Parasite Eggs Research Articles

A lightweight deep-learning model for parasite egg detection in microscopy images.

Intestinal parasitic infections are still a serious public health problem in developing countries, and the diagnosis of parasitic infections requires the first step of parasite/egg detection of samples. Automated detection can eliminate the dependence on professionals, but the current detection algorithms require large computational resources, which increases the lower limit of automated detection. Therefore, we have designed a lightweight deep-learning model, YAC-Net, to achieve rapid and accurate detection of parasitic eggs and reduce the cost of automation. This paper uses the ICIP 2022 Challenge dataset for experiments, and the experiments are conducted using fivefold cross-validation. The YOLOv5n model is used as the baseline model, and then two improvements are made to the baseline model based on the specificity of the egg data. First, the neck of the YOLOv5n is modified to from a feature pyramid network (FPN) to an asymptotic feature pyramid network (AFPN) structure. Different from the FPN structure, which mainly integrates semantic feature information at adjacent levels, the hierarchical and asymptotic aggregation structure of AFPN can fully fuse the spatial contextual information of egg images, and its adaptive spatial feature fusion mode can help the model select beneficial feature and ignore redundant information, thereby reducing computational complexity and improving detection performance. Second, the C3 module of the backbone of the YOLOv5n is modified to a C2f module, which can enrich gradient information, improving the feature extraction capability of the backbone. Moreover, ablation studies are designed by us to verify the effectiveness of the AFPN and C2f modules in the process of model lightweighting. The experimental results show that compared with YOLOv5n, YAC-Net improves precision by 1.1%, recall by 2.8%, the F1 score by 0.0195, and mAP_0.5 by 0.0271 and reduces the parameters by one-fifth. Compared with some state-of-the-art detection methods, YAC-Net achieves the best performance in precision, F1 score, mAP_0.5, and parameters. The precision, recall, F1 score, mAP_0.5, and parameters of our method on the test set are 97.8%, 97.7%, 0.9773, 0.9913, and 1,924,302, respectively. Compared with the baseline model, YAC-Net optimizes the model structure and simplifies the parameters while ensuring the detection performance. It helps to reduce the equipment requirements for performing automated detection and can be used to realize the automatic detection of parasite eggs under microscope images.

Influence of metalloproteinase-3 (-1171 5A>6A) polymorphism on periportal fibrosis in patients with schistosomiasis mansoni, Pernambuco, Brazil

In schistosomiasis mansoni (SM), periportal fibrosis (PPF) arises due to an inflammatory response exacerbated by parasite eggs in the intrahepatic portal space, culminating in the deposition of collagen and extracellular matrix proteins. This fibrosis results from a remodeling process of the extracellular matrix, in which metalloproteinases play a significant role. The study evaluated the association between MMP-3 polymorphism (-1171 5A>6A) (rs 3025058) and sociodemographic factors with PPF in individuals with SM. This is an analytical cross-sectional study involving 242 individuals infected with S. mansoni, of these 122 were diagnosed with hepatosplenic form (HS) and 119 hepatointestinal form (HI), all from the state of Pernambuco, Brazil. Polymerase chain reaction with restriction enzyme digestion (Psyl) was used to determine the MMP-3 polymorphism (-1171 5A>6A). There was a significant association between the male gender and the HS form (OR = 1.7623 95% CI [1.0481-2.9631]; p-value = 0.0439) as well as individuals aged over 41, also had a greater chance of developing this clinical form of the disease (OR = 2.8299; 95% CI [1.5211-5.2650]; p-value = 0.0014), with greater emphasis on individuals over 61 years old (OR= 8.5541; 95% CI [3.6895-19.8326], p-value= 0.0000). There was no statistically significant association between the MMP-3 polymorphism (-1171 5A>6A) between the clinical groups (5A6A CI [0.7144-1.9879] p-value 0.5882 5A5A CI [0.0912-2.9231] p-value 0.7331 5A6A / 5A5A CI [0.6904-1.8937] p-value 0.6949). In conclusion, the results showed no association between the MMP-3 polymorphism (-1171 5A>6A) and the development of PPF. In addition, males, and age over 41 were predictive factors for the HE form of the disease in this Brazilian population.

