Ascaris Larval Infection and Lung Invasion Directly Induce Severe Allergic Airway Disease in Mice.

Ascaris lumbricoides (roundworm) is the most common helminth infection globally and a cause of lifelong morbidity that may include allergic airway disease, an asthma phenotype. We hypothesize that Ascaris larval migration through the lungs leads to persistent airway hyperresponsiveness (AHR) and type 2 inflammatory lung pathology despite resolution of infection that resembles allergic airway disease. Mice were infected with Ascaris by oral gavage. Lung AHR was measured by plethysmography and histopathology with hematoxylin and eosin (H&E) and periodic acid-Schiff (PAS) stains, and cytokine concentrations were measured by using Luminex Magpix. Ascaris-infected mice were compared to controls or mice with allergic airway disease induced by ovalbumin (OVA) sensitization and challenge (OVA/OVA). Ascaris-infected mice developed profound AHR starting at day 8 postinfection (p.i.), peaking at day 12 p.i. and persisting through day 21 p.i., despite resolution of infection, which was significantly increased compared to controls and OVA/OVA mice. Ascaris-infected mice had a robust type 2 cytokine response in both the bronchoalveolar lavage (BAL) fluid and lung tissue, similar to that of the OVA/OVA mice, including interleukin-4 (IL-4) (P < 0.01 and P < 0.01, respectively), IL-5 (P < 0.001 and P < 0.001), and IL-13 (P < 0.001 and P < 0.01), compared to controls. By histopathology, Ascaris-infected mice demonstrated early airway remodeling similar to, but more profound than, that in OVA/OVA mice. We found that Ascaris larval migration causes significant pulmonary damage, including AHR and type 2 inflammatory lung pathology that resembles an extreme form of allergic airway disease. Our findings indicate that ascariasis may be an important cause of allergic airway disease in regions of endemicity.

[ERRORS IN DIAGNOSIS AND TREATMENT OF IMMUNODEPENDENT PATHOLOGY (ORIGINAL CONCEPT)].

Based on many years of experience in 2009, we developed the original concept of a mixed approach to the treatment of infectious diseases in patients. During 2.5 years(from 2013 to June 2015) to have applied for consultative-diagnostic help of 3965 patients who had not verified the primary diagnosis. The basic principle of verification of the pathology of the removal of various causes immunosuppression. Based on our extensive, research and observation was often found in patients ascaridosis (55%) and giardiasis (65%), as a possible cause of immunosuppression. In 13% of patients was found the mucosal candidiasis. Among frequently and chronically ill persons we identified the active forms of Epstein-Barr virus (quantitative polymerase chain reaction in saliva) in 40%. The criterion for assessing performance immunogram was a decrease of two sigmal deviation from the lower age limit. In the study of neutrophil myeloperoxidase content observed decline (< 60%) in 99 (9.7%) of 1015 patients, indicating a fairly common cause of long-term permit infection in the tissues and persistence C. albicans. In the study of lymphocyte subpopulations often demonstrated reduction in the number of natural killer cells (26.7% of subjects), which shows a decline of one of the most important factors of congenital immunity. Among the humoral immune disorders often noted the decrease of total IgG (2.4%) and its subclass IgG1 (22.1%), indicating a significant diagnostic value determination of IgG subclasses it even with normal serum total. Thus, approximately 76% of patients often suffer set of a decrease immunity. Patients developed with mixed infections caused by various bacterial, fungal, viral and protozoan agents and worms. Immunological study of patients should be redynamics after eliminating the causes immunosuppression and sanitation foci of infection. Only multi-level examination of the patient will determine the final diagnosis and adequate treatment.

Ascaris Mouse Model Protocols: Advancing Research on Larval Ascariasis Biology.

Ascariasis, caused by both Ascaris lumbricoides and Ascaris suum, is the most prevalent parasitic disease worldwide, affecting both human and porcine populations. However, due to the difficulties of assessing the early events of infection in humans, most studies of human ascariasis have been restricted to the chronic intestinal phase. Therefore, the Ascaris mouse model has become a fundamental tool for investigating the immunobiology and pathogenesis of the early infection stage referred to as larval ascariasis because of the model's practicality and ability to replicate the natural processes involved. The Ascaris mouse model has been widely used to explore factors such as infection resistance/susceptibility, liver inflammation, lung immune-mediated pathology, and co-infections and, notably, as a pivotal element in preclinical vaccine trials. Exploring the immunobiology of larval ascariasis may offer new insights into disease development and provide a substantial understanding of key components that trigger a protective immune response. This article focuses on creating a comprehensive guide for conducting Ascaris experimental infections in the laboratory as a foundation for future research efforts. © 2024 Wiley Periodicals LLC. Basic Protocol 1: Acquisition and embryonation of Ascaris suum eggs from adult females Alternate Protocol: Cleaning and purification of Ascaris suum from female A. suum uteri Basic Protocol 2: Preparation of Ascaris suum eggs and murine infection Basic Protocol 3: Measurement of larval burden and Ascaris-larva-induced pathogenesis Basic Protocol 4: In vitro hatching and purification of Ascaris L3 larvae Support Protocol: Preparation of crude antigen from Ascaris infectious stages Basic Protocol 5: Ultrastructure-expansion microscopy (U-ExM) of Ascaris suum larval stages.

