Presence of Cryptosporidium parvum and Giardia lamblia in water samples from Southeast Asia: towards an integrated water detection system.

BACKGROUND: Access to clean and safe drinking water that is free from pathogenic protozoan parasites, especially Cryptosporidium parvum and Giardia lamblia that cause gastrointestinal illness in humans, is still an issue in Southeast Asia (SEA). This study is the first attempt to detect the aforementioned protozoan parasites in water samples from countries in SEA, using real-time polymerase chain reaction (qPCR) assays. METHODS: A total of 221 water samples of 10 l each were collected between April and October 2013 from Malaysia (53), Thailand (120), the Philippines (33), and Vietnam (15). A physicochemical analysis was conducted. The water samples were processed in accordance with the US Environmental Protection Agency's methods 1622/1623.1, microscopically observed and subsequently screened using qPCR assays. RESULTS: Cryptosporidium oocysts were detected in treated water samples from the Philippines (1/10), with a concentration of 0.06 ± 0.19 oocyst/L, and untreated water samples from Thailand (25/93), Malaysia (17/44), and the Philippines (11/23), with concentrations ranging from 0.13 ± 0.18 to 0.57 ± 1.41 oocyst/L. Giardia cysts were found in treated water samples from the Philippines (1/10), with a concentration of 0.02 ± 0.06 cyst/L, and in untreated water samples from Thailand (20/93), Vietnam (5/10), Malaysia (22/44), and the Philippines (16/23), with concentrations ranging from 0.12 ± 0.3 to 8.90 ± 19.65 cyst/L. The pathogens C. parvum and G. lamblia were detected using using qPCR assays by targeting the 138-bp fragment and the small subunit gene, respectively. C. parvum was detected in untreated water samples from the Philippines (1/23) and Malaysia (2/44), whilst, G. lamblia detected was detected in treated water samples from the Philippines (1/10) and in untreated water samples from Thailand (21/93), Malaysia (12/44), and the Philippines (17/23). Nitrate concentration was found to have a high positive correlation with (oo)cyst (0.993). CONCLUSION: The presence of (oo)cysts in the water samples means that there is potential risk for zoonotic disease transmission in the studied countries. Detection using qPCR is feasible for quantifying both pathogenic C. parvum and G. lamblia in large water samples.

Occurrence and antimicrobial resistance of pathogenic vibrios isolated from green mussel, Perna viridis L. 1758 in Bacoor Bay, Cavite, Philippines

OBJECTIVES: To determine the occurrence and antimicrobial resistance of pathogenic vibrios from green mussel, Perna viridis in bacoor bay cavite. METHODS: Vibrios were isolated using TCBS agar and confirmed using API Identification kit. The relationship between the density of vibrios and environment parameters such as months, harvest sites and other physico-chemical parameters of water was determined. The isolated vibrios were subjected to antibiotic resistance testing to determine the resistance patterns.RESULTS: Pathogenic vibrios were isolated in all of the 90 mussel samples. Of the 183 vibrio isolates, 75 strains were V. alginolyticus and 53 strains belonged to V. cholerae wherein 18 (34%) of these were V. cholerae serotype O1. Forty nine (49) strains belonged to V. parahaemolyticus and six were V. vulnificus. Using the regression analysis, salinity, water temperature and ph have significant correlation on the density of vibrios (p=0.0309), with regression model, total vibrios = -16990763 + 145858 Salinity + 186808 Temperature + 1037886 pH. In the antimicrobial resistance assay,ampicillin had the highest drug resistance (37.8%) followed by nalidixic acid (10.4%), tetracycline (10.4%) and co-trimoxazole (9.3%).CONCLUSION: Pathogenic vibrios were isolated in mussel of Bacoor, Cavite and can be predicted using salinity, temperature and pH of the water. Thirty six percent (36%) of Vibrio isolates were resistance to ampicillin.

Waterborne parasites: a current status from the Philippines.

BACKGROUND: Despite the amount of awareness created, waterborne disease still poses threat, especially in developing countries. Due to the scarcity of reported data on waterborne parasites, the consumption of unsafe water prolongs. Thus, the occurrences of waterborne parasites from various samples were investigated from one of the Southeast Asian country, the Philippines. METHODS: A total of thirty three samples, each consisting of twelve liters, were collected and processed to obtain the sediment. Ten liters of sample each was processed to detect Cryptosporidium spp. and Giardia spp. using an immunomagnetic separation method prior to enumeration via fluorescence microscope. Meanwhile, the remaining two liters were cultured to detect Acanthamoeba and Naegleria through microscopy examination and polymerase chain reaction (PCR) analysis. RESULTS: Twelve samples (36.4%) from river (5), swimming pool (1), pond (3), rain tank (1), and natural lake (2) were positive for Cryptosporidium spp., 17 (45.5%) samples from river (9), pond (2), swimming pool (1), rain tank (1), and natural lake (4) were positive for Giardia spp. while, 13 (33.3%) samples from river (3), swimming pool (2), pond (2), dispenser (1), well (1), tap (2) and natural lake (2) were positive for Acanthamoeba spp. and 5 (18.2%) samples from river (1), natural lake (1), tap (1), dispenser (1) and mineral (1) were Naegleria spp. positive. Physical parameters such as temperature, conductivity, total dissolved solid (TDS), salinity, dissolved oxygen (DO), pH, and turbidity and chemical parameters such as ammonia, chlorine, fluoride, nitrate and nitrite were also measured. The highest chemical contamination was observed at pond 2. A good correlation was observed between Giardia and nitrite (r = 0.736, p < 0.01) and Giardia and nitrate (r = 0.502, p < 0.01). CONCLUSION: This study was aimed to create greater awareness of parasitic contamination in the water environment in the Philippines and also to act as a platform of the current scenario for policymakers as water pollution is a key health issue in this region.