Suum Eggs Research Articles

Helminth Egg Removal Capacity of UASB Reactors under Subtropical Conditions

This research was conducted to study the anaerobic sludge filtration capacity regarding helminth egg removal in upflow anaerobic sludge blanket (UASB) reactors. Two 25 L lab-scale UASB reactors were operated at an ambient temperature which varied between 17.1 and 28.6 °C. Ascaris suum egg was selected as the model egg considering its similarity in terms of size and morphology to Ascaris lumbricoides, a human pathogen. Ascaris suum eggs were obtained from female parasites of infected pigs. The anaerobic sludge filtration capacity was performed applying upflow velocities between 0.09 and 0.68 m·h−1. Three sludge bed heights in the range of 0.30–0.40 m, 0.50–0.60 m and 0.60–0.70 m were applied. These sludge bed heights corresponded to 19%–25%, 31%–38% and 38%–44% of the total reactor height, respectively. Under the mentioned conditions, the average helminth egg removal efficiency was reciprocally correlated to the imposed upflow velocity. The studied lab-scale reactors reported an average helminth egg removal between 34%–100%, 30%–91% and 34%–56%, when the sludge bed in the UASB reactor was 19%–25%, 31%–38% and 38%–44% of the total reactor height, respectively. The decreased filtration capacity at increasing sludge bed heights might be likely related to biogas production and channeling formation. The average helminth egg removal efficiency in the control experiments performed without any sludge bed, by plain sedimentation, varied between 44% and 66%.

A method to evaluate relative ovicidal effects of soil microfungi on thick-shelled eggs of animal-parasitic nematodes

Thick-shelled eggs of animal-parasitic ascarid nematodes can survive and remain infective in the environment for years. The present study evaluated a simple in vitro method and evaluation scheme to assess the relative effect of two species of soil microfungi, Pochonia chlamydosporia Biotype 10 and Purpureocillium lilacinum Strain 251 (Ascomycota: Hypocreales), on the development and survival of eggs of faecal origin of three ascarid species, Ascaridia galli (chicken roundworm), Toxocara canis (canine roundworm) and Ascaris suum (pig roundworm). Ascarid eggs were embryonated on water agar with or without a fungus, and the resulting viability of the eggs was evaluated on days 7, 14, 21, 28, 35 and 42 post exposure (pe) by observing eggs in situ. On days 7–42 pe, P. chlamydosporia had reduced the viability of A. galli and T. canis eggs by 64–86% and 26–67%. Corresponding reductions for P. lilacinum Strain 251 were only 15–29% and 4–28%. In contrast, A. suum eggs were extremely resistant to both fungi (2–4% reduction). The differences in results are likely due to different morphologies and chemistry of the egg shell of the three ascarid species. The current in vitro method and evaluation criteria allow for a simple, repeatable and non-invasive evaluation of the ovicidal effects of microfungi. This study demonstrates that P. chlamydosporia Biotype 10 may be utilised as a biocontrol agent to reduce A. galli and T. canis egg contamination of the environment.

Ammonia sanitization of blackwater for safe use as fertilizer.

Source-separated blackwater from low-flush toilets contains plant-available nutrients and can be used as a fertilizer. The aim of the study was to evaluate the impact on pathogen inactivation when treating blackwater with urea and/or lime. Blackwater was spiked with Salmonella typhimurium, Escherichia coli O157, Enterococcus faecalis, and Ascaris suum eggs, and treated with urea and/or lime in concentrations up to 0.1% w/w. The bottles were kept in a storage facility (manure slurry tank) for 102 days while monitoring the pathogen concentrations. The treatment time needed to meet the requirement for Salmonella and E. coli reduction could be reduced at least six-fold. The enterococci were more persistent, and only the highest treatment doses had a significantly higher inactivation than the controls. The Ascaris egg viability was only reduced by around 50%, so higher urea/lime doses and/or longer treatment times are required to fulfill the treatment requirements of 3 log10 reductions of parasite eggs.