Interactions between worms and malaria: good worms or bad worms?

In the past decade there have been an increasing number of studies on co-infections between worms and malaria. However, this increased interest has yielded results that have been at times conflicting and made it difficult to clearly grasp the outcome of this interaction. Despite the heterogeneity of study designs, reviewing the growing body of research may be synthesized into some broad trends: Ascaris emerges mostly as protective from malaria and its severe manifestations, whereas hookworm seems to increase malaria incidence. As efforts are made to de-worm populations in malaria endemic areas, there is still no clear picture of the impact these programmes have in terms of quantitative and qualitative changes in malaria.

Immune and genetic aspects of asthma, allergy and parasitic worm infections: evolutionary links.

There are important parallels in the immunobiology of allergy and asthma, and of the human host's response to parasitic worms. Th-2 immune actions with 'weep and sweep' mucosal biology are common to both - pathological in the first and protective in the second. Common up-regulating genetic variants of Th-2 immunity, notably in IL13 and STAT6, predict increased risk of asthma and allergy, but diminished intensity of infection by Ascaris and Schistosoma. Endemic exposures of humans to parasitic worms may have been one evolutionary force selecting for genetic variants that promote asthma and allergy.

Predisposition to ascariasis: patterns, mechanisms and implications.

Ascaris lumbricoides, the human roundworm, is a remarkably infectious and persistent parasite. It is a member of the soil-transmitted helminths or geohelminths and infects in the order of 1472 million people worldwide. Despite, its high prevalence and wide distribution it remains along with its geohelminth counterparts, a neglected disease. Ascariasis is associated with both chronic and acute morbidity, particularly in growing children, and the level of morbidity assessed as disability-adjusted life years is about 10.5 million. Like other macroparasite infections, the frequency distribution of A. lumbricoides is aggregated or overdispersed with most hosts harbouring few or no worms and a small proportion harbouring very heavy infections. Furthermore, after chemotherapeutic treatment, individuals demonstrate consistency in the pattern of re-infection with ascariasis, described as predisposition. These epidemiological phenomena have been identified, in a consistent manner, from a range of geographical locations in both children and adults. However, what has proved to be much more refractory to investigation has been the mechanisms that contribute to the observed epidemiological patterns. Parallel observations utilizing human subjects and appropriate animal model systems are essential to our understanding of the mechanisms underlying susceptibility/resistance to ascariasis. Furthermore, these patterns of Ascaris intensity and re-infection have broader implications with respect to helminth control and interactions with other important bystander infections.

Prevalence of gastrointestinal parasites in bonnet macaque and possible consequences of their unmanaged relocations.

Relocation is one of the mitigating measures taken by either local people or related officers to reduce the human-bonnet macaque Macaca radiata conflict in India. The review on relocations of primates in India indicates that monkeys are unscreened for diseases or gastrointestinal parasites (henceforth endoparasites) before relocation. We collected 161 spatial samples from 20 groups of bonnet macaque across their distribution range in south India and 205 temporal samples from a group in Chiksuli in the central Western Ghats. The isolation of endoparasite eggs/cysts from the fecal samples was by the centrifugation flotation and sedimentation method. All the sampled groups, except one, had an infection of at least one endoparasite taxa, and a total of 21 endoparasite taxon were recorded. The number of helminth taxon (16) were more than protozoan (5), further, among helminths, nematodes (11) were more common than cestodes (5). Although the prevalence of Ascaris sp. (26.0%), Strongyloides sp. (13.0%), and Coccidia sp. (13.0%) were greater, the load of Entamoeba coli, Giardia sp., Dipylidium caninum and Diphyllobothrium sp. were very high. Distant groups had more similarity in composition of endoparasites taxon than closely located groups. Among all the variables, the degree of provisioning was the topmost determinant factor for the endoparasite taxon richness and their load. Temporal sampling indicates that the endoparasite infection remains continuous throughout the year. Monthly rainfall and average maximum temperature in the month did not influence the endoparasite richness. A total of 17 taxon of helminths and four-taxon of protozoan were recorded. The prevalence of Oesophagostomum sp., and Strongyloides sp., and mean egg load of Spirurids and Trichuris sp. was higher than other endoparasite taxon. The overall endoparasite load and helminth load was higher in immatures than adults, where, adult females had the highest protozoan load in the monsoon. The findings indicate that relocation of commensal bonnet macaque to wild habitat can possible to lead transmission of novel endoparasites that can affect their population. Thus, we suggest avoidance of such relocations, however, if inevitable the captured animals need to be screened and treated for diseases and endoparasites before relocations.

