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

BACKGROUND: 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. METHODS: 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. RESULTS: 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. CONCLUSIONS: 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.

Surveillance of Ancylostoma caninum in naturally infected dogs in Quebec, Canada, and assessment of benzimidazole anthelmintics reveal a variable efficacy with the presence of a resistant isolate in imported dogs.

Ancylostoma caninum is a widely prevalent parasitic nematode in dogs across the world. There has been a notable increase in reports of anthelmintic resistance in A. caninum within the United States of America in recent years, which has led us to investigate the potential of this scenario in Canada. The study objectives were to assess the prevalence of A. caninum in two different groups, including a colony of rescued dogs in Canada and three imported Greyhound dogs from USA, and to evaluate the efficacy of two benzimidazole (BZ) anthelmintics against A. caninum, complemented with a molecular genetic analysis adapted to low prevalence. Fecal samples were collected at pre- and post-treatment with fenbendazole for the native shelters-origin group, and a combination of anthelmintic formulations, including the pro-BZ febantel for the USA-origin group. The coprology analyses found several genera of internal parasites. Canine ancylostomiasis was the most prevalent parasitosis with 30.77% in the native group and 100% in the USA group, but with overall low average of A. caninum eggs per gram. Through the fecal egg count reduction test (FECRT), applying a cut-off at 90% as baseline of egg reduction for successful efficacy, BZ showed variable efficacy. Furthermore, molecular analysis confirmed the presence of A. caninum in both groups of dogs and found differences in the genetics linked to BZ resistance on the A. caninum ß-tubulin isotype 1 gene. In the isolate from the native group, both codons 167 and 200 were homozygous without the presence of single nucleotide polymorphism (SNP). In contrast, the selected isolate from the USA group, showed a homozygous allele at position 200 and a heterozygous SNP at position 167. The latter was congruent with the low efficacy in FECRT and agrees with the recent findings of USA A. caninum isolate resistant phenotype to the BZ anthelmintics. The limitations of the study include an overall low eggs-per-gram in both canine groups, and the shortage of additional fecal samples from the USA group, restraining the molecular analysis only to one out of the three Greyhounds. This study provided some insights on the efficacy of BZs against A. caninum and revealed the presence of BZ resistant isolates in imported dogs in Quebec, Canada. All this information should be considered, for choosing the best strategy in the control of A. caninum using anthelmintic drugs.

Evaluation of heating and liming treatments in sand samples artificially contaminated with Ancylostoma spp. eggs.

Ancylostoma spp. are found worldwide. Infected dog and cat feces can contaminate soil in public places. Despite prophylactic measures being available, studies on direct remediation of Ancylostoma-contaminated soils are scarce. This study aimed to determine the impact of heat treatment and liming on the viability of Ancylostoma spp. eggs in artificially contaminated sandy soil. Sterilized sand samples were contaminated with Ancylostoma spp. eggs extracted from infected dogs' feces. Samples were heated (trial I) to 70 °C or 80 °C, then sieved after 24 hours (212, 90, 38, and 25 µm). Larval cultures were assessed for larval development following heat treatment. Five quicklime concentrations (trial II; 50, 30, 20, 10 and 5%) were used to treat sand. The effect of liming on larval cultures was assessed by measuring embryonic development. Filariform larvae were exposed to 20% quicklime (25 °C and 37 °C, 20 min). Heat treatment destroys Ancylostoma spp. eggs and prevents in vitro larval development. Liming at 50, 30, and 20% concentrations made embryonic development impossible. However, filariform larvae treated with 20% lime solution retained their motility. Heating at 70 °C and liming at 20% were sufficient to make Ancylostoma spp. egg embryogenesis impossible in experimentally contaminated sand samples.

Evaluation of Parasight All-in-One system for the automated enumeration of helminth ova in canine and feline feces.

