Interactions between macrophages and helminths.

Macrophages, the major population of tissue-resident mononuclear phagocytes, contribute significantly to the immune response during helminth infection. Alternatively activated macrophages (AAM) are induced early in the anti-helminth response following tissue insult and parasite recognition, amplifying the early type 2 immune cascade initiated by epithelial cells and ILC2s, and subsequently driving parasite expulsion. AAM also contribute to functional alterations in tissues infiltrated with helminth larvae, mediating both tissue repair and inflammation. Their activation is amplified and occurs more rapidly following reinfection, where they can play a dual role in trapping tissue migratory larvae and preventing or resolving the associated inflammation and damage. In this review, we will address both the known and emerging roles of tissue macrophages during helminth infection, in addition to considering both outstanding research questions and new therapeutic strategies.

Soil-transmitted helminth infections associated with wastewater and sludge reuse: a review of current evidence.

OBJECTIVE: To review current evidence on infections related to the concentration of soil-transmitted helminth (STH) eggs in wastewater, sludge and vegetables irrigated with wastewater or grown on sludge-amended soils. METHOD: Search of Web of Science, Science Direct, PubMed and Google Scholar databases for publications reporting on STH egg concentration in wastewater, sludge and vegetables and for epidemiological studies on wastewater/sludge reuse and STH infections. RESULTS: STH egg concentrations were variable but high in wastewater and sludge especially in developing countries. They ranged from 6 to 16 000 eggs/L in wastewater and from 0 to 23 000 eggs/g in sludge and far exceed limits set in the WHO guideline for wastewater/sludge reuse. Numbers of STH eggs on vegetables ranged from 0 to 100 eggs/g. The concentration of STH eggs in wastewater, sludge and vegetables therefore relates to risks of infection through different exposure routes. CONCLUSION: Epidemiological evidence reveals an increased prevalence of STH infections associated with direct exposure to wastewater or sludge (farmers) and consumption of vegetables grown on soil treated with it. This calls for increased efforts to reduce the adverse health impact of wastewater and sludge reuse in line with the WHO multi-barrier approach.

Characterisation of a new, highly effective method for detecting nematode eggs (Ascaris spp., Toxocara spp., Trichuris spp.) in sewage sludge containing flocculants.

Because traditional methods used for sewage sludge parasitological examinations have low sensitivity, a new, highly effective method (own method - OM) was devised. The principle of this method is to eliminate the flocculent effect on the structure of sewage sludge by mechanically damaging floccules in the presence of surfactants and to increase the effectiveness of egg isolation processes in large volumes of liquids. The objective of this study was to estimate the effectiveness of the OM in detecting nematode eggs in sewage sludge samples containing flocculants. In the first stage, the effectiveness of the OM was compared to 4 other methods routinely used in parasitological examinations of dehydrated sewage sludge. Next, method standardisation was performed using sewage sludge samples supplemented with eggs from 3 parasite species (Ascaris suum, Toxocara canis and Trichuris vulpis). The study demonstrated that OM efficiency was 6-65 times greater than other methods, depending on the method and type of detected eggs. Limit of detection (LOD) calculations for the OM were performed on samples supplemented with a known number of parasite eggs resulting in 10, 5 and 3 eggs/50 g of sample for A. suum, T. vulpis and T. canis eggs, respectively. The limits of quantification (LOQ) of the OM were established as 200 eggs/50 g of sample for A. suum and T. vulpis eggs and 50 eggs/50 g of sample for T. canis eggs. The rectilinear regression functions, which determined the relationship between the number of eggs detected in OM measurements and the number of eggs contained in the samples, were characterised by high and statistically significant coefficients of determination (r2). The slopes of the trend lines were 0.3188, 0.3821 and 0.3276, and the intercepts were -11.223, -9.0261 and -23.15 for A. suum, T. canis and T. vulpis eggs, respectively. Method sensitivity, calculated as the slope coefficient of the regression function and expressed as a percentage, ranged from 32% to 38% depending on egg type. The study confirmed that the OM may be applied to quantify parasite eggs in dehydrated sewage sludge containing polyelectrolytes.

