MALAYSIAN JOURNAL OF VETERINARY RESEARCH pages 74-82 • VOLUME 9 NO. 2 JULY 2018 CONTAMINATION OF SOILS WITH TOXOCARA EGGS IN SEVERAL PLAYGROUNDS OF IPOH, PERAK DEBBRA M.1, ZARY S.2, ERWANAS A. I.1, AZIMA LAILI H.1, NURULAINI R.1, ADNAN M.1 AND AZIZAH D.1 1 Parasitology and Haematology Section, Veterinary Research Institute, Jalan Sultan Azlan Shah, Ipoh, Perak, Malaysia. 2 Universiti Sains Malaysia, 11800 Pulau Pinang, Malaysia * Corresponding author: [email protected]. ABSTRACT. Toxocariasis is an important INTRODUCTION cosmopolitan zoonotic disease mainly caused by Toxocara spp., a type of soil- According to Taylor et al. (2016), Toxocara transmitted helminth (STH) on cats and spp. is a group of parasitic STH classified dogs. In this study, 80 soil samples were under the family of Ascarididae and genus of taken from four public playgrounds and six Toxocara. Ascaridoids are among the largest neighbourhood playgrounds in Ipoh, Perak nematodes that affects many vertebrates between October and December 2016 to (mostly the domestic animals) as the adult determine the status of soil contamination stage can cause intestinal unthriftiness to with the eggs of Toxocara spp. Results young animals while their migratory larval showed that 32.5% from the total soil stage can cause pathological consequences. samples were positive with Toxocara spp. Both stages have socio-economic and eggs. Overall, five out of ten of the sampling veterinary significance (Li et al., 2006; Taylor sites were contaminated with Toxocara spp. et al., 2016). eggs. Besides that, the relationship between Toxocariasis (Toxocara infection) is the soil condition and the occurrence of listed as one of the five neglected parasitic the Toxocara spp. eggs in soils were also infections (NPIs) by CDC (Centre of Disease investigated. The findings showed that Control) as they may cause severe illnesses increase of moisture and pH of the soils to humans and yet still being neglected or contributed to the increase of contamination less aware by many people (Woodhall et with Toxocara spp. eggs. Sandy soils were al., 2014). This disease is widely known as found significantly contaminated with the among the most prevalent zoonotic diseases eggs of Toxocara spp. compared to the caused by the infective larval stage of other types of soil. Therefore, appropriate Toxocara spp. of predatory mammals which preventive measures such as treatment of belong to the family Canidae and Felidae soil, regular monitoring and deworming (Okulewicz et al., 2012). This mainly refers to of dogs and cats as well as awareness T. cati and T. canis, which are both known as programmes to the public are important. gastrointestinal parasitic roundworms of cats Keywords: Toxocara spp., STH, and dogs, respectively (Choo et al., 2016). playgrounds, soil, Ipoh Dogs and cats commonly defaecate on soils, therefore the soils of public parks 74 VOLUME 9 NO. 2 JULY 2018 MALAYSIAN JOURNAL OF VETERINARY RESEARCH and playgrounds are important sources of playground (recreational playground) as toxocariasis for humans and other paratenic larger, usually covered with pebbles and (non-definite) hosts (Stojevic et al., 2010). gravel, with sandpits for children to play and Moreover, they were found to be the most few areas barely patched with soil and better widespread and prevalent parasitic zoonotic maintained by local council, in comparison infections in humans (Macpherson, 2013). to the conventional neighbourhood Even though there were studies on playground with exposing soil along the the occurrences (prevalence) of Toxocara perimeter. spp. eggs in Malaysia, no data has been In this study, 80 soil samples found regarding soil contamination with were collected from ten different sites Toxocara spp. eggs in any part of Perak (public playgrounds and neighbourhood or the relationship of soil condition to its playgrounds) between October and occurrence of Toxocara spp. eggs in Malaysia. December 2016 and tested for contamination Therefore, the aim of this study was to (1) by Toxocara spp. eggs. Using a small shovel, investigate the presence Toxocara spp. eight points were randomly selected within eggs in soil samples collected from several the sampling area. At each point, a soil playgrounds in Ipoh, and (2) to determine sample weighing 300 g was taken (Noor the correlation of moisture, pH and texture Azian et al., 2008). Each soil sample was of soil samples with the occurrence of taken from 0 cm to 5 cm depth (upper layer) Toxocara spp. eggs. for the highest occurrence of geohelminth eggs (Mizgajska-Wiktor and Uga, 2006). MATERIALS AND METHOD Each sample was placed in a polyethylene sealed bag (air-tight), appropriately labelled Sample collection and area of the study and then transported to the laboratory for analysis or kept at 4 °C to be processed The present study