Comparison of the complete filtration method using an automated feces analyzer with three manual methods for stool examinations.

Conventional diagnostic techniques using manual methods for stool examination have important limitations. Hence there is a need for improved technologies in routine clinical practice. This study aimed to compare detection rates, agreements, and diagnostic performances for stool examinations in all parameters of the complete filtration method using the Sciendox Feces Analysis System-50 automated feces analyzer with three manual methods, the direct smear, Kato's thick smear, and formalin ethyl concentration techniques. The 252 routine stool samples were examined for parasites, white blood cells (WBCs), red blood cells (RBCs), fat globules, and yeast cells using the four methods indicated above, and the complete filtration detection rates, Cohen's kappa (κ), and diagnostic performances were evaluated and compared. The detection rates of RBCs, fat globules, and yeast cells examined by the complete filtration automated method were comparable to the manual methods, but the detection rates of parasites and WBCs were significantly lower. Most methods detected the same seven parasite species, Ascaris lumbricoides, hookworm, Trichuris trichiura, Strongyloides stercoralis, Entamoeba histolytica/dispar, Blastocystis spp., and Giardia intestinalis. Pairwise agreements between the complete filtration and other methods were good to very good for all parameters showing κ values of 0.74 to 0.89. The diagnostic performances against the combined results showed complete filtration method sensitivities of 70%, 81.82%, 77.27%, 100%, and 95% for parasites, WBCs, RBCs, fat globules, and yeast cells, respectively, while the complete filtration negative predictive values (NPVs) and accuracies showed higher than 95% for all parameters. The complete filtration method using the automated feces analyzer showed high NPVs and accuracies, and good agreements with the three tested manual methods for stool examination in all parameters.

Important Nematodes in Children.

Intestinal nematode infections caused by soil-transmitted helminths (STH), such as the roundworm Ascaris lumbricoides, the whipworm Trichuris trichiura, and the hookworms Ancylostoma duodenale, and Necator americanus, infect more than 1 billion people throughout the world. School-aged children tend to harbor the greatest numbers of intestinal worms, and as a result, experience more adverse health consequences, such as poor growth, anemia, and cognitive decline. Clinicians should maintain a high degree of suspicion in endemic areas when patients present with surgical abdomens, particularly children. Current antihelminthic drugs are moderately effective, but reinfection is possible. Global efforts are needed to eradicate STH infections.

Mobile microscopy and telemedicine platform assisted by deep learning for the quantification of Trichuris trichiura infection.

Soil-transmitted helminths (STH) are the most prevalent pathogens among the group of neglected tropical diseases (NTDs). The Kato-Katz technique is the diagnosis method recommended by the World Health Organization (WHO) although it often presents a decreased sensitivity in low transmission settings and it is labour intensive. Visual reading of Kato-Katz preparations requires the samples to be analyzed in a short period of time since its preparation. Digitizing the samples could provide a solution which allows to store the samples in a digital database and perform remote analysis. Artificial intelligence (AI) methods based on digitized samples can support diagnosis by performing an objective and automatic quantification of disease infection. In this work, we propose an end-to-end pipeline for microscopy image digitization and automatic analysis of digitized images of STH. Our solution includes (a) a digitization system based on a mobile app that digitizes microscope samples using a 3D printed microscope adapter, (b) a telemedicine platform for remote analysis and labelling, and (c) novel deep learning algorithms for automatic assessment and quantification of parasitological infections by STH. The deep learning algorithm has been trained and tested on 51 slides of stool samples containing 949 Trichuris spp. eggs from 6 different subjects. The algorithm evaluation was performed using a cross-validation strategy, obtaining a mean precision of 98.44% and a mean recall of 80.94%. The results also proved the potential of generalization capability of the method at identifying different types of helminth eggs. Additionally, the AI-assisted quantification of STH based on digitized samples has been compared to the one performed using conventional microscopy, showing a good agreement between measurements. In conclusion, this work has presented a comprehensive pipeline using smartphone-assisted microscopy. It is integrated with a telemedicine platform for automatic image analysis and quantification of STH infection using AI models.

Factors associated with soil-transmitted helminths infection in Benin: Findings from the DeWorm3 study.