Comparison of methodologies for enumerating and detecting the viability of Ascaris eggs in sewage sludge by standard incubation-microscopy, the BacLight Live/Dead viability assay and other vital dyes

The aim of this study was to evaluate the Live/Dead BacLight viability kit as a method for enumerating viable eggs of Ascaris suum in sewage sludge as a surrogate for the human roundworm. The number and viability status of eggs of A. suum were accurately measured directly in sewage sludge samples by the BacLight method, compared to the conventional incubation-microscopy procedure. BacLight stains were not toxic to A. suum eggs, in contrast to some conventional vital dyes which disrupted viable eggs. The method was effective for the direct examination of eggs in heavily contaminated samples or seeded sludge containing ∼200 eggs/g DS in sludge with 5% DS content. However, a recovery method would be necessary to examine samples with small numbers of eggs, for instance in sludge from regions where the prevalence of infection with Ascaris lumbricoides is low. The BacLight technique may therefore be an effective alternative to conventional incubation-microscopy for enumerating Ascaris eggs in contaminated field samples or to validate sludge treatment processes by examining decay rates of inoculated A. suum eggs in laboratory simulations. Most field samples would require recovery from an appropriate number of composite samples prior to vital staining.

The reduction rate of invasive Ascaris suum eggs number in slurry and humus layers of selected soil types

Abstract The aim of this study was to determine under laboratory conditions the percentage and rate of reduction in invasive Ascaris suum eggs, as well as the maximal time of invasiveness retaining by them in swine slurry and 3 soil types. A. suum eggs were introduced into perforated perlon bags which then were placed in samples of slurry and in humus layers of the soils: podsolic soil, black earth and browned black earth. The experiment was carried out for 44 weeks at 4 and 20 °C. Finally, at 4 °C a reduction in the number of invasive eggs from 41 % (the humus layer of podsolic soil) to 65 % (the humus layer of black earth) was observed. At 20 °C the fluctuations were smaller and the percentage of elimination ranged from 89 % (slurry) to 96 % (the humus layer of browned black earth).

Antiparasitic efficacy of a novel plant-based functional food using an Ascaris suum model in pigs

Ascaris lumbricoides is the most prevalent soil-transmitted helminth (STH) infection of human beings worldwide. Chemotherapy with synthetic anthelmintics such as albendazole, mebendazole, and pyrantel pamoate is the current method of treatment; however, the emergence of anthelmintic resistance could substantially decrease the efficacy of such treatments and the sustainability of STH control programs. Additionally, benzimidazoles are not recommended for pregnant women or children under age one. A blinded, controlled study was conducted to evaluate the efficacy of two microencapsulated, plant-based essential oil blends, TTN1013 (α-pinene, linalyl acetate, p-cymene, and thymol octanoate) and TTN1014 (α-pinene, linalyl acetate, p-cymene, and thymol acetate) as functional foods against Ascaris suum infection in pigs, an important pathogen that closely resembles human infections with A. lumbricoides. Four groups of 16 female, 21–24 day old, Yorkshire-cross pigs were treated daily with 0.5 or 1.0mg/kg TTN1013, 1.0mg/kg TTN1014, or 1.0mg/kg equivalent of empty capsules, delivered inside a cream-filled sandwich cookie for 14 weeks. Three days after the initiation of daily treatments, pigs were inoculated daily with A. suum eggs for four weeks. Pigs were weighed weekly and fecal egg counts (FEC) were conducted weekly starting five weeks after initial inoculation with A. suum eggs. Fourteen weeks after first infection with eggs, pigs were necropsied and worms were recovered, counted and separated according to sex. TTN1013 administered daily at a dose of 1.0mg/kg yielded a statistically significant reduction in total worm counts (76.8%), female worm counts (75.5%), FEC (68.6%), and worm volume (62.9%) when compared to control group. Reduction of total and female worm numbers and FEC were not significant for TTN1014 or at the 0.5mg/kg dose of TTN1013. All treatments were well-tolerated by all pigs and did not cause any adverse reactions. All pigs remained clinically normal and showed no signs of reduced intestinal health for the duration of treatment. Based on these results, TTN1013 shows promise as a daily supplement to reduce infection burdens of soil transmitted helminths in both pigs and human beings.

