Morphological Observations On Pentatrichomonas Hominis, Enteromonas Hominis and Rodentolepis Nana

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Morphological observations on Pentatrichomonas hominis, Enteromonas

hominis and Rodentolepis nana
Article · January 2010
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TECHNICAL REPORT Table 1 ModiÞed Iron Haematoxylin Parasite Stain
STEP TIME SOLUTION

Morphological Observations on 1 5 min 70% Ethanol
Pentatrichomonas hominis, Enteromo- 2 2 min Running tap water wash
nas hominis and Rodentolepis nana
3 1 min Strong Carbol Fuchsin
Richard S. Bradbury,1,2* Carol R. Males1 and Alan Thomas1
4 1 min Running tap water wash
1
Department of Microbiology and Infectious Diseases, Royal Hobart Hospital,
Hobart, Tasmania, Australia 5 8 min Iron Haematoxylin working solution
2
School of Medicine and Menzies Research Institute, University of Tasmania,
Hobart, Tasmania, Australia 6 1 min Distilled water wash
7 4 min 5% Acetic Acid

ABSTRACT
8 5 min Running tap water wash
The Royal Hobart Hospital is the Tasmanian state reference centre for Parasi-
tology. In this capacity, it receives a large number of specimens for parasitic 9 3 min 70% Ethanol/Ammonia
investigations on an annual basis. The experiences gained in morphological
observations of a number of parasites have been great, and this short report 10 5 min 95% Ethanol
is intended to convey such experiences with regard to three speciÞc intesti-
11 5 min 100% Ethanol
nal parasites, these being Pentatrichomonas hominis, Enteromonas hominis
and Rodentolepis nana. 12 5 min Fresh 100% Ethanol
Ann Australas Coll Trop Med 2010;11:24-25. 13 5 min Xylene
14 5 min Fresh Xylene

