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Pdf Living Amoebae in Household Water, Ohio, USA - 20 Iran J Parasitol: Vol. 12, No. 3, Jul-Sep 2017, pp.413-422 Iran J Parasitol Tehran University of Medical Open access Journal at Iranian Society of Parasitology Sciences Public a tion http:// ijpa.tums.ac.ir http:// isp.tums.ac.ir http:// tums.ac.ir Original Article Assessment of Two Different Drinking Water Treatment Plants for the Removal of Free-living Amoebae, Egypt *Ahmad Z. AL-HERRAWY, Mahmoud A. GAD Dept. of Water Pollution Research, National Research Center, 12622 Dokki, Giza, Egypt Received 11 Nov 2016 Abstract Accepted 15 Mar 2017 Background: The aim of this study was to compare between slow and rapid sand filters for the removal of free-living amoebae during drinking water treatment pro- duction. Keywords: Methods: Overall, 48 water samples were collected from two drinking water Removal, treatment plants having two different filtration systems (slow and rapid sand filters) Efficacy, and from inlet and outlet of each plant. Water samples were collected from Free-living amoebae, Fayoum Drinking Water and Wastewater Holding Company, Egypt, during the Drinking water treat- year 2015. They were processed for detection of FLAs using non-nutrient agar ment plants, (NNA). The isolates of FLAs were microscopically identified to the genus level PCR based on the morphologic criteria and molecularly confirmed by the aid of PCR using genus-specific primers. *Correspondence Results: The percentage of removal for FLAs through different treatment Email: processes reached its highest rate in the station using slow sand filters (83%), while [email protected] the removal by rapid sand filter system was 71.4%. Statistically, there was no signif- icant difference (P=0.55) for the removal of FLAs between the two different drink- ing water treatment systems. Statistically, seasons had no significant effect on the prevalence of FLAs in the two different drinking water treatment plants. Morpho- logical identification of the isolated FLAs showed the presence of 3 genera namely Acanthamoeba, Naegleria, and Vermamoeba (Hartmannella) confirmed by PCR. Conclusion: The appearance of FLAs especially pathogenic amoebae in com- pletely treated drinking water may cause potential health threat although there is no statistical difference between the two examined drinking water filtration systems. 413 Available at: http://ijpa.tums.ac.ir Al-Herrawy & A. Gad: Assessment of Two Different Drinking Water Treatment … Introduction ater contamination is a common (15), but it is clearly ineffective for problem to all over the world (1). acanthamoeba cysts because they can resist The microbial contaminants in- exposure to 50 mg L-1 for 18 h (16) or 100 mg W -1 clude pathogens like bacteria, viruses, and pa- L chlorine for 10 min (17). Two log reduc- rasites such as microscopic protozoa and tion of Naegleria cysts could be achieved by worms. Human and animal wastes knowing or chlorine with CT value 29 mg min /L (18). unknowingly can spread these living organ- The act of producing drinking water free isms (2). Moreover, free-living amoebae from waterborne pathogens is considered the (FLAs) have the ability to survive in diverse main objective of water treatment providers. environments and have been isolated from Because no single treatment process can be soil, different aquatic environments and even expected to remove all of the different types air, indicating the ubiquitous nature of these of pathogens found in water, multiple barriers organisms (3-7). In addition, FLAs have been (pre-chlorination, coagulation, and sedimenta- detected and consequently isolated from hos- tion, filtration and post-chlorination) are de- pitals (like dialysis units, eyewash stations) and sirable. Filtration is a physical removal of or- clinical samples (human lungs tissues, nasal ganisms together with other particulate matter. cavities, corneal biopsies, pharyngeal swabs, Various filtration processes (as rapid and slow skin lesions, brain cerebrospinal and fluid ne- sand filtrations) are used in conventional cropsies) (8-10). Unlike ―true‖ parasites, pa- drinking water treatment plants (19). In Egypt, thogenic FLAs can complete their life cycles although rapid sand filters are widely used in in the environment without entering a human conventional drinking water treatment plants, or animal host. Some of FLAs are pathogenic the slow sand filters are also used but in a for humans (11). Most Acanthamoeba species small scale. have an association with human disease as Therefore, the aim of this study was to granulomatous amoebic encephalitis (GAE), compare between a slow sand filter and a rap- pulmonary and kidney infections, nasopharyn- id sand filter in the corresponding drinking geal, cutaneous lesions, primarily in water treatment plants for removal of FLAs immunocompromised patients. Acanthamoeba and to identify the isolated free-living amoe- species also cause amoebic keratitis in bae. immunocompetent persons. Another species, Balamuthia mandrillaris close relative to Acan- Materials and Methods thamoeba, cause skin and lung infections as well as fatal GAE