Breeding stages affect egg recognition in azure-winged magpies (Cyanopica cyanus)

Egg rejection often involves a cognitive process of recognizing foreign eggs, which can vary not only between species or among different individuals of the same species, but also within the same individual during different breeding stages, leading to markedly different responses to parasitic eggs. We conducted a comparative study in Wuhan, Hubei, and Fusong, Jilin, China, on the recognition and rejection behavior of azure-winged magpies (Cyanopica cyanus) at different breeding stages (pre-egg-laying, one-host-egg, multi-host-egg and early incubation stages). In the Fusong population, there was a significant difference in the rejection rate of model eggs by azure-winged magpies at different stages of the egg-laying period. During the one-host-egg stage, the rejection rate (63.6%) was significantly lower than that during the pre-egg-laying stage (85.7%) and the multi-host-egg stage (100%). The population of azure-winged magpies in Wuhan exhibited a 100% rejection rate towards model eggs during the pre-egg-laying stage. Furthermore, during the incubation stage, azure-winged magpies were able to accurately recognize and reject foreign eggs even when those were in majority. This indicates that azure-winged magpies employ a template-based recognition mechanism rather than relying on discordance mechanism for recognition after the onset of incubation. This study suggests that while azure-winged magpies can truly recognize their own eggs, different breeding stages still influence their rejection response towards parasitic eggs, especially during the pre-egg-laying and egg laying stages.

Comparison of different diagnostic protocols for the detection of Toxocara spp. in faecal samples of cats and dogs

BackgroundToxocara canis and Toxocara cati are parasitic nematodes that occur worldwide. As embryonated Toxocara spp. eggs in the environment pose a zoonotic risk, especially for children, optimal diagnostic approaches are necessary for effective disease response and management, including surveillance. However, little is known about the performance of different diagnostic protocols for detecting Toxocara spp. in the faeces of cats and dogs, hampering movement towards an optimal diagnostic process. This study aimed to compare detection methods, including a newly developed sequential sieving protocol (SF-SSV) and a high-throughput multiplex qPCR-based method to facilitate epidemiological studies.MethodsSpecies-specific Toxocara spp. egg suspensions and canine and feline faecal samples from the field were used to estimate analytical and diagnostic sensitivity of the protocols. The performance of two automated DNA extraction protocols using enzymatic and mechanical lysis were compared by multiplex qPCR, targeting both T. canis and T. cati-specific genomic sequences. All samples were examined by microscopy-based techniques, the sedimentation flotation technique (SF) and a newly developed SF-SSV for the detection, enrichment and purification of parasite eggs. The costs and processing times necessary for all protocols were estimated and compared for both single samples and sets of 100 samples.ResultsTo detect Toxocara spp. eggs, SF-SSV showed the highest analytical sensitivity and a significantly higher diagnostic sensitivity than the DNA detection methods. Mechanical lysis performed better than enzymatic lysis for automated DNA extraction. In automated DNA extraction, 96-well plates performed better than 24-well plates. DNA detection and microscopy-based parasitological methods showed substantial agreement between the results generated by each method. Microscopy-based techniques required the lowest costs and least hands-on time for a single sample. However, when costs and labour were estimated for a set of 100 samples, the DNA detection protocol using 96-well plates for extraction revealed costs similar to SF-SSV and the fastest processing times.ConclusionsSF-SSV was superior in terms of analytical and diagnostic sensitivity for the detection of Toxocara spp. eggs. For larger sets of samples, multiplex qPCR-based DNA detection represents an alternative to microscopy-based methods, based on the possibility of faster sample processing at similar costs to SF-SSV, and the ability to provide species-specific diagnoses.Graphical

Nest box-mounted PIT tag readers reveal cryptic recruits of cavity-nesting waterfowl in south Louisiana

