The objective of this study was to find out the impact of environmental conditions on the survival of intestinal parasites on environmental surfaces commonly implicated in the transmission of these parasites. The study was performed by incubating Cryptosporidium and Giardia (oo)cysts on environmentally relevant surfaces such as brushed stainless steel, formica, ceramic, fabric, and skin. Parallel experiments were conducted using clean and soiled coupons incubated under three temperatures. The die-off coefficient rates (K) were calculated using first-order exponential formula. For both parasites, the fastest die-off was recorded on fabric, followed by ceramic, formica, skin, and steel. Die-off rates were directly correlated to the incubation temperatures and surface porosity. The presence of organic matter enhanced the survivability of the resting stages of test parasites. The decay rates calculated in this study can be used in models for public health decision-making process and highlights the mitigation role of hand hygiene agents in their prevention and control.
This study shows that the ECC-RT-PCR, a new integrated cell culture assay, can be used as a rapid and cost-effective tool for assessing the viability and infectivity of environmental isolates of Giardia sp cysts.
Globally, Giardia is one of the major cause of diarrheal illnesses and rapid diagnostic methods differentiating infectious cysts are critical for developing intervention strategies through contaminated surfaces, food and water. This is currently hampered by lack of in vitro model. We evaluated mRNA expression in trophozoites and their attachment to CaCo2 (C2bb) cell monolayer and changes in trans-cellular resistance as an indicator of Giardia viability and infectivity. Heat shock mRNA in Giardia cysts and variant specific protein (VSP) mRNA in trophozoites were quantified by RT-PCR. When compared with neonatal mice infectivity, the attachment of trophozoites to cell monolayer, expression of VSP and change in the trans-cellular resistance, the infectivity directly correlated with infectivity in neonatal mice. This study highlights the use of molecular method combined with electrophysiological analysis of cell culture (ECC-rtPCR) post-trophozoite's attachment for assessing viability and infectivity of environmental isolates of Giardial cysts.
Globally, Giardia is one of the major cause of diarrheal illnesses and rapid diagnostic methods differentiating infectious cysts are critical for developing intervention strategies through contaminated surfaces, food and water. This is currently hampered by lack of in vitro model. We evaluated mRNA expression in trophozoites and their attachment to CaCo2 (C2bb) cell monolayer and changes in trans-cellular resistance as an indicator of Giardia viability and infectivity. Heat shock mRNA in Giardia cysts and variant specific protein (VSP) mRNA in trophozoites were quantified by RT-PCR. When compared with neonatal mice infectivity, the attachment of trophozoites to cell monolayer, expression of VSP and change in the trans-cellular resistance, the infectivity directly correlated with infectivity in neonatal mice. This study highlights the use of molecular method combined with electrophysiological analysis of cell culture (ECC-rtPCR) post-trophozoite's attachment for assessing viability and infectivity of environmental isolates of Giardial cysts.
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