BACKGROUND: Despite several years of school-based MDA implementation, STH infections remain an important public health problem in Benin, with a country-wide prevalence of 20% in 2015. The DeWorm3 study is designed to assess the feasibility of using community-based MDA with albendazole to interrupt the transmission of STH, through a series of cluster-randomized trials in Benin, India and Malawi. We used the pre-treatment baseline survey data to describe and analyze the factors associated with STH infection in Comé, the study site of the DeWorm3 project in Benin. These data will improve understanding of the challenges that need to be addressed in order to eliminate STH as a public health problem in Benin. METHODS: Between March and April 2018, the prevalence of STH (hookworm spp., Ascaris and Trichuris trichiura) was assessed by Kato-Katz in stool samples collected from 6,153 residents in the community of Comé, Benin using a stratified random sampling procedure. A standardized survey questionnaire was used to collect information from individual households concerning factors potentially associated with the presence and intensity of STH infections in pre-school (PSAC, aged 1-4), school-aged children (SAC, aged 5-14) and adults (aged 15 and above). Multilevel mixed-effects models were used to assess associations between these factors and STH infection. RESULTS: The overall prevalence of STH infection was 5.3%; 3.2% hookworm spp., 2.1% Ascaris lumbricoides and 0.1% Trichuris. Hookworm spp. were more prevalent in adults than in SAC (4.4% versus 2.0%, respectively; p = 0.0001) and PSAC (4.4% versus 1.0%, respectively; p<0.0001), whilst Ascaris lumbricoides was more prevalent in SAC than in adults (3.0% versus 1.7%, respectively; p = 0.004). Being PSAC (adjusted Odds Ratio (aOR) = 0.2, p< 0.001; adjusted Infection Intensity Ratio (aIIR) = 0.1, p<0.001) or SAC (aOR = 0.5, p = 0.008; aIIR = 0.3, p = 0.01), being a female (aOR = 0.6, p = 0.004; aIIR = 0.3, p = 0.001), and having received deworming treatment the previous year (aOR = 0.4, p< 0.002; aIIR = 0.2, p<0.001) were associated with a lower prevalence and intensity of hookworm infection. Lower income (lowest quintile: aOR = 5.0, p<0.001, 2nd quintile aOR = 3.6, p = 0.001 and 3rd quintile aOR = 2.5, p = 0.02), being a farmer (aOR = 1.8, p = 0.02), medium population density (aOR = 2.6, p = 0.01), and open defecation (aOR = 0.5, p = 0.04) were associated with a higher prevalence of hookworm infection. Lower education-no education, primary or secondary school- (aIIR = 40.1, p = 0.01; aIIR = 30.9, p = 0.02; aIIR = 19.3, p = 0.04, respectively), farming (aIIR = 3.9, p = 0.002), natural flooring (aIIR = 0.2, p = 0.06), peri-urban settings (aIIR = 6.2, 95%CI 1.82-20.90, p = 0.003), and unimproved water source more than 30 minutes from the household (aIIR = 13.5, p = 0.02) were associated with a higher intensity of hookworm infection. Improved and unshared toilet was associated with lower intensity of hookworm infections (aIIR = 0.2, p = 0.01). SAC had a higher odds of Ascaris lumbricoides infection than adults (aOR = 2.0, p = 0.01) and females had a lower odds of infection (aOR = 0.5, p = 0.02). CONCLUSION: Hookworm spp. are the most prevalent STH in Comé, with a persistent reservoir in adults that is not addressed by current control measures based on school MDA. Expanding MDA to target adults and PSAC is necessary to substantially impact population prevalence, particularly for hookworm. TRIAL REGISTRATION: ClinicalTrials.gov NCT03014167.

First isolation of Trichuris from wild blue sheep (Pseudois nayaur) in the Helan Mountains, China.

Four adult female worms of Trichuris were isolated from an individual of the wild blue sheep (Pseudois nayaur) inhabiting the Helan Mountains, China, during an epidemiological survey of this wild ruminant. Although there were some differences among the worms in posterior end (rectum) morphology and egg shape, little information regarding species status could be inferred from their morphology. Phylogenetic trees were constructed based on sequences of the ITS1 segment of ribosomal RNA (rRNA), and the sequences of the four Trichuris specimens from wild blue sheep were divided into two distinct lineages (Clade A and Clade B). The two specimens in Clade A were named Genotype I, and had the closest relationship with Trichuris skrjabini; the two specimens in Clade B were named Genotype II and had the closest genetic relationship with a previously described Trichuris sp. In the two Trichuris genotypes identified in the present study, the 18S fragments (261 to 262 bp) of the newly obtained sequences were found to be highly conserved, with merely one insert mutation of a single nucleotide present. The genetic distance based on ITS1 between members of Clade A, composed of two T. skrjabini individuals and two Genotype I individuals, ranged from 0 to 0.0034. These distances are within the intraspecies variation of Trichuris (0-0.0272), suggesting that the Genotype I individuals infesting the wild blue sheep were T. skrjabini. In Clade B, the newly obtained sequences clustered with Trichuris sp. specimens isolated from ruminants (sheep and black goat) with strong support, and the genetic distance ranged from 0.0068 to 0.017, which is also within the intraspecies variation of Trichuris (0-0.0272). However, the genetic distances between the Clade A and Clade B were 0.0442 to 0.0578, which are higher than the intraspecies distances in Trichuris but lower than the interspecies distances (0.102-0.5078). These results implied that Clade A and Clade B most likely represent two subpopulations of T. skrjabini; however, the possibility that Clade A is T. skrjabini and Clade B is a Candidatus Trichuris could not be excluded.

Comparison of the egg recovery rates and limit of detection for soil-transmitted helminths using the Kato-Katz thick smear, faecal flotation and quantitative real-time PCR in human stool.