Survival of Ascaris suum and Ascaridia galli eggs in liquid manure at different ammonia concentrations and temperatures

Eggs of Ascaris suum from pigs are highly resistant and commonly used as a conservative indicator of pathogen inactivation during slurry storage. Eggs of Ascaridia galli, the poultry ascarid, are also known to be highly resistant but the suitability as an indicator of pathogen inactivation has never been tested. Pig slurry has to be stored for several months to inactivate pathogens but chemical treatment of slurry may reduce this time. The suitability of A. galli as an indicator of slurry sanitation was tested by comparing the survival of eggs of A. suum and A. galli in pig slurry. In addition, the effect of urea treatment on inactivation of ascarid eggs in relation to storage time was also tested. Nylon bags with 10,000 eggs of either species were placed in 200ml plastic bottles containing either urea-treated (2%) or untreated pig slurry for up to 120 days at 20°C, 6 days at 30°C, 36h at 40°C or 2h at 50°C. At all the temperatures in both slurry types, A. galli eggs were inactivated at a significantly faster rate (P<0.05) compared to A. suum eggs. For each 10°C raise in temperature from 20°C, T50 (time needed to inactivate 50% of eggs) for both types of eggs was reduced markedly. At all temperatures, viability of eggs of both species was significantly higher (P<0.05) in untreated slurry compared to urea-treated slurry except A. galli eggs at 20°C where no significant difference was detected. In untreated slurry, the levels of pH (6.33–9.08) and ammonia (0.01–1.74mM) were lower (P<0.0001) compared to that of urea-treated slurry (pH: 8.33–9.28 and ammonia 1–13mM). The study demonstrated that A. galli eggs are more sensitive to unfavourable conditions compared to A. suum eggs. The use of A. galli eggs as hygiene indicator may thus be suitable to assess inactivation of pathogens that are more sensitive than A. galli eggs. Addition of urea may markedly reduce the storage time of slurry needed to inactivate A. suum and A. galli eggs.

Effects of Some Pesticides on Development of Ascaris suum Eggs

To evaluate the effects of pesticides to parasite eggs, Ascaris suum eggs were incubated with 5 different pesticides (1:1,500-1:2,000 dilutions of 2% emamectin benzoate, 5% spinetoram, 5% indoxacarb, 1% deltamethrin, and 5% flufenoxuron; all v/v) at 20℃ for 6 weeks, and microscopically evaluated the egg survival and development on a weekly basis. The survival rate of A. suum eggs incubated in normal saline (control eggs) was 90±3% at 6 weeks. However, the survival rates of eggs treated with pesticides were 75-85% at this time, thus significantly lower than the control value. Larval development in control eggs commenced at 3 weeks, and 73±3% of eggs had internal larvae at 6 weeks. Larvae were evident in pesticide-treated eggs at 3-4 weeks, and the proportions of eggs carrying larvae at 6 weeks (36±3%-54±3%) were significantly lower than that of the control group. Thus, pesticides tested at levels similar to those used in agricultural practices exhibited low-level ovicidal activity and delayed embryogenesis of A. suum eggs, although some differences were evident among the tested pesticides.

In vitro evaluation of the ovistatic and ovicidal effect of the cosmopolitan filamentous fungi isolated from soil on Ascaris suum eggs