Ascaris spp. and Capillaria caudinflata infections in captive-bred crested ibis (Nipponia nippon) in China.

Crested ibis (Nipponia nippon), an endan gered native bird, was called the "precious stone" of oriental birds. N. nippon was considered a critically endangered species in the IUCN Red List of Threatened Species and a first-class national protected animal in China. The Chinese government had exerted considerable effort to protect the N. nippon population. An effective approach to increase the number of these birds was captive breeding. However, several pathogens, including parasites, could jeopardize the health of this species. The present study used the fecal flotation method to determine prevalence of intestinal parasites in fresh stool samples by wet mount smearing and iodine staining. Samples were obtained from 63 randomly selected crested ibis bred in Shaanxi Rare Wildlife Rescuing and Breeding Research Center in Zhouzhi County, Xi'an City, Shaanxi Province, China. In the 63 captive individuals, 38 were found positive for intestinal parasites (60.3%, 38/63). Of positive birds, high prevalence of Ascaris spp. (84.2%, 32/38) and Capillaria caudinflata (50.0%, 19/38) were detected. Coccidea (7.8%, 3/38), Fasciolidae (23.7%, 9/38), Blastocystis spp. (15.8%, 6/38), and Entamoeba histolytica (7.8%, 3/38) showed relatively low prevalence rates. This study focuses on the morphological identification of Ascaris spp. and C. caudinflata and their transmission in the N. nippon population. We introduce strategies to improve the breeding management of the birds, enhance their health, and stimulate population productivity.

Parasitological contamination of arable soil in selected regions of Poland - preliminary study.

INTRODUCTION AND OBJECTIVE: The hygienic status of arable soils in most developed countries has been unknown. In the presented study, a preliminary investigation was undertaken to determine the contamination with eggs of parasitic nematodes in the soil of arable fields in Poland. The aim of the study was to determine whether such contamination is common enough to constitute a significant problem and what factors may influence it. MATERIAL AND METHODS: The study was conducted in 5 Polish provinces from autumn 2021 to spring 2022. The provinces differed significantly in terms of the area of agricultural land, agricultural suitability, type of soil, scale of cattle and pig breeding, production of manure and slurry, and the use of manures and organic fertilizers for fertilization. A total of 133 soil samples were collected. Parasitological examination of soil samples was carried out using the PN-Z-19006 method [1], with confirmed high sensitivity. RESULTS: Parasite eggs were found in a total of 67 samples, of which 56 samples contained eggs of roundworms of the genus Ascaris (an average of 3.29 eggs/100 g of soil), 23 contained eggs of whipworms (an average of 1.22 eggs/100 g), and 3 contained eggs of Toxocara (1 egg/100 g). CONCLUSIONS: Differences in the percentage of positive samples were found depending on the period in which the samples were taken. The percentage of positive samples collected in autumn (53.57%) was higher than the percentage of positive samples collected in spring (48.05%). Similarly, the average number of eggs of in positive samples collected in autumn (3.43 eggs/100 g) was higher than the average number of eggs in samples collected in spring (2.90 eggs/100 g). Differences in the percentage of positive samples were also found depending on the region of origin of the samples.

Economic losses associated with respiratory and helminth infections in domestic pigs in Lira district, Northern Uganda.

