Worldwide absence of canonical benzimidazole resistance-associated mutations within ß-tubulin genes from Ascaris.

BACKGROUND: The giant roundworm Ascaris is an intestinal nematode, causing ascariasis by infecting humans and pigs worldwide. Recent estimates suggest that Ascaris infects over half a billion people, with chronic infections leading to reduced growth and cognitive ability. Ascariasis affects innumerable pigs worldwide and is known to reduce production yields via decreased growth and condemnation of livers. The predominant anthelminthic drugs used to treat ascariasis are the benzimidazoles. Benzimidazoles interact with ß-tubulins and block their function, and several benzimidazole resistance-associated mutations have been described in the ß-tubulins of ruminant nematodes. Recent research on ascarids has shown that these canonical benzimidazole resistance-associated mutations are likely not present in the ß-tubulins of Ascaris, Ascaridia or Parascaris, even in phenotypically resistant populations. METHODS: To further determine the putative absence of key ß-tubulin polymorphisms, we screened two ß-tubulin isotypes of Ascaris, highly expressed in adult worms. Using adult and egg samples of Ascaris obtained from pigs and humans worldwide, we performed deep amplicon sequencing to look for canonical resistance-associated mutations in Ascaris ß-tubulins. Subsequently, we examined these data in closer detail to study the population dynamics of Ascaris and genetic diversity within the two isotypes and tested whether genotypes appeared to partition across human and pig hosts. RESULTS: In the 187 isolates, 69 genotypes were found, made up of eight haplotypes of ß-tubulin isotype A and 20 haplotypes of isotype B. Single nucleotide polymorphisms were seen at 14 and 37 positions for ß-tubulin isotype A and isotype B, respectively. No evidence of any canonical benzimidazole resistance-associated mutations was found in either human- or pig-derived Ascaris isolates. There was, however, a difference in the genetic diversity of each isotype and distribution of ß-tubulin genotypes between human- and pig-derived Ascaris. Statistical tests of population differentiation show significant differences (p < 0.001) between pig- and human-derived worms; however, more diversity was seen between worms from different populations than worms from different hosts. CONCLUSIONS: Our work suggests an absence of canonical ß-tubulin mutations within Ascaris, but alternative modes of anthelminthic resistance may emerge necessitating continued genetic scrutiny alongside monitoring of drug efficacy.

Concurrent decreases in the prevalence of wheezing and Ascaris infection among 5-year-old children in rural Bangladesh and their regulatory T cell immunity after the implementation of a national deworming program.

INTRODUCTION: Epidemiological research on the prevalence of asthma and helminthic infections in various countries has led to the hypothesis that helminthic infections protect against asthma by suppressing the host's immune response. This study was conducted to elucidate whether decreased Ascaris infection following a national deworming program was associated with increased recurrent wheezing among rural Bangladeshi children and to test their anti-inflammatory immunity. METHODS: This nested case-control study was conducted from December 2015 to October 2016 in the rural service area of the International Centre for Diarrhoeal Disease Research, Bangladesh. Of the 1800 5-year old children randomly selected for the study, informed consent was obtained from the guardians of 1658 children. Data were collected using a semistructured questionnaire adopted from the International Study of Asthma and Allergies in Childhood and blood samples for the analysis of regulatory T (Treg) cell immune responses and the balance between Th1 and Th2 immunity in Ascaris infections. RESULTS: A total of 145 children were found to have wheezing, yielding a prevalence rate of 8.7%, which was significantly lower than the rate found in 2001 (16.2%, P < .001); Ascaris infection also decreased from 2001 to 2016. The 127 wheezing children who agreed to participate further were compared to 114 randomly selected never-wheezing children. Wheezing had a significant positive association with antibiotic use, history of pneumonia, parents' history of asthma, and Ascaris infection; children with Ascaris infection were twice as likely to have wheezing (adjusted odds ratio = 2.31, P = .053). Flow cytometry found no significant differences in the rates of Th1, Th2, and CD4 + CD25 + CD127low cells by the wheezing group. CONCLUSIONS: Ascaris infection had a positive rather than a negative association with wheezing and the rates of wheezing and Ascaris infections both decreased from 2001 to 2016. These findings undermines the hypothesis that such infections provide protection against asthma.

Risk factors for infection with soil-transmitted helminths during an integrated community level water, sanitation, and hygiene and deworming intervention in Timor-Leste.

