The role of the liver in the migration of parasites of global significance.

Many parasites migrate through different tissues during their life-cycle, possibly with the aim to enhance their fitness. This is true for species of three parasite genera of global importance, Ascaris, Schistosoma and Plasmodium, which cause significant global morbidity and mortality. Interestingly, these parasites all incorporate the liver in their life-cycle. The liver has a special immune status being able to preferentially induce tolerance over immunity. This function may be exploited by parasites to evade host immunity, with Plasmodium spp. in particular using this organ for its multiplication. However, hepatic larval attrition occurs in both ascariasis and schistosomiasis. A better understanding of the molecular mechanisms involved in hepatic infection could be useful in developing novel vaccines and therapies for these parasites.

Co-infections with Plasmodium, Ascaris and Giardia among Rwandan schoolchildren.

OBJECTIVES: Co-infections with Plasmodium, Ascaris and Giardia are common in sub-Saharan Africa but epidemiological and clinical data are rare. We examined factors associated with co-infections and their clinical manifestation among Rwandan schoolchildren. METHODS: Schoolchildren aged 6-10 years attending 12 schools in Huye district, Rwanda, were recruited preceding routine deworming. Data on socioeconomic status (SES) and children's histories were obtained, and children were clinically and anthropometrically examined. Blood and stool samples were collected, and infections with Plasmodium, Ascaris and Giardia were determined by microscopy and PCR assays. RESULTS: Among 878 schoolchildren, Plasmodium, Ascaris and Giardia were present in 22%, 35% and 36%, respectively. Co-infections with two or more parasites were found in 24%; only one-third of the children did not harbour any of the parasites examined. Factors associated with parasite (co-)infections largely overlapped and reflected low SES, in addition to a few specific risk factors. Clinically, most children were asymptomatic but anaemia (38%), underweight (17%), and reported signs and symptoms in the preceding 2 weeks (46%) were common. Many of the reported and assessed signs and symptoms were associated with Plasmodium infection, and co-infection with Ascaris and/or Giardia did basically not modify the clinical picture. One exception was malnutrition, which was pronounced in Ascaris-Giardia co-infection vs. individual mono-infections. CONCLUSIONS: Parasitic co-infections are common in Rwandan schoolchildren, and are associated with a rather silent clinical manifestation that nevertheless may affect school performance and long-term development. School-based health interventions should target such co-infections in an integrated manner.

OBJECTIFS: Les coinfections par Plasmodium, Ascaris et Giardia sont courantes en Afrique subsaharienne, mais les données épidémiologiques et cliniques sont rares. Nous avons examiné les facteurs associés aux coinfections et leurs manifestations cliniques chez les écoliers rwandais. MÉTHODES: Des écoliers âgés de 6 à 10 ans fréquentant 12 écoles du district de Huye au Rwanda ont été recrutés avant le déparasitage de routine. Les données sur le statut socioéconomique (SSE) et les antécédents des enfants ont été obtenues et les enfants ont été examinés cliniquement et anthropométriquement. Des échantillons de sang et de selles ont été recueillis et les infections à Plasmodium, Ascaris et Giardia ont été déterminées par microscopie et par PCR. RÉSULTATS: sur 878 écoliers, Plasmodium, Ascaris et Giardia étaient présents chez 22%, 35% et 36%, respectivement. Des coinfections avec deux parasites ou plus ont été trouvées chez 24%; seul un tiers des enfants n'hébergeait aucun des parasites examinés. Les facteurs associés aux (co)infections parasitaires se chevauchaient largement et reflétaient un faible statut SSE, en plus de quelques facteurs de risque spécifiques. Sur le plan clinique, la plupart des enfants étaient asymptomatiques mais l'anémie (38%), l'insuffisance pondérale (17%) et les signes et symptômes rapportés au cours des deux semaines précédentes (46%) étaient fréquents. De nombreux signes et symptômes rapportés et évalués étaient associés à l'infection au Plasmodium et la coinfection par Ascaris et/ou Giardia n'a fondamentalement pas modifié le tableau clinique. Une exception était la malnutrition, qui était prononcée dans la coinfection Ascaris-Giardia par rapport aux mono-infections individuelles. CONCLUSIONS: Les coinfections parasitaires sont courantes chez les écoliers rwandais et sont associées à une manifestation clinique plutôt silencieuse qui peut néanmoins affecter les performances scolaires et le développement à long terme. Les interventions de santé en milieu scolaire devraient cibler ces coinfections de manière intégrée.

