An aquatic biomonitoring of Giardia cysts and Cryptosporidium oocysts in river water corresponding to five villages situated in three states in peninsular Malaysia was determined. There were 51.3% (20/39) and 23.1% (9/39) samples positive for Giardia and Cryptosporidium (oo)cysts, respectively. Overall mean concentration between villages for Giardia cysts ranged from 0.10 to 25.80 cysts/l whilst Cryptosporidium oocysts ranged from 0.10 to 0.90 oocysts/l. Detailed results of the river samples from five villages indicated that Kuala Pangsun 100% (9/9), Kemensah 77.8% (7/9), Pos Piah 33.3% (3/9) and Paya Lebar 33.3% (1/3) were contaminated with Giardia cysts whilst Cryptosporidium (oo)cysts were only detected in Kemensah (100 %; 9/9) and Kuala Pangsun (66.6%; 6/9). However, the water samples from Bentong were all negative for these waterborne parasites. Samples were collected from lower point, midpoint and upper point. Midpoint refers to the section of the river where the studied communities are highly populated. Meanwhile, the position of the lower point is at least 2 km southward of the midpoint and upper point is at least 2 km northward of the midpoint. The highest mean concentration for (oo)cysts was found at the lower points [3.15 ± 6.09 (oo)cysts/l], followed by midpoints [0.66 ± 1.10 (oo)cysts/l] and upper points [0.66 ± 0.92 (oo)cysts/l]. The mean concentration of Giardia cysts was highest at Kuala Pangsun (i.e. 5.97 ± 7.0 cysts/l), followed by Kemensah (0.83 ± 0.81 cysts/l), Pos Piah (0.20 ± 0.35 cysts/l) and Paya Lebar (0.10 ± 0.19 cysts/l). On the other hand, the mean concentration of Cryptosporidium oocysts was higher at Kemensah (0.31 ± 0.19 cysts/l) compared to Kuala Pangsun (0.03 ± 0.03cysts/l). All the physical and chemical parameters did not show significant correlation with both protozoa. In future, viability status and molecular characterisation of Giardia and Cryptosporidium should be applied to identify species and genotypes/subgenotypes for better understanding of the epidemiology of these waterborne parasites.
We used a combined microscopy-molecular approach to determine the occurrence and identities of waterborne Giardia sp. cysts isolated from 18 separate, 10l grab samples collected from a Malaysian zoo. Microscopy revealed that 17 of 18 samples were Giardia cyst positive with concentrations ranging from 1 to 120 cysts/l. Nine (52.9%) of the 17 cyst positive samples produced amplicons of which 7 (77.8%) could be sequenced. Giardia duodenalis assemblage A (6 of 7) and assemblage B (1 of 7), both infectious to humans, were identified at all sampling sites at the zoo. The presence of human infectious cysts raises public health issues, and their occurrence, abundance and sources should be investigated further. In this zoo setting, our data highlight the importance of incorporating environmental sampling (monitoring) in addition to routine faecal examinations to determine veterinary and public health risks, and water monitoring should be considered for inclusion as a separate element in hazard analysis, as it often has a historical (accumulative) connotation.
Chemical inactivation of microorganisms is a common process widely employed in many fields such as in treatment of water, preservation in food industry and antimicrobial treatments in healthcare. For economy of applications and efficiency of treatment establishment the minimum dosage of breakpoint in the chemical application becomes essential. Even though experimental investigations have been extensive, theoretical understanding of such processes are demanding. Commonly employed theoretical analyses for the inactivation of microorganisms and depletion of chemicals include kinetics expressing the rates of depletion of chemical and microorganisms. The terms chemical demand (x) and specific disinfectant demand (alpha) are often used in theoretical modeling of inactivation. The value of specific disinfectant demand (alpha) has always been assumed to be a constant in these models. Intracellular concentration built up within the cells of the microorganisms during inactivation could lead to possible weakening effects of microorganisms thereby requiring lower doses as disinfection proceeds makes the assumption of constant alpha inaccurate. Model equations are formulated based on these observations co-relating the parameters alpha and x with a progressive inactivation (N/N(0)). The chemical concentration (C) is also presented in terms of the inactivation time (t) and the survival ratio (N/N(0)) for given pH and temperature conditions. The model is examined using experimentally verified Ct data of Giardia Cysts/chlorine system. The respective values of x for different survival ratios were evaluated from the data using MatLab software. Proposed model correlating for the disinfectant demand (x) with the survival ratio (N/N(0)) fits satisfactorily with those evaluated from data. The rate constants for different pH and temperature conditions are evaluated which showed compatibility with the Arrhenius model. The dependence of frequency factors with pH indicated compatibility with accepted models. The Ct values regenerated with the kinetic data shows a very accurate fit with published data.
