Necator Americanus Research Articles

Optimization of 18 S rRNA metabarcoding for the simultaneous diagnosis of intestinal parasites

Recent advancements in next-generation sequencing (NGS) technologies have created new opportunities for comprehensive screening of multiple parasite species. In this study, we cloned the 18 S rDNA V9 region of 11 species of intestinal parasites into plasmids. Equal amounts and concentrations of these 11 plasmids were pooled, and amplicon NGS targeting the 18 S rDNA V9 region was performed using the Illumina iSeq 100 platform. A total of 434,849 reads were identified, and all 11 parasite species were detected, although the number of output reads for each parasite varied. The read count ratio, in descending order, was as follows: Clonorchis sinensis, 17.2%; Entamoeba histolytica, 16.7%; Dibothriocephalus latus, 14.4%; Trichuris trichiura, 10.8%; Fasciola hepatica, 8.7%; Necator americanus, 8.5%; Paragonimus westermani, 8.5%; Taenia saginata, 7.1%; Giardia intestinalis, 5.0%; Ascaris lumbricoides, 1.7%; and Enterobius vermicularis, 0.9%. We found that the DNA secondary structures showed a negative association with the number of output reads. Additionally, variations in the amplicon PCR annealing temperature affected the relative abundance of output reads for each parasite. These findings can be applied to improve parasite detection methodologies and ultimately enhance efforts to control and prevent intestinal parasitic infections.

Low prevalence of soil transmitted helminth infection in Ugandan children hospitalized with severe malaria.

Co-infection by intestinal helminths and Plasmodium spp. may be common in endemic communities. Several studies have identified a relationship between helminth infection, Plasmodium spp. infection and malaria severity. However, the relationship is not well defined, and results are inconclusive. We analyzed 202 stool samples from a cohort of children with severe malaria enrolled in two hospitals in Uganda from 2014-2017 and asymptomatic community children from the same household or neighborhood and enrolled at the same time, all 6 months to 48 months of age. We investigated if intestinal helminth infection modified risk of severe malaria. We extracted nucleic acids from stool and tested them for six helminth species ( Anyclostoma duodenale, Ascaris lumbricoides, Necator americanus, Strongyloides stercolaris, Trichuris trichiura, Shistosoma mansoni ) using highly sensitive quantitative PCR. We found a low prevalence of infection by ≥1 intestinal helminth species in children with severe malaria (5.1%, n=9/177) and community control children (4.0%, n=1/25). Infection by ≥1 of the helminths assessed was not associated with severe malaria (aRR = 1.0, 95% Confidence Interval = 0.82, 1.3, p = 0.78). In 2003 Uganda instituted a national deworming program, with anti-helminth medication provided twice annually to children 6 months to 5 years of age. In these areas of Uganda, the national deworming campaign has been highly successful, as stool-based helminth infection was rare even when using highly sensitive methods of detection and was not a major contributor to risk of severe malaria.

Comparison of multi-parallel quantitative real-time PCRs targeting different DNA regions and detecting soil-transmitted helminths in stool

BackgroundSoil-transmitted helminths infect an estimated 18% of the world’s population, causing a significant health burden. Microscopy has been the primary tool for diagnosing eggs from fecal samples, but its sensitivity drops in low-prevalence settings. Quantitative real-time polymerase chain reaction (qPCR) is slowly increasing in research and clinical settings. However, there is still no consensus on preferred qPCR targets.MethodsWe aimed to compare soil-transmitted helminth (STH) DNA detection methods by testing naïve stool samples spiked with known quantities of STH eggs and larvae. DNA extracts from spiked samples were tested using independent quantitative realtime PCR (qPCR) assays targeting ribosomal or putative non-protein coding satellite sequences.ResultsFor Trichuris trichiura, there was a strong correlation between egg/larvae counts and qPCR results using either qPCR method (0.86 and 0.87, respectively). Strong correlations also existed for A. lumbricoides (0.60 and 0.63, respectively), but weaker correlations were found for Ancylostoma duodenale (0.41 for both assays) and Strongyloides stercoralis (0.48 and 0.65, respectively). No correlation for Necator americanus was observed when testing with either qPCR assay. Both assays had fair-to-moderate agreement across targets when using field-collected stool samples (0.28–0.45, for all STHs), except for S. stercoralis (0.12) with slight agreement.ConclusionsThere is a strong correlation between qPCR results and egg/larvae counts. Our study confirms that qPCR is an effective diagnostic tool, even with low-intensity infections, regardless of the DNA-based diagnostic marker used. However, the moderate agreement between the two different qPCR assays when testing field samples highlights the need to understand the role of these targets in the genome so that the parasite burden can be quantified more accurately and consistently by qPCR.Graphical abstract

Parasitic Contamination of Soil in the Southern United States.

