-old farmer was presented to our clinic with 1-year history of progressive red plaque around the inner canthus. Faint yellow exudation was oozing from the ulceration at the center of plaque. Some scales were also observed on the plaque. b. SEM observations revealed non-apophysate sporangia with pronounced columellae and conspicuous collarette at the base of the columella following sporangiospore dispersal.
The other case is of a 69-year-old female farmer, who presented to our clinic with the history of a progressive purulent granuloma of her left forearm (Fig. 8a) following a fracture of left forearm about 11 months earlier. Broad, nonseparate hyphae were seen in pathologic study with methenamine silver stain (Fig. 8b). Fungal culture revealed white filamentous colonies that were identified as

Figure 8.
a. A 69-year-old female was presented to our clinic with a progressive purulent granuloma of her left forearm. b. Broad nonseparate hyphae were seen in pathologic study (methenamine silver stain, ×200). c-d. The sporangiophores of
3.5. Cutaneous alternariosis
We describe in the following is a rare case of a healthy individual with cutaneous alternariosis due to infection with

Figure 9.
a. Ulcer with an overlying crust on the patient’s skin of left anterior tibia. b. Fungal culture of the tissue revealed dark grey-white colonies with a dark-brown underside. c. SEM observation of slide culture revealed beaked conidia.
3.6. Chromoblastomycosis
Chromoblastomycosis is a chronic fungal infection of the skin and subcutaneous tissue caused by dematiaceous fungi. Common pathogenic fungi are
In this part we describe a case of chromoblastomycosis due to

Figure 10.
a. A 34-year-old male with a 12-year history of a red plaque in the left knee. b. Under SEM observation: dematiaceous hyphae with many well-defined septa, conidiophores, and oval brown spores arranged in a clump could be seen. The surfaces of conidiogenous cells were smooth. Oval spores were arranged around conidiophores.
3.7. Primary laryngeal aspergillosis
Primary laryngeal aspergillosis is a rare opportunistic infection caused by
We describe in the following a case of primary laryngeal aspergillosis due to

Figure 11.
a. A 23-year-old female undergraduate student presented with hoarseness, severe paroxysmal coughing, and tachypnea. Laryngoscopy revealed obvious white plaques on the swollen vocal cords and laryngeal ventricle. b. SEM of the biopsy tissue revealed hyphae branching at 45° angles destroyed the vocal cords tissue.
3.8. Acne
Acne is a chronic inflammatory disease of the sebaceous–pilosebaceous system. It is estimated to affect 9.4% of the global population [17]. Acne is closely related to the combination of genetic and environmental factors, among which
The following is a description of a case of acne in a 24-year-old girl. She suffered recurrent papule and pustuleacne for 6 months (Fig. 12a-b). We removed the follicular plug with sterile hemostatic forceps and observed it through SEM. Under SEM, abundant rod-shaped bacteria were closely spaced in follicular plug tissue (Fig. 12c). Treatment with oral minocyline 50 mg twice a day and topical use of adapalene gel was effective.

Figure 12.
a. A 24-year-old girl who suffered recurrent papule and pustule acne for 6 mouths. b. Under the dermoscopy, the papule was semisphere with tawny follicular plug inside. c. Under SEM, rod-shaped bacteria were closely spaced in follicular plug tissue.
3.9. Pediculosis
Pediculosis is a skin disease caused by arthropods. Its pathogens are three lice species including head louse, crab or pubic louse, and body louse, which cause the
We describe a case of pediculosis on the scalp of a 6-year-old boy caused by the crab louse [23]. The boy was presented to our clinic with the complaint of intense itching of the scalp. There were some small pieces of erythema (in the circle) and a brown dot-like substance on his scalp (Fig. 13a). The dermoscopy revealed a brown parasite (0.9 mm in horizontal axes and 1.2 mm in vertical axes) with two crab-like feet adhered to the scalp (Fig. 13b). Microscopic examination and scanning electron microscope showed the detail of this insect (Fig. 13c-d). Based on these morphological findings, the diagnosis of

Figure 13.
a. There were some small pieces of erythema (in the circle) and a brown dot-like substance on the scalp (arrow). b. The dermoscopy revealed a brown parasite (0.9 mm in horizontal axes and 1.2 mm in vertical axes) with two crab-like feet adhered to the scalp. c. Under the microscope, the parasite was characterized by a flat body like a crab and three pairs of feet in different sizes. There was an area (red box) full of blood in the middle part of the parasite. A large number of short setae (arrow) were noted at the edge of the parasite abdomen. d. The SEM showed a vivid three-dimensional ultrastructure of the parasite: the whole body was composed of three parts including spherical head, chest, and elliptical abdomen; a pair of feelers was noted on the head; the three pairs of feet were in shaped section and curved serrated claws were noted at the end of foot; short setae in different length were not only at the edge of the abdomen but also on the feet.
3.10. Demodiciosis
Demodiciosis is a kind dermatitis caused by
The following is a description of a case of demodicosis due to

