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Analysis of Parasitic Infections in Human and Dogs Using Different Fecalysis

Techniques1

Garcia, Allaine M.
B 1L

02 December 2015

A case report submitted in partial fulfillment of the requirements in ZOO 173 - Introduction to
Parasitology laboratory under Dr. Vachel Gay V. Paller, 1st semester A.Y. 2015 - 2016

ABSTRACT
The prevalence of different parasitic infections in humans and dogs was
determined using the formalin-ether concentration technique (FECT) to
analyze stool samples. Of the 8 human fecal samples observed, 4 tested
positive to parasitic infection, thereby having a parasitic infection prevalence
of 50%. The parasitic infection prevalence in the dog fecal samples is also
50%. The organisms observed in the human fecal samples were Ascaris sp. and
Entamoeba sp. with computed mean intensities of 1.5 and 1, respectively. For
the dog fecal samples observed by the whole class, the species recorded were
Ascaris sp. (mean intensity=3), Trichuris sp. (mean intensity=2.5), Toxocara
canis (mean intensity=18), Ancylostoma sp. (mean intensity=7), Hymenolepis
sp. and Giardia lamblia (both were too many to count). The organisms found
in the dog fecal sample handled by the researcher were Toxocara canis
(epg=180), Ancylostoma sp. (epg=70) Ascaris sp. (epg=30) Trichuris sp.
(epg=30). Moreover, Scotch Tape Method was used to determine the
prevalence of Enterobius vermicularis among children, however, results
showed no infection in 24 samples. Nevertheless, it is noteworthy that the class
observed a 4.17% prevalence of Hymenolepis in the pool of samples.

INTRODUCTION
One of the most frequently administered methods for detecting
intestinal parasites is through stool specimen analysis or fecalysis. It is commonly
done by examining stool samples and checking for the presence of intestinal parasites.
It is particularly effective in detecting parasites since most parasitic species are
expelled out of the body through the feces. Specifically, eggs from adult forms of
parasitic trematodes and nematodes pass out of the hosts body in fecal matter and
these eggs can be easily characterized through certain diagnostic features which can
be easily observed under the microscope. Through fecalysis, proper treatment of
parasitosis may be employed since the parasites can be accurately identified.
Formalin-Ether Concentration Technique (FECT) is one of the
methods used to analyze stool specimens. This method is used instead of direct fecal
smear since it filters and concentrates the formalinized fecal sample in order to allow

the detection of small numbers of organisms that may be missed in direct fecal
smears. It employs sedimentation though centrifugation in order to gather protozoa,
helminth ova and larva in the bottom of the tube. Moreover, ether is used in order to
remove debris and fat from the feces (Garcia & Bruckner, 1997).
Scotch Tape Method is another diagnostic technique specialized in
detecting Enterobius vermicularis (pinworm), which is characterized by D-shaped
eggs. Since gravid female pinworms descend from the large intestine to lay eggs
along the perianal region at night, the method of collection uses the adhesive property
of scotch tape to gather eggs from the perianal region of the subject early in the
morning before defecating or taking a bath (Paller & Reyes, 2014).
This study aims to analyze parasitic infections in humans and dogs
using different fecalysis techniques. Specifically, it aims to:
1. Examine endoparasites of dogs and humans by conducting Formalin-Ether
Concentration technique for stool sample analysis;
2. Determine the prevalence of Enterobius vermicularis among children using
the Scotch Tape Method; and
3. Assess the degree of parasitosis based on the information obtained on parasite
forms and their life cycle.
This study concentrated on the analysis of parasitic infections in
humans and dogs using different fecalysis techniques. It was done last November 16,
2015, Monday, in a laboratory of the Institute of Biological Sciences, University of
the Philippines Los Baos, College, Laguna.

METHODOLOGY
Formalin-Ether Concentration Technique
To determine the presence of parasites in human and dog fecal
samples, the Formalin-Ether Concentration Technique was done. The researchers
collected at least 2 grams of stool sample from 8 humans and 8 dogs and were placed
separately in a labeled screw-capped container with 10ml formalin. At least 1 gram of
the formalinized stool was strained using 3-layered surgical gauze into a centrifuge
tube. Ten (10) ml of 10% formalin and 3ml ether were added into the suspension. The
centrifuge tube was covered and vigorously shook for 1 minute so that the fatty
components of the stool attach to ether. The suspension was centrifuged at 1,500 rpm
for 5 minutes. The ether layer, floated debris and formalin layer were decanted
afterwards leaving only a small quantity of fluid with the precipitate. A drop of the
precipitate was pipette out of the tube and placed onto a glass slide and covered with a
cover slip. Clear nail polish was applied to seal the preparation. The slide was then
examined under the microscope systematically, noting whether the sample is positive
or negative for infection. All species of parasites present in positive samples and the
number of individuals per species encountered were recorded.
Scotch Tape Method
To determine the prevalence of Enterobius vermicularis among
children, the Scotch Tape Method was employed. A total of 24 samples were
collected from children. The collection was done in the morning, before the
individuals took a bath or defecated. To collect a sample, one end of a clear
transparent scotch tape was attached on a glass slide while the other end was attached
to a tongue depressor. The adhesive side of the tape was dabbed gently to the perianal

region of the subject and was mounted on the glass slide. The slides obtained were
systematically examined under the microscope, searching for Enterobius vermicularis
which is characterized by a D-shaped egg as shown in Figure 1. The positivity or
negativity for infection and the number of eggs encountered were recorded.

