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Parasites & Vectors Jul 2023Helminth infections are an important public health problem in humans and have an even greater impact on domestic animal and livestock welfare. Current readouts for...
BACKGROUND
Helminth infections are an important public health problem in humans and have an even greater impact on domestic animal and livestock welfare. Current readouts for anthelmintic drug screening assays are stage development, migration, or motility that can be subjective, laborious, and low in throughput. The aim of this study was to apply and optimize a fluorometric technique using resazurin for evaluating changes in the metabolic activity of Ascaris suum third-stage larvae (L3), a parasite of high economic relevance in swine.
METHODS
Ascaris suum L3 were mechanically hatched from 6- to 8-week embryonated and sucrose-gradient-enriched eggs. Resazurin dye and A. suum L3 were titrated in 96-well microtiter plates, and resazurin reduction activity was assessed by fluorometry after 24 h of incubation. Fluorescence microscopy was used to localize the resazurin reduction site within the larvae. Finally, we exposed A. suum L3 to various stress conditions including heat, methanol, and anthelmintics, and investigated their impact on larval metabolism through resazurin reduction activity.
RESULTS
We show that the non-fluorescent dye resazurin is reduced inside vital A. suum L3 to fluorescent resorufin and released into the culture media. Optimal assay parameters are 100-1000 L3 per well, a resazurin concentration of 7.5 µg/ml, and incubation at 37 °C/5% CO for 24 h. An intact L2 sheath around the L3 of A. suum completely prevents the uptake of resazurin, while in unsheathed L3, the most intense fluorescence signal is observed along the larval midgut. L3 exposed to methanol or heat show a gradually decreased resazurin reduction activity. In addition, 24 h exposure to ivermectin at 0.625 µM, mebendazole at 5 µM, and thiabendazole from 10 to 100 µM significantly decreased larval metabolic activity by 55%, 73%, and 70% to 89%, respectively.
CONCLUSIONS
Together, our results show that both metabolic stressors and anthelmintic drugs significantly and reproducibly reduce the resazurin reduction activity of A. suum L3, making the proposed assay a sensitive and easy-to-use method to evaluate metabolic activity of A. suum L3 in vitro.
Topics: Humans; Animals; Swine; Ascaris suum; Methanol; Anthelmintics; Xanthenes; Ascariasis; Larva
PubMed: 37468906
DOI: 10.1186/s13071-023-05871-5 -
Genome Research Dec 2017Programmed DNA elimination is a developmentally regulated process leading to the reproducible loss of specific genomic sequences. DNA elimination occurs in unicellular... (Comparative Study)
Comparative Study
Programmed DNA elimination is a developmentally regulated process leading to the reproducible loss of specific genomic sequences. DNA elimination occurs in unicellular ciliates and a variety of metazoans, including invertebrates and vertebrates. In metazoa, DNA elimination typically occurs in somatic cells during early development, leaving the germline genome intact. Reference genomes for metazoa that undergo DNA elimination are not available. Here, we generated germline and somatic reference genome sequences of the DNA eliminating pig parasitic nematode and the horse parasite In addition, we carried out in-depth analyses of DNA elimination in the parasitic nematode of humans, and the parasitic nematode of dogs, s. Our analysis of nematode DNA elimination reveals that in all species, repetitive sequences (that differ among the genera) and germline-expressed genes (approximately 1000-2000 or 5%-10% of the genes) are eliminated. Thirty-five percent of these eliminated genes are conserved among these nematodes, defining a core set of eliminated genes that are preferentially expressed during spermatogenesis. Our analysis supports the view that DNA elimination in nematodes silences germline-expressed genes. Over half of the chromosome break sites are conserved between and , whereas only 10% are conserved in the more divergent Analysis of the chromosomal breakage regions suggests a sequence-independent mechanism for DNA breakage followed by telomere healing, with the formation of more accessible chromatin in the break regions prior to DNA elimination. Our genome assemblies and annotations also provide comprehensive resources for analysis of DNA elimination, parasitology research, and comparative nematode genome and epigenome studies.