Water, sanitation and hygiene interventions have been advocated as important complements to deworming programs to improve soil-transmitted helminth control. Evidence for the impact of water, sanitation and hygiene on soil-transmitted helminth infections is mixed, and based mainly on cross-sectional studies. In this study, we assessed associations between individual- and household-level water, sanitation and hygiene variables and soil-transmitted helminth infections, using data collected during the 2 year follow-up study period of the WASH for WORMS randomised controlled trial in Timor-Leste. Data were collected across four surveys, conducted at 6 monthly intervals in 23 communities. We analysed water, sanitation and hygiene and sociodemographic variables as risk factors for infection with Necator americanus, Ascaris spp., and undifferentiated soil-transmitted helminth infection, using generalised linear mixed models to account for clustering at community, household and participant levels. Water, sanitation and hygiene risk factors were examined both concurrently and with a 6 month lag period that coincided with the most recent deworming. The analysis included 2333 participants. Factors associated with N. americanus infection included age group, male sex (adjusted odds ratio (aOR) 3.1, 95% confidence interval (CI) 2.4-4.2), working as a farmer (aOR 1.7, 95% CI 1.2-2.4), and completing secondary school or higher (aOR 0.29, 95% CI 0.16-0.53). Risk factors for Ascaris spp. infection included age group, living in a dwelling with more than six people (aOR 1.6, 95% CI 1.1-2.3), having a tube well or borehole as the household water source (aOR 3.7, 95% CI 1.3-10.8), and using a latrine shared between households 6 months previously (aOR 2.3, 95% CI 1.2-4.3). Handwashing before eating was protective against infection with any soil-transmitted helminth (aOR 0.79, 95% CI 0.65-0.95). In the context of regular deworming, few water, sanitation and hygiene-related factors were associated with soil-transmitted helminth infections. Future research examining the role of water, sanitation and hygiene in soil-transmitted helminth transmission is required, particularly in low transmission settings after cessation of deworming. Identifying improved indicators for measuring water, sanitation and hygiene behaviours is also a key priority.

[STRUCTURAL, MOLECULAR AND FUNCTIONAL FEATURES OF NEMATODE SURFACES AND THE POSSIBILITY OF ANTHELMINTIC DRUGS DEVELOPMENT (REVIEW)].

Helminthiases caused by parasitic nematodes are widespread in different regions of the world. The main adaptation for overcoming adverse conditions is a barrier properties of the cuticle surface structure, which differs from the membrane teguments of trematodes and cestodes. Different types of nematodes have specific structural and biochemical adaptations at different stages of their life cycle. While creating specific areas of habitat and nutrition, some types of parasites change the morphology and functioning of the host tissues. Ascaris suum and Caenorabditis elegans were widely used as model organisms in the study of genetics, biochemistry of nematodes. Studying of biochemistry and molecular biology of structural components of nematode surfaces is important for development of effective and safe anthelmintic drugs. The differences in the structure and functioning of transport enzymes of parasites and humans will help to create effective specific inhibitors and anthelmintic remedies. An important point of application of anthelmintic drugs can serve as inorganic ions transport proteins in the membranes of the surfaces. Glycolipids of cuticle contribute to the evasion from the host immune system, protecting the surface proteins from degradation by proteases. Study of helminth surfaces makes an important contribution to the development of anthelmintic drugs and vaccines, for helminthiasis treat.

Optimization of the synthesis process of an iron oxide nanocatalyst supported on activated carbon for the inactivation of Ascaris eggs in water using the heterogeneous Fenton-like reaction.

An experimental design methodology was used to optimize the synthesis of an iron-supported nanocatalyst as well as the inactivation process of Ascaris eggs (Ae) using this material. A factor screening design was used for identifying the significant experimental factors for nanocatalyst support (supported %Fe, (w/w), temperature and time of calcination) and for the inactivation process called the heterogeneous Fenton-like reaction (H2O2 dose, mass ratio Fe/H2O2, pH and reaction time). The optimization of the significant factors was carried out using a face-centered central composite design. The optimal operating conditions for both processes were estimated with a statistical model and implemented experimentally with five replicates. The predicted value of the Ae inactivation rate was close to the laboratory results. At the optimal operating conditions of the nanocatalyst production and Ae inactivation process, the Ascaris ova showed genomic damage to the point that no cell reparation was possible showing that this advanced oxidation process was highly efficient for inactivating this pathogen.

