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PLoS Neglected Tropical Diseases Apr 2024Infections 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...
BACKGROUND
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.
METHODOLOGY/PRINCIPAL FINDINGS
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.
CONCLUSIONS/SIGNIFICANCE
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.
Topics: Child; Animals; Humans; Artificial Intelligence; Soil; Microscopy; Resource-Limited Settings; Feces; Trichuris; Helminthiasis; Helminths; Ascaris lumbricoides; Ancylostomatoidea; Prevalence
PubMed: 38602896
DOI: 10.1371/journal.pntd.0012041 -
PLoS Neglected Tropical Diseases Apr 2024The World Health Organization emphasizes the importance of integrated monitoring and evaluation in neglected tropical disease (NTD) control programs. Serological assays... (Review)
Review
BACKGROUND
The World Health Organization emphasizes the importance of integrated monitoring and evaluation in neglected tropical disease (NTD) control programs. Serological assays offer a potential solution for integrated diagnosis of NTDs, particularly for those requiring mass drug administration (MDA) as primary control and elimination strategy. This scoping review aims (i) to provide an overview of assays using serum or plasma to detect infections with soil-transmitted helminths (STHs) in both humans and animals, (ii) to examine the methodologies used in this research field and (iii) to discuss advancements in serological diagnosis of STHs to guide prevention and control programs in veterinary and human medicine.
METHODOLOGY
We conducted a systematic search in the Ovid MEDLINE, Embase and Cochrane Library databases, supplemented by a Google search using predefined keywords to identify commercially available serological assays. Additionally, we performed a patent search through Espacenet.
PRINCIPAL FINDINGS
We identified 85 relevant literature records spanning over 50 years, with a notable increased interest in serological assay development in recent years. Most of the research efforts concentrated on diagnosing Ascaris infections in both humans and pigs, primarily using ELISA and western blot technologies. Almost all records targeted antibodies as analytes, employing proteins and peptides as analyte detection agents. Approximately 60% of sample sets described pertained to human samples. No commercially available tests for Trichuris or hookworms were identified, while for Ascaris, there are at least seven different ELISAs on the market.
CONCLUSIONS
While a substantial number of assays are employed in epidemiological research, the current state of serological diagnosis for guiding STH prevention and control programs is limited. Only two assays designed for pigs are used to inform efficient deworming practices in pig populations. Regarding human diagnosis, none of the existing assays has undergone extensive large-scale validation or integration into routine diagnostics for MDA programs.
Topics: Humans; Animals; Swine; Ancylostomatoidea; Trichuris; Ascaris; Soil; Ascaris lumbricoides; Helminths; Feces; Helminthiasis; Prevalence
PubMed: 38574166
DOI: 10.1371/journal.pntd.0012049 -
PLoS Neglected Tropical Diseases Mar 2024Control efforts of soil-transmitted helminthiases rely primarily on large scale administration of anthelminthic drugs. The assessment of drug efficacies and... (Clinical Trial)
Clinical Trial
BACKGROUND
Control efforts of soil-transmitted helminthiases rely primarily on large scale administration of anthelminthic drugs. The assessment of drug efficacies and understanding of drug behavior is pivotal to the evaluation of treatment successes, both in preventive chemo-therapy programs as well as in research of novel treatment options. The current WHO guidelines recommend an interval of 14-21 days between the treatment and follow-up, yet no in-depth analysis of egg excretion patterns of Trichuris trichiura after treatment has been conducted to date.
METHODS
Within the framework of a multi-country trial to assess the efficacy and safety of albendazole-ivermectin combination therapy vs albendazole monotherapy against T. trichiura infections, we conducted a study collecting daily stool samples over the period of 28 days post-treatment in 87 participants in Pak Khan, Lao PDR. Egg counts were derived by duplicate Kato-Katz on-site for T. trichiura, hookworm and Ascaris lumbricoides and stool sample aliquots were subsequently analyzed by qPCR for the detection of T. trichiura infections. Sensitivity and specificity was calculated for each day separately using data derived by Kato-Katz to determine the optimal timepoint at which to assess drug efficacy.
RESULTS
Egg excretion patterns varied across treatment arms. For T. trichiura, only the albendazole-ivermectin treatment led to a considerable reduction in mean egg counts, whereas both treatments reduced hookworm egg counts and A. lumbricoides were cleared in all participants after day 7. For T. trichiura, we found sensitivity to be highest at days 18 and 22 when using egg counts as outcome and days 19 and 24 when using qPCR. Specificity was high (>0.9) from day 14 onwards. For hookworm, the highest sensitivity and specificity were found at days 17 and 25, respectively.
