Suum Eggs Research Articles

40 The gut microbiota modifies host-parasite interactions

Abstract It is well documented that parasitic infections induce a profound change in the structure and function of host gut microbiota, likely due to infection-associated perturbations in the gut microenvironment. However, the mechanisms remain largely unknown. Here we aim to understand how the gut microbiota is modified by host-parasite relationships or vice versa using an integrated multi-omics analysis in a swine-Ascaris infection model. Female pigs (n = 30; 6 to 8 mo of age) were randomly divided into three groups (n = 10 per group). One group served as normal uninfected controls (NU); the remaining 20 pigs were inoculated per os with ~10,000 infective A. suum eggs; and the infection was allowed to progress for 35 d post-inoculation (dpi). At 21 dpi, one of the infected groups (IP) was treated with a single daily dose of an anthelmintic drug, Panacur, for three consecutive days while the other infected pigs remained untreated (IU). At necropsy, intestine tissue, serum and fecal samples were collected for untargeted metabolome analysis using UPLC-MS/MS and the fecal microbiota was characterized using full-length 16S rRNA gene-based sequencing with DADA2. Fecal and serum metabolites were categorized for their origin using MetOrigin. An integrated multi-omics analysis was performed using DIABLO algorithms and NetCoMi networking tools. Our results showed that A. suum worms were well developed in pigs from the IU group, which harbored a significantly reduced microbiota alpha diversity (Simpson, P < 0.005) compared with NU. The infection altered the abundance of at least 17 bacterial species/strains, including multiple butyrate-producing Clostridium spp., such as C. butyricum. The infection also dysregulated 51 fecal metabolites, including microbiota-derived metabolites such as glutaric acid and p-Cresol sulfate, and succinate of mixed origin. Moreover, the anthelmintic treatment was efficacious and eliminated all worms from the gut. Nevertheless, 11 pathways of both host and microbiota origin were significantly enriched in the gut of pigs from the IP group, including peptidoglycan biosynthesis, histidine metabolism (in both feces and serum), and primary bile acid biosynthesis, suggesting the infection-induced alterations may not be transient. In conclusion, the host microbiota modified swine-Ascaris interactions via three different mechanisms. First, microbiota-derived metabolites directly regulate host gene expression, which in turn affects host physiology and immune responses. Second, plasticity of the gut microbiota allows the exploitation of the niche differentiated upon infection, resulting in blossom of certain strains in the gut of post-treated animals. Lastly, the host microbiota is likely one of the key determinants of parasite gut microbiota, indirectly exerting its effect on parasite physiology and nutrition. Further studies aiming to understand reciprocal yet complex relations between the gut microbiota, the host, and parasites (including parasite gut microbiota) will be conducive to the design of next-generation functional anthelmintics.

ASCARIS SUUM EGG RECOVERY FROM SLUDGE SAMPLES AFTER PHASE EXTRACTION.

Some helminth test methods for sanitation samples include a phase extraction step to reduce lipid content and final pellet size before microscopy. Hydrophilic and lipophilic solutions are used to create 2 phases, with a plug of organic material or debris in between, whilst eggs are supposedly compacted at the bottom of the test tube. We tested 10% formalin, acetoacetic buffer, and acid alcohol as the hydrophilic solutions, and ethyl acetate and diethyl ether as the lipophilic solvents for egg recoverability from water, primary sludge, and fatty sludge. Normally, the supernatant and debris plug are discarded and the sedimented pellet of eggs is microscopically examined. We, however, also collected the entire supernatant plus debris plug to determine where eggs were possibly lost. We found that eggs were lost when samples were extracted with 10% formalin + ethyl acetate, 10% formalin + diethyl ether, acetoacetic buffer + ethyl acetate, and acetoacetic buffer + diethyl ether combinations (<50% egg recovery). Acid alcohol + ethyl acetate resulted in 93.2, 89.8, and 57.3% egg recovery in the pellet of water, primary sludge, and fatty sludge, respectively; however, the size of the final pellet was not reduced, defeating the purpose of the extraction step. We thus recommend that this step be excluded.