Gastrointestinal nematode infection is associated with variation in innate immune responsiveness.

Ex vivo monocyte cytokine responses (IL-1beta, TNF-alpha, IL-12p70, IL-10, TGF-beta) to bacterial TLR2 and TLR4 ligands were quantified in 47 gastrointestinal (GI) nematode-exposed children in Pemba Island, Tanzania. Worminess (estimated by faecal egg counts (FEC)) had a positive relationship with pro-inflammatory TNF-alpha and IL-1beta responsiveness to the TLR ligands. In particular, there was a strong significant relationship with TNF-alpha response to TLR4 ligand (LPS). There were no significant associations between regulatory responses (IL-10, TGF-beta) and worminess. These results are consistent with the possibility that GI nematodes modulate innate responses and may indicate a potential mechanism for interactions between GI nematodiasis and important bystander pathogens.

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.

Thermal composting of faecal matter as treatment and possible disinfection method--laboratory-scale and pilot-scale studies.

When using toilets where the urine and faeces are collected separately for reuse as nutrients in agriculture, the collected matter should be disinfected. One way to do this is by thermal composting. Composting of different material mixes was investigated in a laboratory-scale experiment. This showed that the best mixture for dry thermal composting was a mix of faeces, food waste and amendment. The urine was collected separately by use of urine-diverting toilets. A new method was developed to mathematically evaluate and estimate the safety margins of pathogen inactivation during thermal composting. The method is based upon a mathematical calculation of the number of times total inactivation (at least 12log(10) reduction) of the organisms is achieved. In a pilot-scale experiment, the disinfection of a faeces/food waste mix was performed with a calculated safety margin of more than 37 times the total die-off of Enteroviruses and some 550 times that of Ascaris. Thus, well functioning composting seems to be effective for disinfection of faecal matter. To get a high temperature in all of the material, the reactor has to have sufficient insulation. A major disadvantage is the initial need for handling the raw un-disinfected material. The degradation of the organic matter in the compost was almost 75%, resulting in a small final volume that could safely be recycled.

The potential for disinfection of separated faecal matter by urea and by peracetic acid for hygienic nutrient recycling.

No efficient, reliable, and scale independent disinfection methods for toilet waste are available today for safe recycling of plant nutrients. Therefore, two chemical treatment methods, addition of urea or of PAA (a quaternary mixture of 15% peracetic acid, 15% hydrogen peroxide and 30% acetic acid), were evaluated for disinfection of faecal matter.Degradation of the added urea resulted in 30 g of ammonia nitrogen per kilogram of treated matter and a pH increase to approximately 9.3. This produced an efficient disinfection of E. coli, Enterococcus spp., and Salmonella spp. within 3 weeks (>6log(10) reduction) and a reduction of the chemical resistant Salmonella typhimurium 28b phage, corresponding to a decimal reduction within 7.5 days. No viable Ascaris suum eggs were found after 50 days of treatment. No reduction of spore forming Clostridia spp. was observed. Urea treatment proved to be efficient for disinfection of source separated faecal matter in a scale independent method used for safe recycling of nutrients found in the faecal matter.PAA reduced all of the above indicator organisms within 12 h after application. For this faecal material, with a dry matter content of approximately 10%, an addition of 0.5-1% of PAA (active substance, corresponding to 3.3-6.7% of the Proxitane 15 used) was required before no viable organisms were found in the material. However, this was not tested for the A. suum. No viable spore-forming bacteria or phages were detected. A high rate of bacteria regrowth occurred at 0.15% dosage and 5 days of treatment. PAA is an efficient alternative for disinfection of separated faeces if a rapid treatment is needed.

Differential responses of Duroc, Hampshire, and crossbred pigs to a superimposed experimental infection with the intestinal threadworm, Strongyloides ransomi.