Black-bellied Whistling-Ducks (Dendrocygna autumnalis; BBWD) are rapidly expanding northward into the core range of the eastern Wood Duck (Aix sponsa; WODU), yet little is known about BBWD nesting ecology. Typical field methods to study cavity-nesting waterfowl (i.e., weekly nest monitoring) preclude a full understanding of important breeding information, including nest prospecting and parasitic egg laying. To address this, we used subcutaneous passive integrated transponder (PIT) tags embedded in adults and PIT tag readers mounted on nest boxes with the objective to (1) identify individuals that used nest boxes but were not physically captured on a nest, (2) quantify box visitation, and (3) quantify BBWD pair and WODU hen behaviors during the prospecting, laying, and incubation periods. We deployed RFID readers on 40 nest boxes from March–December 2022 in Louisiana with the potential to detect BBWD and WODU marked with PIT tags in 2020–2022. We detected 48 (BBWD n = 26, WODU n = 22) adults of both species via RFID readers, and 33% (n = 16) of individuals (50% of BBWD, n = 12; 14% of WODU, n = 3) were never otherwise recaptured in 2022, meaning that traditional field methods for cavity-nesting waterfowl fail to document a substantial number of birds potentially contributing to the population via parasitism. We also used Bayesian generalized linear models to determine that both species visited a similar number of “new” (< 1 year old) and “old” (> 1 year old) nest boxes (β = 0.66, CI = -0.30, 1.64). However, BBWD preferentially visited (and subsequently nested in) old boxes at a significantly higher rate than WODU (β = 1.32, CI = 0.97, 1.66). Due to the generalist nature and rapid expansion of BBWD, an apparent aversion to newly installed boxes was unexpected, especially since there were several successful WODU nests in the new boxes before BBWD began nesting in 2022. Our study is one of the first to evaluate BBWD nesting behaviors within the core WODU breeding range, and the first to use nest box-mounted PIT tag readers to observe BBWD behavior.

Enhancing agricultural sustainability through optimization of the slaughterhouse sludge compost for elimination of parasites and coliforms

For a sustainable ecology, slaughterhouse sludge must be managed effectively in preview of the parasitic or coliforms’ spill over to the community. In order to determine the effectiveness of a customized biological decomposer solution in lowering the parasitic eggs and coliform bacteria, three composting units (Unit 1, Unit 2, and Unit 3) were treated with its different amounts. Over a period of 60 days, pH, temperature, humidity, number of the parasitic eggs per gram (EPG) of faecal material, viability of eggs, and coliform counts were evaluated. By the fifth day of the composting process, pH had significantly (P < 0.05) increased across all the treatments and then decreased gradually. Also on the 5th day, all three units entered the thermophilic range (> 45 °C), which persisted for 20 days for Unit 3 and 15 days for Units 1 and 2. Humidity levels initially increased significantly (P < 0.05) in all three units (Unit 3 = 71%, Unit 2 = 64%, and Unit 1 = 55%) but then gradually decreased. On day 5, no decrease in EPG in Unit 1 was detected; however, a non-significant (P > 0.05) 12.5% decline in EPG in Unit 2 and Unit 3 was recorded. After that, a significant (P < 0.05) reduction in EPG was observed in all the three treatments until day 25. By day 5, decreased egg viability was significantly (P < 0.05) recorded in Unit 3 (21.43%); in Unit 1 and Unit 2, the decrease was 6.25% and 14.29%, respectively. Additionally, all units showed a significant (P < 0.05) decrease in total coliforms, meeting minimum allowable limit in Unit 2 and 3 on day 10 and on day 15 in Unit 1. The most substantial reduction in faecal coliforms was observed in Unit 3 (from 2.6 log₁₀ to 1.3 log₁₀), followed by Unit 2 (from 2.6 log₁₀ to 1.5 log₁₀), and then Unit 1 (from 2.6 log₁₀ to 1.6 log₁₀). The results of this study support recommendation of advanced composting techniques to eradicate or reduce the abundance of pathogens (parasites and coliforms). Hence, we endorse the value of careful composting procedures in environment-friendly abattoir waste management and agricultural practices through creating pathogen-free, eco-friendly fertilizers to promote both agricultural and environmental sustainability.

Effect of Temperature and Salinity on Survival of Protoscoleces of Hydatid Cyst in Liver In Vitro.