Possibilities of decontaminating organic waste from swine-farming complexes using anaerobic digestion

AbstractAnaerobic digestion with simultaneous production of biogas is one of the most common methods of recycling, processing and decontamination of organic waste to produce alternative fuels. A commonly used indicator of the sanitary safety of the substrate is eggs of Ascaris suum nematodes of pigs, which are characterised by extremely high resistance to environmental factors. The aim is to investigate the effect of the mesophilic mode of anaerobic digestion on the activity of pathogens, particularly Ascaris suum eggs. The eggs of the studied nematode species were placed in a biogas installation at a temperature of 37 °C. The digestate samples with eggs were then taken every 4 days for 28 days, followed by larvae culture to determine the viability of the eggs. The results of our research have shown that the mesophilic regime of anaerobic digestion is an effective method of controlling parasites, but at the same time, it needs to be improved, since only 7.6% of A. suum eggs remained alive after a 1-day stay, about 50% after the week stay and about 9% in the third week stay in the biogas reactor. Thus, further optimisation of anaerobic fermentation in the mesophilic mode can be aimed at improving the suppression of pathogenic activity.

Characterizing Excretory-Secretory Products Proteome Across Larval Development Stages in Ascaris suum.

Ascaris lumbricoides and Ascaris suum are parasitic nematodes that primarily infest the small intestines of humans and pigs, respectively. Ascariasis poses a significant threat to human health and swine health. Understanding Ascaris larval development is crucial for developing novel therapeutic interventions that will prevent ascariasis in both humans and pigs. This study aimed to characterize the excretory-secretory (ES) proteome of different Ascaris suum larval stages (L3-egg, L3-lung, L3-trachea) to identify potential targets for intervention to prevent Ascaris -induced global morbidity. Stage-specific larvae were isolated, cultured in vitro and ES-product was collected. Third-stage Ascaris larvae (L3) were isolated from embryonated eggs (L3-egg), isolated from the lungs of Balb/c mice infected with Ascaris suum eggs at day 8 post infection (L3-lungs) and isolated from the trachea of Balb/c mice infected with Ascaris suum eggs at day 12 post infection (L3-trachea). ES products were obtained by culturing larvae. Proteomic analysis was conducted using liquid chromatography-tandem mass spectrometry (LC-MS/MS) and bioinformatic tools including MaxQuant, Perseus, and Andromeda, following a detailed protocol available on GitHub. The analysis encompassed peptide identification, scoring, and quantification against an organism-specific database, with subsequent quality control, correlation assessment, and differential abundance determination using the Amica algorithm. A total of 58 unique proteins were identified in the ES products. Fourteen proteins were common across all stages, while others were stage-specific. Principal component analysis revealed distinct protein profiles for each stage, suggesting qualitatively different proteomes. Gene ontology analysis indicated stage-specific GO enrichment of specific protein classes, such as nuclear proteins in L3-egg ES products and metabolic enzymes in L3-lung and L3-trachea ES products. This study revealed stage-specific differences in the composition of Ascaris ES products. Further investigation into the functional roles of these proteins and their interactions with host cells is crucial for developing novel therapeutic and diagnostic strategies against ascariasis.

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.

Bayesian estimation of the sensitivity and specificity of coprological and serological diagnostic tests for the detection of Ascaris suum infection on pig farms

Coprological and serological diagnostic tests were compared to define the status of a pig farm with regard to Ascaris suum. On each of the 100 farms in France visited for the study, 10 blood samples were taken from pigs at the end of fattening (at least 22 weeks old) and 20 to 30 faecal samples were taken, depending on the category of animals present on the farm (10 sows, 10 piglets aged 10 to 12 weeks and 10 pigs at the end of fattening, aged at least 22 weeks). A SERASCA® ELISA test (Laboratory of Parasitology, Ghent University) was performed on each blood sample (cut-off 0.5) and a coprological analysis on each faecal sample. A Bayesian approach was used to estimate the sensitivity and specificity of the coprological and serological tests. A farm was considered positive if at least one A. suum egg was observed in the faecal samples. With regard to the serological test, various hypotheses were tested in order to define the number of seropositive animals required to consider a farm positive for A. suum. The coprological test has very good specificity in the search for A. suum, whether 20 or 30 samples are taken per farm. However, even with an increase in the number of samples, the sensitivity of this diagnostic approach is very low (less than 30%). On the other hand, the serological diagnostic method, which consists of taking blood samples from 10 animals at the end of fattening, has good sensitivity and seems better suited to defining the status of a farm with regard to A. suum, provided that a farm is considered seropositive only if two out of 10 samples are positive.