Evaluating the efficacy of a centrifugation-flotation method for extracting Ascaris ova from soil.

BACKGROUND: Soil transmitted helminths (STH) continue to be associated with high burdens of disease, with an estimated 1.45 billion people infected with STH globally. The promotion and construction of latrines is considered the first barrier to prevent transmission of STH. The absence of a reliable method to extract STH ova from soil makes it challenging to examine whether the use of latrines may or may not have an effect on environmental contamination with ova. The present study evaluated the recovery rate of a method developed to extract STH ova from soil. METHODS: The adapted centrifugation and flotation technique was applied to 15 soil types, which were seeded with Ascaris suum ova. Soil type, soil moisture content, soil texture and organic matter content were assessed for each soil sample. RESULTS: The average ova recovery rate was 28.2%, with the recovery rate of the method decreasing with increasing soil moisture content, particle size and organic matter content. The association between recovery rate and organic matter content was statistically significant. CONCLUSIONS: The present study identified a low recovery rate for an adapted centrifugation-flotation method, although this was similar to the recovery rate demonstrated by other methods developed for soil. Soil organic matter content was significantly associated with ova recovery rates.

Contribution of transcription to animal early development.

In mature gametes and during the oocyte-to-embryo transition, transcription is generally silenced and gene expression is post-transcriptionally regulated. However, we recently discovered that major transcription can occur immediately after fertilization, prior to pronuclear fusion, and in the first cell division of the oocyte-to-embryo transition in the nematode Ascaris suum. We postulate that the balance between transcriptional and post-transcriptional regulation during the oocyte-to-embryo transition may largely be determined by cell cycle length and thus the time available for the genome to be transcribed.

A quantitative assessment method for Ascaris eggs on hands.

The importance of hands in the transmission of soil transmitted helminths, especially Ascaris and Trichuris infections, is under-researched. This is partly because of the absence of a reliable method to quantify the number of eggs on hands. Therefore, the aim of this study was to develop a method to assess the number of Ascaris eggs on hands and determine the egg recovery rate of the method. Under laboratory conditions, hands were seeded with a known number of Ascaris eggs, air dried and washed in a plastic bag retaining the washing water, in order to determine recovery rates of eggs for four different detergents (cationic [benzethonium chloride 0.1% and cetylpyridinium chloride CPC 0.1%], anionic [7X 1% - quadrafos, glycol ether, and dioctyl sulfoccinate sodium salt] and non-ionic [Tween80 0.1% -polyethylene glycol sorbitan monooleate]) and two egg detection methods (McMaster technique and FLOTAC). A modified concentration McMaster technique showed the highest egg recovery rate from bags. Two of the four diluted detergents (benzethonium chloride 0.1% and 7X 1%) also showed a higher egg recovery rate and were then compared with de-ionized water for recovery of helminth eggs from hands. The highest recovery rate (95.6%) was achieved with a hand rinse performed with 7X 1%. Washing hands with de-ionized water resulted in an egg recovery rate of 82.7%. This washing method performed with a low concentration of detergent offers potential for quantitative investigation of contamination of hands with Ascaris eggs and of their role in human infection. Follow-up studies are needed that validate the hand washing method under field conditions, e.g. including people of different age, lower levels of contamination and various levels of hand cleanliness.

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

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 labour-intensive 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.

Inactivation of Ascaris eggs in soil by microwave treatment compared to UV and ozone treatment.

This study reports on the effect of microwave radiation for inactivation of Ascaris lumbricoides eggs in 25 g of soil compared to ultraviolet irradiation and ozone expose. Microwave radiation at 700 W with 14% water content (w/w) achieved approximately 2.5 log inactivation of eggs in soil within 60s. On the other hand, UV irradiation at 3 mW cm(-2) with and without shaking soil for 3600 s achieved approximately 0.32 and 0.01 log inactivation of eggs, respectively. In ozone treatment, 0.13 log inactivation of eggs was achieved with 5.8+/-0.7 mg L(-1) of dissolved ozone dose for 30 min in a continuous diffusion reactor. In addition, the inactivation of eggs by three disinfection techniques was conducted in water in order to compare the inactivation efficiency of eggs in soil. The inactivation efficiency of microwave radiation was found to be no significant difference between in soil and water. However, the inactivation efficiency of UV irradiation was significantly increased in water while in ozone expose there was no significant difference between in soil and water. Microwave treatment thus proved to be the most efficient method in controlling A. lumbricoides eggs in soil.