This study sought to quantify direct economic losses due to respiratory and gastrointestinal (GI) helminth infections in domestic pigs in Uganda. In a longitudinal study design with repeated measures, farm visits were made at 2 month intervals from October 2018 to September 2019. Weaner and grower pigs (n = 288) aged 2-6 months from 94 farms were sampled. The pigs were monitored for growth and screened for exposure to four important respiratory pathogens: porcine circovirus type 2 (PCV2), porcine reproductive and respiratory syndrome virus (PRRSv), Mycoplasma hyopneumoniae (M. hyo), Actinobacillus pleuropneumoniae (App) using ELISA tests. Farm management practices were recorded and used to generate management level scores. Treatment expenses incurred were recorded throughout the study. A mixed effects model was fitted to quantify effects of respiratory and helminth infections on average daily weight gains (ADGs), with farm and pig as random effects. Analysis of variance (ANOVA) was used to determine differences in mean treatment costs by farm management standard. Financial losses were estimated from average carcass dressing percentage, ADG reductions during fattening (200 days). Results showed a grower pig in a given farm exposed to PRRSv and Ascaris spp. had significantly lower ADG by 17.10 gr/day and 16.80 grams/day respectively, compared to a similar unexposed pig (p < 0.05). Mean treatment costs per pig declined significantly with increase in management standard scores (MSS), from USD 1.13 per pig in MSS 1 (poor management) farms to USD 0.95 for MSS 3 (better management) farms (p < 0.05). We show that monetary losses due to PRRSv and Ascaris spp. infection amounted to USD 6.6 ± 2.7 and 6.50 ± 3.2 (Mean ± SEM) per pig, respectively during 200 days of fattening. This study strengthens evidence that improving management practices to reduce infections mitigates economic losses. To guide interventions, further studies are required to unravel the full extent of indirect economic losses.

Parasitic Contamination of Fresh Leafy Green Vegetables Sold in Northern Lebanon.

Contaminated, raw or undercooked vegetables can transmit parasitic infections. Here, we investigated parasitic contamination of leafy green vegetables sold in local markets in the Tripoli district, Lebanon, during two consecutive autumn seasons (2020-2021). The study involved the microscopic examination of 300 samples of five different types of vegetables (60 samples per type) and used standardized qualitative parasitological techniques for some protozoa and helminths. The results showed that 16.7% (95% interval for p: 12.6%, 21.4%) (50/300) of the vegetable samples were contaminated with at least one parasite. The most frequently detected parasite was Blastocystis spp. (8.7%; 26/300); this was followed in frequency by Ascaris spp. (3.7%; 11/300). Among the different vegetable types, lettuce (23.3%; 14/60) was the most contaminated, while arugula was the least contaminated (11.7%; 7/60). The statistical analysis did not reveal any significant association between the prevalence of parasitic contamination and the investigated risk factors, which included collection date, vegetable type, market storage status, and wetness of vegetables at the time of purchase (p > 0.05). The high prevalence of parasitic contamination also suggested the potential presence of other microbial pathogens. These findings are important because leafy green vegetables are preferentially and heavily consumed raw in Lebanon. Thus, implementing effective measures that target the farm-to-fork continuum is recommended in order to reduce the spread of intestinal pathogens.

CD4+ T Cell Epitope Identification from Complex Parasite Antigen Mixtures.

Antigen complexity represents a major challenge for scoring CD4+ T cell immunogenicity, a key hallmark of immunity and with great potential to improve vaccine development. In this chapter, we provide a comprehensive picture of a pipeline that can be applied to virtually any complex antigen to overcome different limitations. Antigens are characterized by Mass Spectrometry to determine the available protein sources and their abundances. A reconstituted in vitro antigen processing system is applied along with bioinformatics tools to prioritize the list of candidates. Finally, the immunogenicity of candidate peptides is validated ex vivo using PBMCs from HLA-typed individuals. This protocol compiles the essential information for executing the whole pipeline while focusing on the candidate epitope prioritizing scheme.

Evaluation of the Allplex™ GI-Helminth(I) Assay, the first marketed multiplex PCR for helminth diagnosis.