Modeling the inactivation of ascaris eggs as a function of ammonia concentration and temperature.

Ammonia sanitization is a promising technology for sanitizing human excreta intended for use as a fertilizer in agriculture. Ascaris eggs are the most persistent pathogens regarding ammonia inactivation and are commonly present in fecal sludge in low- and middle-income countries. In this study, a model for predicting ammonia inactivation of ascaris eggs was developed. Data from four previous studies were compiled and analyzed statistically, and a mathematical model for the treatment time required for inactivation was created. The inactivation rate increased with NH3 activity to the power of 0.7. The required treatment time was found to decrease 10-fold for each 16 °C temperature increase. Dry matter (DM) content and pH had no direct effect on inactivation, but had an indirect effect due to their impact on NH3 activity, which was estimated using the Pitzer approach. An additional model giving an approximation of Pitzer NH3 activity but based on the Emerson approach, DM content and total ammonia (NHTot) was also developed. The treatment time required for different log10 reductions of ascaris egg viability can thus easily be estimated by the model as a function of NH3 activity and temperature. The impact on treatment time by different treatment options can then be theoretically evaluated, promoting improvements of the treatment e.g. by adding urea or alkaline agents, or increasing the temperature by solar heating.

[Efficiency of etiologic correction of concomitant ascaridosis in the complex treatment of chronic pancreatitis].

The of antihelmintic preparation albendazole using in the complex treatment of patients with chronic pancreatitis with the concomitant ascaridosis was promote regression of clinical demonstration of basic and concomitant diseases (P < 0.05). Options of coprogram and the structural state of pancreas from data of ultrasonography in marks by Marseille-Cambridge classification of chronic pancreatitis, after the conducted treatment became the better (P < 0.05), that established expedience of the use of albendazole in complex treatment of patients with a chronic pancreatitis with a concomitant ascaridosis.

Efficacy and side effects of albendazole currently in use against Ascaris, Trichuris and hookworm among school children in Wondo Genet, southern Ethiopia.

Monitoring the efficacy of anthelminthic drugs is essential. The objective of this study was to assess the efficacy of a single oral dose of 400mg albendazole (ABZ) against the major soil-transmitted helminth (STH) infection in school children, Wondo Genet, southern Ethiopia. A single fresh stool sample was collected from 298 school children and examined using a duplicate smear of the Kato-Katz method. Children positive for STH infections were treated with single oral dose of 400mg ABZ and re-examined for intestinal helminth infections 21days post-treatment. The participants were interviewed for symptoms related with the drug uptake 24h after ABZ treatment. Children positive for Schistosoma mansoni infections were treated with Praziquantel (40mg/kg of body weight) after an ABZ treatment follow up survey. 51.3%, 49.7%, 44.6% and 88.3% had hookworm, Ascaris lumbricoides, Trichuris trichiura and any intestinal helminth infection, respectively. Cure rates were 97.4% for hookworm, 96.6% for A. lumbricoides and 30.8% for T. trichiura infections. Egg reduction rates (ERRs) were 99.8% for hookworm, 99.9% for A. lumbricoides and 83.1% for T. trichiura infections. Mild and transient symptoms were observed among the participants which were quite frequent. In conclusion, a 400mg single oral dose of ABZ was effective against hookworm and A. lumbricoides but less efficacious against T. trichiura infection. The drug resulted in high ERRs for hookworm, A. lumbricoides and T. trichiura. Administration of the drug in repeated doses or in combination with other drugs might be necessary.

The potential for self-sanitisation of faecal sludge by intrinsic ammonia.

Faecal sludge has the potential to be used as a sustainable fertiliser in agriculture, but the sludge must be sanitised due to its content of pathogenic microorganisms. The intrinsic ammonia from the urine may be sufficient for sanitisation of the sludge if it is not too diluted by flush water or lost by ventilation. To evaluate the potential for this sanitisation method, inactivation of Enterococcus faecalis, Salmonella typhimurium and Ascaris suum eggs during treatment were assessed. The inactivation was studied at different storage temperatures (10-28 °C) and in several sludge mixes with different contents of urine, faeces and flush water, and with ammonia concentrations from 40 to 400 mM. All pathogens were inactivated by the ammonia, and ascaris eggs were the most persistent. Lower flush water volume and higher urine content favoured inactivation, mainly due to increased uncharged ammonia (NH3) concentration. The lag phase in ascaris inactivation was shortened by increasing temperature and NH3 concentration, while post-lag phase inactivation was not influenced by NH3 concentration. Faecal sludge can be sanitised by airtight storage without the use of additives when flush water volumes are sufficiently low. For temperatures of 23-28 °C, a 3 log reduction of ascaris egg viability can be achieved within 1-6 months depending on ammonia concentration and temperature.