BACKGROUND: Monitoring the success of soil-transmitted helminth (STH) control programs relies on accurate diagnosis and quantitative assessment of infection prevalence and intensity. As preventative chemotherapeutic program coverage for STH expands, the necessity of gaining insights into the relative or comparative sensitivities, in terms of limits of detection (LOD) and egg-recovery-rates (ERR) for microscopy and quantitative polymerase chain reaction qPCR-based diagnostic techniques becomes imperative to inform suitability for their intended use for large scale STH monitoring and treatment efficacy studies. METHODOLOGY/PRINCIPAL FINDINGS: The diagnostic performance in terms of ERR and LOD of the Kato-Katz (KK) thick smear technique, sodium nitrate (NaNO3) faecal floatation (FF) and qPCR for the accurate detection and enumeration of STH eggs were calculated and expressed in eggs per gram (EPG), by experimentally seeding parasite-free human faeces with Ascaris spp., Trichuris spp. and Necator americanus eggs representing low, medium and high intensity infections. The efficiency of NaNO3 flotation was also calculated over a range of specific gravities (SpGr) for the optimum recovery of STH eggs. FF of SpGr 1.30 recovered 62.7%, 11% and 8.7% more Trichuris spp., Necator americanus and Ascaris spp. eggs respectively, than the recommended SpGr of 1.20. All diagnostic methods demonstrated strong direct correlation to the intensity of seeded EPG. KK and FF (SpGr 1.30) resulted in significant lower ERRs compared to qPCR (p <0.05). qPCR demonstrated significantly (p <0.05) greater sensitivity with an ability to detect as little as 5 EPG for all three STH, compared to 50 EPG by KK and FF (SpGr 1.30). CONCLUSIONS/SIGNIFICANCE: This study compares the diagnostic parameters in terms of LOD and ERRs of STHs for the KK, FF and qPCR. These results indicate that the diagnostic performance of qPCR assays should be considered by control programs in the phase that aims to seek confirmation of transmission break and cessation of preventive chemotherapy in low-transmission settings, in line with the control targets of the WHO neglected tropical diseases 2030 Roadmap.

Two Helminthic Cases of Human Mummy Remains from Joseon-Period Graves in Korea.

Our previous research on coprolite specimens from the mummies of Joseon Dynasty (1392-1910 CE) has revealed various species of parasite eggs. Herein, we added 2 new helminthic cases of human remains from Joseon-period graves in the Republic of Korea (Korea). The organic materials precipitated on the hip bones of 2 half-mummied cases (Goryeong and Gwangmyeong cases) were collected, rehydrated, and examined by a microscope. In the sample from Goryeong-gun (gun=County), ova of Trichuris trichiura, Clonorchis sinensis, and Metagonimus spp. were detected, and eggs of T. trichiura and A. lumbricoides were found from the sample of Gwangmyeong-si (si=City). By adding this outcome to the existing data pool, we confirm our previous estimates of Joseon-period parasite infection rates. The overall rates of A. lumbricoides, T. trichiura, and C. sinensis decreased dramatically from Joseon to the modern period. In Goryeong mummy specimen, we also found Metagonimus spp. eggs that has rarely been detected in archaeological samples so far.

Epidemiology of soil transmitted helminths and risk analysis of hookworm infections in the community: Results from the DeWorm3 Trial in southern India.

Since 2015, India has coordinated the largest school-based deworming program globally, targeting soil-transmitted helminths (STH) in ~250 million children aged 1 to 19 years twice yearly. Despite substantial progress in reduction of morbidity associated with STH, reinfection rates in endemic communities remain high. We conducted a community based parasitological survey in Tamil Nadu as part of the DeWorm3 Project-a cluster-randomised trial evaluating the feasibility of interrupting STH transmission at three geographically distinct sites in Africa and Asia-allowing the estimation of STH prevalence and analysis of associated factors. In India, following a comprehensive census, enumerating 140,932 individuals in 36,536 households along with geospatial mapping of households, an age-stratified sample of individuals was recruited into a longitudinal monitoring cohort (December 2017-February 2018) to be followed for five years. At enrolment, a total of 6089 consenting individuals across 40 study clusters provided a single adequate stool sample for analysis using the Kato-Katz method, as well as answering a questionnaire covering individual and household level factors. The unweighted STH prevalence was 17.0% (95% confidence interval [95%CI]: 16.0-17.9%), increasing to 21.4% when weighted by age and cluster size. Hookworm was the predominant species, with a weighted infection prevalence of 21.0%, the majority of which (92.9%) were light intensity infections. Factors associated with hookworm infection were modelled using mixed-effects multilevel logistic regression for presence of infection and mixed-effects negative binomial regression for intensity. The prevalence of both Ascaris lumbricoides and Trichuris trichiura infections were rare (<1%) and risk factors were therefore not assessed. Increasing age (multivariable odds ratio [mOR] 21.4, 95%CI: 12.3-37.2, p<0.001 for adult age-groups versus pre-school children) and higher vegetation were associated with an increased odds of hookworm infection, whereas recent deworming (mOR 0.3, 95%CI: 0.2-0.5, p<0.001) and belonging to households with higher socioeconomic status (mOR 0.3, 95%CI: 0.2-0.5, p<0.001) and higher education level of the household head (mOR 0.4, 95%CI: 0.3-0.6, p<0.001) were associated with lower odds of hookworm infection in the multilevel model. The same factors were associated with intensity of infection, with the use of improved sanitation facilities also correlated to lower infection intensities (multivariable infection intensity ratio [mIIR] 0.6, 95%CI: 0.4-0.9, p<0.016). Our findings suggest that a community-based approach is required to address the high hookworm burden in adults in this setting. Socioeconomic, education and sanitation improvements alongside mass drug administration would likely accelerate the drive to elimination in these communities. Trial Registration: NCT03014167.