mostly in healthy children. Naeg- Operational design of a drinking water leria fowleri causes a non-opportunistic primary treatment plant amoebic meningoencephalitis (PAM) in This study was conducted on two different healthy children and young adults. Sappinia drinking water treatment plants (DWTPs) lo- pedata has been reported from a brain infec- cated in Fayoum Drinking Water and Waste- tion in a healthy man (12). Vahlkampfia, water Holding Company, Egypt during 2015. Vannella, and Vermamoeba species have also One DWTP was operated by rapid sand filtra- been isolated from the eye surface of humans tion system, while the other was operated by (13, 14). slow sand filtration system. Moreover, rapid -1 Free and combined chlorine at 10 mg L sand filters required smaller land areas com- was reported to be effective against Hartman- pared to slow sand filters, so they were widely nella vermiformis cysts after 30 min exposure used in large municipal water systems by the Available at: http://ijpa.tums.ac.ir 414 Iran J Parasitol: Vol. 12, No. 3, Jul-Sep 2017, pp.413-422 1920s. Rapid sand filters use relatively coarse, A conventional drinking water treatment sand and other granular media to remove im- plant consists of 4 different steps beginning purities and particles trapped in from the intake water (raw surface water). a flow through the use of chemicals— Raw water from the intake is sucked in pipes typically alum for flocculation. After floccula- having coarse metal sieves with 4cm pore size tion step, the unfiltered water flows through for prevention of coarse objects from getting the filter medium under pumped pressure and entrance with sucked water. The sieved raw the floc material is trapped in the sand matrix. water is pumped to coagulation and precipita- With respect to slow sand, filtration is a proc- tion basins where it is mixed with aluminum ess involving passage of raw water through a sulfate to aid in the flocculation and precipita- sand bed at low velocity (generally less than tion of the debris and microorganisms found 0.4 m/h) compared with 20 m/h in a rapid in raw water. After that, the clear water in the granular media filtration, resulting to substan- top of sedimentation basins is collected and tial particulate removal by physical and bio- passed on sand filters to get rid of the remain- logical mechanisms. These filters work ing microorganisms as well as escaped very through the formation of a gelatinous layer small particles. Filtered water is collected in (or biofilm) named Schmutzdecke. This layer storage tanks where it is injected with chlorine laid at the top few millimeters of the fine sand dose of 2mg/l for disinfection. The disin- layer. Schmutzdecke is formed in the first 10–20 fected water (outlet water) is ready to be d of operation and consists of fungi, bacte- pumped and distributed to the consumers as a ria, protozoa, rotifer and a range of aquatic drinking water (Fig. 1) (21). insect larvae. Schmutzdecke layer provides the effective purification in potable water treat- Water samples ment; sand underlying Schmutzdecke layer pro- Two types of water were collected from the vides the support medium for this biological two previously mentioned DWTPs: raw (inlet) treatment layer. Then water passes through water and treated (outlet) water. Water sam- the hypogeal layer, foreign matter particles are ples (one-liter volume each) were collected trapped in the mucilaginous matrix and so- monthly along on year from each of the two luble organic material is adsorbed. Microor- DWTPs. The samples were separately col- ganisms as the bacteria, fungi, and protozoa lected in autoclavable polypropylene contain- metabolized contaminants (20, 21). ers (one-liter volume) (22). Fig. 1: Operating diagram for a conventional drinking water treatment plant 415 Available at: http://ijpa.tums.ac.ir Al-Herrawy & A. Gad: Assessment of Two Different Drinking Water Treatment … Concentration, cultivation and morpho- GACCATCCGGAGTTCTCG-3) were used logical characterization of freshwater for amplify18S rDNA of Vermamoeba (Hart- amoebae from collected samples mannella) vermiformis (27, 28). Naegleria was Collected water samples were separately identified by genus specific primer (5- concentrated by using the membrane filtration CAAACACCGTTATGACAGGG-3) and (5- technique. Each water sample (One-liter vo- CTGGTTTCCCTTACCTTGCG-3) (28). In lume) was filtered through a nitrocellulose addition, species-specific primer was used to membrane filter (0.45µm pore size and 47mm confirm the presence of Naegleria fowleri (5- in diameter) by using a stainless steel holder GTGAAAACCTTTTTTCCATTTACA-3) and (5- connected with a suction pump. The mem- AAATAAAAGATTGACCATTTGAAA-3) brane of each filtered water sample was face (29). Amplification of DNA was performed to face inverted on the surface of non-nutrient using Maxima™ Hot Start Green PCR Master agar medium seeded with living E. coli bacteria Mix (Thermo Fisher Scientific Inc, Waltham, and incubated at 30 °C for two week with dai- MA, USA) according to the manufacturer ma- ly microscopic examination using the inverted nual.
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