The ovicidal activity of seven fungal strains: Acremonium alabamense, Alternaria chlamydospora, Cladosporium herbarum, Fusarium solani, Paecilomyces variotii, Paecilomyces viridis and Penicillium verruculosum isolated from urban soil samples from Poland was determined in vitro. The fungal mycelium was co-cultured with Ascaris suum eggs on plates with 2% water–agar for 28 days. Eggs exposed and unexposed (control) to fungal mycelium were observed weekly by light microscopy and the percentage of malformed eggs were determined. The eggs were classified according to following parameters: type 1 – biochemical and physiological effect without morphological damage to the eggshell; type 2 – lytic effect with morphological alteration of the eggshell and embryo; type 3 – lytic effect with morphological alteration of eggshell and embryo with hyphal penetration and internal egg colonization. All examined species of fungi extended embryogenesis, but the retardation of embryonic development was varied and depended on the species. A. alabamense, A. chlamydospora and P. verruculosum exhibited very high inhibitory activity on A. suum egg development. The fungus-exposed eggs revealed morphological alternations in all stages of embryogenesis. Isolates of F. solani, P. variotii and P. viridis showed hyphal penetration and internal colonization of A. suum eggs (type 3 effect). No appressoria were produced and simple hyphal penetrations were most commonly observed. A. alabamense and P. verruculosum demonstrated morphological destruction, with eggshell destruction. The remaining fungi showed type 1 effect. The results demonstrated that examined strains of F. solani, P. variotii and P. viridis may be considered to be potential limiting factors of parasitic geohelminth populations.

The risk to human health related to disposal of animal wastes to soil — microbiological and parasitical aspects

Abstract The study was conducted to investigate the microbiological and parasitical risk related to the disposal of animal manure to soil by storage of raw pig slurry at temperatures 4 °C, 20 °C and 42 °C for 115 days. Plate counts of Salmonella typhimurium and number of devitalized non-embryonated model Ascaris suum eggs were determined on days 0, 7, 12, 22, 32, 40, 55, 90 and 115 of storage. At the same intervals level of selected physico-chemical parameters were determined. Microbiological examination showed that S. typhimurium survived in the slurry for less than 115 days at 4 °C and less than 90 days at 20 °C and 42 °C. Devitalization of A. suum eggs increased with temperature and time of storage, but complete devitalization was not achieved even after 115 days at 42 °C. Physico-chemical parameters showed changes related to decomposition processes, but did not allow us to draw definite conclusion regarding their influence on devitalization of pathogens. The results indicate potential risk to human food chain that can be prevented by strict observation of legislative provisions and appropriate treatment of animal manure.

Analysis of the accuracy and precision of the McMaster method in detection of the eggs of Toxocara and Trichuris species (Nematoda) in dog faeces

The aim of this study was to determine the accuracy and precision of McMaster method with Raynaud's modification in the detection of the eggs of the nematodes Toxocara canis (Werner, 1782) and Trichuris ovis (Abildgaard, 1795) in faeces of dogs. Four variants of McMaster method were used for counting: in one grid, two grids, the whole McMaster chamber and flotation in the tube. One hundred sixty samples were prepared from dog faeces (20 repetitions for each egg quantity) containing 15, 25, 50, 100, 150, 200, 250 and 300 eggs of T. canis and T. ovis in 1 g of faeces. To compare the influence of kind of faeces on the results, samples of dog faeces were enriched at the same levels with the eggs of another nematode, Ascaris suum Goeze, 1782. In addition, 160 samples of pig faeces were prepared and enriched only with A. suum eggs in the same way. The highest limit of detection (the lowest level of eggs that were detected in at least 50% of repetitions) in all McMaster chamber variants were obtained for T. canis eggs (25-250 eggs/g faeces). In the variant with flotation in the tube, the highest limit of detection was obtained for T. ovis eggs (100 eggs/g). The best results of the limit of detection, sensitivity and the lowest coefficients of variation were obtained with the use of the whole McMaster chamber variant. There was no significant impact of properties of faeces on the obtained results. Multiplication factors for the whole chamber were calculated on the basis of the transformed equation of the regression line, illustrating the relationship between the number of detected eggs and that of the eggs added to the'sample. Multiplication factors calculated for T. canis and T. ovis eggs were higher than those expected using McMaster method with Raynaud modification.