Figure 1. The characteristic D-shaped eggs of Enterobius vermicularis: (A) Scotch Tape
Method preparation; and (B) higher magnification.

Data Analysis
The prevalence, mean intensity and fecal egg count in eggs per gram
(epg) was determined in order to quantify and help assess the degree of parasitosis.
The prevalence of parasitic infections was determined using the formula:

While the mean intensity of a particular parasite was computed using the formula:

The fecal egg count in eggs per gram (epg) was also determined using the formula:

RESULTS AND DISCUSSION


Class Data
Results showed that 4 out of 8 samples for both humans and dogs
tested positive for parasitic infection, thus giving a 50% prevalence of parasitic
infection for both dogs and humans. The organisms observed in the human fecal
samples were Ascaris sp. and Entamoeba sp. The computed mean intensities for these
organisms were 1.5 and 1, respectively. As seen in Table 1, the parasitic species
recorded from the dog stool samples were Ascaris sp. with a computed mean intensity
of 3, Trichuris sp. (mean intensity=1.5), Toxocara canis (mean intensity=18),
Ancylostoma sp. (mean intensity=7), Hymenolepis sp. and Giardia lamblia. The mean
intensities for the last two recorded species were not computed because too many
individual were encountered to count.
Table 1. The computed mean intensities of parasitic infection present in the
human and dog samples of the class.
Organisms
observed

No. if individuals per


species

No. of infected samples

Mean Intensity

Human

Dog

Human

Dog

Human

Dog

Ascaris
Trichuris

6
0

3
3

1
2

1.5
-

3
1.5

Toxocara

18

18

Ancylostoma

Giardia

TNTC

Entamoeba

Hymenolepis

TNTC

Dog Fecal Sample


The sample handled by the researcher was that of a dogs. The stool
sample handled was dark brown to green in color and was intensely odorous. The
sample was collected from a street in front of a residence situated in Sta. Fe
Subdivision, Brgy. Batong Malake, Los Baos, Laguna. The dog is owned by the said
residence and was observed to freely roam around the subdivision. The subject also
sleeps outdoors and does not frequently take a bath. An adult worm, which was
identified as a Trichuris species, was found in the fresh stool sample. The organisms
found in the dog fecal sample (Figure 2) after employment of FECT were Toxocara
canis with a computed fecal egg count of 180 epg, Ancylostoma sp. (epg=70), Ascaris
sp. (epg=30), Trichuris sp. (epg=30). This implies that the subjects liberty to roam
around the area and interact with other dogs and the subjects way of living greatly
affects the acquisition of the parasite. This also implies that the dog is a possible
source of infection to other dogs and humans as well. Thus, it is important that the
owner of the subject be properly educated about the transmission of parasitic
infections because they have a responsibility in the prevention and control of the
spread of diseases caused by parasites.

Figure 2. Photographs of parasitic species found from the dog fecal sample: (A)
Ancylostoma; (B) Trichuris; (C) Ascaris; and (D) Toxocara.

Scotch Tape Method

Twenty-four (24) samples were observed in total for this part of the
exercise but not one tested positive for the presence of Enterobius vermicularis.
However, one sample tested positive for Hymenolepis. The negative result may be a
false negative but it may also be accounted by the proper hygiene practice of the
family where the samples were collected. The sample collected by the researcher, for
example, came from a 1 year-old boy who plays outside frequently but with constant
adult supervision. The parents and guardians of the subject see to it that the subject is
well groomed right after playing outside, thus, the subject has a good hygienic
practice. The subject is from a family with average income. The residence of the
subject has screen doors and windows which keep insects and other biological and
mechanical vectors of diseases off of the house.

SUMMARY AND CONCLUSION

The prevalence of parasitic infections in humans and dogs was


determined. Formalin-Ether Concentration Technique was used in order to assess the
degree of parasitosis in humans and dogs. It was computed that the prevalence of
parasitic infection for both dogs and humans is 50%, that is, out of 8 samples, 4
individuals tested positive for parasitic infection. Based on the fecal egg count
obtained from the fecalysis of a dog sample done by the researcher, it can be
concluded that the subject is heavily parasitized by Toxocara canis (180 epg). This
indicates the problem of controlling the spread of the parasite since the subject
defecates outdoors. Twenty-four (24) samples tested negative for the presence of
Enterobius vermicularis as observed using the Scotch Tape Method. This indicates
that either there was error while collecting the samples thus giving a false negative
result or the samples collected all came from subjects with good hygiene practices.

REFERENCES
Garcia, L. S., & Bruckner, D. A. (1997). Diagnostic Medical Parasitology (3rd ed.).
Washington DC: ASM Press.
Mergo Jr., J. C., & Crites, J. L. (1986, June). Prevalence, Mean Intensity, and Relative
Density of Lintaxine Cokeri Linton 1940 (Monogenea: Heteraxinidae) on
Freshwater drum (Aplodinotus grunniens) in Lake Erie (1984). The Ohio
Journal of Science, 86(3), 101-105. Retrieved November 29, 2015, from
http://kb.osu.edu/dspace/handle/1811/23139
Paller, V. G., & Reyes, R. C. (2014). Laboratory Manual in Parasitology. Laguna:
ABD, IBS, CAS, UPLB.