In both spring and fall, 12 Duroc, 12 Hampshire, and 12 Duroc times Hampshire F1 weanling pigs all reared under the same management were fed in pens of 3 to slaughter weights. Three Duroc and 4 Hampshire boars, essentially unrelated within breed, were used in sampling the breeds. Swine herd management allowed pig infection with Strongyloides ransomi and Ascaris suum, but neither clinical nor subclinical parasitism was evident in the herd. Pigs were percutaneously exposed by pens within breed and season, half to none (control) and half to 3,000,000 (exposed) S. ransomi infective larvae, Breed, treatment, and seasons were prominent sources of variation in pig response. Breeds failed to respond alike to parasitism in respect to experimental periods and exposure levels. This interaction response (P smaller than 0.01) showed that S. ransomi egg production increased rapidly for all breed groups but decreased quicker and greater in Durocs, slowest and least in Hampshires, with cross breds intermediate in these respects. The 2.0830 for mean of log A. suum EPG from exposed Durocs was near double that of control Durocs but the mean for exposed Hampshires was less than half that for controls; crossbreds tended to be intermediate in this respect. Daily gains of 0.70 and 0.73 for Durocs and crossbreds were similar (P greater than 0.10) but averaged 11.7% more (P smaller than 0.05) than the 0.64 kg for Hampshires and gains by control pigs were 20.6% above (P greater than 0.01) that of exposed pigs. Exposed pigs required more feed per kg of gain (P greater than 0.05) than control pigs (3.60 vs. 3.33 kg). Comparison of relative gains and feed efficiences of control and exposed pigs among and within breed groups supported the position that a superimposed exposure of 3,000,000 S. ransomi larvae was more severe for Hampshires, intermediate for crossbreds, and least severe for Durocs.

Experimental infection of domestic swine with Baylisascaris procyonis from raccoons.

Six 8-week-old ascarid-naive pigs which were experimentally infected with 72,000 embryonated Baylisascaris procyonis eggs of raccoon origin developed lesions limited to the intestines and liver. Intestinal lesions consisted of multifocal areas of inflammation by macrophages, eosinophils, and lymphocytes in the mucosa and submucosa, in association with Baylisascaris larvae; similar lesions were seen in the mesenteric lymph nodes. Typical white, granulation type, multifocal interstitial hepatitis ("milk-spots"), 1 to 5 mm in diameter, were seen in the livers by 7 days, with resolution by 47 days. Microscopically, these consisted of multifocal areas of marked periportal and interlobular edema, and influx of eosinophils, and large intralobular aggregates of eosinophils. At 47 days, hundreds to thousands of small white granulomas were seen on the serosa of the intestines; microscopically, they were discrete collections of macrophages, lymphocytes, and eosinophils in the submucosa and muscle layers surrounding nonviable remnants of Baylisascaris larvae. Larvae or lesions were not seen in other tissues, including the brain. These experiments indicated that B procyonis will undergo limited migration in swine and can produce typical white spots in the liver. The larvae were killed by cellular reactions in the intestinal wall and liver, and, unlike the situation in most other animals infected with this parasite, no somatic migration or CNS disease occurred after infection.

Experimentally induced cerebrospinal nematodiasis in rabbits (Oryctolagus cuniculus).

To determine if the rabbit can act as an intermediate host for ascarids from certain carnivorous animals (skunks, pigs, dogs), 12 rabbits were infected with Ascaris columnaris, Ascaris suum, or Toxocara canis. Clinical signs, microscopic lesions, and morphologic features of ascarids with naturally occurring and experimentally induced A columnaris infections were identical. Of the 3 induced parasitic infections, A columnaris caused the most severe lesions in the central nervous system (CNS). It appears that rabbits may potentially act as intermediate hosts for A columnaris.

Determination of the inactivation rate of Ascaris eggs in wastewater stabilization pond sludge using dialysis chambers and sludge cores.

The inactivation rate of Ascaris eggs was studied in the sludge layer of a primary, facultative wastewater stabilization pond located in Mexico City. Two independent methods, sludge cores and dialysis chambers, were used, to determine the inactivation rates through which a comprehensive picture of the inactivation was gained. The dialysis chambers provided a detailed picture of the initial inactivation (14 months) at one location in the pond, whereas the sludge cores provided less precise information about the inactivation rate at several locations and over the entire lifetime of the pond (10 years). The inactivation curve was characterized by an initial lag phase, a period of roughly first-order inactivation, and a tailing region. During the first year, 50 to 60% of the eggs were inactivated, after which the rate decreased. Although the observed, initial first-order rate constant was greater than 0.002 d(-1), the average, long-term rate constant was closer to 0.001 d(-1).