Topics: Alternative Splicing; Animals; Ascaridoidea; Ascaris suum; Chromosome Breakage; Chromosome Breakpoints; DNA, Helminth; Evolution, Molecular; Female; Genome; Germ-Line Mutation; Male; Molecular Sequence Annotation; Nematoda; RNA, Helminth; Repetitive Sequences, Nucleic Acid; Sequence Analysis, DNA; Sex Chromosomes; Telomere; Toxocara canis; Transcriptome
PubMed: 29118011
DOI: 10.1101/gr.225730.117 -
PLoS Neglected Tropical Diseases Apr 2020The scientific community has recently summarized the desired characteristics for diagnostic tools across the different phases of a soil-transmitted helminth (STH) mass... (Comparative Study)
Comparative Study
Evaluation of copromicroscopy and serology to measure the exposure to Ascaris infections across age groups and to assess the impact of 3 years of biannual mass drug administration in Jimma Town, Ethiopia.
BACKGROUND
The scientific community has recently summarized the desired characteristics for diagnostic tools across the different phases of a soil-transmitted helminth (STH) mass drug administration (MDA) program. Although serology meets some of the desired criteria, there is a scarcity of data on baseline serological profiles in human populations, both prior to and during MDA programs.
METHODS
In this study, we compared the copromicroscopic and the serological infection profiles in 600 school-aged children (SAC) and 600 adults at the advent of the MDA program in Jimma Town, Ethiopia. The serological profiles were examined by two ELISAs that measure IgG4 responses to the Ascaris suum haemoglobin antigen (AsHb) and a somatic extract of lung stage larvae (AsLungL3). Three years into the MDA program, we sampled another group of 600 SAC from the same schools to assess the reduction in prevalence and intensity of Ascaris infections measured by copromicroscopy and serology.
PRINCIPAL FINDINGS
Prior to the start of MDA, copromicroscopy revealed an Ascaris prevalence of 31.0% and a mean fecal egg count of 2,919 eggs per gram (EPG) in SAC. Following three years of biannual treatment, the prevalence reduced to 13.2% (57.8% reduction) and the mean fecal egg count to 1,513 EPG (48.1% reduction). This reduction was also reflected in the serological results. The seroprevalence reduced with 40.9% and 27.4% and the mean optical density ratio reduced with 44.2% and 38.2% as measured by the AsHb or AsLungL3 ELISA respectively. We also showed that, despite a decreasing coproprevalence, seroprevalence to Ascaris increased with age.
CONCLUSIONS
This study is the first to provide IgG4 response profiles of an endemic population to two different A. suum antigens. The results suggest that exposure to the infectious stages of Ascaris reaches beyond SAC alone. Furthermore, it highlights the possible use of serological assays to monitor changes in STH exposure during MDA programs.
Topics: Adolescent; Adult; Aged; Aged, 80 and over; Animals; Antibodies, Helminth; Antigens, Helminth; Ascariasis; Ascaris suum; Child; Child, Preschool; Cross-Sectional Studies; Drug Monitoring; Enzyme-Linked Immunosorbent Assay; Ethiopia; Feces; Female; Humans; Immunoglobulin G; Male; Mass Drug Administration; Microscopy; Middle Aged; Prevalence; Serologic Tests; Treatment Outcome; Young Adult
PubMed: 32282815
DOI: 10.1371/journal.pntd.0008037 -
Frontiers in Veterinary Science 2022Parasite infection is a common problem in organic pig production, which can compromise health and growth of pigs, threaten food safety of pork products, and cause...
Parasite infection is a common problem in organic pig production, which can compromise health and growth of pigs, threaten food safety of pork products, and cause economic losses to organic farmers. To develop management strategies for controlling parasites, we evaluated intestinal parasite infection in pigs at different ages and of different sexes, and investigated whether parasite infection influences growth performance and carcass traits in a cross-sectional study. Fecal samples were collected from pigs ( = 298) raised under near-organic standards during nursery, growing, finishing, and gestating phases for analysis of fecal egg counts (FEC) of , and spp. eggs were not detected in the feces of nursery pigs. Eggs of were found in 45%, 74%, and 0% of fecal samples of growing pigs, finishing pigs, and gestating sows, respectively, after false-positive adjustment ( < 0.001). Mean FEC of was higher in infected finishing pigs than in infected growing pigs [2,502 vs. 724 eggs per gram (epg), < 0.001]. No differences in percent of positive samples or FEC of were detected between sexes. Growth performance and carcass traits were not different between non-infected pigs and those infected with . All pigs ( = 32) examined at slaughter had white spots on the liver, and 78% harbored worms. eggs were not detected in any fecal samples. Eggs of spp. were found in 7%, 0%, 1%, and 9% of fecal samples of nursery pigs, growing pigs, finishing pigs, and gestating sows, respectively, with a maximum FEC of 40 epg in all age groups. These results indicate was the predominant parasite infecting growing and finishing pigs in the herds studied. To control infection, future research should investigate the efficacy of treating pigs with organically approved anthelmintics during the growing phase of production.