BACKGROUND:  Digital imaging combined with deep-learning-based computational image analysis is a growing area in medical diagnostics, including parasitology, where a number of automated analytical devices have been developed and are available for use in clinical practice. METHODS: The performance of Parasight All-in-One (AIO), a second-generation device, was evaluated by comparing it to a well-accepted research method (mini-FLOTAC) and to another commercially available test (Imagyst). Fifty-nine canine and feline infected fecal specimens were quantitatively analyzed by all three methods. Since some samples were positive for more than one parasite, the dataset consisted of 48 specimens positive for Ancylostoma spp., 13 for Toxocara spp. and 23 for Trichuris spp. RESULTS: The magnitude of Parasight AIO counts correlated well with those of mini-FLOTAC but not with those of Imagyst. Parasight AIO counted approximately 3.5-fold more ova of Ancylostoma spp. and Trichuris spp. and 4.6-fold more ova of Toxocara spp. than the mini-FLOTAC, and counted 27.9-, 17.1- and 10.2-fold more of these same ova than Imagyst, respectively. These differences translated into differences between the test sensitivities at low egg count levels (< 50 eggs/g), with Parasight AIO > mini-FLOTAC > Imagyst. At higher egg counts Parasight AIO and mini-FLOTAC performed with comparable precision (which was significantly higher that than Imagyst), whereas at lower counts (> 30 eggs/g) Parasight was more precise than both mini-FLOTAC and Imagyst, while the latter two methods did not significantly differ from each other. CONCLUSIONS: In general, Parasight AIO analyses were both more precise and sensitive than mini-FLOTAC and Imagyst and quantitatively correlated well with mini-FLOTAC. While Parasight AIO produced lower raw counts in eggs-per-gram than mini-FLOTAC, these could be corrected using the data generated from these correlations.

Retrospective analysis of canine fecal flotation and coproantigen immunoassay hookworm positive results in Greyhounds and other dog breeds.

Recent studies demonstrated that Greyhounds are commonly infected with Ancylostoma caninum and these infections have been shown to be resistant to anthelmintics. This study evaluated samples submitted to a commercial reference laboratory (IDEXX Laboratories) for canine fecal flotation zinc sulfate centrifugation and coproantigen immunoassay between January 1, 2019, and July 30, 2023 for evidence that Greyhounds were more often positive for Ancylostoma spp. (hookworms) compared to other breeds. The purpose of the study was to determine if Greyhounds were more likely to be hookworm-positive compared to other breeds, if Greyhounds on preventives with efficacy against hookworm infections are more likely to test positive than other breeds, if their infections take longer to resolve, to estimate how long this takes and to assess whether the proportion of hookworm positive tests for all breeds is increasing over time. Records of 25,440,055 fecal results were obtained representing 17,671,724 unique dogs. Of these, 49,795 (∼0.3%) were Greyhounds. The overall odds ratio (OR) of 15.3 (p < 0.001) suggests that Greyhounds are at significantly higher risk than other breeds for hookworm positive float findings, and the OR of 14.3 (p < 0.001) suggests significantly higher risk for hookworm antigen positive results. The median time to negative testing event from the Turnbull distribution estimate was in the interval of 1-2 days for other breeds and 71-72 days for Greyhounds. These results provide evidence that anthelmintic resistant A. caninum strains may be having population-level impacts on the frequency and duration of infections in Greyhounds. The findings have broader health implications beyond Greyhounds as MADR A. caninum strains could spread to other breeds and even pet owners.

Zoonotic Ancylostoma ceylanicum Infection in Coyotes from Guanacaste Conservation Area, Costa Rica, 2021.

Ancylostoma ceylanicum is the second most common hookworm infecting humans in the Asia-Pacific region. Recent reports suggest presence of the parasite in the Americas. We report A. ceylanicum infections in coyotes from the Guanacaste Conservation Area, Costa Rica. Our findings call for active surveillance in humans and animals.

Effect of Soil Temperature on Canine Soil-Transmitted Nematodes in Kandy District with the First Record of Hookworm, Ancylostoma tubaeforme from Sri Lanka.

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, χ2 = 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.

A Case of Zoonotic Ancylostoma ceylanicum Infection in a Suburban Area of Selangor, Malaysia.