Parasites are generally associated with lower income countries in tropical and subtropical areas. Still, they are also prevalent in low-income communities in the southern United States. Studies characterizing the epidemiology of parasites in the United States are limited, resulting in little comprehensive understanding of the problem. This study investigated the environmental contamination of parasites in the southern United States by determining each parasite's contamination rate and burden in five low-income communities. A total of 499 soil samples of approximately 50 g were collected from public parks and private residences in Alabama, Louisiana, Mississippi, South Carolina, and Texas. A technique using parasite floatation, filtration, and bead-beating was applied to dirt samples to concentrate and extract parasite DNA from samples and detected via multiparallel quantitative polymerase chain reaction (qPCR). qPCR detected total sample contamination of Blastocystis spp. (19.03%), Toxocara cati (6.01%), Toxocara canis (3.61%), Strongyloides stercoralis (2.00%), Trichuris trichiura (1.80%), Ancylostoma duodenale (1.42%), Giardia intestinalis (1.40%), Cryptosporidium spp. (1.01%), Entamoeba histolytica (0.20%), and Necator americanus (0.20%). The remaining samples had no parasitic contamination. Overall parasite contamination rates varied significantly between communities: western Mississippi (46.88%), southwestern Alabama (39.62%), northeastern Louisiana (27.93%), southwestern South Carolina (27.93%), and south Texas (6.93%) (P <0.0001). T. cati DNA burdens were more significant in communities with higher poverty rates, including northeastern Louisiana (50.57%) and western Mississippi (49.60%) compared with southwestern Alabama (30.05%) and southwestern South Carolina (25.01%) (P = 0.0011). This study demonstrates the environmental contamination of parasites and their relationship with high poverty rates in communities in the southern United States.

Genomic analysis of Strongyloides stercoralis and Strongyloides fuelleborni in Bangladesh.

About 600 million people are estimated to be infected with Strongyloides stercoralis, the species that causes most of the human strongyloidiasis cases. S. stercoralis can also infect non-human primates (NHPs), dogs and cats, rendering these animals putative sources for zoonotic human S. stercoralis infection. S. fuelleborni is normally found in old world NHPs but occasionally also infects humans, mainly in Africa. Dogs in southeast Asia carry at least two types of Strongyloides, only one of which appears to be shared with humans ("dog only" and "human and dog" types). For S. stercoralis with molecular taxonomic information, there is a strong sampling bias towards southeast and east Asia and Australia. In order to extend the geographic range of sampling, we collected human and dog derived Strongyloides spp. and hookworms from two locations in Bangladesh and subjected them to molecular taxonomic and genomic analysis based on nuclear and mitochondrial sequences. All hookworms found were Necator americanus. Contrary to earlier studies in Asia, we noticed a rather high incidence of S. fuelleborni in humans. Also in this study, we found the two types of S. stercoralis and no indication for genetic isolation from the southeast Asian populations. However, we found one genomically "dog only" type S. stercoralis in a human sample and we found two worms in a dog sample that had a nuclear genome of the "dog only" but a mitochondrial genome of the "human and dog" type. S. fuelleborni may play a more prominent role as a human parasite in certain places in Asia than previously thought. The introgression of a mitochondria haplotype into the "dog only" population suggests that rare interbreeding between the two S. stercoralis types does occur and that exchange of genetic properties, for example a drug resistance, between the two types is conceivable.

Correlating Cases of Soil-Transmitted Helminths and Remote Sensing Covariates: A Case Study Utilizing 2015, 2017, and 2019 Metadata of Camarines Sur, Philippines.