PubMed: 35677931
DOI: 10.3389/fvets.2022.911561 -
The Cochrane Database of Systematic... Jun 2022It is estimated that 1.5 billion people are infected with soil-transmitted helminths (STHs) worldwide. Re-infection occurs rapidly following deworming, and interruption... (Meta-Analysis)
Meta-Analysis Review
BACKGROUND
It is estimated that 1.5 billion people are infected with soil-transmitted helminths (STHs) worldwide. Re-infection occurs rapidly following deworming, and interruption of transmission is unlikely without complementary control efforts such as improvements in water, sanitation, and hygiene (WASH) access and behaviours.
OBJECTIVES
To assess the effectiveness of WASH interventions to prevent STH infection.
SEARCH METHODS
We used standard, extensive Cochrane search methods. The latest search date was 19 October 2021.
SELECTION CRITERIA
We included interventions to improve WASH access or practices in communities where STHs are endemic. We included randomized controlled trials (RCTs), as well as trials with an external control group where participants (or clusters) were allocated to different interventions using a non-random method (non-RCTs). We did not include observational study designs. Our primary outcome was prevalence of any STH infection. Prevalence of individual worms was a secondary outcome, including for Ascaris lumbricoides, Trichuris trichiura, hookworm (Ancylostoma duodenale or Necator americanus), or Strongyloides stercoralis. Intensity of infection, measured as a count of eggs per gram of faeces for each species, was another secondary outcome.
DATA COLLECTION AND ANALYSIS
Two review authors independently reviewed titles and abstracts and full-text records for eligibility, performed data extraction, and assessed risk of bias using the Cochrane risk of bias assessment tool for RCTs and the EPOC tool for non-RCTs. We used a random-effects meta-analysis to pool study estimates. We used Moran's I² statistic to assess heterogeneity and conducted subgroup analyses to explore sources of heterogeneity. We assessed the certainty of the evidence using the GRADE approach.
MAIN RESULTS
We included 32 studies (16 RCTs and 16 non-RCTs) involving a total of 52,944 participants in the review. Twenty-two studies (14 RCTs (16 estimates) and eight non-RCTs (11 estimates)) reported on our primary outcome, prevalence of infection with at least one STH species. Twenty-one studies reported on the prevalence of A lumbricoides (12 RCTs and 9 non-RCTs); 17 on the prevalence of T trichiura (9 RCTs and 8 non-RCTs); 18 on the prevalence of hookworm (10 RCTs and 8 non-RCTs); and one on the prevalence of S stercoralis (1 non-RCT). Sixteen studies measured the intensity of infection for an individual STH type. Ten RCTs and five non-RCTs reported on the intensity of infection of A lumbricoides; eight RCTs and five non-RCTs measured the intensity of infection of T trichiura; and eight RCTs and five non-RCTs measured the intensity of hookworm infection. No studies reported on the intensity of infection of S stercoralis. The overall pooled effect estimates showed that the WASH interventions under study may result in a slight reduction of any STH infection, with an odds ratio (OR) of 0.86 amongst RCTs (95% confidence interval (CI) 0.74 to 1.01; moderate-certainty evidence) and an OR of 0.71 amongst non-RCTs (95% CI 0.54 to 0.94; low-certainty evidence). All six of the meta-analyses assessing individual worm infection amongst both RCTs and non-RCTs had pooled estimates in the preventive direction, although all CIs encapsulated the null, leaving the possibility of the null or even harmful effects; the certainty of the evidence ranged from very low to moderate. Individual studies assessing intensity of infection showed mixed evidence supporting WASH. Subgroup analyses focusing on narrow specific subsets of water, sanitation, and hygiene interventions did very little to elucidate which interventions might be better than others. Data on intensity of infection (e.g. faecal egg count) were reported in a variety of ways across studies, precluding the pooling of results for this outcome. We did not find any studies reporting adverse events resulting from the WASH interventions under study or from mass drug administration (MDA).