CONCLUSIONS
Based on our study, the ideal time period to assess drug efficacy for soil-transmitted helminth infections would be between day 18 and 24. The current WHO recommendation of 14 to 21 days is likely to yield acceptable outcome measures for soil-transmitted helminth infections.
TRIAL REGISTRATION
NCT03527732.
Topics: Animals; Humans; Albendazole; Ancylostomatoidea; Anthelmintics; Feces; Helminthiasis; Ivermectin; Soil; Trichuriasis; Trichuris
PubMed: 38517907
DOI: 10.1371/journal.pntd.0012073 -
Acta Parasitologica Jun 2024Soil-transmitted nematodes (STNs) are widespread in tropical and subtropical regions, particularly where the communities are socio-economically challenged. We...
PURPOSE
Soil-transmitted nematodes (STNs) are widespread in tropical and subtropical regions, particularly where the communities are socio-economically challenged. We investigated the effect of soil temperature on the prevalence and intensity of STN infection in free-roaming dogs.
METHODS
Fresh faecal samples collected from free-roaming dogs in Digana and Pussellawa town areas in the Kandy District, Sri Lanka, were microscopically analysed for canine STNs. Soil temperature was measured at each sampling site. Highly prevalent canine hookworm Ancylostoma, was further studied using PCR and sequencing, followed by phylogenetic analysis.
RESULTS
The soil temperature ranged between 28 and 31 °C (mean = 29.79 °C) and 18-21 °C (mean = 19.52 °C) in Digana and Pussellawa, respectively, showing a significant difference in the two sites (Students t-test t = 1.68, p < 0.0001). Of the total 44 dogs sampled, 41 (93.2%) were positive for STNs. During microscopic analysis, five nematodes: Ancylostoma spp., Capillaria sp., Strongyloides sp., Toxocara canis, and Trichuris sp., were identified. Ancylostoma species (93.2%) were the most prevalent, followed by Strongyloides sp. (22.7%) and Toxocara canis (15.9%). Infection prevalence of Strongyloides sp. was higher in Digana (40.9%) compared to that in Pussellawa (4.5%; Chi-square test, χ = 8.28, p = 0.004) and also the infection intensity from Digana (EPG = 8.02 ± 20.2) compared to that from Pussellawa (0.45 ± 2.1; Mann Whitney U test, p = 0.006). Amplicons (ITS1-5.8S-ITS2) of the expected size for A. caninum, and A. tubaeforme were produced. An A. caninum sequence reported here (OQ101719) illustrated the highest similarity of 99.2% to one of the local sequences (MZ707153) upon pairwise comparison.
CONCLUSION
Digana, with a higher soil temperature than Pussellawa, had a significantly higher prevalence and infection intensity, particularly Strongyloides sp. This study also signifies the first molecular identification of hookworm species A. tubaeforme in Sri Lanka.
Topics: Animals; Dogs; Soil; Sri Lanka; Dog Diseases; Feces; Temperature; Ancylostoma; Phylogeny; Prevalence; Ancylostomiasis; Nematoda
PubMed: 38514604
DOI: 10.1007/s11686-024-00829-8 -
The Lancet. Infectious Diseases Jul 2024
Topics: Humans; Hookworm Infections; Animals; Vaccines; Ancylostomatoidea
PubMed: 38513685
DOI: 10.1016/S1473-3099(24)00145-2 -
The Lancet. Infectious Diseases Jul 2024A human hookworm vaccine is being developed to protect children against iron deficiency and anaemia associated with chronic infection with hookworms. Necator americanus... (Randomized Controlled Trial)
Randomized Controlled Trial
Safety and immunogenicity of the co-administered Na-APR-1 and Na-GST-1 hookworm vaccines in school-aged children in Gabon: a randomised, controlled, observer-blind, phase 1, dose-escalation trial.
BACKGROUND
A human hookworm vaccine is being developed to protect children against iron deficiency and anaemia associated with chronic infection with hookworms. Necator americanus aspartic protease-1 (Na-APR-1) and N americanus glutathione S-transferase-1 (Na-GST-1) are components of the blood digestion pathway critical to hookworm survival in the host. Recombinant Na-GST-1 and catalytically inactive Na-APR-1 (Na-APR-1[M74]) adsorbed to Alhydrogel were safe and immunogenic when delivered separately or co-administered to adults in phase 1 trials in non-endemic and endemic areas. We aimed to investigate the safety and immunogenicity of these antigens in healthy children in a hookworm-endemic area.