Molecular biology has been gaining more importance in parasitology. Recently, a commercial multiplex PCR assay detecting helminths was marketed: the Allplex™ GI-Helminth(I) Assay. It targets Ancylostoma spp., Ascaris spp., Enterobius vermicularis, Hymenolepis spp., Necator americanus, Strongyloides spp., Taenia spp. and Trichuris trichiura, but also the two most common microsporidia genera in human health, i.e. Enterocytozoon spp. and Encephalitozoon spp. This study aimed to evaluate and compare the Allplex™ GI-Helminth(I) Assay to classical diagnostic methods, based on a cohort of 110 stool samples positive for helminths (microscopy) or for microsporidia (PCR). Samples were stored at -80 °C until analysis by the Allplex™ GI-Helminth(I) Assay. False-negatives were re-tested with bead-beating pretreatment. Without mechanical lysis, concordance and agreement between microscopy and Allplex™ GI-Helminth(I) Assay ranged from 91% to 100% and from 0.15 to 1.00, respectively depending on the target. Concordance was perfect for Taenia spp. (n = 5) and microsporidia (n = 10). False-negative results were observed in 54% (6/13), 34% (4/11) and 20% (7/35) of cases, for hookworms, E. vermicularis and Strongyloides spp. detection, respectively. For these targets, pretreatment improved the results, but only slightly. Trichuris trichiura detection was critically low without pretreatment, as only 9% (1/11) of the samples were positive, but detection reached 91% (10/11) with bead-beating pretreatment. Mechanical lysis was also needed for Ascaris spp. and Hymenolepis spp. to reduce false-negative results from 1/8 to 1/21, respectively, to none for both. Overall, with an optimized extraction process, the Allplex™ GI-Helminth(I) Assay allows the detection of numerous parasites with roughly equivalent performance to that of microscopy, except for hookworms.

TITLE: Évaluation du kit Allplex™ GI-Helminth(I) Assay, la première PCR multiplex commercialisée pour le diagnostic des helminthes. ABSTRACT: La biologie moléculaire a maintenant une place importante en parasitologie. Le kit Allplex™ GI-Helminth(I) Assay est le premier panel multiplex commercialisé détectant des helminthes : Ancylostoma spp., Ascaris spp., Enterobius vermicularis, Hymenolepis spp., Necator americanus, Strongyloides spp., Taenia spp. et Trichuris trichiura, mais également les deux genres de Microsporidies les plus fréquents en santé humaine, Enterocytozoon spp. et Encephalitozoon spp. Cette étude a comparé la PCR Allplex™ GI-Helminth(I) Assay aux techniques diagnostiques usuelles, sur une banque préservée à −80 °C, comprenant 110 échantillons de selles positifs à helminthes (microscopie) ou à microsporidies (PCR). Les faux négatifs ont été retestés après prétraitement par broyage en billes. Sans lyse mécanique, la concordance et l'accord entre la microscopie et le test Allplex™ GI-Helminth(I) Assay variaient respectivement de 91 % à 100 % et de 0,15 à 1,00, selon la cible. La concordance était parfaite pour Taenia spp. (n = 5) et les microsporidies (n = 10). Des faux négatifs ont été observés pour la détection des ankylostomes, E. vermicularis et Strongyloides spp. à des taux respectifs de 54 % (6/13), 34 % (4/11) et 20 % (7/35). Pour ces cibles, le prétraitement a peu amélioré les résultats. La détection de T. trichiura était défectueuse sans prétraitement, avec 9 % (1/11) de positifs, mais a atteint 91 % (10/11) après prétraitement par broyage en billes. La lyse mécanique était également nécessaire pour Ascaris spp. et Hymenolepis spp. pour réduire les faux négatifs de 1/8 et 1/21, respectivement, à aucun pour les deux. Au total, avec une optimisation de l'étape d'extraction, le test Allplex™ GI-Helminth(I) Assay permet la détection de nombreux parasites avec des performances proches de celles de la microscopie, excepté pour les ankylostomes.

Prevalence and distribution of soil-transmitted helminth infection in free-roaming dogs in Bali Province, Indonesia.