[Diversity of soil fauna in corn fields in Huang-Huai-Hai Plain of China under effects of conservation tillage].

An investigation was made on the abundance and diversity of soil fauna in the corn fields under conventional and conservation tillage in Huang-Huai-Hai Plain of China. The abundance and diversity of soil fauna were higher at corn maturing (September) than at its jointing stage (July), and higher at jointing stage under conservation tillage than under conventional tillage. Soil fauna mainly distributed in surface soil layer (0-10 cm), but still had a larger number in 10-20 cm layer under conservation tillage. The individuals of acari, diptera, diplura, and microdrile oligochaetes, especially those of acari, were higher under conservation tillage than under conventional tillage. At maturing stage, an obvious effect of straw-returning under conservation tillage was observed, i. e., the more the straw returned, the higher the abundance of soil fauna, among which, the individuals of collembola, acari, coleopteran, and psocoptera, especially those of collembolan, increased significantly. The abundance of collembola at both jointing and maturing stages was significantly positively correlated with the quantity of straw returned, suggesting that collembola played an important role in straw decomposition and nutrient cycling.

Multiplex PCR on single unembryonated Ascaris (roundworm) eggs.

A sensitive and inexpensive method for DNA isolation and amplification by polymerase chain reaction (PCR) from single unembryonated Ascaris sp. eggs is described. The resistant shell of single eggs was crushed mechanically and PCR applied to the crude egg contents without any further purification steps. The ITS1 region of the rDNA and three regions of the mtDNA could be successfully amplified. Using two primer sets, it was possible to amplify the rDNA and mtDNA simultaneously in one single reaction. The ability to perform PCR on single unembryonated eggs may result in better and more precise species identification of eggs recovered from faecal material, environmental samples and possibly archaeological samples. In addition, single egg PCR makes it possible to perform population genetic studies without having to recover adult worms by deworming or autopsy.

Helminth eggs inactivation efficiency by faecal sludge dewatering and co-composting in tropical climates.

This study investigates helminth eggs removal and inactivation efficiency in a treatment process combining faecal sludge (FS) dewatering and subsequent co-composting with organic solid waste as a function of windrow turning frequency. Fresh public toilet sludge and septage mixed at a 1:2 ratio were dewatered on a drying bed. Biosolids with initial loads of 25-83 helminth eggs/g total solids (TS) were mixed with solid waste as bulking material for co-composting at a 1:2 volume ratio. Two replicate sets of compost heaps were mounted in parallel and turned at different frequencies during the active composting period: (i) once every 3 days and (ii) once every 10 days. Turning frequency had no effect on helminth eggs removal efficiency. In both setups, helminth eggs were reduced to <1 viable egg/g TS, thereby complying with the WHO guidelines 2006 for the safe reuse of FS.

Status of intestinal helminthic infections of borderline residents in North Korea.

The present authors investigated intestinal parasitic infections among North Korean residents and refugees in China in 2003. The Kato-Katz method was applied to 236 residents and soldiers in a town on the North Korea-China border and to 46 people at a refugee camp in China. Only eggs of Ascaris and Trichuris were detected, with egg positive rates of 41.1% and 37.6%, respectively. The total egg positive rate was 55.0% and most of those who were egg positive were only lightly infected. Women of 61.2% and men of 53.1% were egg positive. The refugees from rural areas showed higher egg positive rates than those from urban areas. The present investigation confirmed high prevalence of soil-transmitted intestinal helminths in rural borderline areas of North Korea.

Studies on the interaction between Salmonella enterica ser. Typhimurium and intestinal helminths in pigs.