Human density, economic level and frequency of canine helminths in Buenos Aires.

The aim of this study was to explore through cross-sectional study the variation in the prevalence of parasitic helminths in canine faeces collected from green spaces of Buenos Aires according to the human density (HD) and economic level (EL) in the surroundings. HD and EL were considered as independent variables with three categories each. Twenty public squares (one hectare of surface) were randomly selected for each existing combination of the two independent variables. Ten random samples of fresh canine faeces were obtained in each square and analysed for helminths by the sedimentation and flotation techniques. The prevalence for each of the species was analysed using generalized linear models (GLM). The prevalence was modelled with a binomial error distribution and a logit link function. Helminth eggs were detected in 45 out of the 200 (22.5%) faecal samples collected and in 18 of the 20 green spaces sampled. The species observed were Ancylostoma caninum (13% of samples), Trichuris vulpis (8%) and Toxocara canis (4.5%). The GLM indicated that the prevalence of A. caninum in the slum areas (very high HD and very low EL) was higher than that in the other areas studied. However, the HD seemed to contribute more than the EL to the variations in the prevalence of A. caninum in faecal samples. The GLM showed no differences in the prevalence of the other parasite species for the different levels of the independent variables.

A 3-year retrospective analysis of canine intestinal parasites: fecal testing positivity by age, U.S. geographical region and reason for veterinary visit.

BACKGROUND: Canine life stage is a key factor in parasite prevalence as clinical signs associated with parasitism are more common in pups. In adult dogs, health status and geographical region may also play a role in parasite prevalence. The purpose of this study was to evaluate fecal test results using zinc sulfate flotation by centrifugation combined with fecal antigen testing for hookworms (Ancylostoma spp. Uncinaria stenocephala), ascarids (Toxocara canis, Toxascaris spp., Baylisascaris spp.) and whipworms (Trichuris vulpis) sorted by age, geographical region and veterinary visit type. METHODS: A retrospective sample of intestinal parasite panels submitted to IDEXX Laboratories from 1,626,104 individual dogs were selected from the continental USA from 1 January 2017 to 31 December 2019. These data contain results from fecal exams performed using zinc sulfate flotation by centrifugation paired with coproantigen immunoassay results for hookworms, ascarids, whipworms and Giardia (Fecal Dx® with Giardia coproantigen immunoassay plate). For paired testing, if either the coproantigen assay or flotation test was positive, the sample was considered to be positive. Data were summarized by age category, U.S. Census Bureau geographical region (Northeast, South, Midwest, West) and veterinary visit type. Visit types were subdivided into Wellness Visits and Other Clinical Visits in which a fecal sample was submitted. RESULTS: In dogs presenting for either Wellness Visits or Other Clinical Visits in which Giardia testing was included, Giardia had the highest positivity (combined results for microscopy and coproantigen: 12.2 and 10.8%, respectively), followed by hookworms (combined microscopy and coproantigen: 4.1 and 4.2%, respectively), ascarids (combined microscopy and coproantigen: 2.5 and 1.7%, respectively) and whipworms (combined microscopy and coproantigen: 1.1 and 1.4%, respectively). When all test results were pooled together, pups aged 2-6 months were observed to have the highest proportion of positive results by either microscopy or coproantigen immunoassay regardless of clinical visit type. Parasite positivity varied by geographical region. Regardless of visit type, age or geographical region, the coproantigen method was observed to find a higher proportion of positive test results than microscopy in Giardia, ascarids, hookworms and whipworms. CONCLUSIONS: The Fecal Dx® coproantigen immunoassay combined with the zinc sulfate flotation by centrifugation method uncovers a higher number of positive hookworm, ascarid and whipworm infections than zinc sulfate flotation alone in both pups and adult dogs across all geographical regions of the USA regardless of visit type.

α-Gal specific-IgE prevalence and levels in Ecuador and Kenya: Relation to diet, parasites, and IgG4.