Soil-transmitted helminth (STH) is a group of helminths that are considered to be neglected tropical diseases (NTDs) and, globally, affect more than 1.5 to 2.6 million people yearly. Depending on the species, they can be acquired by ingesting embryonated eggs from contaminated matter or by skin penetration. Most species of STH are found in the tropics, such as the Philippines. Despite the Mass Drug Administration (MDA), the cases of STH infection continue to rise in the country. Surveys from the Research Institute of Tropical Medicine (RITM) indicate that a high prevalence of STH (Ascaris lumbricoides, Trichuris trichiura, and Necator americanus) was primarily observed in the provinces of the country, such as in Camarines Sur. To correlate remote sensing covariates such as Normalized Difference Vegetation Index (NDVI) and Normalized Difference Built-up Index (NDBI) - to STH-infected cases of the 37 municipalities of Camarines Sur. The available public health record of STH cases from 2015 to 2019 were calculated using the Quantum Geographic Information System (QGIS)and correlated using Pearson Correlation Coefficient. The results showed that infection was higher in children than adults, and A. lumbricoides caused 60% of infection. No correlation of indices with infection cases during 2015 and 2017 was observed; however, 2019 showed a moderate strength (p = 0.037) in correlation. This indicates that infection relied not mainly on vegetation and urbanization but on additional environmental factors and non-environmental variables.

Pharmacotherapy Study Of Albendazole And Mebendazole For Soil Transmitted Helminths Infections In Children

Soil Transmitted Helminths (STH) infection is a nematode infection of the intestine that requires soil for its transmission. The worms that cause Soil Transmitted Helminths infection include; Ascaris lumbricoides, Trichuris trichiura, hookworms (Necator americanus and Ancylostoma duodenale. The incidence of STH infection is quite high with the highest spread of STH infection in the child age group. Treatment of cases of STH infection is by administering antihelmintic drugs or worming drugs. Although there are four The drugs recommended by WHO, only the two drugs most often used in the treatment of STH infections, namely albendazole and mebendazole. Both drugs have been used for more than 30 years, but cases of STH infection still have a high incidence. That is why this literature review was written aims to describe the efficacy of the two drugs for STH infection in children so that their effectiveness can be assessed.The research method used was a literature review, a literature search was sourced from Proquest, Google scholar, Pubmed, Elsevier and Europe PMC. keywords Albendazole, Mebendazole, Soil Transmitted Helminths Infections, Children. There are 10 selected literatures to be studied in this literature review. The results showed that albendazole had satisfactory efficacy in STH infections caused by Ascaris lumbricoides and hookworms, but reduced efficacy in Trichuris trichiura worms. Mebendazole has satisfactory efficacy in STH infections caused by Ascaris lumbricoides and Trichuris trichiura, but reduced efficacy in hookworms. Albendazole and mebendazole does not cause side effects.

Prevalence and intensity of soil-transmitted helminth infections among school-aged children in five districts in Uganda.

Soil-transmitted helminth (STH) infections, commonly caused by roundworms (Ascaris lumbricoides), whipworms (Trichuris trichiura), and hookworms (Necator americanus and Ancylostoma duodenale), were widespread among Ugandan schoolchildren in the late 1990s and early 2000s. Since 2003, the Ugandan Ministry of Health has administered biannual preventive chemotherapy to children aged 1-14 years to control these infections. Twenty years after the program's inception, there is scant data to show the long-term impact of these national deworming efforts. To estimate the prevalence and intensity of STH infections among 10-14-year-old primary school children, school-based, cross-sectional surveys were conducted in November 2023 across five districts (Kamwenge, Sheema, Adjumani, Lamwo, and Zombo). Sixty-five children from five schools per district were selected for inclusion. Fecal egg counts were determined using the Kato-Katz microscopy technique, performed in duplicate by trained laboratory technicians. The survey findings revealed a high prevalence of any STH infection in Kamwenge District (21.2%, 95% confidence limits (CL): 5.7%, 36.6%), while the remaining four districts exhibited lower prevalences, ranging from 0.4% (95% CL: 0.0%, 1.2%) in Adjumani District to 5.6% (95% CL: 0.0%, 11.4%) in Sheema District. The prevalence of moderate-to-heavy-intensity infections was below 1% across all districts. A. lumbricoides was identified infrequently. Hookworm infections were primarily identified in the western districts of Kamwenge and Sheema, while T. trichiura infections were common only in Kamwenge District. Hookworm and T. trichiura infections were uncommon in the northern districts of Adjumani, Lamwo, and Zombo. These surveys suggest that morbidity due to STH infections among schoolchildren may be well controlled in these five districts, as evidenced by low moderate-to-heavy-intensity infection prevalence. However, the prevalence of any intensity infection remains elevated in some districts, indicating the need for continued preventive chemotherapy distribution. A reduction from biannual treatment may be warranted in four districts, per World Health Organization recommendations.

