Assessment of viability of the nematode eggs (Ascaris, Toxocara, Trichuris) in sewage sludge with the use of LIVE/DEAD Bacterial Viability Kit
More details
Hide details
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
Jacek Karamon   

Department of Parasitology and Invasive Diseases, National Veterinary Research Institute in Pulawy, 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 Ascaris spp., Trichuris spp. and Toxocara 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 Ascaris spp., Toxocara spp. and Trichuris spp. 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 (Ascaris and Trichuris) and green-blue (Toxocara). 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 Toxocara, Ascaris and Trichuris eggs occurring in the sludge.
Jimenez B. Helminths (worms) eggs control in wastewater and sludge. International Symposium on New Directions in Urban Water Management; Paris 2007.
Othman AA. Therapeutic battle against larval toxocariasis: Are we still far behind? Acta Trop. 2012; 124(3): 171–178.
Jagannath P M, Venkataramana N K, Rao S A, Naik A L, Shivakumar S K, Saktepar A et al. Recurrent cerebral larva migrans: A case report and review of literature. J Pediatr Neurosci. 2009; 4(1): 36–40.
Acar A, Oncul O, Cavuslu S, Okutan O, Kartaloglu Z. Case report: Loffler’s syndrome due to Ascaris lumbricoides mimicking acute bacterial community-acquired pneumonia. Turkiye Parazitol Derg. 2009; 33(3): 239–241.
Hefny A F, Saadelin Y A, Abu-Zidan F M. Management algorithm for intestinal obstruction due to ascariasis: a case report and review of the literature. Ulus Travma Acil Cerrahi Derg. 2009; 15(3): 301–305.
Bahu Mda G, Baldisserotto M, Custodio C M, Gralha C Z, Mangili A R. Hepatobiliary and pancreatic complications of ascariasis in children: a study of seven cases. J Pediatr Gastroenteral Nutr. 2001; 33(3): 271–275.
Baldisserotto M. Trichuriasis colitis detected by Doppler sonography. Pediatr Radiol 2010; 40: S95–97.
Khuroo MS, Khuroo NS. Trichuris dysentery syndrome: a common cause of chronic iron deficiency anemia in adults in an endemic area (with videos). Gastrointest Endosc. 2010; 71(1):200–204.
Stephenson L S, Holland C V, Cooper E S. The public health significance of Trichuris trichura. Parasitology 2000; 121: 73–95.
Bowman DD, Little MD, Reimers RS. Precision and accuracy of an assay for detecting Ascaris eggs in various biosolid matrices. Water Res. 2003; 37: 2063–2072.
Gaspard P G, Wiart J, Shwartzbrod J. Urban sludge reuse in agriculture: waste treatment and parasitological risk. Bioresource Technol. 1995; 52, 37–40.
Zdybel J, Cencek T, Karamon J. Comparison of the efficiency of different parasitological diagnostic methods used in analysis of dehydrated sewage sludges. 2 nd Congress of the European Association of Veterinary Laboratory Diagnosticians (EAVLD); Kazimierz Dolny, Poland 2012.
Gaspard P, Schwartzbrod J. Irrigation with waste water: parasitological analysis of soil. Zentralbl Hyg Umweltmed. 1993; 193 (6): 513–520.
Johnson P W, Dixon R, Ross A D. An in vitro test for assessing the viability of Ascaris suum eggs exposed to various sewage treatment processes. Int J Parasitol. 1998; 28: 627–633.
Papajová I, Juriš P, Szabová E, Venglovský J, Sasáková N, Šefčíková H, Martinez J, Gáboň T. Decontamination by anaerobic stabilisation of the environment contaminated with enteronematode eggs Toxocara canis and Ascaris suum. Bioresource Technology. 2008; 99: 4966–4971.
Chapalamadugu KC, Busboom JR, Nelson ML, Hancock DD, Tang J, Jasmer DP. Taenia taeniaeformis: Effectiveness of staining oncospheres is related to both temperature of treatment and molecular weight of dyes utilized. Vet Parasitol. 2008; 151: 203–211.
de Victorica J, Galvan M. Preliminary testing of rapid coupled methodology for quantitation/viability determination of helminth eggs in raw and treated wastewater. Water Res. 2003; 37: 1278–1287.
Taghi-Kilani, Gyurek LL, Millard PJ, Finch GR, Belosevic M. Nucleic Acid Stains as Indicators of Giardia muris Viability Following Cyst Inactivation. Int J Parasitol. 1995; 26: 637–646.
Schupp DG, Erlandsen SL. Determination of Giardia muris cyst viability by differential interference contrast, phase or brightfield of microscopy. J Parasitol. 1987; 73(4): 723–729.
Boulos L, Prevost M, Barbeau B, Coallier J, Desjardins R. LIVE/ DEAD BacLight: application of a new rapid staining method for direct enumeration of viable and total bacteria in drinking water. J Microbiol Meth. 1999; 37: 77–86.
Queric N D, Soltwedel T, Arntz WE.Application of rapid direct viable count method to deep-sea sediment bacteria. J Microbiol Meth. 2004; 57: 351–367.
Ivanova IA, Kambarev S, Popova RA, Naumovska EG, Markoska KB, Dushkin CD. Determination of Pseudomonas putida live cells with classic cultivation and staining with “LIVE/DEAD Baclight Bacterial Viability Kit”; Second Balkan Conference on Biology; 21–23 May 2010; Bulgaria; Diagnosis Press, 2010.