was conducted within (Mohd Zain et al., 2014) within 24 hours. the area of Ipoh, the capital city of Perak, Malaysia (Figure 1). By random assignment, Isolation of Toxocara spp. eggs from soil ten sampling locations which consists of samples six neighbourhood playgrounds and four public playgrounds in good condition, Toxocara spp. eggs was recovered from soil easy to access, and not isolated or far from using the washing method devised by Dada publics or residences. According to Alonso (1979) and modified by Ruiz De Ybáñez et et al. (2001), public playgrounds were the al. (2000) who recorded a recovery rate categorized as playgrounds that placed of 99.91%, with a minor modification in within public parks or recreational areas, this study. At least one confirmed egg of while the neighborhood playgrounds were Toxocara spp. was needed for the sample categorized as playgrounds that placed to be recorded as positive. Briefly, 1 g of within private or housing areas. Meanwhile, evenly mixed soil sample was weighed O’Lorcain (1994) categorised the adventure before washing with distilled water by 75 MALAYSIAN JOURNAL OF VETERINARY RESEARCH VOLUME 9 NO. 2 JULY 2018 Figure 1: Maps of study area. Parts of Peninsular Malaysia showing Perak (red pinned) and its neighboring states (left), this study was conducted within Ipoh area (hatched), (right). Figure 2: Three forms of Toxocara spp. eggs were detected in soil samples. The recovered soil samples observed under 400x magnification by using CX31 Olympus compound microscope. The non-embryonated egg (left), embryonated egg (middle), ruptured egg (right). However, the occurrence of pitted surface of each egg shells was still visible (all pictures by Debbra, 2016). 76 VOLUME 9 NO. 2 JULY 2018 MALAYSIAN JOURNAL OF VETERINARY RESEARCH centrifugation. The recovery of Toxocara spp. Paller and de Chavez (2014) and “feeling“ eggs was carried out by simple floatation analysis by Lesikar (2005). Briefly, each soil and followed by microscopic examination sample was kneaded into a bolus before at 100x and 400x magnification. The egg of performing texture “feeling” analysis and Toxocara spp. was identified based on their then categorised into four types based on typical morphology (Uga et al., 2000), while their texture namely sandy, silty, loamy and the size was determined using an ocular clayey respectively. eyepiece micrometer. Data analysis Determination of soil condition analysis All the results obtained were analysed Soil pH analysis using the IBM Statistical Package for Social Science® (version 22, released 2013). It was For soil pH examination, data was collected assumed that the data for this study were using a method suggested by Rayment suitable for non-parametric statistical tests and Lyons (2011). Briefly, soil pH analysis (Flynn, 2011). was carried out using 2 g each of air-dried soil sample. Water was added and the RESULTS AND DISCUSSION suspension stirred at 15 rpm for 1 hour. Its pH was taken after calibration with acid and In this study, five out of ten selected alkaline buffers. playgrounds in Ipoh were contaminated with Toxocara spp. eggs. Table 1 shows Soil moisture content analysis samples from sites E, A, C, G and D with 87.5%, 75.0%, 62.5%, 62.5% and 37.5% Soil moisture content was obtained using positive respectively. Samples from sites B, the method by Paller and de Chavez (2014). F, H, I and J were all recorded as negative. Briefly, the weight of a jar was tared. The pre- Table 2 shows that the contamination rate dried weight (w0) of each soil sample in the for the neighbourhood playgrounds (43.8% jar was taken before drying in the oven at from 48 soil samples) was higher than the 125 °C. The dried weight (w1) of the soil taken public playgrounds (15.6% from 32 soil and the moisture content of the sample was samples). By applying Pearson’s chi-squared determined using Eq. 1 test, there was a significant difference in the occurence of Toxocara spp. eggs between (1) the neighborhood playgrounds and the � � 2 � public playgrounds ( = 6.923, df = 1, p = � � � � ��� 0.009). Overall, 32.5% of the 80 soil samples Soil texture analysis from ten selected playgroundsχ in this study were contaminated. Soil texture was categorised into four The mean value for moisture content types by referring to the method used by and pH of the soil samples are shown in 77 MALAYSIAN JOURNAL OF VETERINARY RESEARCH VOLUME 9 NO. 2 JULY 2018 Table 1: The contamination of soil samples with Toxocara spp. eggs in current study. Playground category Sampling Site No. of samples (n) Positive samples Percentage of positive (%) Site A 8 6 75.0 Site B 8 0 0.0 Site C 8 5 62.5 Neighborhood Site D 8 3 37.5 Site E 8 7 87.5 Site F 8 0 0.0 Site G 8 5 62.5 Site H 8 0 0.0 Public Site I 8 0 0.0 Site J 8 0 0.0 Table 2: Contamination of soil samples with Toxocara spp eggs.