BACKGROUND: IgE to α-Gal is a cause of mammalian meat allergy and has been linked to tick bites in North America, Australia, and Eurasia. Reports from the developing world indicate that α-Gal sensitization is prevalent but has been little investigated. OBJECTIVE: We sought evidence for the cause(s) of α-Gal sensitization and lack of reported meat allergy among children in less developed settings in Ecuador and Kenya. METHODS: IgE to α-Gal and total IgE were assessed in children from Ecuador (n = 599) and Kenya (n = 254) and compared with children with (n = 42) and without known (n = 63) mammalian meat allergy from the southeastern United States. Information on diet, potential risk factors, and helminth infections was available for children from Ecuador. IgG4 to α-Gal and antibodies to regionally representative parasites were assessed in a subset of children. RESULTS: In Ecuador (32%) and Kenya (54%), α-Gal specific IgE was prevalent, but levels were lower than in children with meat allergy from the United States. Sensitization was associated with rural living, antibody markers of Ascaris exposure, and total IgE, but not active infections with Ascaris or Trichuris species. In Ecuador, 87.5% reported consuming beef at least once per week, including 83.9% of those who had α-Gal specific IgE. Levels of α-Gal specific IgG4 were not high in Ecuador, but were greater than in children from the United States. CONCLUSIONS: These results suggest that in areas of the developing world with endemic parasitism, α-Gal sensitization is (1) common, (2) associated with Ascaris exposure, and (3) distinguished by a low percentage of specific/total IgE compared with individuals with meat allergy in the United States.

Profile of endoparasites in dairy cattle in the microregion of São João del-Rei, state of Minas Gerais, Brazil / [Perfil de endoparasitas em bovinos leiteiros na microrregião de São João del-Rei, Minas Gerais, Brasil]

The present study provides the first evaluation of the endoparasite profile in dairy cattle from the microregion of São João del-Rei, Minas Gerais, Brazil, including seasonal variations in parasitic infections. Fecal samples were collected directly from rectum of 123 dairy cows and 160 calves on 15 farms. These were kept in insulated boxes and sent for immediate analysis. Using eggs per gram of feces (EPG) we found the following prevalences: Strongyloidea 43.5%, Moniezia sp. 7.6% and both Strongyloides sp. and Trichuris spp. 2.2%, in dairy cows. In calves, the prevalence was Strongyloidea 50.9%, Moniezia sp. 5.6% and Strongyloides sp. 1.9%. The third-stage larvae recovered from coprocultures from cows and calves consisted mainly of Cooperia spp., Haemonchus spp. and Ostertagia spp. EPG counts were significantly higher in calves. Regarding seasonality, the EPG counts and prevalence of Moniezia sp. tended to be higher in the dry period, whereas those of Strongyloidea were higher in the rainy season. Evidence suggests that parasite control and prophylaxis were inadequately managed on the farms studied here. The present parasitological profile correlates with those found by other similar studies, indicating that the main problem is still the inadequate management of parasite control and prophylaxis by farmers.(AU)

O presente estudo representa a primeira avaliação do perfil de endoparasitos em bovinos leiteiros da microrregião de São João del-Rei, Minas Gerais, Brasil, e das variações sazonais das infecções parasitárias. Foram avaliadas amostras fecais de 123 vacas e 160 bezerros de 15 propriedades rurais, coletadas diretamente do reto, mantidas em caixas térmicas isoladas e analisadas imediatamente. A contagem de ovos por grama de fezes (OPG) revelou prevalências de 43,5% de Strongyloidea, 7,6% de Moniezia sp. e 2,2% de Strongyloides sp. e Trichuris spp. em vacas leiteiras. As prevalências de OPG em bezerros foram 50,9% de Strongyloidea, 5,6% de Moniezia sp. e 1,9% de Strongyloides sp. As larvas do terceiro estádio recuperadas de coproculturas de vacas e bezerros foram principalmente de Cooperia spp., Haemonchus spp. e Ostertagia spp. OPG dos parasitos foram significativamente maiores em bezerros. Em relação à sazonalidade, OPG e a prevalência de Moniezia sp. tenderam a ser maiores durante a estação seca, enquanto OPG de Strongyloidea foram maiores na estação chuvosa. Com base nos resultados, é possível concluir que o manejo antiparasitário nas propriedades estudadas não é realizado de maneira plenamente satisfatória. O presente perfil parasitológico se assemelha a outros encontrados em estudos semelhantes, indicando que o maior problema ainda é o manejo antiparasitário inadequado realizado nas propriedades de bovinos leiteiros.(AU)

Gastrointestinal parasite burden in 4th-5th c. CE Florence highlighted by microscopy and paleogenetics.