Study of helminth eggs (Ascaris suum) inactivation by anaerobic digestion and electrochemical treatment

Background The use of insufficiently treated wastewater or Faecal sludge in agriculture raises concerns because of the pathogen content. Helminth eggs (HE) are one of the most crucial pathogens for ensuring public health and safety. Widely used disinfection treatment methods do not guarantee the complete inactivation of helminth eggs. The current study evaluated the effectiveness of anaerobic digestion and electrochemical process on helminth (Ascaris suum) egg inactivation. Methods Lab-scale biochemical methane potential (BMP) assay was conducted by spiking A. suum eggs in a serum bottle. Total solid (TS), volatile solid (VS), pH, biogas production and its composition, and volatile fatty acids (VFA) were analyzed along with A. suum inactivation every third day for the initial 15 days and fifth day for 45 days. In the second set of experiments, a hypochlorite (4700 ppm) solution was generated by electrolysis of aqueous NaCl solution in a membrane-less electrochemical cell. The hypochlorite was diluted (940, 470, 235, and 156ppm) in wastewater, spiked with A. suum eggs and then examined for inactivation at regular intervals. Results The results of the anaerobic digestion treatment documented 98% inactivation of A. suum eggs (0.15 eggs/mL) in 35 days and remained at 0.14 eggs/mL until day 45. Correlation analysis revealed a positive relationship between non-viable eggs and pH and a negative relationship with all the other parameters. Electrochemical treatment achieved 10% inactivation at 940 ppm concentration in 24h. Conclusions This study revealed that the inactivation of A. suum eggs by anaerobic digestion or electrochemical treatment is a combined effect of more than one parameter.

Performance evaluation of protocols for Taenia saginata and Ascaris suum egg recovery from the house fly’s gastrointestinal tract and exoskeleton

BackgroundThe synanthropic house fly (Musca domestica) can potentially contribute to the mechanical spread of eggs of Taenia and Ascaris spp. in the environment and between hosts. However, the absence of validated protocols to recover eggs hampers an in-depth analysis of the house fly's role in parasite egg transmission.MethodsThe gastrointestinal tract and exoskeleton of euthanized house flies were spiked with Taenia saginata eggs. The performance of several recovery protocols, in terms of both the recovery rate and ease-of-use, was (microscopically) evaluated and compared. These protocols employed steps such as washing, maceration, filtration, flotation and both passive and centrifugal sedimentation. The final validated protocols were subsequently evaluated for the recovery of Ascaris suum eggs.ResultsThe final protocol validated for the recovery of T. saginata eggs from the house fly’s gastrointestinal tract involved homogenization in phosphate-buffered saline and centrifugation at 2000 g for 2 min, yielding a recovery rate of 79.7%. This protocol required 6.5 min to perform (which included 1.5 min of hands-on time) and removed large debris particles that could hinder the differentiation of eggs from debris. Similarly, the final protocol validated for the recovery of T. saginata eggs from the fly’s exoskeleton involved washing by vortexing for 2 min in Tween 80 (0.05%), 15 min of passive sedimentation and centrifugation at 2000 g for 2 min, yielding a recovery rate of 77.4%. This protocol required 20.5 min to perform (which included 3.5 min of hands-on time) and successfully removed debris. The same protocols yielded recovery rates of 74.2% and 91.5% for the recovery of A. suum eggs from the fly’s gastrointestinal tract and exoskeleton, respectively.ConclusionsEffective, simple and easy-to-use protocols were developed and validated for the recovery of T. saginata and A. suum eggs from the house fly’s gastrointestinal tract and exoskeleton. These protocols can be applied to investigate the importance of flies as parasite egg transmitters in laboratory and field settings.Graphical

Effect of Selected Entomopathogenic Fungal Species on Embryonic Development of Ascaris suum (Nematoda).