Concomitant infections with helminths and bacteria may affect the course and the resulting disease outcome of the individual infections. Salmonella, Oesophagostomum, Trichuris and Ascaris coexist naturally in pig herds in Denmark, and possible interactions were studied. Pigs in one experiment were trickle infected with low or moderate dose levels of Oesophagostomum spp. and challenge infected with S. Typhimurium. In another experiment, pigs were inoculated with S. Typhimurium followed by a challenge exposure to either Oesophagostomum, Trichuris or Ascaris. Enhancement of the Salmonella infection was not demonstrated in either experiment. The helminth effect on the pigs was modest and may explain the lack of influence on the Salmonella infection. A previous experiment with a larger Oesophagostomum infection level resulted in enhancement of the S. Typhimurium infection. A dose dependency of the interaction is therefore suggested. However, the relatively high worm burdens in the present study suggest that infection with these common pig helminths does generally not influence the course of concurrent S. Typhimurium infections under natural conditions.

Status of intestinal helminthic infections of borderline residents in North Korea

The present authors investigated intestinal parasitic infections among North Korean residents and refugees in China in 2003. The Kato-Katz method was applied to 236 residents and soldiers in a town on the North Korea-China border and to 46 people at a refugee camp in China. Only eggs of Ascaris and Trichuris were detected, with egg positive rates of 41.1% and 37.6%, respectively. The total egg positive rate was 55.0% and most of those who were egg positive were only lightly infected. Women of 61.2% and men of 53.1% were egg positive. The refugees from rural areas showed higher egg positive rates than those from urban areas. The present investigation confirmed high prevalence of soil-transmitted intestinal helminths in rural borderline areas of North Korea.

Potential regrowth and recolonization of salmonellae and indicators in biosolids and biosolid-amended soil.

This study evaluated the potential for conversion of Class B to Class A biosolids with respect to salmonellae and fecal coliforms during solar drying in concrete lined drying beds. Anaerobically (8% solids) and aerobically (2% solids) digested Class B biosolids were pumped into field-scale drying beds, and microbial populations and environmental conditions were monitored. Numbers of fecal coliforms and salmonellae decreased as temperature and rate of desiccation increased. After 3 to 4 weeks, Class A requirements were achieved in both biosolids for the pathogens and the indicators. However, following rainfall events, significant increase in numbers was observed for both fecal coliforms and salmonellae. In laboratory studies, regrowth of fecal coliforms was observed in both biosolids and biosolid-amended soil, but the regrowth of salmonellae observed in the concrete-lined drying beds did not occur. These laboratory studies demonstrated that pathogens decreased in numbers when soil was amended with biosolids. Based on serotyping, the increased numbers of salmonellae seen in the concrete lined drying beds following rainfall events was most likely due to recolonization due to contamination from fecal matter introduced by animals and not from regrowth of salmonellae indigenous to biosolids. Overall, we conclude that the use of concrete-lined beds created a situation in which moisture added as rainfall accumulated in the beds, promoting the growth of fecal coliforms and salmonellae added from external sources.

Multi-trophic level interactions in a cassava-maize mixed cropping system in the humid tropics of West Africa.

Multi-trophic level interactions in a mixed crop, involving cassava and maize, were studied in derived-savanna in Benin, West Africa. Two trials were planted, one during the short rainy season two months before onset of the dry season and one during the long rainy season in spring. Key pests under study on maize were the noctuid Sesamia calamistis Hampson and the pyralids Eldana saccharina Walker and Mussidia nigrivenella Ragonot, and on cassava, the exotic mealybug, Phenacoccus manihoti Matile-Ferrero and its encyrtid parasitoid Apoanagyrus lopezi De Santis. Both crops received insecticide treatments to assess the crop loss by a pest species. On maize, intercropping with cassava reduced egg and immature numbers of S. calamistis by 67 and 83%, respectively, as a result of reduced host finding by the ovipositing adult moth and of higher egg parasitism by Telenomus spp. Both trials showed similar effects on maize yields: on insecticide-treated maize, intercropping with cassava reduced maize yields by 9-16%, while on untreated maize the net effect of reduced pest density and increased plant competition resulted in zero yield differences; yield losses were lower in inter- compared to monocropped maize. For cassava, cropping system had no effect on parasitism by A. lopezi. Yield differences between mono- and intercropped cassava depended on time of harvest: they were large at the beginning and zero at final harvest. Land equivalent ratios were mostly > 1.5 indicating that a maize/cassava mixed crop, protected or unprotected, considerably increased the productivity per unit area of land.

Ascaris egg profiles in human faeces: biological and epidemiological implications.