Hydatid cyst is the metacestode stage of Echinococcus granulosus that occurs in herbivores and humans as intermediate hosts by consuming parasite eggs through forage and vegetables. Carnivores, as definitive hosts, become infected by consuming infected vesicles of herbivores. The most effective treatment for a hydatid cyst is surgical operation. Inactivating E. granulosus protoscoleces through heating, cooling, or chemicals such as sodium chloride can be considered an effective method for controlling hydatidosis in both humans and animals. The main objective of this study was to evaluate the effect of different temperatures and salinity conditions on the survival of Echinococcus granulosus protoscoleces. For this purpose, 50 g of infected liver (in triplicate) was separately treated with different temperatures (+10°C, +50°C, +60°C, +72°C, and -20°C) and concentrations of sodium chloride (5%, 10%, 15%, and 20%) for 3, 6, 12, 24, 48, and 72 h. Additionally, 50 g of infected liver was stored separately in the refrigerator (+4°C) as a control group. The survival rate of the protoscoleces was evaluated by staining with 1% eosin under a light microscope. The results showed that the protoscoleces were significantly affected, with 100% mortality at -20°C after 0.5 h, and complete death at +72°C, +60°C, +50°C, and +10°C after 1, 1.5, 3, and 24 h, respectively (p < 0.005). Similarly, the protoscoleces in the liver mass survived at 5% NaCl after 3 h but died at 10% after 24 h, at 15% after 12 h, and at 20% after 6 h. It is concluded that exposing the liver infected with protoscoleces hydatid cyst to a temperature of -20°C and a sodium chloride concentration of 10% for 24 h is suitable for inactivating the protoscoleces.

High voltage pulsed electric field: A novel method for killing parasitic eggs on the surface of raisins

Aiming to address the issue of stored grain pests easily breeding during the process of dried fruits in Xinjiang, this study proposes a method and a device for killing raisin parasitic eggs based on a high-voltage pulsed electric field. A one-way test and a Box-Behnken central combination test were conducted to investigate the effects of high-voltage pulsed electric field strength and frequency on the unhatched rate and larval survival rate of Plodia interpunctella eggs on raisin surfaces. The experimental results were qualitatively and quantitatively analyzed using biooptical microscope observation and incubation at constant temperature and humidity post-treatment. The findings revealed that with an output voltage of 22.8 kV, the delivery speed of 0.024 m/s, and the electric field frequency of 3.8 Hz, the unhatched rate of the eggs was 68.14 % while the survival rate of the larvae was 20.36 %. These results can provide new insights for both theoretical development and system implementation regarding the use of high voltage pulsed electric fields for eliminating raisin surface eggs, as well as providing valuable academic references for field crop diseases and pests control strategies.

Inverse Correlation of Th2-Specific Cytokines with Hepatic Egg Burden in S. mansoni-Infected Hamsters.

Schistosomiasis, a parasitic disease caused by Schistosoma spp., affects more than 250 million people worldwide. S. mansoni in particular affects the gastrointestinal tract and, through its eggs, induces a Th2 immune response leading to granuloma formation. The relationship between egg load and immune response is poorly understood. We investigated whether the quantity of parasitic eggs influences the immune response in S. mansoni-infected hamsters. The hepatic and intestinal egg load was assessed, and cytokine expression as well as the expression of three major egg-derived proteins were analyzed in monosex- and bisex-infected animals by qRT-PCR. Statistical correlations between egg load or egg-derived factors Ipse/alpha-1, kappa-5, and omega-1, and the immune response were analyzed in liver and colon tissue. Surprisingly, no correlation of the Th1 cytokines with the hepatic egg load was observed, while the Th2 cytokines Il4, Il5, and Il13 showed an inverse correlation in the liver but not in the colon. A longer embryogenesis of the parasitic eggs in the liver could explain this correlation. This conclusion is supported by the lack of any correlation with immune response in the colon, as the intestinal passage of the eggs is limited to a few days.