INTRODUCTION: The animal hookworm, Ancylostoma ceylanicum, is a dominant hookworm species of dogs and cats. However, it has increasingly been found infecting humans in Southeast Asia. PURPOSE: We report an autochthonous case of A. ceylanicum in a suburban area of Selangor, Malaysia. A 66-year-old Indian lady who is an avid gardener presented with chronic diarrhea of 4 months' duration. METHODS: The patient was examined clinically and colonoscopy was performed. Adult parasites obtained via colonoscopy were subjected to microscopy and molecular investigations. RESULTS: Clinical examinations were unremarkable, and blood investigation revealed normochromic normocytic anemia. Stool occult blood was positive but negative for ova, cyst and adult parasites. Colonoscopy performed showed multiple diverticulae and worm infestation from the terminal ileum to sigmoid colon. Morphological examination on the adult worms showed the specific characteristics of Ancylostoma species. Molecular investigations further confirmed the nematode as Ancylostoma ceylanicum. She was treated with albendazole 400 mg daily for 3 days with symptomatic improvements sustained 3 months later. It is suspected that the patient had ingested or contacted soil contaminated with filariform larvae while gardening. CONCLUSION: Information on the A. ceylanicum infection in humans, especially in urban and suburban areas, is limited, necessitating further epidemiological and clinical studies.

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.

Ancylostoma ceylanicum infections in humans in Vietnam.

Ancylostoma ceylanicum is recognized as the only zoonotic hookworm species that is able to mature into adult stage in the human intestine. While human infections caused by this hookworm species have been reported from neighboring countries and this hookworm is prevalent in dogs in Vietnam, human infection has never been reported in Vietnam. The present study, therefore, aimed to identify human infections with A. ceylanicum in Vietnam. A total of 526 fecal samples from the residents in Long An Province were collected and the presence of hookworm eggs was detected by the Kato-Katz method. The results indicated that the overall prevalence of human hookworm infection was 85/526 (16.2%). After filter paper culture, 3rd stage larvae were successfully obtained from 48 egg-positive samples. The larvae were identified for their species using semi-nested PCR-RLFP on the cox1 gene. As a result, two hookworm species were confirmed; single species infections with Necator americanus or A. ceylanicum, and mixed infections with both species were found in 47.9%, 31.3%, and 20.8% of the samples, respectively.

Endoparasite prevalence and infection risk factors among cats in an animal shelter in Estonia.

Cats are important hosts for different zoonotic parasites that can be hazardous to human health. To date, few studies have attempted to identify the factors affecting parasitic infections in shelter animals. This study aims to analyse the presence of endoparasites in shelter cats in Tartu, Estonia, and identify factors affecting endoparasite prevalence and intensity. The risk factors considered were age, location (urban vs rural cats) and time spent in shelter. In total, 290 faecal samples were collected from cats at an animal shelter in 2015-2016 and investigated for endoparasites using the concentration flotation technique. In total, 138 shelter cats (47.6%) were infected with endoparasites and their overall prevalence was: Toxocara cati (36.6%), Cystoisospora spp. (12.4%), Taeniidae gen. sp. (4.1%), Toxoplasma gondii/Hammondia hammondi (3.4%), Eucoleus aerophilus (2.1%), Cryptosporidium spp. (2.1%), Ancylostoma sp. (0.7%) and Giardia sp. (0.7%). Coinfections occurred in 38 cats (13.1%) most frequently of T. cati and Cystoisospora spp. (4.5%), Cystoisospora spp. and T. gondii/H. hammondi (2.1%). Where species identification of cestode and nematode samples was not possible according to morphology, genetic analysis of the mitochondrial cox1 gene was carried out. DNA was successfully analysed for 6 out of 13 samples that required genetic identification, revealing Ancylostoma tubaeforme in one nematode sample and Hydatigera taeniaeformis in five cestode samples. Cats from rural areas had significantly higher endoparasite prevalence than cats from urban areas. Helminth prevalence decreased to some extent due to anthelmintic treatment in cats available for adoption (held ≥15 days in the shelter), whereas the prevalence of infection with protists increased significantly in these animals. It is important to note that the analysis revealed lower infection intensity for quarantine cats (held 1-14 days in the shelter) compared with cats available for adoption. The relatively high prevalence of endoparasites (including zoonotic) in shelter cats ready for adoption suggests that current anthelminthic procedures require improvements.