Prevalence of Strongyloides stercoralis and other helminths in four districts of Madagascar

BackgroundEstimation of prevalence of Strongyloides stercoralis infection is required in endemic areas, in order to identify areas in need of control programmes. Data on prevalence of strongyloidiasis in Madagascar are scant. Aim of this work was to estimate prevalence of S. stercoralis in four districts of Madagascar.MethodsFecal and serum samples collected in the context of a previous study on schistosomiasis were tested with S. stercoralis real-time PCR and serology, respectively. A multiplex real-time PCR for Ascaris lumbricoides, Ancylostoma duodenalis, Necator americanus, and Trichuris trichiura was done on fecal samples collected in the areas demonstrating higher prevalence of strongyloidiasis. Comparisons between proportions were made using Fisher exact test, with false discovery rate correction used for post-hoc comparisons. A multivariable Firth logistic regression model was used to assess potential risk factors for S. stercoralis infection.ResultsOverall, 1775 serum samples were tested, of which 102 of 487 (20.9%) and 104 of 296 (35.2%) were serological-positive in Marovoay and in Vatomandry districts (both coastal areas), respectively, compared to 28 of 496 (5.6%) and 30 of 496 (6.1%) in Tsiroanomandidy and in Ambositra districts (both highlands), respectively (adj. p < 0.001). PCR for S. stercoralis was positive in 15 of 210 (7.1%) and in 11 of 296 (3.7%) samples from Marovoay from Vatomandry, respectively, while was negative for all samples tested in the other two districts. High prevalence of A. lumbricoides (45.9%), hookworm (44.6%) and T. trichiura (32.1%) was found in Vatomandry. In the multivariable analysis, strongyloidiasis was associated with hookworm infection. Hookworm infection was also associated with male sex and lower education level.ConclusionsS. stercoralis prevalence proved higher in coastal areas compared to highlands. Different climatic conditions may explain this distribution, along with previous rounds of anthelminthics distributed in the country, which may have reduced the parasite load in the population. The high prevalence of the other soil-transmitted helminths (STH) in Vatomandry was unexpected, given the good coverage with benzimidazole in control campaigns. Further studies are needed to explore the risk factors for STH and S. stercoralis infections in Madagascar, in order to align with the WHO recommendations.

Effects of household concrete floors on maternal and child health - the CRADLE trial: a randomised controlled trial protocol.

Early life soil-transmitted helminth infection and diarrhea are associated with growth faltering, anemia, impaired child development, and mortality. Exposure to fecally contaminated soil inside the home may be a key contributor to enteric infections, and a large fraction of rural homes in low-income countries have soil floors. The objective of this study is to measure the effect of installing concrete floors in homes with soil floors on child soil-transmitted helminth infection and other maternal and child health outcomes in rural Bangladesh. The Cement-based flooRs AnD chiLd hEalth (CRADLE) trial is an individually randomised trial in Sirajganj and Tangail districts, Bangladesh. Households with a pregnant woman, a soil floor, walls that are not made of mud will be eligible, and no plan to relocate for 3 years. We will randomise 800 households to intervention or control (1:1) within geographic blocks of 10 households to account for strong geographic clustering of enteric infection. Laboratory staff and data analysts will be blinded; participants will be unblinded. We will install concrete floors when the birth cohort is in utero and measure outcomes at child ages 3, 6, 12, 18, and 24 months. The primary outcome is prevalence of any soil-transmitted helminth infection (Ascaris lumbricoides, Necator americanus, or Trichuris trichiura) detected by qPCR at 6, 12, 18, or 24 months follow-up in the birth cohort. Secondary outcomes include household floor and child hand contamination with E. coli, extended-spectrum beta-lactamase producing E. coli, and soil-transmitted helminth DNA; child diarrhea, growth, and cognitive development; and maternal stress and depression. Study protocols have been approved by institutional review boards at Stanford University and the International Centre for Diarrheal Disease Research, Bangladesh (icddr,b). We will report findings on ClinicalTrials.gov, in peer-reviewed publications, and in stakeholder workshops in Bangladesh. NCT05372068, pre-results.