The study of ancient parasites, named paleoparasitology, traditionally focused on microscopic eggs disseminated in past environments and archaeological structures by humans and other animals infested by gastrointestinal parasites. Since the development of paleogenetics in the early 1980s, few paleoparasitological studies have been based on the ancient DNA (aDNA) of parasites, although such studies have clearly proven their utility and reliability. In this paper, we describe our integrative approach for the paleoparasitological study of an ancient population from Florence in Italy, dated to the 4th-5th c. CE. The first stage consisted in the study of sediment samples from the pelvic area of 18 individuals under light microscopy. This allowed us to detect Ascarid-type eggs belonging very probably to the human-infesting roundworm Ascaris lumbricoides. Ten subsamples were selected corresponding to five individuals, and we extracted their whole DNA following sediment aDNA protocols. A targeted approach allowed us to detect two nematodes and one trematode aDNA fragments, namely Ascaris sp., Trichuris trichiura, and Dicrocoelium dendriticum. Among the five individuals tested for microscopic eggs and aDNA, three of them showed the remains of eggs (only Ascarid-type), but all of them tested positive to the presence of at least one parasite aDNA. Microscopic diagnosis first guided our research for the selection of promising samples while the targeted aDNA approach significantly improved our knowledge in terms of parasitic diversity and frequency in this population subgroup. These results enabled us to discuss the possible impact of latent parasitism in this past population at the time of an epidemic, as suggested in Florence. In particular, the singular case of D. dendriticum detection is discussed in light of the present-day scarcity of genuine human infections. Nevertheless, actual infections are known in the paleoparasitological record, and food habits may have led to false parasitism in this historical context. aDNA leaching from overlying strata may also explain this detection. This study strongly pleads for a systematic integrative approach combining microscopy and aDNA in paleoparasitology.

An unpleasant souvenir: Endoscopic finding of Trichuris trichiura (Nematoda: Trichuridae).

Whipworms are responsible for up to 500 million cases of trichuriasis worldwide, with higher endemicity in tropical and sub-tropical countries. In non-endemic countries, trichuriasis can be accidentally diagnosed upon colonoscopy, often in the presence of negative microscopy. Here, we describe an incidental diagnosis of trichuriasis in an HIV patient residing in a non-endemic area (i.e., Turin, Italy), six months after his return from Antigua. The species-level diagnosis was made thanks to PCR-based molecular identification of Trichuris sp. following optical microscopy detection. Overall, this case highlights the importance of improving parasitic diseases diagnosis through cutting-edge clinical and laboratory diagnostic tools alongside advanced training of specialists in the area of parasitology.

Occurrence of gastrointestinal parasites in dogs and cats domiciliated in Santos, SP, Brazil / Ocorrência de parasitos gastrintestinais em cães e gatos domiciliados em Santos, SP, Brasil

Abstract Helminths and protozoa are major causes of diseases in domestic animals, and many can also cause infections in humans. Knowledge of intestinal parasitoses affecting domestic animals is important for the implementation of appropriate preventive measures. The objective of this study was to evaluate the occurrence of gastrointestinal parasites in fecal samples of dogs and cats attended at the Veterinary Hospital of the Metropolitan University of Santos, SP, Brazil. We also attempted to determine whether such infection was associated with sex, age, or the presence of diarrhea. We analyzed 100 fecal samples: 85 from dogs and 15 from cats. Among the dogs, 31.8% of the samples were positive, and to 40.0% among the cats. Infection was not associated with sex or age. However, among the dogs, parasitism showed a significant association with the presence of diarrhea (P = 0.013). The helminths Ancylostoma spp. and the protozoa Giardia duodenalis were the most frequent parasites in this research. Although they present unknown species and assemblages, they are parasites with a zoonotic potential of great importance in public health. Therefore, it is essential that pets are properly diagnosed and treated against gastrointestinal parasitic infection to prevent the spread of diseases.

Resumo As enfermidades causadas por helmintos e protozoários representam uma das principais causas de doenças em animais domésticos, e muitos desses parasitos podem causar infecções em seres humanos. O conhecimento das enteroparasitoses que acometem os animais domésticos é de suma importância para que medidas preventivas adequadas sejam implementadas. O objetivo do presente trabalho foi avaliar a frequência de ocorrência de parasitos gastrointestinais em amostras de fezes de cães e gatos atendidos no Hospital Veterinário da Universidade Metropolitana de Santos, bem como sua associação com o sexo, a idade e a presença de diarreia. Do total das amostras de cães analisadas, 31,8% estavam positivas, em relação aos gatos, e 40% apresentaram positividade. Não houve associação entre o sexo e a idade, porém, foi observada associação significativa entre a presença da parasitose e da diarreia (p=0,013) entre os cães. O helminto Ancylostoma spp. e o protozoário Giardia duodenalis foram os mais frequentes na pesquisa. Embora apresentem espécies e "assemblages" desconhecidas, são parasitos de potencial zoonóticos de grande importância em saúde pública. Assim, é essencial que os animais de companhia sejam corretamente diagnosticados e tratados contra infecções parasitárias gastrintestinais para evitar a propagação de doenças.

Helminth infections among rural schoolchildren in Southern Ethiopia: A cross-sectional multilevel and zero-inflated regression model.