Prevalence of schistosomiasis among school children at Esuekyir community in the Central Region of Ghana

Schistosomiasis, an endemic neglected tropical disease in areas with poor sanitation, causes physical and mental defects in both children and adults. Various strategies, especially drug administration for morbidity control, have been implemented to combat the disease in Ghana and globally. Despite these efforts, schistosomiasis remains prevalent in Ghana, negatively impacting children's academic performance, growth, and overall quality of life. This study aimed to determine the prevalence of schistosomiasis in school children at Esuekyir, a peri-urban community in Ghana. A cross-sectional study using simple random sampling technique to select participants and collect stool and urine samples from 246 school children in Esuekyir was adopted. Microscopy of urine and stool samples was performed involving urine sedimentation and stool formol-ether sedimentation techniques to analyse for parasite eggs. Questionnaires were developed to help detect risk factors that expose these children to the disease. The prevalence of urogenital schistosomiasis in children at Esuekyir was 15.45% while that of intestinal schistosomiasis was 6.957.0%. There was one case of co-infection of urogenital and intestinal schistosomiasis from a 13 year old primary student. Children in primary school had higher risks of infection due to their activities around the water body. There was a significant association between class groups and urogenital schistosomiasis (p-value = 0.042). The presence of schistosomiasis in school children highlights the importance of targeted interventions and public health initiatives in addressing this specific disease condition especially in primary school children. Findings from the research revealed a higher prevalence of urogenital schistosomiasis in the study population as compared to intestinal schistosomiasis.

Deep humoral profiling coupled to interpretable machine learning unveils diagnostic markers and pathophysiology of schistosomiasis.

Schistosomiasis, a highly prevalent parasitic disease, affects more than 200 million people worldwide. Current diagnostics based on parasite egg detection in stool detect infection only at a late stage, and current antibody-based tests cannot distinguish past from current infection. Here, we developed and used a multiplexed antibody profiling platform to obtain a comprehensive repertoire of antihelminth humoral profiles including isotype, subclass, Fc receptor (FcR) binding, and glycosylation profiles of antigen-specific antibodies. Using Essential Regression (ER) and SLIDE, interpretable machine learning methods, we identified latent factors (context-specific groups) that move beyond biomarkers and provide insights into the pathophysiology of different stages of schistosome infection. By comparing profiles of infected and healthy individuals, we identified modules with unique humoral signatures of active disease, including hallmark signatures of parasitic infection such as elevated immunoglobulin G4 (IgG4). However, we also captured previously uncharacterized humoral responses including elevated FcR binding and specific antibody glycoforms in patients with active infection, helping distinguish them from those without active infection but with equivalent antibody titers. This signature was validated in an independent cohort. Our approach also uncovered two distinct endotypes, nonpatent infection and prior infection, in those who were not actively infected. Higher amounts of IgG1 and FcR1/FcR3A binding were also found to be likely protective of the transition from nonpatent to active infection. Overall, we unveiled markers for antibody-based diagnostics and latent factors underlying the pathogenesis of schistosome infection. Our results suggest that selective antigen targeting could be useful in early detection, thus controlling infection severity.

Archaeoparasitology and pathoecology of Buchta Nakhodka, a medieval settlement in the far north of Western Siberia, dating from the 13th century

Buchta Nakhodka, which was inhabited by the Northern Samoyeds, is one of the few sites in the far north of Western Siberia that has been explored archaeologically and archaeoparasitologically. Many factors complicate research work in this region; therefore, any additional study of known sites can provide valuable data. Here, we report the results of soil samples from the cultural layer taken during the excavation of structures of the medieval settlement of Buchta Nakhodka. Six types of human and animal parasite eggs were found in the studied samples. The extent of contamination and number of parasite species varied, depending on the sampling location. The analysis of research results in the historical and archaeological context enabled the reconstruction of certain nutritional habits of the population of Buchta Nakhodka in the 13th century and such activities as hunting, fishing, and dog breeding, as well as an evaluation of the level of hygiene and possible population movements and trade links with neighbouring settlements.

Detection of parasitic forms in feces of Chelonoidis carbonarius (Linnaeus, 1766) from Brazilian Cerrado.