The aim of the study was to evaluate the potential of using five selected species of entomopathogenic fungi (Beauveria bassiana, B. brongniartii, Conidiobolus coronatus, Isaria fumosorosea, and Metarhizium robertsii) in the bioregulation of the dispersive stages of the parasitic nematode-Ascaris suum. Experimental cultures of each of the selected entomopathogenic fungi, as well as a control culture without fungi, were incubated with A. suum eggs at 26 °C for 28 days. Development of the A. suum eggs was observed using a light microscope on the 7th, 14th, 21st, and 28th days of incubation. The API-ZYM® test was used to determine, semiquantitatively, the activity of 19 hydrolytic enzymes from the entomopathogenic fungi. The cytotoxicity of the fungi was determined using tetrazole salt MTT. It was found that none of the five tested strains of entomopathogenic fungi showed an ovicidal effect, and none of them colonized the A. suum egg shells. However, ovistatic activity was observed mainly until the 14th day of incubation by I. fumosorosea, M. robertsii, and B. bassiana. In the MTT test, M. robertsii showed moderate cytotoxicity, while the other species showed low cytotoxicity. Among the strains tested, I. fumosorosea showed the highest spectrum of hydrolase production (13 out of 19 enzymes gave a positive reaction from 3 to 5; 20-40 nM or more). The absence of morphological changes in the A. suum egg shells suggests that the antagonistic effect of the studied entomopathogenic fungi may be due to their cytotoxicity, associated with the production of secondary metabolites-toxins (M. robertsii) and enzymatic activity (I. fumosorosea).

PSIV-3 Biofumigation to Reduce Swine Parasite Contamination on Organic Pastures

Abstract Parasite infection is a common problem in organic pig production. Pastures grazed by infected pigs can become sources of parasite transmission because parasite eggs can survive in soil for several years. Biofumigation is a method used by farmers to reduce plant pathogens and nematodes in soil by incorporating plant residues into the soil that rapidly release toxic isothiocyanates. This study explored whether grazing crops with biofumigation properties affects parasite fecal egg counts of pigs managed organically, and subsequent parasite eggs in organic pasture soil after biofumigation. A study was conducted using 66 pigs (BW = 72.6 kg ± 21.2 SD) that were raised organically in two blocks. Within each block, pigs were housed in four pens (8 to 9 pigs/pen) and provided feed ad libitum in a hoop barn. Each pen had access to a pasture (0.5 acres). Each pasture was subjected to one of four management treatments: control pastures (Ladino clover) or biofumigation pastures (Rapeseed), each with or without incorporating plant residues. Each pasture was split into four paddocks (9.1m × 10.7m) and pigs were given access to each paddock for one week. Fecal samples (n = 198) were collected from each pig before, two and four weeks after grazing. Triple soil samples (n = 480) were collected from each paddock before grazing, immediately after pigs were removed from pasture paddocks, and weekly thereafter for three weeks. Fecal and soil samples were analyzed for Ascaris suum, Trichuris suis, and Oesphagostomum spp. eggs. Data were analyzed using a mixed model with pen or paddock as the experimental unit. Fecal egg counts of the three parasite species did not differ between pigs that grazed on pastures with different management treatments. All pigs had drecreased Ascaris suum (P &amp;lt; 0.01) and Trichuris suis (P &amp;lt; 0.01) fecal egg counts after four weeks of grazing compared with before grazing. Both clover and rapeseed pastures without incorporating plant residue had decreased Ascaris suum egg counts in the soil compared with incorporating plant residue (both P &amp;lt; 0.05). Rapeseed pastures without incorporating plant residue had reduced (P &amp;lt; 0.05) Trichuris suis egg counts in the soil compared with clover pastures but did not differ from rapeseed pasture with incorporated plant residue. Across pasture management treatments, Ascaris suum and Trichuris suis eggs in the soil were reduced (P &amp;lt; 0.05) two and three weeks after pigs were removed from pastures compared with before, immediately after, and one week after pigs were removed. These results suggest that grazing pastures crops with biofumigation properties did not affect fecal egg counts of pigs. While the number of parasite eggs decreased overtime in pasture soil, biofumigation may not be an effective way to reduce swine parasite eggs in organic pastures.

Comparison of the effectiveness of various parasitological methods in detecting nematode eggs in different types of soil.