BACKGROUND AND AIM: Several free-roaming dogs can be easily found in the public areas of Bali. They go out in search of foods and friends and defecate everywhere. In general, these groups of dogs do not receive good healthcare from their owners and are generally threatened by some disease-causing organisms, especially helminths. This study was conducted to identify and measure the prevalence of soil-transmitted helminths (STHs) that cause infection in free-roaming dogs in Bali Province, Indonesia. MATERIALS AND METHODS: A total of 1611 fresh dog fecal samples were collected from all areas of Bali Province and subjected to qualitative fecal examination using flotation techniques to obtain STH eggs. RESULTS: The incidence of STHs in free-roaming dogs was 38.36%. However, three types of STHs were identified, including Ancylostoma spp., Ascaris spp., and Trichuris spp. Ancylostoma spp. had the highest prevalence of 37.8%, followed by Ascaris spp. and Trichuris spp. at 6.02% and 0.87%, respectively. Multiple infections of these worms were also recorded. The polyparasitism prevalence of Ancylostoma spp. and Ascaris spp. was 3.85%, followed by that of Ancylostoma spp. and Trichuris spp. at 0.5% and that of Ascaris spp. and Trichuris spp. at 0.06%. There were no altitude-wise differences in the prevalence of STH infection. CONCLUSION: The prevalence of STHs was high in free-roaming dogs. This finding necessitates more serious attention as it affects both animal and public health.

Epidemiology of infections with zoonotic pig parasites in Enugu State, Nigeria.

Pig production is an important component of animal agriculture in Nigeria but zoonotic parasites constrain the production and make pork unsafe for human consumption. This epidemiological study was therefore conducted, to ascertain the prevalence and determinants of Taenia solium cysticerci and Ascaris spp infections in pigs in Enugu State. Presence of cysticerci was determined by post-mortem examination of pig carcasses while pig faeces were coprologically examined for presence of Ascaris spp egg, according to standard parasitological protocol. Structured questionnaire was used to elicit information on pig farmers' involvement in practices predisposing to parasitic infections and deworming progammes in 107 piggeries surveyed. Cysticerci were detected in 3.51% (12/342) of pig carcasses inspected. Overall prevalence of Ascaris spp. infections were 38.3% (41/107) and 14.3% (63/441) at farm and individual pig levels respectively. Major determinants of the infection and the percentage of farmers involved were: eating on duty (66.4%), open defecation (35.5%) and semi-extensive husbandry system (62.3%). Others are non-availability of deworming programme (54.4%), rearing pigs of different ages together (41.9%), non-disinfection of feeding or water troughs (41.2%) and early weaning (54.3%). Cognizant of the zoonotic potentials of Ascaris spp and Taenia solium cysticerci, the prevalence found for the parasites are significant from public health and food safety points of view. This underscores the need for cost-effect control measures against the parasites, using a coordinated One Health approach; in order to boost pig production, limit spread of the zoonoses and hence, the public health consequences thereof.

Parasitological contamination with eggs Ascaris spp., Trichuris spp. and Toxocara spp. of dehydrated municipal sewage sludge in Poland.

The objective of the present study was to evaluate the contamination of sewage sludge produced by municipal waste treatment plants in Poland by viable eggs of intestinal parasites of the genera Ascaris, Toxocara and Trichuris (ATT). Ninety-two municipal, mechanical-biological sewage treatment plants located within Poland were selected. These plants belonged to types of agglomerations: group 0 (large), group 1 (medium), group 2 (smaller) and group 3 (small). Samples were collected at the final stage of sewage treatment after the addition of flocculent to sludge, followed by dehydration. The samples were examined by a method adjusted to examine sewage sludge dehydrated using polyelectrolytes. The viability of the isolated eggs was evaluated based on incubation in a moist chamber. Live eggs of intestinal nematodes were found in 99% of samples. Most samples were contaminated by the eggs of Ascaris spp. (95%) and Toxocara spp. (96%). However, Trichuris spp. eggs were detected in 60% of samples. The mean number of eggs in 1 kg of dry mass (eggs/kg d.m.) was 5600 for Ascaris, 3700 for Toxocara and 1100 for Trichuris. The highest number of ATT eggs was detected in samples from sewage treatment plants located in south-eastern and central Poland. The highest number of ATT eggs was found in sewage sludge produced in large sewage treatment plants (agglomeration Groups 0 and 1), with mean values of 15,000 and 8900 eggs/kg d.m. The present study is the first parasitological investigation conducted on a large number of samples (92 samples) taken from various types of municipal sewage treatment plants located throughout Poland (16 regions) after the common introduction of polyelectrolytes during sewage sludge dehydration. The results of this study indicate that sludge produced in municipal sewage treatment plants is highly contaminated with parasite eggs.