Assessment of viability of the nematode eggs ([i]Ascaris[/i], [i]Toxocara[/i], [i]Trichuris[/i]) in sewage sludge with the use of LIVE/DEAD Bacterial Viability Kit

Jacek Karamon 1,  
Tomasz Cencek 1,  
Department of Parasitology and Invasive Diseases, National Veterinary Research Institute in Pulawy, Poland
Department of Water and Soil Safety, Institute of Rural Health in Lublin, Lublin, Poland
Ann Agric Environ Med 2014;21(1):35–41
Sewage sludges from wastewater treatment plants may contain live parasite eggs, which can be a source of humans and animals infection. According to the current rules, parasitological examination includes detection of the [i]Ascaris [/i]spp., [i]Trichuris [/i]spp. [i]and Toxocara [/i]spp. eggs and estimation of their viability. The viability assessment based only on the incubation and observation of isolated egg is long and imprecise. The aim of this study was to develop sensitive and less labourintensive methods for assessing viability of [i]Ascaris [/i]spp[i]., Toxocara [/i]spp[i]. and Trichuris [/i]spp[i].[/i] eggs in sewage sludge. For this purpose, LIVE/DEAD Kit was used. Firstly, the possibility of distinguishing between live and dead eggs in water was assessed. Secondly, an appropriate amount of dyeing mixture needed to distinguish the live and dead eggs in the sewage sludge was determined using experimentally enriched samples and naturally contaminated samples of sludge. Eggs were isolated from the samples by own method which was a combination of flotation and sedimentation, preceded by a long mixing. After the last stage of the procedure, sediment containing the eggs of parasites was stained by LIVE/DEAD kit according to the manufacturer instructions, but with the use of different variants of dyes mixture concentration. The investigation showed that live and dead eggs of these three parasites could be differed by this method with the use of proper concentration of dyes. Live eggs were stained in green ([i]Ascaris and Trichuris[/i]) and green-blue ([i]Toxocara[/i]). However, all types of dead eggs were red coloured. The study demonstrated that after some modifications (resulted from the nature of the samples) the LIVE/DEAD kit is useful for assessing the viability of [i]Toxocara[/i], [i]Ascaris[/i] and [i]Trichuris[/i] eggs occurring in the sludge.