Natural fertilizers, sewage sludge, digestates, as well as organic fertilizers produced on their basis, can become a source of parasitological contamination of cultivated land. High concentration of invasive forms of parasites in the soil may pose a threat to human and animal health. Therefore, it is necessary to control the hygienic condition of fertilizers and fertilized soils with particular emphasis on parasites. The aim of the study was to compare the effectiveness of methods commonly used for parasitological examination of soil with own methods which were used to develop the standards. The study was carried out using samples of sandy soil (SS), horticultural mix soil (HS) and peat-based substrate (PS). Each sample was spiked with 100 dyed Ascaris suum eggs and examined with the use of 6 methods: Vasilkova, Dada, Quinn, and 3 methods according to the Polish Standards (PN-19000, PN- 19005, PN-19006). For each variant, 8 repetitions were made. The largest number of A. suum eggs were found with PN-19006 (mean number of detected eggs was 21.25, 46.50, 23.00 for HS, SS, PS, respectively. Slightly lower results were obtained using PN-19005 - the mean number eggs was 21.25, 36.00, 16.75, respectively. On the other hand, the mean number of A. suum eggs found with the Dada method was about 2-3 times lower than with the PN-19006 - 15.75, 22.50, 6.50 for HS, SS, PS soil, respectively. Other methods were much less effective. PN-19006 method turned out to be the most effective in detecting A. suum eggs. This method can be used for parasitological examination of soils and can be the basis for developing a system of methods dedicated to testing different types of soils for the presence of nematode eggs.

Study of helminth eggs (Ascaris suum) inactivation by anaerobic digestion and electrochemical treatment

Background: The use of insufficiently treated wastewater or faecal sludge in agriculture raises concerns because of the pathogen content. Helminth eggs are one of the most crucial pathogens for ensuring public health and safety. Widely used disinfection treatment methods do not guarantee the complete inactivation of helminth eggs. The current study evaluated the effectiveness of anaerobic digestion and electrochemical process on helminth (Ascaris suum) egg inactivation. Methods: Lab-scale biochemical methane potential (BMP) assay was conducted by spiking A. suum eggs in a serum bottle. Total solid (TS), volatile solid (VS), pH, biogas production and its composition, and volatile fatty acids (VFA) were analyzed along with A. suum inactivation every third day for the initial 15 days and fifth day for 45 days. In the second set of experiments, a hypochlorite (4700 ppm) solution was generated by electrolysis of aqueous NaCl solution in a membrane-less electrochemical cell. The hypochlorite was diluted (940, 470, 235, and 156ppm) in wastewater, spiked with A. suum eggs and then examined for inactivation at regular intervals. Results: The results of the anaerobic digestion treatment documented 98% inactivation of A. suum eggs (0.15 eggs/mL) in 35 days and remained at 0.14 eggs/mL until day 45. Correlation analysis revealed a positive relationship between non-viable eggs and pH and a negative relationship with all the other parameters. Electrochemical treatment achieved 10% inactivation at 940 ppm concentration in 24h. Conclusions: This study revealed that the inactivation of A. suum eggs by anaerobic digestion or electrochemical treatment is a combined effect of more than one parameter.

Avaliação da eficácia da solução hidroalcoólica de própolis com glicina na redução conjunta de microrganismos e ovos de helmintos em vegetais de folhas verdes

Abstract The main goal of this study was to evaluate the effectiveness of a propolis hydroalcoholic solution with glycine on the reduction of microorganisms and helminth eggs removal in lettuce leaves. Three experimental groups were evaluated in triplicate or replicates: microbiological (ME), parasitological (PE), and microbiological and parasitological (MPE). Before the experiments, the initial concentration of the native microbiota was assessed by counting mesophilic aerobes, molds and yeasts, using conventional microbiological techniques. Lettuce leaves were washed in tap water and subjected to different immersion treatments: propolis hydroalcoholic solution (PHS), propolis hydroalcoholic solution at pH 5.5 (PHS2), and propolis hydroalcoholic solution with glycine (PHS-glycine). In ME, propolis hydroalcoholic solution combined or not with glycine, was evaluated on the reduction of native microbiota. Residual microbiological contamination was assessed. In the PE, the effect of PHS2 and PHS-glycine was evaluated on the removal of a known number of Ascaris suum eggs on leaves artificially contaminated. In MPE, PHS-glycine was evaluated on the reduction of native microbiota and removal of eggs. A reduction of more than 2 log CFU/g was observed in ME. In PE, PHS-glycine removed 37.22% of eggs. In MPE, the PHS-glycine reduced 3.3 CFU/g of mesophilic aerobes, 5.0 log CFU/g of fungi, and 35.53% of eggs from vegetables. The results highlighted the solution’s efficiency in reducing or eliminating different contaminants in vegetables and reinforced the need for the development of hygiene methods that do not harm human health and are reproducible at the household level.