Ammonia inactivation of Ascaris ova in ecological compost by using urine and ash.

Viable ova of Ascaris lumbricoides, an indicator organism for pathogens, are frequently found in feces-derived compost produced from ecological toilets, demonstrating that threshold levels of time, temperature, pH, and moisture content for pathogen inactivation are not routinely met. Previous studies have determined that NH(3) has ovicidal properties for pathogens, including Ascaris ova. This research attempted to achieve Ascaris inactivation via NH(3) under environmental conditions commonly found in ecological toilets and using materials universally available in an ecological sanitation setting, including compost (feces and sawdust), urine, and ash. Compost mixed with stored urine and ash produced the most rapid inactivation, with significant inactivation observed after 2 weeks and with a time to 99% ovum inactivation (T(99)) of 8 weeks. Compost mixed with fresh urine and ash achieved a T(99) of 15 weeks, after a 4-week lag phase. Both matrices had relatively high total-ammonia concentrations and pH values of >9.24 (pK(a) of ammonia). In compost mixed with ash only, and in compost mixed with fresh urine only, inactivation was observed after an 11-week lag phase. These matrices contained NH(3) concentrations of 164 to 173 and 102 to 277 mg/liter, respectively, when inactivation occurred, which was below the previously hypothesized threshold for inactivation (280 mg/liter), suggesting that a lower threshold NH(3) concentration may be possible with a longer contact time. Other significant results include the hydrolysis of urea to ammonia between pH values of 10.4 and 11.6, above the literature threshold pH of 10.

Use of Moringa oleifera seed extracts to reduce helminth egg numbers and turbidity in irrigation water.

Water from wastewater-polluted streams and dug-outs is the most commonly used water source for irrigation in urban farming in Ghana, but helminth parasite eggs in the water represent health risks when used for crop production. Conventional water treatment is expensive, requires advanced technology and often breaks down in less developed countries so low cost interventions are needed. Field and laboratory based trials were carried out in order to investigate the effect of the natural coagulant Moringa oleifera (MO) seed extracts in reducing helminh eggs and turbidity in irrigation water, turbid water, wastewater and tap water. In medium to high turbid water MO extracts were effective in reducing the number of helminth eggs by 94-99.5% to 1-2 eggs per litre and the turbidity to 7-11 NTU which is an 85-96% reduction. MO is readily available in many tropical countries and can be used by farmers to treat high turbid water for irrigation, however, additional improvements of water quality, e.g. by sand filtration, is suggested to meet the guideline value of ≤ 1 helminth egg per litre and a turbidity of ≤ 2 NTU as recommended by the World Health Organization and the U.S. Environmental Protection Agency for water intended for irrigation. A positive correlation was established between reduction in turbidity and helminth eggs in irrigation water, turbid water and wastewater treated with MO. This indicates that helminth eggs attach to suspended particles and/or flocs facilitated by MO in the water, and that turbidity and helminth eggs are reduced with the settling flocs. However, more experiments with water samples containing naturally occurring helminth eggs are needed to establish whether turbidity can be used as a proxy for helminth eggs.

Inactivation of Ascaris eggs in water using sequential solar driven photo-Fenton and free chlorine.