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.

Further evaluation and validation of the VETSCAN IMAGYST: in-clinic feline and canine fecal parasite detection system integrated with a deep learning algorithm.

BACKGROUND: Fecal examinations in pet cats and dogs are key components of routine veterinary practice; however, their accuracy is influenced by diagnostic methodologies and the experience level of personnel performing the tests. The VETSCAN IMAGYST system was developed to provide simpler and easier fecal examinations which are less influenced by examiners' skills. This system consists of three components: a sample preparation device, an automated microscope scanner, and analysis software. The objectives of this study were to qualitatively evaluate the performance of the VETSCAN IMAGYST system on feline parasites (Ancylostoma and Toxocara cati) and protozoan parasites (Cystoisospora and Giardia) and to assess and compare the performance of the VETSCAN IMAGYST centrifugal flotation method to reference centrifugal and passive flotation methods. METHODS: To evaluate the diagnostic performance of the scanning and algorithmic components of the VETSCAN IMAGYST system, fecal slides were prepared by the VETSCAN IMAGYST centrifugal flotation technique with pre-screened fecal samples collected from dogs and cats and examined by both an algorithm and parasitologists. To assess the performance of the VETSCAN IMAGYST centrifugal flotation technique, diagnostic sensitivity and specificity were calculated and compared to those of conventional flotation techniques. RESULTS: The performance of the VETSCAN IMAGYST algorithm closely correlated with evaluations by parasitologists, with sensitivity of 75.8-100% and specificity of 93.1-100% across the targeted parasites. For samples with 50 eggs or less per slide, Lin's concordance correlation coefficients ranged from 0.70 to 0.95 across the targeted parasites. The results of the VETSCAN IMAGYST centrifugal flotation method correlated well with those of the conventional centrifugal flotation method across the targeted parasites: sensitivity of 65.7-100% and specificity of 97.6-100%. Similar results were observed for the conventional passive flotation method compared to the conventional centrifugal flotation method: sensitivity of 56.4-91.7% and specificity of 99.4-100%. CONCLUSIONS: The VETSCAN IMAGYST scanning and algorithmic systems with the VETSCAN IMAGYST fecal preparation technique demonstrated a similar qualitative performance to the parasitologists' examinations with conventional fecal flotation techniques. Given the deep learning nature of the VETSCAN IMAGYST system, its performance is expected to improve over time, enabling it to be utilized in veterinary clinics to perform fecal examinations accurately and efficiently.

Field application of a novel multiplex qPCR assay reveals the occurrence of the zoonotic hookworm Ancylostoma braziliense in Nigerian dogs.

A number of gastrointestinal parasites have been reported to infect dogs in Nigeria, some of which have zoonotic potential. Of these, hookworms are the most prevalent, with both Ancylostoma caninum and Uncinaria stenocephala reported in the country. In this study, we subjected 203 hookworm microscopy-positive samples of the 885 individual faecal samples collected from dogs in Nigeria to a recently developed multiplex qPCR for the detection and characterisation of canine hookworm species. The qPCR demonstrated a diagnostic sensitivity of 98% (95% CI 95-99.4) allowing the detection of A. caninum and A. braziliense in 81.3% (165/203, 95% CI 75.3-86.1) and 51.2% (104/203, 95% CI 44.4-58) of the microscopy-positive faecal samples of dogs from Nigeria, respectively and 34.5% (70/203, 95% CI 28.3-41.3) of mixed infections with both hookworm species. The finding of A. braziliense is particularly worrisome given this is a well-known agent of persistent cutaneous larva migrans, commonly referred to as "creeping eruptions" in humans. Although this parasite has been diagnosed in locals and in people travelling in Nigeria suffering from dermatological illnesses, this represents the first molecular identification of A. braziliense in its canine reservoir in the country. These results update the occurrence and distribution of hookworm species affecting dogs in Nigeria highlighting the suitability of the newly developed multiplex qPCR assay as a high-throughput tool for the surveillance of zoonotic hookworms, globally.