Intestinal roundworm infection in organic and alternative swine farms in the USA

Swine parasites are a challenge for organic and alternative pig farmers and represent suboptimal animal health because use of chemical prophylaxis is not recommended. There is no comprehensive study of how prevalent gastrointestinal parasites are in organic or alternative pig farms across the USA. This study investigates prevalence, abundance, and intensity measures for three intestinal roundworm parasites (Ascaris suum, Oesophagostomum spp. and Trichuris suis) on organic and alternative swine farms by quantifying their eggs in pig feces, pastured soil, and bedding. Nine farms raising pigs in bedded facilities across four states (Iowa, Minnesota, Pennsylvania, and Wisconsin) were sampled, and analysis of parasite egg counts from feces indicate that 88.9%, 55.5%, and 44.4% of farms were infected with A. suum, Oesophagostomum spp. and T. suis, respectively. Prevalence, mean abundance, and mean intensity of eggs per gram (EPG) in feces for A. suum and T. suis were higher in growing and finishing pigs, whereas they were higher for Oesophagostomum spp. in finishing pigs and sows. Prevalence and abundance with A. suum eggs in feces were higher in organic certified farms, whereas abundance of Oesophagostomum spp. and T. suis was lower than those in alternative farms. Fewer eggs were recovered from soils than bedding samples, but their presence in both substrates confirmed that dispersal stages occur in farms. Overall, our results indicate that intestinal parasite infection across organic and alternative swine farms in the USA is common, and not surprising given that few options to control infection within organic regulations are available.

Modulation of the porcine intestinal microbiota in the course of Ascaris suum infection

BackgroundThe porcine roundworm Ascaris suum impairs feed conversion and weight gain, but its effects on intestinal microbiota remain largely unexplored.MethodsModulation of the intestinal microbiota was assessed in pigs that were infected once with 10,000 A. suum eggs and pigs that received a trickle infection (1000 eggs/day over 10 days), compared with a non-infected control group. Six pigs each were sacrificed per group at days 21, 35 and 49 post-infection (p.i.). Faecal samples taken weekly until slaughter and ingesta samples from different intestinal compartments were subjected to next-generation sequencing of the bacterial 16S rRNA gene.ResultsThe results revealed marked differences between the single- and the trickle-infected group. Single infection caused a remarkable but transient decrease in microbial diversity in the caecum, which was not observed in the trickle-infected group. However, an increase in short-chain fatty acid-producing genera in the caecum on day 21 p.i., which shifted to a decrease on day 35 p.i., was common to both groups, possibly related to changes in excretory–secretory products following the parasite’s final moult. Faecal microbial interaction networks were more similar between the single-infected and control group than the trickle-infected group. In addition, a lower degree of similarity over time indicated that A. suum trickle infection prevented microbiota stabilization.ConclusionsThese different patterns may have important implications regarding the comparability of experimental infections with natural scenarios characterized by continuous exposure, and should be confirmed by further studies.Graphical

The Effects of Allium sativum L., Artemisia absinthium L., Cucurbita pepo L., Coriandrum sativum L., Satureja hortensis L. and Calendula officinalis L. on the Embryogenesis of Ascaris suum Eggs during an In Vitro Experimental Study.