Sequential helminth egg inactivation using a solar driven advanced oxidation process (AOP) followed by chlorine was achieved. The photo-assisted Fenton process was tested alone under different H(2)O(2) and/or Fe(II) concentrations to assess its ability to inactivate Ascaris suum eggs. The effect of free chlorine alone was also tested. The lowest egg inactivation results were found using Fe(II) or H(2)O(2) separately (5 and 140 mmol L(-1), respectively) in dark conditions, which showed about 28% inactivation of helminth eggs. By combining Fe(II) and H(2)O(2) at the same concentrations described earlier, 55% of helminth egg inactivation was achieved. By increasing the reagent's concentration two-fold, 83% egg inactivation was achieved after 120 min of reaction time. Process efficiency was enhanced by solar excitation. Using solar disinfection only, the A. suum eggs inactivation reached was the lowest observed (58% egg inactivation after 120 min (120 kJ L(-1))), compared with tests using the photo-Fenton process. The use of the photo-Fenton reaction enhanced the process up to over 99% of egg inactivation after 120 kJ L(-1) when the highest Fe(II) and H(2)O(2) concentration was tested. Practically no effect on the helminth eggs was observed with free chlorine alone after 550 mg min L(-1) was used. Egg inactivation in the range of 25-30% was obtained for sequential processes (AOP then chlorine) using about 150 mg min L(-1).

Viability of Ascaris and other helminth genera non larval eggs in different conditions of temperature, lime (pH) and humidity.

Helminth eggs are the pathogens most resistant to inactivation during sludge and wastewater treatment. For this reason, in several regulations and the WHO guidelines for wastewater and excreta reuse for agriculture and aquaculture they are considered as indicators of the performance of the treatment process. Conditions required to inactivate helminth eggs, notably Ascaris lumbricoides, are recommended in the literature, but in practice these have not always proven effective, not only for Ascaris but also other genera of helminth eggs. The objective of this research was to study the inactivation of a high total content of non larval Ascaris and other genera of helminth eggs of medical importance to developing countries under controlled conditions of (a) temperature (30 °C to 80 °C) and humidity (80, 90 and 95%) and (b) lime doses (15 and 20% of CaO w/w dry basis) and humidity (90 and 80%), using different contact times in both cases. The inactivation data obtained for different genera of non larval helminth eggs is presented. Results showed that there is a combination of conditions (temperature, pH and humidity) that is optimal for inactivation. To completely inactivate any genera of non larval helminth eggs: (a) a temperature above 70 °C and 80% humidity for a duration of 120 min; and, (b) a 20% CaO dose (pH 12.5) and a humidity level of 80% for a duration of 8 months are needed. With regard to the resistance of different genera of helminth eggs, Ascaris, Toxocara and Taenia, in that order, were the most resistant, while the most sensitive were Trichuris and Hymenolepis. For most of the conditions tested Ascaris showed the highest resistance, probably due to the chemical arrangement of its membrane.

The actions of Caenorhabditis elegans neuropeptide-like peptides (NLPs) on body wall muscle of Ascaris suum and pharyngeal muscle of C. elegans.

The actions of six Neuropeptide-like Peptides (NLPs) were investigated on Ascaris suum dorsal body wall muscle and Caenorhabditis elegans pharyngeal muscle. The NLPs tested on A. suum muscle, viz, NLP-1A, NLP-2A and B, had no direct effect on the muscle but all reversibly reduced acetylcholine (ACh)-induced contractions of the muscle, NLP-2A being the most potent, with an IC50 value of 78 microM. NLP-2A also reversibly reduced ACh-induced depolarizations of A. suum muscle resting membrane potential. NLP-1A, NLP-2A, NLP-3C and NLP-10A all increased pharyngeal pumping in C. elegans, NLP-8A weakly inhibited pumping while NLP-2B was inactive. NLP-10A induced a concentration-dependent increase in pumping activity with a threshold of 10-100 nM. Since genes encoding NLPs (nlp genes) are expressed in C. elegans pharyngeal neurons it is likely these peptides play a role in feeding behaviour.

The effects of temperature, pH, and ammonia concentration on the inactivation of Ascaris eggs in sewage sludge.

The reported inactivation of Ascaris eggs during alkaline sludge stabilization is highly variable. The objective of our research was to better understand the sources of this variability by quantifying the effects of temperature, pH, and ammonia concentration on the inactivation of indigenous Ascaris eggs in wastewater sludge. Primary sludge was supplemented with ammonia (0, 1000, and 5000 mg/l NH(3)-N) and Ca(OH)(2) and incubated in sealed bottles across the range of temperatures (20, 30, 40, and 50 degrees C) and pH (7 and 12) that may be encountered during treatment. Changes in egg viability over time were fit to a two-parameter kinetic model (shoulder and first-order region); to compare treatment conditions, the time for 99% inactivation (t(99)) was also calculated. Each 10 degrees C increase in temperature caused a significant decrease in t(99) at every pH and ammonia concentration tested. At 50 degrees C, the effect of temperature was dominant, such that no effect of pH or ammonia was observed. At 30 and 40 degrees C, raising the pH from 7 to 12 decreased t(99), but at 20 degrees C no pH effect was seen over 80 d (very little inactivation occurred). At 20, 30, and 40 degrees C, the addition of ammonia dramatically decreased t(99). The effect of pH could not be completely separated from that of ammonia, as the unamended sludge samples contained 100-200mg/l indigenous ammonia. Because temperature, pH, and ammonia all contributed to Ascaris egg inactivation, it is essential that these parameters are measured and accounted for when assessing the effectiveness of alkaline stabilization. Furthermore, inactivation by ammonia could be exploited to improve the effectiveness of alkaline sludge stabilization.