Pathogen and indicator inactivation in source-separated human urine heated by the sun

Source-separation of urine enables energy-efficient recycling of large quantities of high quality plant nutrients from the wastewater system to food production. To manage the risk of diseases transmission the urine should be sanitised, which is commonly done by storage. The sanitisation of urine at three ambient exposures, resulting in fluctuating storage temperatures, was studied in 10 L plastic jerry cans in Kampala, Uganda. Follow-up studies using similar temperature cycles were performed under laboratory conditions. Daily mean mid-point temperature in jerry cans in full sun, adjacent to a wall and indoors was 24, 24 and 22 °C (amplitude 7.5, 4.7 and 1.2 °C), respectively. In urine stored at full sun (total ammonia nitrogen 4 ± 1.5 g L−1, pH 9) Escherichia coli, Salmonella and Enterococcus spp. (initially 105–106cfu mL−1) were not detected (&amp;lt;101cfu mL−1) after 11 h, 14 h and 50 d, respectively. No viable Ascaris suum eggs were found after 40 d. The t90 for enterobacteria phage MS2, coliphage Φx 174 and Salmonella typhimurium phage 28B in sun-exposed cans was 8.2, 37 and 55 d respectively. Fluctuating temperatures in combination with ammonia gave faster inactivation of bacteria and Ascaris suum eggs than the same constant mean temperature.

A Role for Eosinophils in the Intestinal Immunity against Infective Ascaris suum Larvae

The aim of this study was to explore the mechanisms of resistance against invading Ascaris suum larvae in pigs. Pigs received a low dose of 100 A. suum eggs daily for 14 weeks. This resulted in a >99% reduction in the number of larvae that could migrate through the host after a challenge infection of 5000 A. suum eggs, compared to naïve pigs. Histological analysis at the site of parasite entry, i.e. the caecum, identified eosinophilia, mastocytosis and goblet cell hyperplasia. Increased local transcription levels of genes for IL5, IL13, eosinophil peroxidase and eotaxin further supported the observed eosinophil influx. Further analysis showed that eosinophils degranulated in vitro in response to contact with infective Ascaris larvae in the presence of serum from both immune and naïve animals. This effect was diminished with heat-inactivated serum, indicating a complement dependent mechanism. Furthermore, eosinophils were efficient in killing the larvae in vitro when incubated together with serum from immune animals, suggesting that A. suum specific antibodies are required for efficient elimination of the larvae. Together, these results indicate an important role for eosinophils in the intestinal defense against invading A. suum larvae.

Biological interactions between soil saprotrophic fungi and Ascaris suum eggs

The in vitro effect of saprotrophic soil fungi on the embryonic development of Ascaris suum was evaluated. The fungi tested were Aspergillus fumigatus, Aspergillus terreus, Penicillium citrinum, Penicillium expansum, Fusarium oxysporum and Trichothecium roseum, isolated from children's recreation areas in the city of Lodz (Poland). Each species was co-cultured with A. suum egg suspension (6×103eggs/ml) at 25±2°C for 60 days. Each day, 100 eggs were randomly collected and their developmental stage was classified macroscopically. Additionally, at days 4, 7, 14, 28, 42 and 60 of incubation, the viability and the percentage of eggs with morphological altered embryo/larva were determined in each sample. Microscopic examination revealed that exposure of eggs to the mycelium of examined fungi inhibited embryogenesis of A. suum. All control culture eggs reached L2 larval stage after 26 days of incubation, while the experimental cultures did so after 32–51 days, depending on the fungal species. Three species were found to exhibit very high inhibitory activity on A. suum egg development: A. terreus, P. expansum and F. oxysporum. Embryopathies and non-viable embryos/larvae were observed significantly more frequently in the eggs co-cultured with fungal species than in control cultures. The fungus-exposed eggs revealed morphological alternations in the early zygotic cleavage, blastula, gastrula and larval stages. After 60 days of incubation with mycelia of P. expansum, A. terreus and F. oxysporum, the mortality of the larvae reached 55.3–60.3%. P. expansum and F. oxysporum showed hyphal penetration and internal egg colonization of A. suum eggs.

