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Polytechnic University of the Philippines Sta. Mesa, Manila College of Science Department of Biology

Prevalence of Geohelminths in Soils Samples from the Polytechnic University of the Philippines
Fanoga, Maria Geraldine A., Legatub, Franco Rigor M., Mendoza, Charmaine B., Tuazon, Jessel Y. BS Biology 4-1

Abstract.

Geohelminths are a group of parasitic nematode worms causing infection through contact with parasite eggs or larvae that thrive in the warm and moist soil. Nematode larvae was extracted and observed using the Baermann Funnel Extraction technique. Results of the study showed high prevalence of hookworm larvae 62% and Strongyloides spp. 46%. High prevalence of these nematode larvae suggests that soil samples obtained from the PUP Main Campus is possibly polluted with fecal samples and has high rate of nematode larval contamination.

constant health problem among poorly developed areas in the Philippines despite the efforts in developing the countrys health care system. The disease is very common in the poorest areas in the country where fecal contamination due to improper waste disposal is one of the serious environmental problems (Hoffman et al. 2008). It has greatly

arasitic

infections

have

been

of the Philippines Institute of Health and Department of Health in 2005, the prevalence rate of soil- transmitted helminthiasis in the Philippines reached to 66%. Overcrowding, lack of personal and environmental hygiene, polluted water and the remote and agricultural areas with humid environment make it all conducive for egg survival, fecal parasite growth 2004). Common helminthes that can be found in the soil are the eggs and larvae of and transmission (Belizario,

affected the health and socio- economic status of individuals and communities. Based on a study made by the University

Polytechnic University of the Philippines, Department of Biology, 2012 Fanoga, M. G., Legatub, F. R., Mendoza, C., Tuazon, J.

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Ascaris lumbricoides, Trichuris trichiura, Strongyloides stercoralis and Necator americanus. These helminthes could either be parasitic or free-living when observed. Heavy worm burden of such parasites could lead to morbidity and could develop into severe diseases. Such infections could lead to growth age group anemia, and are malnutrition, Although poor all

to determine the prevalence and risk factors of the common soil transmitted parasites on the soil samples obtained from the different sites in the Polytechnic University of the Philippines Campus, Sta. Mesa Manila.

Methodology. Study area and Sample collection Site Number 1 2 3 4 5 6 7 8 9 10 Site Name Chapel Oval Gym NALRC Main Building Mabini Shrine Lagoon Laboratory High School Open Court Linear Park

development and among others. susceptible to such parasitic infections, school- aged children are particularly at risk of acquiring the infection due to increased opportunity of ingesting the eggs of the parasite while playing in soil that is contaminated with human feces (Belizario, 2004). Much more disturbing is that these parasitic infections are highly associated with poor growth, reduced physical activity and impaired learning ability among children of developing countries (Cooper, 2008). The infection is promoted by poor hygienic habits such as indiscriminate disposal of human and animal feces. Other risk factors include lack of safe water source, overcrowding poverty, geophagia, failure to wear foot wears, having pools of water/sewage around houses (Chukwuma et al., 2009). This study aims

Table 1. Designation of Sampling Areas in the Polytechnic University of the Philippines

Random sampling was performed and ten sampling areas were designated inside the Polytechnic University of the Philippines. Each sampling area was subdivided into 5 sampling points and ten grams of soil sample were obtained per sampling point

Polytechnic University of the Philippines, Department of Biology, 2012 Fanoga, M. G., Legatub, F. R., Mendoza, C., Tuazon, J.

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Baermann Funnel Extraction

up supporting a glass funnel and rubber tubing and a pinch clamp was attached to the bottom of the funnel. A collection beaker or centrifuge tube was placed underneath (figure 1). Wire gauze or nylon filter was placed over the top of the funnel, followed by a pad consisting of two layers of gauze. Then the pinch clamp at the bottom of the tubing was closed and the funnel was filled with lukewarm tap water (about 45OC) until it just soaks the gauze padding. The soil sample (well broken up) was placed on the gauze and was covered with a piece of filter paper. The apparatus was allowed to stand for 2 hours. Then 10 ml of fluid was drawn off into the beaker or centrifuge tube by releasing the pinch clamp; the fluid was centrifuged for 2 minutes at 400-500 rpm. The sediment as well as the supernatant was examined under the microscope under the LPO and HPO for motile larvae.