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.

Epidemiological aspects of Ancylostoma spp. infection in naturally infected dogs from São Paulo state, Brazil.

Ancylostoma spp. are the most prevalent canine nematode parasites in Brazil. Despite their widespread parasitism in juvenile dogs, infections may occur regardless of host age. Although eosinophilia is a frequent finding in experimental infections, it is uncommon in naturally infected dogs. This study aimed to evaluate the prevalence of Ancylostoma spp. and the eosinophil blood counts (EBC) in naturally infected dogs, with or without comorbidities, admitted to the Veterinary Teaching Hospital of the São Paulo State University (UNESP), School of Veterinary Medicine and Animal Science, Botucatu campus, São Paulo state, Brazil, from 2009 to 2018. All retrospective data were gathered from veterinary medical records (VMR); diagnosis of Ancylostoma infection required the identification of eggs in fecal samples by the simple flotation test (SFT). Ancylostoma-infected animals were evaluated for other intestinal parasitic coinfections (IPC) by either the SFT or the centrifugal-flotation test. Dogs free of any gastro-intestinal parasites were prospectively included in control group (Group C). Ancylostoma-infected animals were defined: Ancylostoma spp. only intestinal parasite infection (Group A), Ancyslostoma spp. with concurrent IPC (Group B), Ancylostoma spp. only intestinal parasite infection with concurrent systemic disorders (Group D), and Ancylostoma spp. with both IPC and concurrent systemic disorders (Group E). The overall prevalence of Ancylostoma spp. was 12.1% (207/1715), that was decreased from 2014 to 2018 (9.7%) relative to the 2009-to-2013 period (13.9%). Prevalence was not significantly different between dogs <1-year-old (10.7% [51/478]) and ≥ 1 year-old (11.7% [130/1109]). IPC was observed in 45.4% (93/205) of the animals positive for Ancylostoma spp., while dogs <1 year old experienced IPC more often (58.8% [30/51]) than dogs ≥1 year old (38.5% [50/130]) (P = 0.02). Group A (n = 35) exhibited median EBC of 1.05 × 109/L, and an eosinophilia ratio of 34.3% that was significantly higher (P < 0.05) than Group C (0.45 × 109/L and 4.1%, respectively). Both variables did not differ in Group B (n = 20), D (n = 39) or E (n = 36) in comparison to Group C (P > 0.05). By ROC curve analysis, only Group A generated a significant area under the curve (0.72). With EBC cutoff of 0.85 × 109 eosinophils/L, sensitivity and specificity were 65.7% and 70.8%, respectively. Eosinophil counts alone may be helpful in raising suspicion of an Ancylostoma spp. infection if further intestinal parasites and concurrent disorders are absent. However, local prevalence data and epidemiological findings should also be evaluated, since eosinophilia is less frequently observed with Ancylostoma spp. infections in the presence of comorbidities.

Molecular identification of Ancylostoma ceylanicum in the Philippines.

Hookworms are some of the most widespread of the soil-transmitted helminths (STH) with an estimated 438.9 million people infected. Until relatively recently Ancylostoma ceylanicum was regarded as a rare cause of hookworm infection in humans, with little public health relevance. However, recent advances in molecular diagnostics have revealed a much higher prevalence of this zoonotic hookworm than previously thought, particularly in Asia. This study examined the prevalence of STH and A. ceylanicum in the municipalities of Palapag and Laoang in the Philippines utilizing real-time polymerase chain reaction (PCR) on stool samples previously collected as part of a cross-sectional survey of schistosomiasis japonica. Prevalence of hookworm in humans was high with 52.8% (n = 228/432) individuals positive for any hookworm, 34.5% (n = 149/432) infected with Necator americanus, and 29.6% (n = 128/432) with Ancylostoma spp; of these, 34 were PCR-positive for A. ceylanicum. Considering dogs, 12 (n = 33) were PCR-positive for A. ceylanicum. This is the first study to utilize molecular diagnostics to identify A. ceylanicum in the Philippines with both humans and dogs infected. Control and elimination of this zoonotic hookworm will require a multifaceted approach including chemotherapy of humans, identification of animal reservoirs, improvements in health infrastructure, and health education to help prevent infection.