Field studies on endoparasites of Thoroughbred foals on seven farms in central Kentucky in 2004.

Fecal samples (n=1,584) for a parasite study were collected from Thoroughbred foals (n=349), 28 to 330 days old, on seven farms in central Kentucky during 49 monthly trips from May to October, November, or December 2004. The main purpose was to determine possible drug resistance of ascarids, also of strongyles, to ivermectin (IVM) and other commercially available compounds [fenbendazole, moxidectin (MOX), oxibendazole (OBZ), and pyrantel pamoate]. In addition, interest was on prevalence of foal parasites. Qualitative data were obtained by recording presence or absence of ascarid and strongyle eggs in feces from 7 to 25 days after treatment of some of the foals. None of the compounds completely eliminated ascarid eggs in feces of all foals posttreatment. Activity on ascarids was significantly the highest for OBZ and lowest for IVM. Activity on strongyles was significantly higher for IVM and MOX than for the other compounds. Prevalence (mean percentage of foals infected) was recorded for eggs of Parascaris equorum (39%), of strongyles (32%), and of Strongyloides westeri (2%) and oocysts of Eimeria leuckarti (28%) in feces of foals. One or more foals had infections of P. equorum on six farms (86%), strongyles on seven farms (100%), S. westeri on two farms (29%), and E. leuckarti on six farms (86%). The oldest foal infected with E. leuckarti was 301 days of age. Comparison was made on the prevalence of parasites in foals on five of the same farms in the 2004 study, which were part of a similar 14-farm project in 2003 (Lyons and Tolliver in Parasitol Res (2004) 92:400-404).

Concomitant parasitism: bancroftian filariasis and intestinal helminths and response to albendazole.

Subjects in an disease-endemic area in Orissa, India concomitantly infected with filariasis and intestinal helminths had significantly lower intensity of filarial infection in comparison with those who were infected only with filariasis. Administration of albendazole resulted in a significant decrease in the prevalence of filarial antigenemia in subjects concomitantly infected with intestinal helminths, but produced little change in this infection measure in subjects infected only with Wuchereria bancrofti. These results indicate that intestinal helminths could play a role in the anti-filarial activity of albendazole, most probably by depressing filarial infection intensity in co-infected individuals. Confirmation of these findings in a larger cohort may yield important new insights regarding the role of using albendazole in the ongoing intervention programs for the control of lymphatic filariasis.

Liming as an advanced treatment for sludge sanitisation: helminth eggs elimination--Ascaris eggs as model.

The presence of helminth eggs (Ascaris eggs) in sewage sludge may constitute a sanitary risk when used as agricultural fertiliser. Sanitisation of sewage sludge can be achieved by treatment with quick lime, a process that destroys sludge pathogens in two ways: pH increase and temperature rise. Among the pathogens of epidemiological relevance, Ascaris eggs are the most resistant to liming, and, hence, may serve as indicators of hygienic quality of biosolids. This research aims at defining, between 50 degrees C and 60 degrees C, the time required in the case of limed sludge to obtain a product with a negligible level of viable Ascaris eggs. To achieve this objective, investigations on inactivation kinetics of Ascaris eggs were conducted in the following products: contaminated milk of lime; naturally contaminated sludge treated with slaked lime and heat; naturally contaminated sludge treated with quick lime; and sludge treated at full scale with quick lime. For the inactivation kinetics where a negligible level of Ascaris eggs was reached, the inactivation threshold was determined. Depending on the experimental situation, the inactivation threshold period was found to fluctuate between 5 and 75 min at 55 degrees C and between 1 and 8 min at 60 degrees C.