Chelonoidis carbonarius is host to a great diversity of endoparasites. This research aimed to identify and taxonomically classify the parasitic forms of endoparasites found in the feces of C. carbonarius using coproparasitological methods. In total, 38 fecal samples were collected from animals captured in the municipalities of Cuiabá (n = 23) and Jataí (n = 15) in the Brazilian Ceraado. Fecal samples were submitted to at least one of the following techniques: Willis-Mollay flotation technique, zinc sulfate centrifugal flotation technique, and spontaneous sedimentation of Hoffman, and Pons, and Janer. Overall, 86.85% (33/38) of animals were positive in at least one of the diagnostic techniques. Of the 38 samples used, 50% of the animals were captive and 50% of the animals were free-living. Cysts, oocysts, and parasitic eggs were identified, as follows: Chapiniella spp. (65.78%; n = 24), Sauricola spp. (31.57%; n = 12), Ascaroidea (15.78%; n = 6), Oxyuridae (2.63%; n = 1), Trematoda (2.63%; n = 1), Acanthocephala (2.63%; n = 1), Balantidium spp. (39.47%; n = 15), Eimeria spp. (21.05%; n = 8), and Nyctotherus spp. (2.63%; n = 1). The study showed that both captive and free-living C. carbonarius are often parasitized by different endoparasite species, with nematode eggs being the most common.

MARKER REGIONS OF THE GENOME FOR MOLECULAR DIAGNOSTICS OF OPISTHORCHIIDAE FAMILY

Infectious diseases caused by representatives of the Opisthorhidae family rank 8th in terms of health concern in the world among all food parasites. According to FAO data, studies of representatives of the Opisthorhidae family rank first and range from 0.66 to 88.7%. When performing a PCR analysis, a molecular marker plays an important role. In our studies, ribosomal genome clusters and mitochondrial genes were used as molecular markers. In eukaryotic organisms, ribosomal genes are presented in the form of clusters located in groups on different chromosomes. Each cluster includes three ribosomal genes (18S, 5.8S and 28S), separated by spacer regions. The ribosomal gene cluster consists of a transcribed region, internal transcribed spacers (ITS - Internal Transcribed Spacer) and flanking external transcribed spacers (ETS - Eternal Transcribed Spacer) The ITS noncoding sequence differs greatly among closely related organisms and is therefore used to classify parasites at the genus, species, and subspecies level. The ribosomal nuclear gene 5.8S forms the complete transcribed internal spacer region together with the molecular markers ITS1 and ITS2. This region of the genome is used in species studies of 19 families of digenetic flukes. Mitochondrial genes are also one of the most commonly used mtDNA gene fragments in the molecular identification of the family Opisthorchiidae. The advantage of using COX in research is the high degree of mutation, which makes it possible to differentiate closely related species The use of these genome regions makes it possible to quickly and accurately identify pathogens of the family Opishorchiidae, even for such individuals with practically indistinguishable morphological stages of the life cycle, such as parasite eggs and capsules in fish muscles. Therefore, in our studies, primers were used for the nuclear regions of ribosomal DNA ITS1, ITS2 and mitochondrial genes COX1, COX3.

MARKER REGIONS OF THE GENOME FOR MOLECULAR DIAGNOSTICS OF OPISTHORCHIIDAE FAMILY

Infectious diseases caused by representatives of the Opisthorhidae family rank 8th in terms of health concern in the world among all food parasites. According to FAO data, studies of representatives of the Opisthorhidae family rank first and range from 0.66 to 88.7%. When performing a PCR analysis, a molecular marker plays an important role. In our studies, ribosomal genome clusters and mitochondrial genes were used as molecular markers. In eukaryotic organisms, ribosomal genes are presented in the form of clusters located in groups on different chromosomes. Each cluster includes three ribosomal genes (18S, 5.8S and 28S), separated by spacer regions. The ribosomal gene cluster consists of a transcribed region, internal transcribed spacers (ITS - Internal Transcribed Spacer) and flanking external transcribed spacers (ETS - Eternal Transcribed Spacer) The ITS noncoding sequence differs greatly among closely related organisms and is therefore used to classify parasites at the genus, species, and subspecies level. The ribosomal nuclear gene 5.8S forms the complete transcribed internal spacer region together with the molecular markers ITS1 and ITS2. This region of the genome is used in species studies of 19 families of digenetic flukes. Mitochondrial genes are also one of the most commonly used mtDNA gene fragments in the molecular identification of the family Opisthorchiidae. The advantage of using COX in research is the high degree of mutation, which makes it possible to differentiate closely related species The use of these genome regions makes it possible to quickly and accurately identify pathogens of the family Opishorchiidae, even for such individuals with practically indistinguishable morphological stages of the life cycle, such as parasite eggs and capsules in fish muscles. Therefore, in our studies, primers were used for the nuclear regions of ribosomal DNA ITS1, ITS2 and mitochondrial genes COX1, COX3.