METHODS
This was a randomised, controlled, observer-blind, phase 1, dose-escalation trial, conducted in a clinical research centre, in 60 children aged six to ten years in Lambaréné, a hookworm-endemic region of Gabon. Healthy children (determined by clinical examination and safety laboratory testing) were randomised 4:1 to receive co-administered Na-GST-1 on Alhydrogel plus Na-APR-1(M74) on Alhydrogel and glucopyranosyl lipid A in aqueous formulation (GLA-AF), or co-administered ENGERIX-B hepatitis B vaccine (HBV) and saline placebo, injected into the deltoid of each arm. Allocation to vaccine groups was observer-masked. In each vaccine group, children were randomised 1:1 to receive intramuscular injections into each deltoid on two vaccine schedules, one at months 0, 2, and 4 or at months 0, 2, and 6. 10 μg, 30 μg, and 100 μg of each antigen were administered in the first, second, and third cohorts, respectively. The intention-to-treat population was used for safety analyses; while for immunogenicity analyses, the per-protocol population was used (children who received all scheduled vaccinations). The primary outcome was to evaluate the vaccines' safety and reactogenicity in healthy children aged between six and ten years. The secondary outcome was to measure antigen-specific serum IgG antibody levels at pre-vaccination and post-vaccination timepoints by qualified ELISAs. The trial is registered with ClinicalTrials.gov, NCT02839161, and is completed.
FINDINGS
Between Jan 23 and Oct 3, 2017, 137 children were screened, of whom 76 were eligible for this trial. 60 children were recruited, and allocated to either 10 μg of the co-administered antigens (n=8 for each injection schedule), 30 μg (n=8 for each schedule), 100 μg (n=8 for each schedule), or HBV and placebo (n=6 for each schedule) in three sequential cohorts. Co-administration of the vaccines was well tolerated; the most frequent solicited adverse events were mild-to-moderate injection-site pain, observed in up to 12 (75%) of 16 participants per vaccine group, and mild headache (12 [25%] of 48) and fever (11 [23%] of 48). No vaccine-related serious adverse events were observed. Significant anti-Na-APR-1(M74) and anti-Na-GST-1 IgG levels were induced in a dose-dependent manner, with peaks seen 14 days after the third vaccinations, regardless of dose (for Na-APR-1[M74], geometric mean levels [GML]=2295·97 arbitrary units [AU] and 726·89 AU, while for Na-GST-1, GMLs=331·2 AU and 21·4 AU for the month 0, 2, and 6 and month 0, 2, and 4 schedules, respectively). The month 0, 2, and 6 schedule induced significantly higher IgG responses to both antigens (p=0·01 and p=0·04 for Na-APR-1[M74] and Na-GST-1, respectively).
INTERPRETATION
Co-administration of recombinant Na-APR-1(M74) and Na-GST-1 to school-aged Gabonese children was well tolerated and induced significant IgG responses. These results justify further evaluation of this antigen combination in proof-of-concept controlled-infection and efficacy studies in hookworm-endemic areas.
FUNDING
European Union Seventh Framework Programme.
Topics: Humans; Male; Child; Female; Gabon; Necator americanus; Animals; Hookworm Infections; Antigens, Helminth; Antibodies, Helminth; Glutathione Transferase; Single-Blind Method; Vaccines; Immunogenicity, Vaccine
PubMed: 38513684
DOI: 10.1016/S1473-3099(24)00104-X -
Medical Science Monitor : International... Mar 2024BACKGROUND Ancylostoma spp., including A. duodenale, A. braziliense, A. caninum, and A. ceylanicum, are hookworms that are transmitted from infected soil and by contact...
BACKGROUND Ancylostoma spp., including A. duodenale, A. braziliense, A. caninum, and A. ceylanicum, are hookworms that are transmitted from infected soil and by contact with domestic animals and rodent hosts, and can cause systemic disease and cutaneous larva migrans. The objective of this study was to describe the ecoepidemiology of Ancylostoma caninum and Ancylostoma spp. in urban-marginal sectors and in rural sectors located in Ecuador. MATERIAL AND METHODS Through addressed sampling, a total of 498 domestic dogs and 40 synanthropic rodents were analyzed via the following coproparasitic methods: direct, flotation, sedimentation with centrifugation using saline (egg identification), modified Baermann (larval identification), and morphometric methods (confirmation). A total of 236 people were surveyed, and a clinical analysis was performed via physical examination. The environmental variables were obtained through reports from the INAMHI of Ecuador and the use of online environmental programs. Through surveys, data related to social determinants were obtained. Epidemiological indicators (prevalence, morbidity, and mortality) were obtained through microbial analysis and surveys. RESULTS A total of 250 domestic dogs were diagnosed with Ancylostoma caninum (50, 20%), and 41 were diagnosed with Ancylostoma spp. (8.23%). One synanthropic rodent (2.5%) was positive for A. caninum. In the clinical analysis, 3 patients were identified as positive (1.27%) for cutaneous larva migrans (CLM). Likewise, environmental variables and social determinants influence the transmission, prevalence, and nature of parasitism by hookworm. CONCLUSIONS People, domestic dogs, and rodents were infected with these parasites. Consequently, there is a risk of ancylostomiasis and cutaneous larvae migrans spreading.