Although the prevalence of helminths infection among schoolchildren is known, there has been little progress in the application of count model for modelling the risk factors of helminths egg. Only a few studies applied multilevel analysis to explore the variation in helminths prevalence across schools and classes. This study aimed to assess the prevalence, intensity of helminths infection, and identify risk factors at the individual-, household-, and school-level among schoolchildren in Southern Ethiopia. Using multistage random sampling, we recruited 864 students in the Wonago District. We applied multilevel-logistic and zero-inflated negative binomial regression models (ZINB). Risk factors were concentrated at the individual level; school-level and class-level variables explained less than 5% of the variance. The overall helminths prevalence was 56% (479/850); Trichuris trichiura prevalence was 42.4% (360/850); and Ascaris lumbricoides prevalence was 18.7% (159/850). The rate of any helminths increased among thin children (AOR: 1.73 [95% CI: (1.04, 2.90]), anemic (AOR: 1.45 [95% CI: 1.04, 2.03]), mothers who had no formal education (AOR: 2.08 [95% CI: 1.25, 3.47]), and those in households using open containers for water storage (AOR: 2.06 [95% CI: 1.07, 3.99]). In the ZINB model, A. lumbricoides infection intensity increased with increasing age (AOR: 1.08 [95% CI: 1.01, 1.16]) and unclean fingernails (AOR: 1.47 [95% CI: 1.07, 2.03]). Handwashing with soap (AOR: 0.68 [95% CI: 0.48, 0.95]), de-worming treatment [AOR: 0.57 (95% CI: 0.33, 0.98)], and using water from protected sources [AOR: 0.46 (95% CI: 0.28, 0.77)] were found to be protective against helminths infection. After controlling for clustering effects at the school and class levels and accounting for excess zeros in fecal egg counts, we found an association between helminths infection and the following variables: age, thinness, anemia, unclean fingernails, handwashing, de-worming treatment, mother's education, household water source, and water storage protection. Improving hygiene behavior, providing safe water at school and home, and strengthening de-worming programs is required to improve the health of schoolchildren in rural Gedeo.

Epidemiology of soil-transmitted helminth infections in Semarang, Central Java, Indonesia.

Soil-transmitted helminth (STH) infections are endemic in Indonesia. However, prevalence data for many parts of the country are incomplete. The aim of this study was to determine human STH prevalence and knowledge and practices relating to STH risk behaviour, to provide a current view of the status of STH infection in rural communities in Central Java. A cross-sectional survey of 16 villages was conducted in Semarang, Central Java in 2015. Demographic and household data together with information about knowledge and practices relating to STH and hygiene were elicited through face-to-face interviews. Stool samples were collected and examined using the flotation method. Children (aged 2-12 years) also had their haemoglobin (Hb) levels, height and weight data collected, and BMI estimated. Data were analysed using univariate logistic regression analysis. A total of 6,466 individuals with a mean age of 33.5 years (range: 2-93) from 2,195 households were interviewed. The overall prevalence of STH was 33.8% with Ascaris lumbricoides (roundworm) the predominant nematode identified (prevalence = 26.0%). Hookworm and Trichuris trichiura (whipworm) were found in 7.9% and 1.8% of participants, respectively. Females were at increased odds of infection with A. lumbricoides (adjusted OR 1.14, 95% CI [1.02-1.29], p = 0.02). Adults in age groups 51-60 and over 60 years had the highest odds of being infected with hookworm (adjusted OR 3.01, 95% CI [1.84-4.91], p<0.001 and adjusted OR 3.79, 95% CI [2.30-6.26], p<0.001, respectively) compared to 6-12 year olds. Farmers also had higher odds of being infected with hookworm (adjusted OR 2.36, 95% CI [1.17-4.76], p = 0.02) compared to other occupation categories. Poverty (OR 2.14, 95% CI [1.77-2.58], p<0.001), overcrowding (OR 1.35, 95% CI [1.27-1.44], p<0.001), goat ownership (OR 1.61, 95% CI [1.10-2.41], p = 0.02) and the presence of dry floor space in the home (OR 0.73, 95% CI [0.58-0.91], p = 0.01) were all household factors significantly associated with an increased odds of infection. Infection with STH was not significantly associated with the gastrointestinal illness (p>0.05), BMI or Hb levels; however, one third of all 2-12 year olds surveyed were found to be anaemic (i.e. Hb concentrations below 110g/l or 115g/l for children under 5 and 5 years or older, respectively), with a greater proportion of school-age children at risk. Knowledge and behaviour related to hygiene and gastrointestinal diseases varied widely and were generally not associated with STH infection. The study revealed that STH infection remains endemic in Central Java despite ongoing deworming programs. Current control efforts would benefit from being re-evaluated to determine a more effective way forward.

Hand hygiene intervention to optimize helminth infection control: Design and baseline results of Mikono Safi-An ongoing school-based cluster-randomised controlled trial in NW Tanzania.