Introduction
Beginning in 2002, Tasmania has become the new home for many refugees, 15 - Mount slide
predominantly from Sudan and Sierra Leone and Myanmar; though a small
Method employed by the Microbiology department of the Royal Hobart Hospital (strong carbol
number are emigrants from other African, Middle Eastern and Asian coun-
fuchsin and iron-haematoxylin stains are prepared in-house)
tries. This has resulted in the presentation of a number of unusual parasitic
infections at the RHH Microbiology laboratories. Included in these presen-
tations have been examples of infection with the commensal protozoa; E. Figure 1 Trophozoites Pentatrichomonas hominis
hominis and P. hominis. Whilst these organisms are harmless to the human
host, they are not normal gut ßora, and their presence signiÞes exposure
to faecally contaminated water or food. It is therefore important that these
organisms be reported when observed in clinical laboratories, as they may
act as sentinels for more infections with pathogenic parasites also spread
by the faecal-oral route. Giardia intestinalis will often be shed intermittently
in patient’s faeces, with more than Þve stool specimens requiring examina-
tion prior to detection of infection.1 The presence of commensal protozoa
transmitted via the same route may therefore alert the treating physician to
continue investigations for pathogenic organisms, despite there being unde-
tected in the submitted stool specimen.
A large number of cases of infection with the intestinal cestode; R. nana
(formerly Hymenolepis nana) have also been reported in this refugee popula-
tion. R. nana is a veriÞed pathogen in humans. The clinical presentation of
symptoms is worm burden dependent, and infection may be asymptomatic
or present with symptoms of anorexia, abdominal pain and diarrhoea1.
Methods
Faecal Concentration
Three separate Sodium-acetate Acetic-acid Formalin (Para-Pak SAF - Merid- slide (Remel, Lenexa, KS) and in-house prepared Cryptosporidium species
ian Bioscience, Cincinnati, OH) preserved faecal samples were collected from control slide was included in each batch of slides stained. Microscopy of the
each patient and submitted for parasitological investigation to the Microbiol- permanent smear at x1000 (bright light) was performed using the “battle-
ogy Laboratory, RHH. Specimens were homogenised and 2-5 mL (depending ments” method.
on viscosity) of each added to separate plastic Evergreen faecal concentrate
tubes (Evergreen, Los Angeles, CA) This volume was made up to 10 mL with Results and Discussion
sterile saline then tube capped and mixed. Tubes were centrifuged at 500g The human commensal ßagellate protozoan; P. hominis (Figure 2) is easily
for 10 minutes and the excess supernatant discarded. Smears for permanent identiÞed by its distinctly pyriform shape, round nucleus, anterior ßagella
staining (Table 1) were prepared by mixing one drop of the sediment with and distinct pointed axostyle, which extends beyond the posterior end.2 In
approximately ½ drop of Mayer’s albumin (Meridian Bioscience, Cincinnati, saline preparations, the trophozoites have a distinctive jerky motility, often
OH) on a glass microscope slide. These slides were then allowed to air dry described as resembling a “man trapped in a plastic bag”. This protozoan is
for at least 10 min. Prepared smears were stained using the modiÞed iron- uncommon, and stains poorly in permanent stains,1 and thus is often over-
haematoxylin method described in Table 1. A commercial protozoan control looked when present. No cyst stage of this organism exists.3
Figure 2 Cysts and trophozoites Enteromonas hominis Figure 3 Egg of Rodentolepis nana (saline preparation)
The commensal protozoan E. hominis (Figure 3) morphologically resembles During the course of this work, an observation was made regarding the
Endolimax. nana. These two organisms may be differentiated by the smaller modiÞed acid fast nature of the oncosphere of R. nana eggs (Figures 3, 4).
size of E. hominis compared to E. nana (with marked size overlap). Cysts of E. This previously unreported phenomenon is universally observed in eggs of
hominis are usually 6-8 įm by 4-6 įm, whilst trophozoites usually measure this species when stained with the modiÞed iron-haematoxylin stain method.
3-6 įm by 4-10 įm.3 Trophozoites of E. hominis possess a distinctive Þnely This observation lends itself to improved screening for R. nana in faeces for
vacuolated cytoplasm.3 There is a predominance of binucleate forms (note epidemiological studies. As few normal elements of faeces are modiÞed acid
that uninucleate and quadrinucleate forms are seen), and trophozoites often fast, screening for R. nana eggs could be performed by preparing a thick
taper to a point at one end.2 The authors have also noted when observing smear of faeces and staining with a modiÞed acid fast stain, followed by
E. hominis that the nuclei appear smaller than those of E. nana and that the miscroscopic screening on low power for the modiÞed acid fast oncosphere
trophozoites are more “amoeboid” in appearance (having a greater numbers of these eggs. Such a method; when performed on concentrated faecal sam-
of cytoplasmic blebs and protuberances than are seen in the trophozoites E. ples; would allow a higher sensitivity than a faecal concentration technique
nana). It is suggested that human colonisation with this organism is under- followed by screening of two microscope cover-slips alone.
reported, due to its misidentiÞcation as E. nana.
Figure 4 ModiÞed iron-haermatoxylin stain of Rodentolepis nana egg*
*Note: modiÞed acid-fast oncosphere
*Corresponding author
References Richard S. Bradbury
Department of Microbiology and Infectious Diseases,
1. Garcia L. Diagnostic Medical Parasitology. Washington, DC: ASM Press, 2007.
2. Ash LR, Orihel TC. Atlas of Human Parasitology. Chicago: ASCP Press, 1997.
Royal Hobart Hospital, Liverpool Street,
3. Beaver PC, Jung RC, Wayne E. Clinical Parasitology. 9th Ed. Philadelphia: Lea and Febiger, 1984. Hobart, Tasmania, Australia
Email: [email protected]
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See discussions, stats, and author profiles for this publication at: https://www.researchgate.

Morphological observations on Pentatrichomonas hominis, Enteromonas

Article · January 2010

Richard Stewart Bradbury

Vector Borne Disease Control Provramme, Mandalay, Myanmar View project

The user has requested enhancement of the downloaded file.

STEP TIME SOLUTION

7 4 min 5% Acetic Acid

Ann Australas Coll Trop Med 2010;11:24-25. 13 5 min Xylene

14 5 min Fresh Xylene

Figure 4 ModiÞed iron-haermatoxylin stain of Rodentolepis nana egg*

*Note: modiÞed acid-fast oncosphere

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