Anthroponotic and Zoonotic Hookworm DNA in an Indigenous Community in Coastal Ecuador: Potential Cross-Transmission between Dogs and Humans.

Humans can be infected with anthroponotic (Ancylostoma duodenale and Necator americanus) and with zoonotic (Ancylostoma ceylanicum, A. caninum, A. braziliense, and Uncinaria stenocephala) hookworms from dogs. Anthroponotic species are usually thought not to infect dogs. We used the internal transcribed spacer-1 (ITS1) gene in a quantitative PCR to detect anthroponotic and zoonotic hookworm species in fecal samples from 54 children and 79 dogs living in an indigenous community in tropical Northwestern Ecuador. Hookworm DNA was detected in 59.3% of children and 92.4% of dogs. Among samples from children, zoonotic hookworms were detected in 24.1% (A. ceylanicum 14.8%, A. caninum 11.1%, and A. braziliense 1.9%), whilst in dog samples, anthroponotic species were detected in 19.0% (N. americanus 12.4% and A. duodenale 6.3%). Sanger sequencing was performed successfully on 60 qPCR-positive samples (16 from children and 44 from dogs), and consensus sequences were obtained with >98% homology to GenBank references for hookworm spp. Phylogenetic analysis showed a close relationship between anthroponotic and zoonotic Ancylostoma species and no heterogeneity between A. duodenale and A. caninum; in human samples, we found A. ceylanicum but not A. braziliense sequences and we were unable to identify N. americanus in the dog samples. No infections with U. stenocephala were detected. Our data provide evidence for high rates of hookworm infections in indigenous children and dogs in a marginalized rural setting in coastal Ecuador. We also found evidence for potential cross-transmission of hookworm spp. between humans and dogs that represent a potential domestic reservoir for zoonotic and anthroponotic hookworms.

Controlled human hookworm infection remodels plasmacytoid dendritic cells and regulatory T cells towards profiles seen in natural infections in endemic areas

Hookworm infection remains a significant public health concern, particularly in low- and middle-income countries, where mass drug administration has not stopped reinfection. Developing a vaccine is crucial to complement current control measures, which necessitates a thorough understanding of host immune responses. By leveraging controlled human infection models and high-dimensional immunophenotyping, here we investigated the immune remodeling following infection with 50 Necator americanus L3 hookworm larvae in four naïve volunteers over two years of follow-up and compared the profiles with naturally infected populations in endemic areas. Increased plasmacytoid dendritic cell frequency and diminished responsiveness to Toll-like receptor 7/8 ligand were observed in both controlled and natural infection settings. Despite the increased CD45RA+ regulatory T cell (Tregs) frequencies in both settings, markers of Tregs function, including inducible T-cell costimulatory (ICOS), tumor necrosis factor receptor 2 (TNFR2), and latency-associated peptide (LAP), as well as in vitro Tregs suppressive capacity were higher in natural infections. Taken together, this study provides unique insights into the immunological trajectories following a first-in-life hookworm infection compared to natural infections.