Figure 1. Diagram of the Baermann apparatus used for recovery of larval-stage nematodes

The Baermann technique uses a special apparatus and relies on the principle that active larvae will migrate out of a fecal specimen that has been placed on a wire mesh covered with several layers of gauze (Baermann, 1917; Garcia, 2001). Larvae migrate through gauze into the water and settle to the bottom of the funnel, where they can be collected and examined (Watson and AlHafidh, 1957). Besides being useful for diagnosis from stool specimens directly or after enhancement by culture, this technique can be used by epidemiologist to examine soil specimens for larvae. Baermann funnel technique was carried out to extract nematode larval stages from soil. Briefly, a clamp was set-

Polytechnic University of the Philippines, Department of Biology, 2012 Fanoga, M. G., Legatub, F. R., Mendoza, C., Tuazon, J.

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Results.
Table 1. Prevalence of Nematode Larvae in three soil samples in Polytechnic University of the Philippines vicinity

prevalence

since

out

samples

examined shown the species.

Number of Sample 1 2 3

Hookworm 100% 100% 100%

Strongyloides sp. 0% 100% 100%

The values enclosed in each species in Table 2 the calculated prevalence with its confidence limits as given in the next column. The calculated values lies in

Table 2. Overall Prevalence of Nematode Larvae In Ten Soil Sample Sites in Polytechnic University of the Philippines vicinity

Category Hookworm Strongyloides sp.

Prevalence 100% ( 1.000) 67% (0.667)

between the computed 95% confidence, thus accepted.

Confidence Limits at 95%: Hookworm (0.2924 to 1.0000); Strongyloides sp. (0.0942 to 0.9916)

Table 3. Frequency Types of Nematode Larvae Stages (N=54)

A total of three samples were observed during the experiment. Table 1 shows the prevalence of Nematode larvae in three soil samples that were examined. As indicated, hookworms were present in all soil a sample thus corresponds to a 100% percentage prevalence in Table 2. Strongyloides sp., had 67% percentage

Table 3 shows the types of nematode larvae found with their frequencies. Out of total 54 parasite population, 31 were hookworms, 8 were in filariform stage and 23 were in Rhabditiform. For the Strongyloides sp., 5 were in filariform while 18 were in Rhabditiform. Other or the free living larvae were not included since those were not verified if they are parasites or not.

Polytechnic University of the Philippines, Department of Biology, 2012 Fanoga, M. G., Legatub, F. R., Mendoza, C., Tuazon, J.

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120% 100% Prevalence (%) 80% Hookworm 60% 40% 20% 0% 1 2 3 Number of Sample
Figure 1.Figure 2. Strongyloides sp.,of Nematode Larvae Frequency Distribution rhabditiform stage

Strongyloides sp.
Figure 3. Hookworm, rhabditiform stage

Figure shows the actual images of parasites examined and extracted using the Baermann Funnel Extraction Technique. Discussions. The results of the study revealed geohelminth larval contamination in the soil samples obtained from the Polytechnic University of the Philippines and this comprised of hookworm with 80% overall prevalence and Strongyloides species larvae with 60% overall prevalence. Eight out of 10 sampling areas specifically, 30 sampling points (60% of total sampling points) were positive for the presence hookworm larvae were while positive 6 sampling areas or of approximately 23 (46%) sampling points

Figure 1 shows the frequency distribution of nematode larvae. Hookworm (in blue bar) obtained the highest number since it is prevalent in all three samples observed. Also, Strongyloides sp., shown to be present in two samples. Figures 3 and 4 shows the actual images of hookworm and Strongyloides sp., seen during the course of observation.

Figure 2. Strongyloides sp., rhabditiform stage

for

contamination

Strongyloides species larvae. Due to the

Polytechnic University of the Philippines, Department of Biology, 2012 Fanoga, M. G., Legatub, F. R., Mendoza, C., Tuazon, J.