Gastrointestinal nematodes in German outdoor-reared pigs based on faecal egg count and next-generation sequencing nemabiome data

BackgroundThere is a higher risk for nematode infections associated with outdoor-reared pigs. Next to Ascaris suum, Oesophagostomum dentatum and Trichuris suis, there is the potential of infections with other nodular worm species, Hyostrongylus rubidus, Stongyloides ransomi and Metastrongylus spp. lungworms. Next-generation sequencing methods describing the nemabiome have not yet been established for porcine nematodes.MethodsFLOTAC was used for faecal egg counts of porcine gastrointestinal nematodes and lungworms in piglets, fatteners and adults individually. A nemabiome analyses based on ITS-2 gene region metabarcoding was used to differentiate strongyle species. Additionally, questionnaire data was analysed using mixed-effect regression to identify potential risk factors associated with parasite occurrences and egg shedding intensity.ResultsOn 15 of 17 farms nematode eggs were detected. Ascaris suum, strongyles and T. suis were detected on 82%, 70% and 35% of the 17 farms, respectively. Lungworms were detected on one out of four farms with access to pasture. Strongyloides ransomi was not detected. 32% (CI 28–36%), 27% (24–31%), 5% (4–7%) and 3% (0.9–8%) of the samples where tested positive for strongyles, A. suum, T. suis and lungworms, respectively. The nemabiome analysis revealed three different strongyle species, with O. dentatum being the most common (mean 93.9%), followed by O. quadrispinulatum (5.9%) and the hookworm Globocephalus urosubulatus (0.1%). The bivariate and multivariate risk factor analyses showed among others that cleaning once a week compared to twice a week increased the odds significantly for being infected with A. suum (OR 78.60) and strongyles (2077.59). Access to pasture was associated with higher odds for A. suum (43.83) and strongyles (14.21). Compared to shallow litter systems, deep litter and free range systems resulted in significant higher odds for strongyles (85.74, 215.59, respectively) and T. suis (200.33, 623.08).ConclusionsInfections with A. suum, O. dentatum, O. quadrispinulatum, T. suis, Metastrongylus spp. and G. urosubulatus are present in German outdoor-reared pigs. This is the first report of G. urosubulatus in domestic pigs in Europe. Metabarcoding based on the ITS-2 region is a suitable tool to analyse the porcine nemabiome. Furthermore, management practices have the potential of reducing the risk of parasite infections.

Papaya Seed Extract Combined Lauryl Glucoside Against Human Parasites Contaminated in Fresh Vegetables

Background: The prevalence of parasites in Thailand, primarily transmitted through contaminated vegetables, poses a significant public health concern. Ensuring the safety of natural produce from both parasites and chemical residues necessitates robust sanitation measures. Objective: This study aims to develop effective vegetable-washing solutions using papaya seed extract with potent anti-parasitic properties contamination in fresh vegetables. Methods: Fresh vegetables contaminated with Strongyloides stercoralis larvae, Ascaris lumbricoides, and Taenia spp. eggs were subjected to various treatments: distilled water, tap water, vinegar (5%), papaya seeds extract (200 mg/L), lauryl glucoside (1%) from coconut oil, papaya seeds extract combined with lauryl glucoside, and Albendazole as a standard drug. Immersion for 5 minutes was followed by parasite observation using the sedimentation method over 60 minutes, examining inhibitory effects on parasite larvae movement. Results: Papaya seed extract achieved the most rapid cessation within 10 minutes for larvae movement, followed by vinegar and papaya seed extract combined with lauryl glucoside. Furthermore, papaya seed extract combined with lauryl glucoside exhibited the highest efficacy in the sedimentation larval and parasite eggs, followed by lauryl glucoside solution and papaya seed extract, surpassing the control group. Conclusion: Papaya seeds extract immobilized larvae, while lauryl glucoside effectively eliminated parasite eggs from vegetables. Natural and organic compounds derived from papaya and coconut oil exhibit inherent diversification in their properties. These compounds originate from natural sources, showcasing distinct characteristics attributed to their organic nature. These results indicate the potential for developing natural extract-based products as future vegetable-washing solutions, emphasizing their significance against parasitic efficacy.