A Review of Traditional Anthelmintic Approaches: Exploring Medicinal Plants and Polyherbal Formulations for Effective and Safe Gastrointestinal Parasite Treatment

Abstract: Gastrointestinal parasites have become the most common reason for infection in humans, affecting many people in the world. Helminthic infections pose a large threat to public health in developing countries. The most common parasites of helminths in humans are whipworms(Trichuristrichiura), roundworms(Ascarislumbricoides), tapeworms(Taenia solium), and hookworms (Ancylostoma duodenale and Necator americanus). In the current scenario, synthetic medicines are found to be effective in curing the infection but these medicines produce side-effects that impair the effectiveness of the treatment. Fortunately, in the traditional system of medicines various medicinal plants have been mentioned for their anthelmintic activities, which are relatively free from side effects and are used by ethnic groups worldwide. In the 21st century, an ideal anthelmintic agent should be cost-effective, provide a high percentage of cure, have a broad spectrum of action, and free from toxicity to the host. However, none of the modern marketed medicines fulfill these requirements. Therefore, in this view, several researchers have undertaken various in vivo and in vitro studies to scrutinize folklore medicinal plants for their proclaimed anthelmintic activity. In thisreview, we have summarized some scientifically approved polyherbal formulations and Indian medicinal plants documented for anthelmintic activity, which can be further studied to achieve novel herbal lead molecules from them by isolating their principal compounds.

Soil-Transmitted Helminth Infections in Shanghai, China: Surveillance from 2014 to 2023.

Soil-transmitted helminthiasis remains a notable health problem in developing countries. In 1990 in Shanghai, a city in eastern China, 47% of the population was affected. Due to a series of comprehensive approaches, the prevalence decreased to 0.5% in 2009. We collected 10-year surveillance data to assess the epidemic situation of soil-transmitted helminth (STH) infections in Shanghai. Stool samples and questionnaires from participants were collected. The Kato-Katz technique was used to detect infections with Ascaris lumbricoides, hookworm (Ancylostoma duodenale and Necator americanus), and Trichuris trichiura. From 2014 to 2023, a total of 77,685 participants were screened for parasitic infections, and the overall prevalence of STH infections decreased from 0.1% to 0% in Shanghai. Of 77,685 participants, 25 (0.03%) were positive for intestinal helminths, with the most common parasite being A. lumbricoides (72.0%). Generally, elderly participants from rural areas with lower education levels were more likely to be infected with STHs. A total of 2,914 questionnaires were collected, and most respondents (72.6%) had good knowledge about the transmission routes of STHs. However, 12.3% of participants did not know the damage to health caused by STHs, and 19.3% had no idea of how to prevent infection. These data demonstrate the current state of STH infections in Shanghai. The results suggest that various comprehensive measures should be encouraged, continuously implemented, and strengthened accordingly so that STH elimination can be integrated into the Healthy China initiative by 2030.

Public engagement for the conduct of a controlled human infection study testing vaccines against Necator americanus (hookworm) in areas of active hookworm transmission in Brazil.

This is a qualitative action research that includes the development of an educational intervention with residents of a rural area in Minas Gerais, Brazil, where there is continuous natural transmission of the human pathogen Necator americanus ("hookworm"). In this area, it is intended to carry out a proposed phase 3 vaccine clinical trial in the future to test the efficacy of hookworm vaccines using controlled human infection. Two data collection strategies were used: an educational intervention and a focus group. The participants' perceptions showed distinct perspectives on CHIS. On one side, they recognized that the investigation is essential for the community, but on the other side, they thought that there would be resistance to its conduct by fear of infection. The idea that the study would generate a benefit for the greater good, contributing to the prevention of hookworm infection, was clearly stated. The participants perceived that the study offered concrete risks that could be reduced by constant monitoring by the researchers. They also mentioned the importance of access to information and the positive influence those who express interest in participating in the study can exert in the community. In relation to comprehension the participants memorized the information, mobilized it to explain everyday situations and created strategies to disseminate the study and engage the community in its development. By repeating and making sense of the information, the participant not only assimilates the knowledge transmitted, but also creates new knowledge. We concluded that an educational process of discussion and dialogue around participants' perceptions about the CHIS, promotes understanding and allows ways to disseminate information about the research to be collectively created.

The effect of novel aromatic heterocycle substituted aminamidine derivatives on Necator americanus.

