MAY 22, 2018 ZOOLOGY 173 – INTRODUCTION TO PARASITOLOGY | 2ND SEMESTER 2017-2018

Qualitative and Quantitative Methods for the Detection of

Endoparasites in Human (Homo sapiens sapiens) and Dog (Canis
lupus familiaris)
Castillo, John Michael1
1
Animal Biology Division, Institute of Biological Sciences, University of the Philippines Los
Baños

To know the prevalence of endoparasites in humans and dogs,

fecalysis and scotch tape methods were used. A total of 6 human fecal
samples and 5 dog samples were collected for a period of 2 weeks from
different sources around Los Baños, Laguna. The former procedure
requires the addition of formalin and ether. Filtration and centrifugation
then proceed. There have been 3 fecal samples that show the presence
of parasites, one with Toxocara, and the other two with hookworms. To
quantitatively measure the prevalence of parasites certain parameters
were computed. There is 16.67% prevalence rate of Toxocara in contrast
with 33.33% at hookworm. There is a computed 300 egg count and 3
mean intensity for hookworm while 7000 egg count and 14 mean
intensity counts in hookworm. Meanwhile, to check if there is a presence
of Enterobius vermiclaris, the scotch tape method is utilized, all samples
tested negative for the test. Sanitation, personal hygiene, financial
status, and other related information were gathered to know if there is a
relation with the occurrence of the said parasites.

Keywords: Enterobius, Fecal, parasites, Toxocara

I. INTRODUCTION experiment conducted we focused mainly in

Parasites have been regarded as one of
the most dangerous organism in the world
because of its high reproductive rate and the
ability to co-evolved with the host. It poses a
serious threat to the public health because of
its easy transmission and its ability to defy the
hosts’ immune response. Clinically there have
been a lot of diseases brought by bacteria,
viruses and even parasites. Some people are
confused with which causes which disease, so
a reliable identification is needed to properly
address the corresponding disease.
the medical parasites. It is indeed a
prerequisite to have a familiarity with the
morphology of such parasites because this will
aid in efficient and accurate description of the
parasites. Meanwhile to those parasites that
cannot be detected using fecalysis other
techniques will be used. Scotch tape method
is mainly used to detect pinworm (Enterobius
vermicularis) eggs deposited around the
perianal region at night by the gravid female
from the large intestines.
II. OBJECTIVE

In the Philippines, there has been an 1. Conduct qualitative and quantitative

alarming >50% prevalence rate of soil-
transmitted helminths (STHs) and around
47.7% infected with protozoans (Baldo,
Belizario, De Leon, Kong, &Chung, 2014).
Stool analysis or fecalysis is widely used
especially in the medical field. For the
laboratory techniques for
endoparasite examination; and
2. Assess the degree of parasitosis of
the subject based on the information

_____________________________________________________________________________________
MAY 22, 2018 ZOOLOGY 173 – INTRODUCTION TO PARASITOLOGY | 2ND SEMESTER 2017-2018

obtained on parasite forms and their The study can be used in determining the
life cycles. parasites that are of public health concern and
causes NTDs using simple techniques
III. SIGNIFICANCE OF THE STUDY especially to the areas which do not have
access to high-end diagnostic institutions.

IV. MATERIALS AND METHODS required to use face mask and gloves to avoid
infection. It is advised that it must be done
Two techniques were employed to early in the morning before taking a bath or
qualitatively and quantitatively detect defacation.
endoparasites in human and dog subjects.