Figure 14.
a. A 28-year-old man came to our clinic because of itching, multiple erythema, papules, pustules lesions on the nose and cheek. b. SEM revealed a
References
- 1.
Ellabib MS, Agaj M, Khalifa Z, Kavanagh K. Trichophyton violaceum is the dominant cause of tinea capitis in children in Tripoli, Libya: Results of a two year survey.Mycopathologia . 2001; 153: 145-147. - 2.
Yu J, Li R, Bulmer G. Current topics of tinea capitis in China. Jpn J Med Mycol . 2005; 46: 61-66. - 3.
Zhuang K, Ran X, Lei S, Zhang C, Lama J, Ran Y. Scanning and transmission electron microscopic observation of the parasitic form of Trichophyton violaceum in the infected hair from tinea capitis.Scanning . 2014; 36: 465-470. - 4.
Chen S, Ran Y, Dai Y, Lama J, Hu W, Zhang C. Administration of oral itraconazole capsule with whole milk shows enhanced efficacy as supported by scanning electron microscopy in a child with tinea capitis due to Microsporum canis .Pediatric Dermatology . 2015; 32:e312–e313 - 5.
Rubenstein RM, Malerich SA. Malassezia (Pityrosporum ) folliculitis.J Clin Aesthet Dermatol . 2014; 7: 37-41. - 6.
Hald M, Arendrup MC, Svejgaard EL, Lindskov R, Foged EK, Saunte DM. Evidence-based Danish guidelines for the treatment of Malassezia -related skin diseases.Acta Derm Venereol . 2015; 95: 12-19. - 7.
Renati S, Cukras A, Bigby M. Pityriasis versicolor. BMJ . 2015; 350: 1394-1400. - 8.
Kaushik A, Pinto HP, Bhat RM, Sukumar D, Srinath MK. A study of the prevalence and precipitating factors of pruritus in pityriasis versicolor. Ind Dermatol Online J . 2014; 5: 223-224. - 9.
Kyriakis KP, Terzoudi S, Palamaras I, Pagana G, Michailides C, Emmanuelides S. Pityriasis versicolor prevalence by age and gender. Mycoses . 2006; 49: 517-518. - 10.
Kang D, Jiang X, Wan H, Ran Y, Hao D, Zhang C. Mucor irregularis Infection around the inner canthus cured by amphotericin B: A case report and review of published literatures.Mycopathologia . 2014; 178: 129-133. - 11.
Lu W, Lu J, Ran Y, Lin Z, Wan H, Cui F, Cao L, Pan N, Song X, Chen J, Wanf Y, Yu M. Cutaneous and subcutaneous coinfection by Lichtheimia corymbifera andCandida parapsilosis : a case report.Chin J Dermatol . 2012; 45(10):727-730. (in Chinese) - 12.
Anaissie EJ, Bodey GP, Rinaldi MG. Emerging fungal pathogens. Eur J Clin Microbiol Infect Dis . 1989; 8: 323-330. - 13.
Hu W, Ran Y, Zhuang K, Lama J, Zhang C. Alternaria arborescens infection in a healthy individual and literature review of cutaneous alternariosis.Mycopathologia . 2015; 179: 147-152. - 14.
Krzyściak PM, Pindycka-Piaszczyńska M, Piaszczyński M. Chromoblastomycosis. Adv Dermatol Allergol/Postepy Dermatologii i Alergologii . 2014;31(5):310-321. doi:10.5114/pdia.2014.40949. - 15.
Ran Y, Yang B, Liu S, Dai Y, Pang Z, Fan J, Bai H, Liu S. Primary vocal cord aspergillosis caused by Aspergillus fumigatus and molecular identification of the isolate.Med Mycol . 2008; 46: 475-479. - 16.
Ran Y, Li L, Cao L, Dai Y, Wei B, Zhao Y, Liu Y, Bai H, Zhang C. Primary vocal cord aspergillosisand scanning electron microscopical observation of the focus of infection. Mycoses 2011;54:e634-637. - 17.
Ran Y, Lu Y, Cao L. Primary laryngeal aspergillosis related to oral sex? A case report and review of the literature. Med Mycol Case Rep . 2013; 2: 1-3. - 18.
Tan JK, B Ran Y, Lu Y, Cao L.hate K. A global perspective on the epidemiology of acne. Br J Dermatol . 2015; 172; Suppl 1: 3-12. - 19.
Antiga E, Verdelli A, Bonciani D, Bonciolini V, Caproni M, Fabbri P. Acne: a new model of immune-mediated chronic inflammatory skin disease. G Ital Dermatol Venereol . 2015; 150: 247-254. - 20.
Yu Y, Champer J, Garbán H, Kim J. Typing of Propionibacterium acnes : a review of methods and comparative analysis.Br J Dermatol . 2015; 172: 1204-1209. - 21.
David C, Flinders, Peter DS. Pediculosis and scabies. Am Fam Physician . 2004;69: 341-348. - 22.
Ko CJ, Elston DM. Pediculosis. J Am Acad Dermatol . 2004, 50: 1-12. - 23.
Gunning K, Pippitt K, Kiraly B, Sayler M. Pediculosis and scabies: treatment update. Am Fam Physician. 2012; 15: 535-541. - 24.
Ran Y, Feng X, Zhuang K, Zhang C. Dermatoscopy, microscopy and scanning electron microscopy diagnosed scalp pediculosis pubis in a child. J Clin Dermatol . 2014;43(12):725-726. (in Chinese) - 25.
Rusiecka-Ziółkowska J, Nokiel M, Fleischer M. Demodex – an old pathogen or a new one? Adv Clin Exp Med . 2014; 23: 295-298. - 26.
Elston CA, Elston DM. Demodex mites. Clin Dermatol . 2014; 32: 739-743. - 27.
Rufli T, Mumcuoglu Y. The hair follicle mites Demodex folliculorum andDemodex brevis – biology and medical importance: a review.Dermatologica . 1981;162:1-11.