Ascaris suum is present in traditionally managed indoor pig herds and in industrialized farms, especially in older fatteners and sows. The increasing resistance to common antihelminthic drugs redirected research towards alternative and traditional therapies, which also include medicinal plants. This study comparatively evaluated the in vitro antiparasitic effects of Allium sativum L., Artemisia absinthium L., Cucurbita pepo L., Coriandrum sativum L., Satureja hortensis L. and Calendula officinalis L. against A. suum egg hatching and larval development. A. suum eggs were sampled from randomized fecal specimens collected from traditionally raised swine. The egg suspension (ES, 12 × 103/mL) was divided into two controls (C) (1C—1 mL ES + 1 mL distilled water, 2C—five plates of 1 mL ES + 1 mL ethanol of 70%, 35%, 17.5%, 8.75%, and 4.375%, respectively) and six experimental groups, and placed in 3 mL cell plates. The experimental groups (EG, 1–6) included ES + each alcoholic plant extract (10%, 5%, 2.5%, 1.25%, 0.625%). Both C and EG were performed in quintuplicate. All variants were incubated at 27 °C for a total of 21 days. A. suum eggs were examined after 2, 14 (L1), and 21 (L2/L3) days of incubation. The efficacy of all tested plant extracts increased with concentration. Anti-embryogenic effects on A. suum eggs were expressed by all plants. A superior influence was observed in A. sativum L., A. absinthium L., C. pepo L. and S. hortensis L. extracts, at all concentrations tested. A. sativum L. and A. absinthium L. extracts showed the strongest antihelminthic activity, while C. sativum L. and C. officinalis L. were the weakest ascaricids. Future in-depth phytochemical studies are required to identify the compounds responsible for the anthelminthic properties of these plant species.

Distribution of Gastrointestinal Parasitic Infection in Domestic Pigs in the Republic of Korea: Nationwide Survey from 2020-2021.

This study aimed to examine the distribution of gastrointestinal parasitic infections in domestic pigs in the Republic of Korea. From May 2020 to October 2021, 364 pig fecal samples were collected from 75 farms in 7 Provinces and microscopically examined. A total of 170 (46.7%) pigs were infected with at least one of the following parasites: Balantioides coli, strongyles, Ascaris suum, Trichuris suis, and coccidia. By parasite species, B. coli, strongyles, A. suum, T. suis, and coccidia oocysts or eggs were detected in 144 (39.6%), 24 (6.6%), 14 (3.8%), 4 (1.1%), and 1 (0.3%) samples, respectively. One hundred fifty-four, 15, and 1 cases showed single, double, and triple infections, respectively. Of the swine fecal samples from 75 farms, 69 specimens (92.0%) were infected with 1 or more parasites. All surveyed farms across the country exhibited a positive rate of over 30%, among which the highest positive rate was 65.0% in Chungcheongnam-do, and Jeollabuk-do was followed by 61.9%. Winter showed a statistically lower prevalence than other seasons. This study showed that gastrointestinal parasites are prevalent in pigs in Korea, although the diversity of parasites is low.

Occurrence of Intestinal Parasites and Its Impact on Growth Performance and Carcass Traits of Pigs Raised Under Near-Organic Conditions.

Parasite infection is a common problem in organic pig production, which can compromise health and growth of pigs, threaten food safety of pork products, and cause economic losses to organic farmers. To develop management strategies for controlling parasites, we evaluated intestinal parasite infection in pigs at different ages and of different sexes, and investigated whether parasite infection influences growth performance and carcass traits in a cross-sectional study. Fecal samples were collected from pigs (n = 298) raised under near-organic standards during nursery, growing, finishing, and gestating phases for analysis of fecal egg counts (FEC) of Ascaris suum, Trichuris suis, and Oesophagostomum spp. Ascaris suum eggs were not detected in the feces of nursery pigs. Eggs of Ascaris suum were found in 45%, 74%, and 0% of fecal samples of growing pigs, finishing pigs, and gestating sows, respectively, after false-positive adjustment (P < 0.001). Mean FEC of Ascaris suum was higher in infected finishing pigs than in infected growing pigs [2,502 vs. 724 eggs per gram (epg), P < 0.001]. No differences in percent of Ascaris suum positive samples or FEC of Ascaris suum were detected between sexes. Growth performance and carcass traits were not different between non-infected pigs and those infected with Ascaris suum. All pigs (n = 32) examined at slaughter had white spots on the liver, and 78% harbored Ascaris suum worms. Trichuris suis eggs were not detected in any fecal samples. Eggs of Oesophagostomum spp. were found in 7%, 0%, 1%, and 9% of fecal samples of nursery pigs, growing pigs, finishing pigs, and gestating sows, respectively, with a maximum FEC of 40 epg in all age groups. These results indicate Ascaris suum was the predominant parasite infecting growing and finishing pigs in the herds studied. To control A. suum infection, future research should investigate the efficacy of treating pigs with organically approved anthelmintics during the growing phase of production.