The efficacy of current drugs against hookworms at a single dose is highly variable across regions, age groups and infection intensity. Extensive and repeated use of these drugs also leads to potential drug resistance. Therefore, novel drugs are required for sustained disease control. Novel aromatic heterocycle substituted aminamidine derivatives (AADs) were synthesized based on tribendimine (TBD), and their in vivo potency against Necator americanus was tested. The efficacy of the AADs was tested in male hamsters. Oral and IV pharmacokinetic parameters were determined in male Sprague-Dawley rats. The proteomic profiles of N. americanus samples treated with AADs were compared using tandem mass tag-based quantitative proteomic analyses. Most AADs exhibited better anthelmintic activity than TBD at a single oral dose. Compound 3c exhibited improved solubility (>50×), and the curative dose was as low as 25 mg/kg. Similar to TBD, 3c was rapidly metabolized after oral administration and transformed into p-(1-dimethylamino ethylimino)aniline (dADT), an active metabolite against intestinal nematodes. dADT from 3c had better pharmacokinetic profiles than that from TBD and achieved an oral bioavailability of 99.5%. Compound 3c possessed rapid anthelmintic activity, clearing all worms within 24 h after an oral dose of 50 mg/kg. Quantitative proteomic analysis indicated that it might be related to ATP metabolism and cuticle protein synthesis. Compound 3c is a novel and promising compound against N. americanus in vivo.

Modulation of intestinal epithelial permeability by chronic small intestinal helminth infections.

Increased permeability of the intestinal epithelial layer is linked to the pathogenesis and perpetuation of a wide range of intestinal and extra-intestinal diseases. Infecting humans with controlled doses of helminths, such as human hookworm (termed hookworm therapy), is proposed as a treatment for many of the same diseases. Helminths induce immunoregulatory changes in their host which could decrease epithelial permeability, which is highlighted as a potential mechanism through which helminths treat disease. Despite this, the influence of a chronic helminth infection on epithelial permeability remains unclear. This study uses the chronically infecting intestinal helminth Heligmosomoides polygyrus to reveal alterations in the expression of intestinal tight junction proteins and epithelial permeability during the infection course. In the acute infection phase (1 week postinfection), an increase in intestinal epithelial permeability is observed. Consistent with this finding, jejunal claudin-2 is upregulated and tricellulin is downregulated. By contrast, in the chronic infection phase (6 weeks postinfection), colonic claudin-1 is upregulated and epithelial permeability decreases. Importantly, this study also investigates changes in epithelial permeability in a small human cohort experimentally challenged with the human hookworm, Necator americanus. It demonstrates a trend toward small intestinal permeability increasing in the acute infection phase (8 weeks postinfection), and colonic and whole gut permeability decreasing in the chronic infection phase (24 weeks postinfection), suggesting a conserved epithelial response between humans and mice. In summary, our findings demonstrate dynamic changes in epithelial permeability during a chronic helminth infection and provide another plausible mechanism by which chronic helminth infections could be utilized to treat disease.

Diagnosis of soil-transmitted helminth infections with digital mobile microscopy and artificial intelligence in a resource-limited setting.

Infections caused by soil-transmitted helminths (STHs) are the most prevalent neglected tropical diseases and result in a major disease burden in low- and middle-income countries, especially in school-aged children. Improved diagnostic methods, especially for light intensity infections, are needed for efficient, control and elimination of STHs as a public health problem, as well as STH management. Image-based artificial intelligence (AI) has shown promise for STH detection in digitized stool samples. However, the diagnostic accuracy of AI-based analysis of entire microscope slides, so called whole-slide images (WSI), has previously not been evaluated on a sample-level in primary healthcare settings in STH endemic countries. Stool samples (n = 1,335) were collected during 2020 from children attending primary schools in Kwale County, Kenya, prepared according to the Kato-Katz method at a local primary healthcare laboratory and digitized with a portable whole-slide microscopy scanner and uploaded via mobile networks to a cloud environment. The digital samples of adequate quality (n = 1,180) were split into a training (n = 388) and test set (n = 792) and a deep-learning system (DLS) developed for detection of STHs. The DLS findings were compared with expert manual microscopy and additional visual assessment of the digital samples in slides with discordant results between the methods. Manual microscopy detected 15 (1.9%) Ascaris lumbricoides, 172 (21.7%) Tricuris trichiura and 140 (17.7%) hookworm (Ancylostoma duodenale or Necator americanus) infections in the test set. Importantly, more than 90% of all STH positive cases represented light intensity infections. With manual microscopy as the reference standard, the sensitivity of the DLS as the index test for detection of A. lumbricoides, T. trichiura and hookworm was 80%, 92% and 76%, respectively. The corresponding specificity was 98%, 90% and 95%. Notably, in 79 samples (10%) classified as negative by manual microscopy for a specific species, STH eggs were detected by the DLS and confirmed correct by visual inspection of the digital samples. Analysis of digitally scanned stool samples with the DLS provided high diagnostic accuracy for detection of STHs. Importantly, a substantial number of light intensity infections were missed by manual microscopy but detected by the DLS. Thus, analysis of WSIs with image-based AI may provide a future tool for improved detection of STHs in a primary healthcare setting, which in turn could facilitate monitoring and evaluation of control programs.