Clinical helminthiases in Thailand border regions show elevated prevalence levels using qPCR diagnostics combined with traditional microscopic methods.

BACKGROUND: Under-regulated national borders in Southeast Asia represent potential regions for enhanced parasitic helminth transmission and present barriers to helminthiasis disease control. METHODS: Three Thailand border regions close to Myanmar, Laos and Cambodia were surveyed for clinical parasitic helminth disease. In-field microscopy was performed on stools from 567 individuals. Sub-samples were transported to Bangkok for molecular analysis comprising three multiplex qPCR assays. RESULTS: The overall helminth infection prevalence was 17.99% as assessed by Kato-Katz and 24.51% by qPCR. The combined prevalence of the two methods was 28.57%; the most predominant species detected were Opisthorchis viverrini (18.34%), hookworm (6.88%; Ancylostoma spp. and Necator americanus), Ascaris lumbricoides (2.29%) and Trichuris trichiura (1.76%). CONCLUSIONS: These data demonstrate the value of molecular diagnostics for determining more precise prevalence levels of helminthiases in Southeast Asia. Availability of such accurate prevalence information will help guide future public health initiatives and highlights the need for more rigorous surveillance and timely intervention in these regions.

Evaluation of the VETSCAN IMAGYST: an in-clinic canine and feline fecal parasite detection system integrated with a deep learning algorithm.

BACKGROUND: Fecal examination is an important component of routine companion animal wellness exams. Sensitivity and specificity of fecal examinations, however, are influenced by sample preparation methodologies and the level of training and experience of personnel who read fecal slides. The VETSCAN IMAGYST system consists of three components: a sample preparation device, a commercially available scanner, and an analysis software. The VETSCAN IMAGYST automated scanner and cloud-based, deep learning algorithm, locates, classifies, and identifies parasite eggs found on fecal microscopic slides. The main study objectives were (i) to qualitatively evaluate the capabilities of the VETSCAN IMAGYST screening system and (ii) to assess and compare the performance of the VETSCAN IMAGYST fecal preparation methods to conventional fecal flotation techniques. METHODS: To assess the capabilities of VETSCAN IMAGYST screening components, fecal slides were prepared by the VETSCAN IMAGYST centrifugal and passive flotation techniques with 100 pre-screened fecal samples collected from dogs and cats and examined by both the algorithm and parasitologists. To determine the diagnostic sensitivity and specificity of the VETSCAN IMAGYST sample preparation techniques, fecal flotation slides were prepared by four different techniques (VETSCAN IMAGYST centrifugal and passive flotations, conventional centrifugal flotation, and passive flotation using OVASSAY® Plus) and examined by parasitologists. Additionally, required sample preparation and scanning times were estimated on a subset of samples to evaluate VETSCAN IMAGYST ease-of-use. RESULTS: The algorithm performance of the VETSCAN IMAGYST closely matched that of the parasitologists, with Pearson's correlation coefficient (r) ranging from 0.83-0.99 across four taxa of parasites, Ancylostoma, Toxocara, Trichuris and Taeniidae. Both VETSCAN IMAGYST centrifugal and passive flotation methods correlated well with conventional preparation methods on all targeted parasites (diagnostic sensitivity of 75.8-100%, specificity of 91.8-100%, qualitative agreement between methods of 93.8-94.5%). Sample preparation, slide scan and image analysis were completed within 10-14 min by VETSCAN IMAGYST centrifugal and passive flotations, respectively. CONCLUSIONS: The VETSCAN IMAGYST scanning system with the VETSCAN IMAGYST sample preparation methods demonstrated a qualitative match in comparison to the results of parasitologists' examinations with conventional fecal flotation techniques. The VETSCAN IMAGYST is an easy-to-use, next generation qualitative and possibly quantitative diagnostic platform that brings expert clinical results into the hands of veterinary clinics.