Comparison of different detection methods for Ascaris suum infection on Austrian swine farms

BackgroundAscaris suum, the large roundworm of pigs, is one of the economically most important pig parasites worldwide. In Austria it is commonly diagnosed by monitoring livers for milk spots at the slaughterhouse and intravital diagnosis (flotation for detection of fecal egg shedding). Recently, serological diagnosis based on the detection of specific antibodies with an ELISA (SERASCA®) with high sensitivity has been developed. To introduce and evaluate serology for A. suum screening in Austrian pigs, blood (for serology) (n = 177) and feces (for copromicroscopy) (n = 177) were taken from randomly selected slaughter pig batches from 18 farms at a slaughterhouse in Lower Austria. In addition, livers presented at slaughter (n = 844; max. 70/farm) were evaluated for milk spots.ResultsOverall, 19% of the livers were milk spot-positive (22% of those with complete diagnostic evaluations). Thirteen percent of the fecal samples contained A. suum eggs, while 69% of the blood samples were serologically positive. Despite we did not determine the sensitivity of the ELISA specifically, results ouf our study confirmed the high sensitivity of the ELISA, which was claimed by the manufacturer prior to our work (sensitivity: liver assessment: 23.5–27.0%; copromicroscopy: 8.5–9.0%; ELISA: 99.5%), and a high percentage of A. suum infections that remained undetected by standard liver assessment.ConclusionsThis suggests that the current method of roundworm diagnostics is insufficient and antibody detection at the end of the fattening period should be established as the standard procedure.

Changes in porcine nutrient transport physiology in response to Ascaris suum infection

BackgroundThe roundworm Ascaris suum is one of the parasites with the greatest economic impact on pig farming. In this context, lower weight gain is hypothesized to be due to decreased nutrient absorption. This study aims at characterizing the effects of A. suum infection on intestinal nutrient transport processes and potential molecular mechanisms.MethodsThree groups of six piglets each were infected orally (10,000 embryonated A. suum eggs) in a single dose (“single infection”). Another three groups were infected orally (1000 embryonated eggs) for 10 consecutive days (“trickle infection”). Animals were necropsied 21, 35 and 49 days post-infection (dpi). Three groups served as respective controls. The Ussing chamber technique was applied for the functional characterization of small intestinal tissues [short-circuit currents (Isc) as induced by glucose, alanine and peptides; 3H-glucose net flux rates; tissue conductance (Gt)]. Transcription and expression levels of relevant cytokines and nutrient transporters were evaluated (qPCR/western blot).ResultsPeptide- and alanine-induced changes in Isc were significantly decreased in the jejunum and ileum of the trickle-infected group at 49 dpi and in the ileum of the single-infected group at 49 dpi. No significant differences regarding glucose transport were observed between the Ascaris-infected groups and the control group in Ussing chamber experiments. Transcription levels of the glucose and peptide transporters as well as of selected transcription factors (transcription of signal transducer and activator of transcription 6 [STAT6] and hypoxia-inducible factor 1-alpha [Hif-1α]) were significantly increased in response to both infection types after some periods. The transcription of interleukins 4 and 13 varied between decrease and increase regarding the respective time points, as did the protein expression of glucose transporters. The expression of the peptide transporter PepT1 was significantly decreased in the ileal single-infected group at 35 dpi. Hif-1α was significantly increased in the ileal tissue from the single-infected group at 21 dpi and in the trickle-infected group at 35 dpi. The expression levels of Na+/K+-ATPase and ASCT1 remained unaffected.ConclusionsIn contrast to the current hypothesis, these results indicate that the nutrient deprivation induced by A. suum cannot be explained by transcriptional or expression changes alone and requires further studies.Graphical abstract