Concentrations and inactivation of Ascaris eggs and pathogen indicator organisms in wastewater stabilization pond sludge.

During treatment in wastewater stabilization ponds (WSPs) many pathogens, in particular helminth eggs, are concentrated in the sludge layer. Because periodic removal of the sludge is often required, information is needed on the concentrations and inactivation of pathogens in the sludge layer to evaluate the public health risk they pose upon removal of the sludge. In this paper, previous reports on the sludge concentrations of various pathogen indicator organisms and helminth eggs are reviewed and results from our own recent experiments are reported. The advantages and disadvantages of several methods for studying inactivation in the sludge layer are discussed, as well as implications for the management of WSP sludge. In our recent experiments, which were conducted at three WSPs in central Mexico, sludge cores, dialysis chambers, and batch experiments were used to measure the inactivation rates of fecal coliform bacteria, fecal enterococci, F+ coliphage, somatic coliphage, and Ascaris eggs. The first-order inactivation rate constants were found to be approximately 0.1, 0.1, 0.01, 0.001, and 0.001 d(-1), respectively. The concentrations of all the organisms were found to vary both vertically and horizontally in the sludge layer; therefore, to determine the maximum and average concentration of organisms in the sludge layer of a WSP, complete sludge cores must be collected from representative locations throughout the pond.

Ozonation by-products issued from the destruction of microorganisms present in wastewaters treated for reuse.

This work demonstrates the reaction of ozone on the amino acids comprising the covering layer of resistant micro-organisms. A secondary aim was to check the byproducts generated when ozone was applied to synthetic samples (such as Vibrio cholerae NO 01 WFCC-449, Salmonella typhi ATTC-6539, faecal coliforms and Ascaris suum). The ozone was applied at a concentration of 18.4 mgO3/min at pH 3, for different lengths of time. In the case of bacteria, results showed that, at 8 minutes, the number was reduced to the level of the Official Mexican Standards set for treated water destined for irrigation purposes (1,000 MPN/100 mL). Excellent correlation coefficients (0.95 to 0.99) were obtained for microbial concentrations versus ozone contact time. Destruction times required for 100% removal of the initial bacteria population varied between 2 and 14 minutes, while Ascaris suum required 1 hour. When Gram-negative bacteria die due to the effects of ozone, cellular lysis and the liberation of endotoxins (biodegradable) were observed. The ozonation of amino acids in the shell of Ascaris suum eggs, leads to the formation of aldehydes, such as formaldehyde and acetaldehyde, in low concentrations (0.0003 and 0.0005 microg/mL respectively). These levels are not hazardous to human health.

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.

[Trypsin and alpha-amylase activities in the pancreas of guinea pigs. II. Effect of per os and intra-intestinal infection with invasive ova of Ascaris suum]. / Aktywnosc trypsyny i alfa-amylazy w trzustce swinek morskich. II. Wplyw zarazenia per os i dojelitowo inwazyjnymi jajami ascaris suum.

The guinea pigs were infected orally or intraintestinally (after laparotomy) with 5000 invasive eggs of Ascaris suum. On the 7th day after infection the invasion was controlled by lung's weight, degree of congestion, and number of larvae in the lungs. The activity of amylase was determined according to Fennel method, trypsin to Anson method. The activities of both enzymes in pancreas of orally infected animals (after laparotomy or not) were lower, and relative weight of lungs of these animals was higher than in control animals. In pancreas of guinea pig with invasive material introduced into its, the activities of amylase and trypsin were higher than in control animals, but only the activity of trypsin differed significantly.

[Trypsin and alpha-amylase activity in the pancreas of guinea pigs. IV. Comparison of the effects of administration of the mixtures of volatile organic acids, invasive ova of Ascaris suum and their homogenates]. / Aktywnosc trypsyny i alfa-amylazy w trzustce swinek morskich. IV. Porównanie wplywu podawania mieszaniny lotnych kwasów organicznych, inwazyjnych jaj Ascaris suum i ich homogenatów.

The activity of alpha-amylase in the experimental groups of guinea pigs was lower than in control animals. The decreased level of activity of trypsin was noted (p less than 0.05) also in infected animals, and in treatment with organic acids. In the group of guinea pigs, which was administrated homogenized eggs of parasite, the activity of enzyme was a little higher than in control animals. The infection of animals was associated with increase of relative weight of lungs. The guinea pigs treatment with organic acids had also a higher weight of spleen.