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fact that the adult stages of these worms reside in the intestine, the presence of the larval stages in soil is indicative of faecal pollution (Chukwuma et al., 2009). The results of the study for the presence of hookworm and Strongyloides species larvae was supported by the study done by Brown in 2007 that Strongyloides spp. and hookworms are the most common nematodes found in soils of tropical and subtropical regions. A rapid, reliable and highly efficient method for the extraction of viable nematodes from soil is of particular importance not only for field diagnostics but also laboratory research. Nematodes are most frequently extracted using modifications of the Baermann funnel (Walker and Wilson, 1960; Thorne, 1961; Hooper, 1990), live bait (Fan and Hominick, 1991) or centrifugal flotation techniques (Byrd et al., 1966; Saunders and All, 1982). The mechanical separation of

funnel method could only recover live nematodes; however, it takes longer. Moreover, the study obtained soil samples in places wherein students tend to spend more of time in direct contact with soil. This would give the students and other school personnel the awareness to avoid direct soil contact in such places and be able to educate them of the risk factors associated with Strongyloides infection. spp. and hookworm

Conclusion. Soil samples in the Polytechnic University in the Philippines are contaminated with geohelminth larvae and possible fecal pollution. Hookworms Strongyloides species, are the most

prevalent nematode larvae followed by

nematodes from soil by the Baermann funnel is on nematode motility, while the live bait method relies on infectivity and the flotation relies on the physical properties of the juvenile. The Baermann

Recommendations. Increase the amount of soil

samples to be examined. Increase the no. of sampling points per sampling area.

Polytechnic University of the Philippines, Department of Biology, 2012 Fanoga, M. G., Legatub, F. R., Mendoza, C., Tuazon, J.

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Conduct parallel studies for the detection and extraction of nematode larval stages in soil. Characterize the soil samples for supplemental information for the prevalence on larvae.

Responses to Ascaris Lumbricoides, but not to Aeroallergens, in Children from Rural Communities in the Tropics. The Journal of Infectious Diseases, 182:12071213. DOI: 10.1086/315830 Dewang Deng, Allan Zipf, Y. Tilahun, G.C.Sharma, J. Jenkins and K. Lawrence. 2008. An improved method for the extraction of nematodes using iodixanol (OptiPrep ). African Journal of Microbiology Research Vol.(2) pp. 167-170.

References. Baermann, G. 1917. Eine einfache Methode zur Auffindung vor Ankylostomum (Nematoden) Larven in Erdproben. Meded. Geneesk Laborat. Weltever Feestbundel, p. 41. Belizario, Vicente and De Leon, Winifreda. (2004). The Philippine Textbook of Medical Parasitology. 2nd ed. Bethony, F.,, S. Brooker, M. Albonico, S. Geiger, A. Loukas, D. Diemert and P.Hotez. 2006. Soil-transmitted helminth infections: ascariasis, trichuriasis, and hookworm. Vol 367. Chukwuma, M.C., I.M. Ekejindu, N.R. Agbakoba, D.A. Ezeagwuna, I.C. Anaghalu and D.C. Nwosu, 2009. The Prevalence and Risk Factors of Geohelminth Infections among Primary School Children in Ebenebe Town, Anambra State, Nigeria. Middle-East Journal of Scientific Research 4 (3): 211-215. Cooper, C. (2008). Repeated Treatments with Albendazole Enhance Th2

Garcia, L.S. 2001. Diagnostic Medical Parasitology, 4th ed., p. 786-795. ASM Press, Washington, DC. Hoffman, H., et al. (2008). Hookworm, Ascaris lumbricoides infection and polyparasitism associated with poor cognitive performance in Brazilian schoolchildren. Tropical Medicine & International Health: TM & IH. 13 (8), 994-1004. Muznebin, F., H. Khanum and A. Hossain, 2007. Incidence of Nematode Infections Among the Children Brought to ICDDR, B Hospital, Dhaka, Bangladesh. J. bio-sci. 15: 159-164. Truant, A., S. Elliot, M. Kelly, and J. Smith, 1981. Comparison of Formalin-Ethyl Ether Sedimentation, Formalin-Ethyl Acetate Sedimentation and Zinc Sulfate Flotation Techniques for Detection of Intestinal Parasites.

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Journal of Clinical Microbiology, May 1981, p. 882-884 Vol. 13, No. 5. Watson,J.M., and R. Al-Hafidh. 1957. A modification of Baermann funnel technique and its use in establishing the infection potential of human hookworm carriers. Ann. Trop. Med. Parasitol. 51:15-16.

WHO (2005) Deworming for health and development. Reports on the third global meeting of the partners for parasite control. Geneva: WHO.

Polytechnic University of the Philippines, Department of Biology, 2012 Fanoga, M. G., Legatub, F. R., Mendoza, C., Tuazon, J.