Radiation hygienization of cattle and swine slurry with high energy electron beam

The research was carried out to assess the efficiency of radiation hygienization of cattle and swine slurry of different density using the high energy electron beam based on the inactivation rate of Salmonella ssp, Escherichia coli, Enterococcus spp and Ascaris suum eggs. The experiment was conducted with use of the linear electron accelerator Elektronika 10/10 in Institute of Nuclear Chemistry and Technology in Warsaw. The inoculated slurry samples underwent hygienization with high energy electron beam of 1, 3, 5, 7 and 10kGy. Numbers of reisolated bacteria were determined according to the MPN method, using typical microbiological media. Theoretical lethal doses, D90 doses and hygienization efficiency of high energy electron beam were determined. The theoretical lethal doses for all tested bacteria ranged from 3.63 to 8.84kGy and for A. suum eggs from 4.07 to 5.83kGy. Salmonella rods turned out to be the most sensitive and Enterococcus spp were the most resistant to electron beam hygienization. The effectiveness or radiation hygienization was lower in cattle than in swine slurry and in thick than in thin one. Also the species or even the serotype of bacteria determined the dose needed to inactivation of microorganisms.

Viability of Ascaris suum eggs in stored raw and separated liquid slurry

Separation of pig slurry into solid and liquid fractions is gaining importance as a way to manage increasing volumes of slurry. In contrast to solid manure and slurry, little is known about pathogen survival in separated liquid slurry. The viability of Ascaris suum eggs, a conservative indicator of fecal pollution, and its association with ammonia was investigated in separated liquid slurry in comparison with raw slurry. For this purpose nylon bags with 6000 eggs each were placed in 1 litre bottles containing one of the two fractions for 308 days at 5 °C or 25 °C. Initial analysis of helminth eggs in the separated liquid slurry revealed 47 Ascaris eggs per gramme. At 25 °C, egg viability declined to zero with a similar trend in both raw slurry and the separated liquid slurry by day 308, a time when at 5 °C 88% and 42% of the eggs were still viable in separated liquid slurry and raw slurry, respectively. The poorer survival at 25 °C was correlated with high ammonia contents in the range of 7.9-22.4 mM in raw slurry and 7.3-23.2 mM in liquid slurry compared to 3.2-9.5 mM in raw slurry and 2.6-9.5 mM in liquid slurry stored at 5 °C. The study demonstrates that at 5 °C, A. suum eggs have a higher viability in separated liquid slurry as compared to raw slurry. The hygiene aspect of this needs to be further investigated when separated liquid slurry is used to fertilize pastures or crops.

Ineffectiveness of a quaternary ammonium salt and povidone-iodine for the inactivation of Ascaris suum eggs

Two commonly used disinfectants, a quaternary ammonium salt and povidone-iodine, were tested for effectiveness against unembryonated Ascaris suum eggs. The quaternary ammonium salt (alkyl dimethyl benzyl ammonium chloride) had no effect on the Ascaris eggs (10 minutes and 22°C) when compared with the controls in water with egg viabilities of 88.8% ± 3.3% and 86.9% ± 6.2%, respectively. An additional quaternary ammonium salt, 2.5% benzethonium chloride, also had no effect. Phenol (5%) and cresol (3%) completely inactivated the eggs. Povidone-iodine at 100%, 50%, 10%, and 1% had no effect on the eggs at exposures of 5, 15, 30, 60, or 120 minutes (22°C) compared with the water controls.

Inactivation Kinetics ofCryptosporidium parvumOocysts in a Swine Waste Lagoon and Spray Field