Improvement of the Diagnostic Method for the Detection of Parasite Eggs Using the Formalin Ethyl Acetate Concentration Technique.

The formalin-ethyl acetate (FEA) concentration method is commonly used in routine clinical practice to detect parasite eggs in feces. This procedure involves extraction of oil with the organic solvent ethyl acetate (EA), which reduces fecal sediment and provides a cleaner background for microscopic analysis. However, clinically, some sediment failed to float after EA treatment. Hexane, commonly used in the food oil extraction from oilseeds did not float the feces. Gas chromatography-mass spectrometry (GC-MS) analysis showed that neither the amount of the oil nor the classes of the oil determined was differed whether hexane or EA was used to float the feces. Oil red, Bodipy and Calcofluor staining showed that the unabsorbed oil droplets in the fecal sediment were trapped within the leaf structure. HCl or acetic acid was added to see if the acid residue could dissolve the cellulose of the leaf to promote the bulk float. Our result showed that the fecal bulk contained the loosened mesophyll cell wall. The addition of acid residues improved fecal bulk float. The proximity of cellulose fiber to EA, but not hexane, may enhance the efficacy of oil extraction from cellulose. This is the first report that the interaction of cellulose with ethyl acetate in fecal solution has an effect on bulk float. This study improves the understanding of fecal bulk flotation and may assist in the visualization of parasite eggs in clinical practice with non-floating fecal samples in the FEA concentration method.

Crossroad between the sylvatic and domestic cycles of Mesocestoides litteratus: mesocestodiasis in dogs adopted from shelters in Türkiye.

This study aimed to report the presence of Mesocestoides litteratus in dogs adopted from shelters in Türkiye. Gravid segments were examined microscopically in the faeces of dogs from different shelters located in Ankara and Kirikkale provinces in the central region of Türkiye. Then, genomic DNA obtained from these segments, a 446-bp fragment of the mitochondrial cytochrome C oxidase subunit 1 gene, and a 350-bp fragment of mitochondrial 12S rRNA were amplified and sequenced. BLASTn search was performed. During light microscopic examination, an egg-filled paruterine organ was observed in the middle part of the segment. Thin-shelled, oval, 35-µm-diameter parasite eggs containing an oncosphere with three pairs of hooklets were observed. The gravid segments were determined as Mesocestoides spp. based on the appearance of the typical paruterine organ. PCR results supported our diagnosis; moreover, according to the BLAST results, it was detected that the species infecting two dogs was 98.01-100% similar to M. litteratus. Praziquantel-containing medication was administered to the infected dogs at a dosage of 5mg/kg. Foxes act as the final host of M. litteratus and the parasite is prevalent in wildlife; however, these animals may disperse the parasite in urban life. Veterinarians need to be made more aware of this parasite, especially if the dogs are owned from shelters.

Gut microbiota metabolically mediate intestinal helminth infection in zebrafish.

Intestinal helminth parasites (IHPs) impact human health globally and interfere with animal health and agricultural productivity. While anthelmintics are critical to controlling parasite infections, their efficacy is increasingly compromised by drug resistance. Recent investigations suggest the gut microbiome might mediate helminth infection dynamics. So, identifying how gut microbes interact with parasites could yield new therapeutic targets for infection prevention and management. We conducted a study using a zebrafish model of parasitic infection to identify routes by which gut microbes might impact helminth infection outcomes. Our research linked the gut microbiome to both parasite infection and to metabolites in the gut to understand how microbes could alter parasite infection. We identified a metabolite in the gut, salicylaldehyde, that is putatively produced by a gut microbe and that inhibits parasitic egg growth. Our results also point to a class of compounds, N-acyl-ethanolamines, which are affected by changes in the gut microbiome and are linked to parasite infection. Collectively, our results indicate the gut microbiome may be a source of novel anthelmintics that can be harnessed to control IHPs.