Since 2 morphological forms (fertilized and unfertilized) of egg can be produced by Ascaris, infected humans can release in their faeces fertilized eggs only (FEO), unfertilized eggs only (UEO) or both fertilized and unfertilized eggs (FUE) (designated herein as the 3 different egg profiles). Epidemiologically, fertilized eggs are of significance as they enable effective transmission of the parasite. This study, for the first time, characterizes the Ascaris egg profiles in human faeces in an endemic region of China, explores possible host- and parasite-factors related to these profiles, and discusses the biological and epidemiological implications of the findings. The 3 egg profiles were recorded throughout the study period of 2 years, and the overall percentages of people with FEO, FUE and UEO profiles were approximately 41-47%, 32-42% and 17-21%, respectively. The overall number of unfertilized eggs for the entire population accounted for approximately 6-9% of all eggs excreted. The different Ascaris egg profiles showed no correlation to host gender, but they did relate to age and worm burden of the host and to the sex ratio and developmental status of the parasite. While an annual universal anthelmintic treatment resulted in some fluctuation in the values of individual egg profiles, the general features of these profiles remained similar throughout the study period. The findings of this study should have significant implications for understanding transmission patterns of Ascaris and for the implementation of control measures against ascariasis in endemic regions.

Inactivation of E. coli and helminth eggs in aerobic and anaerobic effluents using UV radiation.

This paper evaluates the performance of a simplified bench-scale UV-photoreactor used to inactivate Escherchia coli and eggs of Ascaris lumbricoides. The photoreactor consisted of a tubular unit constructed with PVC tube, 100 mm diameter and 45 cm total height, with a low-pressure mercury lamp adapted in the centre of the tube. The reactor was tested to disinfect the effluent from a trickling filter and from an UASB reactor, both fed with domestic sewage. The results showed an excellent performance of the photoreactor, with very high E. coli inactivation efficiencies being observed for the aerobic effluent (in the range of 4 to 5 log-units, for doses varying from 50.7 to 13.6 mW x s x cm(-2)) and also for the effluent from the UASB reactor (usually above 4 log-units, for doses of 20.3 and 13.6 mW x s x cm(-2)). In relation to the inactivation of helminth eggs, it was observed that UV radiation significantly affected the development of eggs of Ascaris lumbricoides, with the better results being obtained for radiation times of 40 and 60 seconds (doses of 13.6 and 20.3 mW x s x cm(-2), respectively), when approximately 65% of the eggs remained in the stage of single cell and only 9 to 10% were able to fully develop to the stage of motile larva.

The effect of repeated chemotherapy on age-related predisposition to Ascaris lumbricoides and Trichuris trichiura.

This study examines the persistence of predisposition to Ascaris lumbricoides and Trichuris trichiura during repeated chemotherapy in an urban community in Kuala Lumpur, Malaysia. Significant predisposition was observed over 2 periods of reinfection with and without age-standardization of data. Analysis of different age groups indicated that predisposition was most strongly detectable in the younger age classes. The intensities of infection with both parasites were strongly correlated at each cycle of intervention, suggesting that individuals were similarly predisposed to both species.

Response to repeated inoculations with Ascaris suum eggs in pigs during the fattening period. I. Studies on worm population kinetics.

This experimental study on pigs was designed to simulate natural, long-term exposure to Ascaris suum under modern management conditions. Parasite kinetics were followed in pigs receiving A. suum eggs as repeated trickle inoculations at two dose levels beginning at a body weight of 25 kg until their slaughter at 90 kg (baconers). In pigs inoculated twice weekly with 500 eggs, there was an initial marked rise in the numbers of hepatic milk spots, but as early as around week 6 after the start of inoculations and until week 16, at which time the last pigs were slaughtered, the numbers of spots diminished drastically. In pigs receiving only 25 eggs twice weekly, low and moderately fluctuating numbers of spots were seen throughout the experiment. Larvae recoverable from the livers and lungs were observed mainly during the beginning of the experiment. Before patency, immature intestinal worms were found in moderate numbers that showed a rough positive correlation with the dose levels, but at the time at which adult worms started to appear, immature parasites could practically no longer be found. In all, only 10 of 40 pigs harbored adults, and 4 of these 10 pigs harbored 80% of the total worm population. The results show that acquired dose-dependent host responses to A. suum play an important role in regulating the worm population along the migratory route of the parasite and that the final burden of worms in the small intestine is dose-dependent and highly variable.