Because of outbreaks of cryptosporidiosis in humans, some Cryptosporidium spp. have become a public health concern. Commercial swine operations can be a source of this protozoan parasite. Although the species distribution of Cryptosporidium is likely dominated by Cryptosporidium suis , a fraction may be comprised of other Cryptosporidium species infectious to humans such as Cryptosporidium parvum . To better understand the survival dynamics of Cryptosporidium spp., oocysts associated with swine operations, 2 experiments were performed to determine die-off rates of C. parvum oocysts in a swine waste lagoon (2009 and 2010) and its spray field (2010 and 2011). Sentinel chambers containing a lagoon effluent suspension of C. parvum oocysts were submerged in the lagoon, and triplicate chambers were removed over time; oocysts were extracted and assayed for viability. For comparative purposes, inactivation rates of Ascaris suum eggs contained in sentinel chambers were also determined. For 2 spray field experiments, air-dried and sieved surface soil was placed in sentinel chambers, hydrated, and inoculated with a lagoon effluent suspension of C. parvum oocysts. Sentinel chambers and control oocysts in PBS contained in microcentrifuge tubes were buried 1.5 cm below the soil surface in 3 blocks. Triplicate chambers and controls were removed over time; oocysts were extracted and assayed for viability. Based on the first order decay equation, days to reach 99% die-off (T(99)) were determined. T(99)-values determined for the 2 lagoon experiments were 13.1 and 20.1 wk, respectively. A T(99)-value for C. parvum in the spray field was significantly longer at 38.0 wk than the control oocysts in PBS at 29.0 wk. The waste lagoon and spray field system of manure management at this large-scale farrowing operation appeared to reduce the load of C. parvum oocysts before they can be hydrologically transported off the operation and reduces their likelihood of contaminating surface waters and threatening public health.

Quantitative Evaluation of Viability- and Apoptosis-Related Genes in Ascaris suum Eggs under Different Culture-Temperature Conditions

Ascaris suum eggs are inactivated by composting conditions; however, it is difficult to find functional changes in heat-treated A. suum eggs. Here, unembryonated A. suum eggs were incubated at 20℃, 50℃, and 70℃ in vitro, and the gene expression levels related to viability, such as eukaryotic translation initiation factor 4E (IF4E), phosphofructokinase 1 (PFK1), and thioredoxin 1 (TRX1), and to apoptosis, such as apoptosis-inducing factor 1 (AIF1) and cell death protein 6 (CDP6), were evaluated by real-time quantitative RT-PCR. No prominent morphological alterations were noted in the eggs at 20℃ until day 10. In contrast, the eggs developed rapidly, and embryonated eggs and hatched larvae began to die, starting on day 2 at 50℃ and day 1 at 70℃. At 20℃, IF4E, PFK1, and TRX1 mRNA expression was significantly increased from days 2-4; however, AIF1 and CDP6 mRNA expression was not changed significantly. IF4E, PFK1, and TRX1 mRNA expression was markedly decreased from day 2 at 50℃ and 70℃, whereas AIF1 and CDP6 mRNA expression was significantly increased. The expressions of HSP70 and HSP90 were detected for 9-10 days at 20℃, for 3-5 days at 50℃, and for 2 days at 70℃. Taken together, incremental heat increases were associated with the rapid development of A. suum eggs, decreased expression of genes related to viability, and earlier expression of apoptosis-related genes, and finally these changes of viability- and apoptosis-related genes of A. suum eggs were associated with survival of the eggs under temperature stress.

Effect of Temperature on Embryonation of Ascaris suum Eggs in an Environmental Chamber

The influence of temperature on the development and embryonation of Ascaris suum eggs was studied using coarse sand medium in an environmental chamber with 50% humidity. The time required for development and embryonation of eggs was examined under 3 different temperature conditions, 5℃, 25℃, and 35℃. A. suum eggs did not develop over 1 month at the temperature of 5℃. However, other temperature conditions, 25℃ and 35℃, induced egg development to the 8-cell-stage at days 5-6 after incubation. All eggs examined developed to the 8-cell stage at day 6 after incubation in the sand medium at 25℃. The higher temperature, 35℃, slightly accelerated the A. suum egg development compared to 25℃, and the development to the 8-cell stage occurred within day 5 after incubation. The formation of larvae in A. suum eggs at temperatures of 35℃ and 25℃ appeared at days 17 and 19 after incubation, respectively. These findings show that 35℃ condition shortens the time for the development of A. suum eggs to the 8-cell-stage in comparison to 25℃, and suggest the possibility of accelerated transmission of this parasite, resulting from global warming and ecosystem changes.