A catchment-scale investigation of the prevalence of Cryptosporidium and Giardia in the Kuang River Basin was carried out during the dry and rainy seasons. Water samples were collected from the Kuang River and its tributaries as well as a major irrigation canal at the study site. We also investigated the prevalence of gastrointestinal parasitic infection among dairy and beef cattle hosts. Cryptosporidium and/or Giardia were detected in all the rivers considered for this study, reflecting their ubiquity within the Kuang River Basin. The high prevalence of Cryptosporidium/Giardia in the upper Kuang River and Lai River is of a particular concern as both drain into the Mae Kuang Reservoir, a vital source of drinking-water to many local towns and villages at the research area. We did not, however, detected neither Cryptosporidium nor Giardia were in the irrigation canal. The frequency of Cryptosporidium/Giardia detection nearly doubled during the rainy season compared to the dry season, highlighting the importance of water as an agent of transport. In addition to the overland transport of these protozoa from their land sources (e.g. cattle manure, cess pits), Cryptosporidium/Giardia may also be re-suspended from the streambeds (a potentially important repository) into the water column of rivers during storm events. Faecal samples from dairy and beef cattle showed high infection rates from various intestinal parasites - 97% and 94%, respectively. However, Cryptosporidium and Giardia were only detected in beef cattle. The difference in management style between beef (freeranging) and dairy cattle (confined) may account for this disparity. Finally, phylogenetic analyses revealed that the Cryptosporidium/Giardia-positive samples contained C. ryanae (non-zoonotic) as well as Giardia intestinalis assemblages B (zoonotic) and E (non-zoonotic). With only basic water treatment facilities afforded to them, the communities of the rural area relying on these water supplies are highly at risk to Cryptosporidium/Giardia infections.
This study is the first report on the occurrence of Giardia and Cryptosporidium (oo)cysts in recreational rivers water from Malaysia. It was carried out in water samples at two rivers, 'Sungai Congkak' and 'Sungai Batu', located in Selangor State. The occurrence of both Giardia lamblia and Cryptosporidium parvum (oo)cysts was higher in Sungai Congkak (50% or 15/30 and 10% or 3/30 respectively) than Sungai Batu (16% or 5/30 and 3.3% or 1/30 respectively). The mean density of cysts/L was 0.72 in Sungai Congkak and 0.023 in Sungai Batu, and that of oocysts/L was 0.023 in Sungai Congkak and 0.0033 in Sungai Batu, showing that the occurrence of Giardia was higher and more frequent than Cryptosporidium in both rivers. Sungai Congkak also showed higher faecal coliforms count (ranging from 0.48x10³ to 73x10³ CFU/100 mL) than Sungai Batu (0.41x10³ to 16x10³ CFU/100 mL). On the other hand, the Giardia and Cryptosporidium (oo)cysts and faecal coliforms were more concentrated at the downstream station, followed by midstream and upstream stations which might be due to human factors where settlements and recreation areas were located around and between midstream and downstream stations. The (oo)cysts and faecal coliforms also increased during public holidays due to the significantly higher number of visitors (bathers) compared with the week days. All the parameters (physical, faecal coliforms and rainfall) did not show consistent significant correlation (based on r values of Pearson correlation analysis) with both protozoa, therefore these parameters are not suitable as indicator for the presence of Giardia and Cryptosporidium (oo)cysts in both rivers.
A study to determine the contribution of Giardia cysts and Cryptosporidium oocysts from cattle farms was carried out at the Langat Basin. This study investigated the contribution of cattle farms, located near Sungai Langat and Sungai Semenyih, towards river contamination with these cysts and oocysts. The findings showed that out of 24 samples of water taken from Sungai Semenyih, 4.2% was positive for Giardia cysts with a concentration of 1.3 cysts/L and 20.8% were positive with Cryptosporidium oocysts with a range of 0.7 - 2.7 oocysts/L. At Sungai Langat, from the 43 samples taken, 23.3% were positive for Giardia cysts with a range of 1.5 - 9 cysts/L whereas 11.6% were positive with Cryptosporidium oocysts with a range of 2.5 - 240 oocysts/L. Isolation of cysts and oocysts in bovine faecal materials revealed that 14.6% of faecal samples were positive for Giardia cysts which had a range of 75 - 1.3x104 cysts/g and 25% were positive for Cryptosporidium oocysts with a range of 50 - 3.9x105 oocysts/g. From the cattle wastewater, 98% were positive with oocysts and 6.7% with cysts. The concentrations were between 20 - 3.1x103 oocysts/mL for Cryptosporidium and 4 - 75 cysts/mL for Giardia. Given that the prevalence of Cryptosporidium and Giardia are high amongst the cattle and the positive findings of the (oo)cysts in the river samples, it could be deduced that there is a very high possibility of the cattle farms contaminating the river with Giardia cysts and Cryptosporidium oocysts. Viability study of Cryptosporidium oocysts in the surrounding soil and pond within the cattle farm showed that the viability of Cryptosporidium oocysts decreased with time. It was estimated that it will take 52 days for all the oocysts from both environment to be non-viable. With a viability rate of approximately 2 months in a cattle farm setup, river water contaminated with Cryptosporidium oocysts has a high chance of acting as an agent of transmission. As cattle farms are also inhabited by the owners and their families, this problem may pose a threat to humans (e.g. children) especially if they are dependent on the river water as their source of water for their daily activities.