INTRODUCTION: Soil transmitted helminths (STH) can affect over 50% of children in some parts of Tanzania. Control measures involve annual deworming campaigns in schools, but re-infection is rapid. This paper presents the design and baseline survey results of an ongoing school-based cluster-randomised controlled trial in Kagera region, NW Tanzania. The trial aims to determine whether the effect of routine deworming on the prevalence of Ascaris lumbricoides and Trichuris trichiura infections among school aged children can be sustained when combined with a behaviour change intervention promoting handwashing with water and soap. METHODS: As part of the trial, a total of 16 schools were randomised to receive the intervention (N = 8) or as controls (N = 8). Randomisation was stratified per district and restricted to ensure pre-trial STH prevalence was balanced between study arms. The combination intervention to be tested comprises class-room based teacher-led health education, improvement of handwash stations, coloured nudges to facilitate handwashing and parental engagement sessions. The impact evaluation involves two cross-sectional surveys conducted at baseline and endline. The objectives of the baseline survey were: (i) to confirm whether the deworming campaign was successful, and identify and treat students still infected about 2 weeks after deworming, (ii) to document any baseline differences in STH prevalence between trial arms, and (iii) to assess handwashing behaviours, and access to water and sanitation at school and home. We randomly sampled 35 students per class in Grades 1-6 (an average of 200 children per school), stratified to ensure equal representation between genders. Assenting students were interviewed using a structured questionnaire and asked to provide a stool specimen. RESULTS: Results of the baseline survey conducted about 2 weeks after deworming shows balanced demographic and STH prevalence data across trial arms. We observed a low prevalence of ascariasis (< 5%) as expected; however, the prevalence of trichuriasis was still about 35% in both arms. CONCLUSION: The randomisation procedure was successful in achieving a balanced distribution of demographic characteristics and helminth infections between trial arms. The intervention is being rolled out. The current deworming treatment regimen may need to be revised with regards to the treatment of trichuriasis.

Serious limitations of the current strategy to control Soil-Transmitted Helminths and added value of Ivermectin/Albendazole mass administration: A population-based observational study in Cameroon.

BACKGROUND: Soil-transmitted helminth (STH) infections remain a public health concern in sub-Saharan Africa. School-based mass drug administration (MDA) using the anthelminthic drug Mebendazole/Albendazole have succeeded in controlling morbidity associated to these diseases but failed to interrupt their transmission. In areas were filarial diseases are co-endemic, another anthelminthic drug (Ivermectin) is distributed to almost the entire population, following the community-directed treatment with ivermectin (CDTI) strategy. Since Ivermectin is a broad spectrum anthelmintic known to be effective against STH, we conducted cross-sectional surveys in two health districts with very contrasting histories of Ivermectin/Albendazole-based PC in order to investigate whether CDTI might have contributed in STH transmission interruption. METHODOLOGY: Cross-sectional surveys were conducted in two health districts with similar socio-environmental patterns but with very contrasting CDTI histories (Akonolinga health district where CDTI was yet to be implemented vs. Yabassi health district where CDTI has been ongoing for two decades). Stool samples were collected from all volunteers aged >2 years old and analyzed using the Kato-Katz technique. Infections by different STH species were compared between Akonolinga and Yabassi health districts to decipher the impact of Ivermectin/Albendazole-based MDA on STH transmission. PRINCIPAL FINDINGS: A total of 610 and 584 participants aged 2-90 years old were enrolled in Akonolinga and Yabassi health districts, respectively. Two STH species (Ascaris lumbricoides and Trichuris trichiura) were found, with prevalence significantly higher in Akonolinga health district (43.3%; 95% CI: 38.1-46.6) compared to Yabassi health district (2.5%; 95% CI: 1.1-5.1) (chi-square: 90.8; df: 1; p < 0.001). CONCLUSION/SIGNIFICANCE: These findings (i) suggest that Mebendazole- or Albendazole-based MDA alone distributed only to at-risk populations might not be enough to eliminate STH, (ii) support the collateral impact of Ivermectin/Albendazole MDA on A. lumbricoides and T. trichiura infections, and (iii) suggest that Ivermectin/Albendazole-based PC could accelerate STH transmission interruption.

Trichuris trichiura isolated from Macaca sylvanus: morphological, biometrical, and molecular study.

BACKGROUND: Recent studies have reported the existence of a Trichuris species complex parasitizing primate. Nevertheless, the genetic and evolutionary relationship between Trichuris spp. parasitizing humans and Non-Human Primates (NHP) is poorly understood. The hypothesised existence of different species of Trichuris in primates opens the possibility to evaluate these primates as reservoir hosts of human trichuriasis and other putative new species of whipworms. RESULTS: In this paper, we carried out a morphological, biometrical and molecular study of Trichuris population parasitizing Macaca sylvanus from Spain based on traditional morpho-biometrical methods, PCA analysis and ribosomal (ITS2) and mitochondrial (cox1 and cob) DNA sequencing. Morphological results revealed that Trichuris sp. from M. sylvanus is Trichuris trichiura. Ribosomal datasets revealed that phylogenetic relationships of populations of Trichuris sp. from M. sylvanus were unresolved. The phylogeny inferred on mitochondrial datasets (partitioned and concatenated) revealed similar topologies; Thus, phylogenetic trees supported the existence of clear molecular differentiation between individuals of Trichuris sp. from M. sylvanus appearing in two different subclades. CONCLUSIONS: Based on morphological parameters, biometrical measurements, and molecular sequence analysis, we conclude that the whipworms isolated from M. sylvanus were T. trichiura. Further, the evolutionary relationship showed that these worms belonged to two genotypes within the T. trichiura lineage. Since T. trichiura is of public health importance, it is important to carry out further studies to improve the understanding of its hosts range, evolution and phylogeography.