AUTHORS' CONCLUSIONS
Whilst the available evidence suggests that the WASH interventions under study may slightly protect against STH infection, WASH also serves as a broad preventive measure for many other diseases that have a faecal oral transmission route of transmission. As many of the studies were done in addition to MDA/deworming (i.e. MDA was ongoing in both the intervention and control arm), our data support WHO recommendations for implementation of improvements to basic sanitation and adequate access to safe water alongside MDA. The biological plausibility for improved access to WASH to interrupt transmission of STHs is clear, but WASH interventions as currently delivered have shown impacts that were lower than expected. There is a need for more rigorous and targeted implementation research and process evaluations in order that future WASH interventions can better provide benefit to users. Inconsistent reporting of the intensity of infection underscores the need to define the minimal, standard data that should be collected globally on STHs to enable pooled analyses and comparisons.
Topics: Animals; Ascaris lumbricoides; Humans; Hygiene; Observational Studies as Topic; Sanitation; Soil; Water
PubMed: 35726112
DOI: 10.1002/14651858.CD012199.pub2 -
Pathogens and Global Health Sep 2022The role of various parasitic infections in the occurrence of appendicitis is illustrated through cases recorded all over the world. The purpose of the current study was... (Meta-Analysis)
Meta-Analysis
The role of various parasitic infections in the occurrence of appendicitis is illustrated through cases recorded all over the world. The purpose of the current study was to estimate the global prevalence of parasite infestation (other than ) in appendectomy specimens.In the setting of the PRISMA guidelines, multiple databases (Science Direct, Scopus, Web of Science, PubMed, and Google Scholar) were explored in articles published until 28 September 2020. Totally, 62 studies (106 datasets) with 77, 619 participants were included in the analysis.The pooled prevalence of parasites in appendectomy samples was as follows; 0.012% (95% CI; 0.004-0.025) for , 0.004% (95% CI; 0.001-0.009) for , 0.025% (95% CI; 0.007-0.052) for , 0.002% (95% CI; 0.001-0.005) for spp., 0.061% (95% CI; 0.020-0.122) for and 0.034% (95% CI; 0.018-0.056) for .Our results demonstrated that the risk of appendicitis may increase in the presence of helminth and protozoan infections. As such, the most cases of parasites in appendectomy specimens were reported in developing countries. Regular screening plans for diagnosis, treatment and prevention are needed for prevention of parasitic infection as well as parasitic associated appendicitis, especially in endemic regions of the world.
Topics: Animals; Appendicitis; Ascaris lumbricoides; Feces; Humans; Intestinal Diseases, Parasitic; Parasites; Prevalence; Public Health
PubMed: 34842078
DOI: 10.1080/20477724.2021.2008701 -
Transactions of the Royal Society of... Jan 2017The morbidity due to Ascaris lumbricoides and Trichuris trichiura is caused by infections of moderate and heavy intensity while hookworm infections of all intensities... (Meta-Analysis)
Meta-Analysis Review
The morbidity due to Ascaris lumbricoides and Trichuris trichiura is caused by infections of moderate and heavy intensity while hookworm infections of all intensities are recognized to cause morbidity. This study aims to evaluate the effect of repeated rounds of preventive chemotherapy on the proportion of soil-transmitted helminth (STH) infections causing morbidity. We identified studies from 17 countries, reporting changes in the proportion of STH infection causing morbidity between baseline and follow-up. In the studies identified, the average proportion of individuals with STH infections of moderate and heavy intensity was of 14% at baseline and was on average reduced to 2% by the intervention (i.e., 85% reduction). There was an average reduction of 73% after the first year of treatment, which reached almost 80% after 5 years and over 95% in 10 years of deworming interventions. The reduction in hookworm prevalence was 57% after 12 months reaching 78% after 5 years. We consider the results presented in this study especially useful for decision makers as it demonstrates the effectiveness of preventive chemotherapy in reducing STH prevalence and morbidity. We encourage the implementation of deworming programs to achieve the goal, set by WHO for 2020, to eliminate STH morbidity in children.