A. Stool Sample Collection b. Slide Examination

Before conducting the experiment, the
students were paired into two gathers 1
human and 1 dog stool. All the necessary
information was listed from the owner of the
subject, address, age, hygienic practices, etc.
Moreover, the color, smell and consistency of
the stool were also considered. The samples
were then placed in a vial with 10% formalin.
B. Formalin-Ether Concentration
Technique
Before the addition of formalin-ether, the
stool was homogenized first by shaking the
vial. Then, the solution was filtered in a 3-
layered surgical gauze to obtain at least 1
gram of formanilized stool. The residue was
discarded together with the gauze immediately
after the filtration. The filtrate was then added
with 3-ml ether and then the solution was
shaken vigorously for 1 minute. Then, the
solution was then subjected in centrifugation
for 5 minutes at 1,500 rpm. Using a stick, the
solution was decanted to gather the precipitate
found at the bottom of the tube. A drop of
precipitate was transferred onto a glass slide
and examined under the microscope. All data
obtained was recorded into Table 10.1
Eggs of Enterobius vermicularis was
examined systematically under the
microscope, which was then confirmed by
the laboratory instructor. The result was
recorded for the parasite egg in Table 10.1
After all the qualitative measures, the
following formulas were used for
quantitative detection and measures:
𝑛𝑜. 𝑜𝑓 𝑖𝑛𝑓𝑒𝑐𝑡𝑒𝑑 𝑠𝑎𝑚𝑝𝑙𝑒𝑠
𝑃𝑟𝑒𝑣𝑎𝑙𝑒𝑛𝑐𝑒 (%) =
𝑡𝑜𝑡𝑎𝑙 𝑛𝑜. 𝑜𝑓 𝑠𝑎𝑚𝑝𝑙𝑒𝑠
𝑀𝑒𝑎𝑛 𝑖𝑛𝑡𝑒𝑠𝑖𝑡𝑦
𝑛𝑜. 𝑜𝑓 𝑝𝑎𝑟𝑎𝑠𝑖𝑡𝑒𝑠 𝑜𝑓 𝑎 𝑠𝑝𝑒𝑐𝑖𝑚𝑒𝑛
=
𝑡𝑜𝑡𝑎𝑙 𝑛𝑜. 𝑜𝑓 𝑖𝑛𝑓𝑒𝑐𝑡𝑒𝑑 𝑠𝑎𝑚𝑝𝑙𝑒𝑠
𝐸𝑔𝑔𝑠 𝑝𝑒𝑟 𝑔𝑟𝑎𝑚 (𝑒𝑝𝑔)
𝑡𝑜𝑡𝑎𝑙 𝑣𝑜𝑙𝑢𝑚𝑒
= × 𝑒𝑔𝑔 𝑐𝑜𝑢𝑛𝑡
2𝑔 × 0.1𝑚𝐿

1. Scotch Tape Method

a. Collection

Using a scotch tape attached in a glass

slide, the perianal region of the subject was
then gently dabbed in the adhesive side of the
tape. During the collection, the students were

_____________________________________________________________________________________
MAY 22, 2018 ZOOLOGY 173 – INTRODUCTION TO PARASITOLOGY | 2ND SEMESTER 2017-2018

Figure 10.2 Hookworm egg 2 found in dog

stool sample 3.

Table 10. 1 shows all the results for

V. RESULTS AND DISCUSSION the qualitative measurements for the
detection of endoparasites in the human and
Table 10. 1. Parasites observed in each of the dog samples. Three dog samples were
stool samples. positive of endoparasites, Toxocara and
Pair Dog stool Human stool Hookworms. The first type as seen in Figure
1 - 10.1 has 7 in count that were present, while
Toxocara (7) there are one and five hookworms in the 2nd
and 3rd sample (Figure 10.2).
2 -
There are 6 hookworm’s egg from 2
Hookworm (1)
dog fecal sample. There is 50% prevalence of
parasites. The colorless, thin shelled
3 - appearance and visible embryonic cleavages
Hookworm (5) of egg is the diagnostic feature of hookworm.
The ingestion of the parasite eggs in the soil is
4 - - probably due to the access of dogs on the
ground. Moreover, since the dogs were not yet
‘dewormed’ and is not introduced to a
- - veterenarian, there is a high relation that the
5 dogs have poor hygienic practices adding to
the probable cause of parasites. We can also
deduce from the table that the dogs might be
6 infected by Ancylostoma caninum, which is the
infective larva and it usually infects canids.
Hookworms as their name suggests have
hook-like mouthparts for attachment to the
lining of the intestinal wall (Ward, n.d.). They
cause life-threatening anemia, most common
in puppies but can still occur in adult dogs.
Results showed a 33 % commonness of stool
samples, implying that 2 out of 6 dogs may be
Figure 10.1 Toxocora egg 1 found in dog positive of hookworm, with a mean intensity of
stool sample 1 3 probably due to it just still being a puppy, and
300 eggs in estimate are present in one gram
of dog stool. Meanwhile, there is a presence of
Toxocara eggs in sample no. 1 with 7 counts,
17% of commoness in stool samples.