Toward the elimination of NTDs: application of cost-effective and sensitive molecular environmental surveillance tools—a pilot study

Neglected tropical diseases (NTDs) affect over a billion people worldwide. The 2021–2030 NTD road map calls for innovative and highly efficient interventions to eliminate or significantly reduce the burden of NTDs. These include sensitive and cost-effective diagnostic techniques for disease surveillance. Environmental surveillance has been employed effectively in this regard to measure and track infectious diseases such as polio on a population-wide scale. In this study, environmental surveillance was used as a cost-effective tool for the detection of soil-transmitted helminths (STHs) in Accra, Ghana, in an area that is populated by urban vegetable farmers. The activities of urban farmers expose them to the risk of STH infection, as well as impact the transmission in urban areas since leafy vegetables could carry infective stages of STHs. A total of 32 wastewater samples were collected from eight points on the Nima Creek (the main source of irrigation for the farmers) over a 7-week period. Real-time PCR and melt peak analysis were used to screen four STHs (i.e., Ascaris lumbricoides, Necator americanus, Ancylostoma duodenale, and Trichuris trichiura). This study revealed that A. lumbricoides (17 out of 32 wastewater samples, 53.3%) was the most prevalent STH, followed by A. duodenale (31.2%), T. trichiura (21.9%), and N. americanus (12.5%). Environmental surveillance helps in the detection of the types of STH pathogens circulating within the community and in the design of mass drug administration (MDA) strategies. This surveillance technique can also provide preliminary information for environmental modifications to help reduce STH transmission in line with the One Health approach recommended in the 2021–2030 NTD road map.

Erythrocyte and iron profiles in soil-transmitted helminth-infected children in a rural area in Banten, Indonesia

BACKGROUND Soil-transmitted helminth (STH) infection, frequently caused by Ascaris lumbricoides, Trichuris trichiura, and hookworms (Ancylostoma duodenale and Necator americanus), often gives rise to malabsorption of nutrients that form hemoglobin (Hb) thereby causing anemia. This study aimed to know the difference in erythrocyte profile in children with and without STH infections. METHODS This cross-sectional study involved 205 children from two Panimbang Jaya elementary schools in Pandeglang, Banten, Indonesia. Blood and stool samples were collected from a previous study conducted from November 2021 to May 2022. Erythrocyte parameters were Hb concentration, erythrocyte count, mean corpuscular volume (MCV), mean corpuscular Hb (MCH), and red cell distribution width (RDW). The iron profile included serum iron, total iron-biding capacity (TIBC), and ferritin. Worm infestation was detected by direct stool microscopical examination. Statistical analysis was performed using SPSS software version 20. RESULTS The prevalence of STH infection in Pandeglang was 44.4%, primarily characterized by mild intensity STH infection (79%). The identified STH species were A. lumbricoides, T. trichiura, and combination of both. The median differences between erythrocyte count, MCV, and MCH, and the mean differences of TIBC and serum iron were not statistically significant (p = 0.388, 0.098, and 0.057, and p = 0.304 and 0.455). However, children with STH infection had lower Hb (12.57 versus 12.95 g/dl) and ferritin (19.60 versus 30.57 µg/dl) levels but higher RDW (13.20 versus 13.10%). CONCLUSIONS A high prevalence of STH infection was identified among schoolchildren, but their erythrocyte profiles were similar regardless of STH infection status.