Topics: Ancylostomatoidea; Animals; Ascariasis; Ascaris lumbricoides; Chemoprevention; Child; Female; Global Health; Helminthiasis; Hookworm Infections; Humans; Male; Soil; Trichuriasis; Trichuris
PubMed: 28340144
DOI: 10.1093/trstmh/trx011 -
PLoS Neglected Tropical Diseases May 2016Soil-transmitted helminths (STH) have acute and chronic manifestations, and can result in lifetime morbidity. Disease burden is difficult to quantify, yet quantitative... (Meta-Analysis)
Meta-Analysis Review
BACKGROUND
Soil-transmitted helminths (STH) have acute and chronic manifestations, and can result in lifetime morbidity. Disease burden is difficult to quantify, yet quantitative evidence is required to justify large-scale deworming programmes. A recent Cochrane systematic review, which influences Global Burden of Disease (GBD) estimates for STH, has again called into question the evidence for deworming benefit on morbidity due to STH. In this narrative review, we investigate in detail what the shortfalls in evidence are.
METHODOLOGY/PRINCIPAL FINDINGS
We systematically reviewed recent literature that used direct measures to investigate morbidity from STH and we critically appraised systematic reviews, particularly the most recent Cochrane systematic review investigating deworming impact on morbidity. We included six systematic reviews and meta-analyses, 36 literature reviews, 44 experimental or observational studies, and five case series. We highlight where evidence is insufficient and where research needs to be directed to strengthen morbidity evidence, ideally to prove benefits of deworming.
CONCLUSIONS/SIGNIFICANCE
Overall, the Cochrane systematic review and recent studies indicate major shortfalls in evidence for direct morbidity. However, it is questionable whether the systematic review methodology should be applied to STH due to heterogeneity of the prevalence of different species in each setting. Urgent investment in studies powered to detect direct morbidity effects due to STH is required.
Topics: Anemia; Animals; Anthelmintics; Ascariasis; Ascaris lumbricoides; Chronic Disease; Cost of Illness; Helminthiasis; Humans; Prevalence; Soil
PubMed: 27196100
DOI: 10.1371/journal.pntd.0004566 -
Revista Brasileira de Parasitologia... 2023The aim of the present study was to assess morphologic and genetic data on ascariasis in swine (Sus scrofa domesticus) and humans in low-resource rural and periurban...
The aim of the present study was to assess morphologic and genetic data on ascariasis in swine (Sus scrofa domesticus) and humans in low-resource rural and periurban communities in the state of Piauí, Brazil. Our cross-sectional survey included 100 fecal samples obtained from swine and 682 samples from humans. Fifteen pigs were necropsied. Human and porcine fecal samples were examined to identify Ascaris eggs. Parasites obtained in the swine necropsies were studied using scanning electron microscopy (SEM), and the mitochondrial gene encoding the cytochrome oxidase 1 (cox1) enzyme was partially amplified and sequenced for molecular taxonomy and phylogenetic analyses. The overall prevalence of Ascaris eggs in the swine fecal samples was 16/100 (16%). No Ascaris eggs were identified in the human fecal samples. SEM of six worms recovered from pigs demonstrated morphological characteristics of A. suum. Cox1 sequences were compatible with A. suum reference sequences. Original and reference (GenBank) nucleotide sequences were organized into clusters that did not segregate the parasites by host species or and region. The largest haplogroups were dominated by haplotypes H01, H02 and H31. In the communities studied, there was no epidemiological evidence of the zoonotic transmission of ascariasis at the human-swine interface.
Topics: Humans; Animals; Swine; Ascaris suum; Ascariasis; Phylogeny; Brazil; Cross-Sectional Studies; Ascaris; Swine Diseases
PubMed: 37851714
DOI: 10.1590/S1984-29612023057 -
Emerging Infectious Diseases Apr 2024We describe a case of a 2-year-old child who expelled a single adult female Ascaris lumbricoides worm. The patient is from a rural county in Mississippi, USA, with no...
We describe a case of a 2-year-old child who expelled a single adult female Ascaris lumbricoides worm. The patient is from a rural county in Mississippi, USA, with no reported travel outside of the United States. The caregivers in the home practice good sanitation. Exposure to domestic pigs is the likely source of infection.
Topics: Swine; Adult; Animals; Humans; Female; Child, Preschool; Mississippi; Ascariasis; Ascaris lumbricoides; Sus scrofa; Travel
PubMed: 38526329
DOI: 10.3201/eid3004.240176