The presence of many Toxocora than

hookworms can be attributed to its
susceptability in varying environmental
conditions. Toxocara eggs are more resistant
can survive for almost 3-4 weeks in soils
(Harris-Linton, 2001). Moreover, it has simpler
life cycle compared to hookworm (hookworm
has to molt three times), hence, lesser number
of eggs develop in the intestine.

_____________________________________________________________________________________
MAY 22, 2018 ZOOLOGY 173 – INTRODUCTION TO PARASITOLOGY | 2ND SEMESTER 2017-2018

Table 2. Computed parameters for For the scotch tape method, no

quantitative detection of parasites samples showed presence of E.
vermicularis. Pinworms are very easy to
Mean Prevalenc Egg transmit, being a group disease due to its
Intensit e s very light egg that can be passed by
y per inhalation, oral transmission, and
gra autoinfection, requires all of the household
m members to receive medications such as
(epg mebendazole and albendazole. An infection
) also requires cleaning all of the household
Toxocara 14 16.67% 700 items from sheets to furniture, that could
possibly have eggs. Prevention can be done
Hookwor 3 33.33% 300 by often washing of the household items and
m most importantly of hands.
Pinworm 0 0% 0
s
VI. SUMMARY AND CONCLUSION

Human subjects tests negative for Qualitative detection of parasites in

the presence of Pinworm. Basically, dog and human were done using formalin-
pinworms have distinctly D-shaped ether concentration. On the other hand,
(elongated, ovoid, and flattened on one Scotch tape method was used to determine
side), 45-60 long by 25-30 wide, two layers the presence of E. vermicularis. The
of eggshells, an outer think hyaline fecalysis showed that two out of the six dog
albuminous shell and an inner embryonic samples were positive for hookworm while
lipoidal membrane, containing an one out of six is positive for Toxocara. On the
embryonated larvae. Negative result other hand, no human subject was positive
suggests that the subject has good hygenic for helminths, protozoans, nor pinworm. This
practices and have been dewormed could be attributed with the good hygiene of
regularly. However, wrong preparation of the the human subjects. However, if ever
sample can also be attributed to the negative infected, there are still numerous ways to
result. treat it, that is why is it important to properly
diagnose the parasite before giving the
Table 3. Presence of E. vermicularis in three respective appropriate treatment. There are
replicates also several ways to prevent acquiring
endoparasites, an example is simply good
Pair 1st 2nd 3rd hygiene.
No Replicate Replicate Replicate
1 0 0 0

2 0 0 0 VII. RECOMMENDATIONS

3 0 0 0 Necessary information should be strictly

noted. In addition, the procurement of
4 0 0 0 samples should be randomized to fully
represent the population. Lastly, to further
5 0 0 0
assess the prevalence of the parasites,
6 0 0 0 previous study can be used to compare the
results of the recent studies.

_____________________________________________________________________________________
MAY 22, 2018 ZOOLOGY 173 – INTRODUCTION TO PARASITOLOGY | 2ND SEMESTER 2017-2018

VIII. LITERATURE CITED

Baldo, E. Belizario, V., De Leon, W., Kong,

H., Chung, D.-L. (2004). Infection
status of intestinal parasites in
children living in residential
institutions in Metro Manila, The
Philippines, The Korean Journal of
Parasitology, 42(2), 67-70.

Giorgi, Anna. 2016. Pinworm Infection

accessed on
https://www.healthline.com/health/pi
nworms#risk-factors

Mandell, Gerald L., John E. Bennett, and

Raphael Dolin. Mandell, Douglas,
and Bennett’s Principles and
Practice of Infectious Diseases.
Philadelphia, PA: Churchill
Livingstone/Elsevier, 2010

Paller, Rachel Gay V., and Reyes, Roberto

C. 2014. Laboratory Manual in
Parasitology.

Ward, Edward. n.d. Hookworm Infection in

Dogs accessed on
https://vcahospitals.com/know-your-
pet/hookworm-infection-in-dogs

_____________________________________________________________________________________