Topics: Humans; Animals; Dogs; Ancylostoma; Larva Migrans; Ecuador; Prevalence; Larva; Feces
PubMed: 38500436
DOI: 10.12659/MSM.943931 -
Parasites, Hosts and Diseases Feb 2024Ancylostoma ceylanicum is a zoonotic soil-derived nematode that parasitizes the intestines of humans and animals (dogs and cats), leading to malnutrition and...
Ancylostoma ceylanicum is a zoonotic soil-derived nematode that parasitizes the intestines of humans and animals (dogs and cats), leading to malnutrition and iron-deficiency anemia. Helminth parasites secrete calreticulin (CRT), which regulates or blocks the host's immune response. However, no data on A. ceylanicum calreticulin (Ace-CRT) are available. We investigated the biological function of recombinant Ace-CRT (rAce-CRT). rAce-CRT showed reliable antigenicity and stimulated the proliferation of mouse splenocytes and canine peripheral blood mononuclear cells. Quantitative reverse-transcription PCR assays revealed that rAce-CRT primarily promoted the expression of T helper 2 cytokines, particularly IL-13, in canine peripheral blood lymphocytes. rAce-CRT inhibited complement-mediated sheep erythrocyte hemolysis in vitro. Our findings indicate that Ace-CRT plays an immunomodulatory role and may be a promising candidate molecule for a hookworm vaccine.
Topics: Humans; Animals; Dogs; Cats; Mice; Sheep; Ancylostoma; Calreticulin; Cat Diseases; Leukocytes, Mononuclear; Dog Diseases; Immunity
PubMed: 38443772
DOI: 10.3347/PHD.23108 -
Parasitology Research Mar 2024Soil-transmitted helminth (STH) infections inflict disability worldwide, especially in the poorest communities. Current therapeutic options against STHs show limited...
Soil-transmitted helminth (STH) infections inflict disability worldwide, especially in the poorest communities. Current therapeutic options against STHs show limited efficacy, particularly against Trichuris trichiura. The empirical management of patients coming from high-prevalence areas has been suggested for non-endemic areas. This study aimed to describe the management of STH infections in a non-endemic setting using an individualised approach. We performed a retrospective, descriptive study of all patients up to 16 years of age with STH infections attended at an international health unit in a non-endemic area (2014-2018), including all T. trichiura, Necator americanus, Ancylostoma duodenale, and Ascaris lumbricoides infections diagnosed using a formol-ether concentration technique and direct visualisation. Patients were treated according to current international guidelines. Sixty-one stool samples from 48 patients testing positive for STHs were collected, with 96% (46/48) reporting a previous long-term stay in endemic areas. Cure rates with 3-day benzimidazole regimens were 72% for T. trichiura, 40% for hookworms, and 83% for A. lumbricoides. The results were not influenced by any reinfection risk due to the study being performed in a non-endemic area. Patients coming from STH-endemic areas should be evaluated with appropriate diagnostic tools and followed up until cure control results. Cure rates in our cohort were moderate to low, similar to those published in studies in endemic areas. The efficacy of current treatment options is insufficient to recommend a specific empirical approach in high-income countries' healthcare systems.
Topics: Humans; Child; Animals; Global Health; Retrospective Studies; Helminthiasis; Ascariasis; Ancylostoma
PubMed: 38441714
DOI: 10.1007/s00436-024-08171-8 -
Parasitology International Jun 2024Follicular larva migrans (FLM) is a rare and atypical clinical presentation of hookworm-related cutaneous larva migrans (HrCLM). FLM is characterized clinically by... (Review)
Review
Follicular larva migrans (FLM) is a rare and atypical clinical presentation of hookworm-related cutaneous larva migrans (HrCLM). FLM is characterized clinically by follicular, round, small, erythematous papules that are sometimes topped by vesicles or pustules. These lesions are usually located on the abdomen, back, buttocks and thighs and are accompanied by more or less severe pruritus. Some typical and/or short and fragmented tracks may also be visible. FLM is more resistant to anti-helminthic drugs than classical HrCLM: this is likely due to the deep location of larvae in hair follicles. We present two cases of FLM and a review of the literature.
Topics: Animals; Larva Migrans; Anthelmintics; Ancylostomatoidea; Larva
PubMed: 38428565
DOI: 10.1016/j.parint.2024.102872