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For the majority of intestinal parasites, real-time PCR-based diagnosis outperforms microscopy. However, the data for Trichuris trichiura have been less convincing and most comparative studies have been performed in populations with low prevalence. This study aims to improve detection of T. trichuria DNA in human stool by evaluating four sample preparation methods. Faecal samples (n = 60) were collected at Flores island, Indonesia and examined by microscopy. Aliquots were taken and a bead-beating procedure was used both on directly frozen stool and on material preserved with 96% ethanol. PCR on frozen samples showed 40% to be positive for T. trichiura, compared with 45% positive by microscopy. The percentage positive increased when using ethanol preservation (45·0%), bead-beating (51·7%) and a combination (55·0%) and all three methods showed significantly higher DNA loads. The various procedures had a less pronounced effect on the PCR results of nine other parasite targets tested. Most prevalent were Ascaris lumbricoides (≈60%), Necator americanus (≈60%), Dientamoeba fragilis (≈50%) and Giardia lamblia (≈12%). To validate the practicality of the procedure, bead-beating was applied in a population-based survey testing 910 stool samples. Findings confirmed bead-beating before DNA extraction to be a highly efficient procedure for the detection of T. trichiura DNA in stool.

Background The rapid growth in off-leash dog parks provides opportunity for canine socialization activities but carries risk of exposure to intestinal parasites. This study assessed the prevalence of these infections in dogs visiting off-leash dog parks. Methods Fresh defecations were collected from dogs visiting parks in 30 metropolitan areas across the USA. Samples were analyzed by coproantigen immunoassay (CAI) (Fecal Dx® and Giardia Test, IDEXX Laboratories, Inc.) and zinc sulfate centrifugal flotation (CF). Owners responded to a questionnaire on their dog’s signalment and use of heartworm/intestinal parasite control medications (HWCM). Results Samples were examined from 3006 dogs, 87.9% aged at least 12 months, visiting 288 parks. At least one intestinal parasite was detected in 622 (20.7%) samples, nematodes in 263 (8.8%), with hookworms, whipworms and ascarids in 7.1, 1.9 and 0.6% of samples, respectively. A sample positive for one or more intestinal parasites was found in 245 (85.1%) parks, with nematodes found in 143 (49.7%). Combined, CAI and CF detected 78.4% more intestinal nematode infections than CF alone. Hookworm and whipworm infections were detected in all age groups, but ascarids were only detected in dogs less than 4 years-old. Approximately 42% of dogs aged less than 1 year were positive for nematodes or Giardia . Based on owner reports, HWCM was current for 68.8% of dogs, dogs previously diagnosed with intestinal parasitism were more likely to be receiving a HWCM than those without such history, and a significantly lower ( P  = 0.0003) proportion of dogs receiving a HWCM were positive for intestinal nematodes compared with those not on such medication. Conclusions Intestinal parasites, the most common of which were Giardia , Ancylostoma caninum and Trichuris vulpis , were found in 20% of dogs and 85% of dog parks across the USA. Enhanced detection of canine intestinal parasitism was achieved by combining CF and CAI. Canine intestinal parasites are common across the USA and dog health can be improved by regular testing of fecal samples and routine administration of medications effective against the most common infections.

Background Intestinal parasitic infections are still one of the major health concerns in developing countries. Monitoring of intestinal parasitic infection and associated risk factors are essential for intervention strategies. Therefore, the aim of this study was to assess the prevalence of intestinal parasitic infection and associated risk factors among students at Dona Berber primary school, Bahir Dar, Ethiopia. Methods School based cross-sectional study was conducted among students at Dona Berber primary school from October 2015 to June 2016. Three hundred fifty nine students were involved in the study by providing stool specimens and detailed personal information. Students were selected by stratified and systematic random sampling method. Fresh stool samples were collected from each student and processed by formal-ether fecal concentration technique. Data were analyzed using SPSS version 20.0 statistical software and p value <0.05 were used as statistically significant. Results Among the 359 students participated in the study, 235 (65.5%) were infected by one or more intestinal parasites. The rates of single and double parasitic infections among students were 49.6% and 16.2%, respectively. The most prevalent parasite detected in the study was E. histolytica/dispar (24.5%) followed by hookworm (22.8%). Among the different variables assessed in the study, family size of 6 (AOR = 4.90; 95% CI, 2.03–11.83), irregularly shoe wearing habit (AOR = 2.85; 95% CI, 1.53–5.32) and unclean finger nail (AOR = 3.68; 95% CI, 1.87–7.26) were independently predict intestinal parasitic infections. Student drinking well water (AOR = 2.51; 95% CI, 2.30–4.86) and unclean finger nail (AOR = 4.42; 95% CI, 2.55–7.65) were strongly associated with E. histolytica/dispar infection. Likewise, irregular shoe wearing habit (AOR = 14.13; 95% CI, 7.06–28.29) was strongly associated with hookworm infections. Conclusion High prevalence of intestinal parasitic infection among the study participants demands improvement of health education, environmental sanitation and quality of water sources.

Dientamoeba fragilis (D. fragilis) is an intestinal parasite frequently detected in humans with abdominal pain and diarrhoea, but it is also commonly found in asymptomatic subjects. Hence its clinical relevance is often disputed. The introduction of polymerase chain reaction (PCR) is a versatile and sensitive diagnostic technique for the detection of intestinal parasites, and in some Western world countries PCR has almost completely replaced microscopic diagnostics. PCR has however resulted in an increase in the number of D. fragilis-positive patients. The disputed pathogenic nature of this intestinal parasite and an apparent increase in the incidence of patients with positive PCR results have renewed the discussions between clinicians and microbiologists on how to deal with an infected patient. Moreover, treatment guidelines differ throughout the world which makes it difficult for clinicians to choose an optimal therapeutic regimen.

Introduction: Intestinal parasites are known to cause infection in humans worldwide, with higher prevalence in low- and middle- incoming countries. Children are greatly affected leading to malnutrition and subsequently to physical and cognitive development impairment. Despite the scale and importance of this issue, there are few studies conducted in Mozambique concerning parasitic intestinal infections in hospitalized children. To our knowledge this is the first published report with data on this subject from Northern Mozambique. Methodology: A cross-sectional study was conducted in 2012 and 2013 in 831 children, attending the Central Hospital of Nampula in Northern Mozambique. One single stool sample was obtained from each child. Socio-demographic and clinical data were also obtained. Parasitological analysis of feces was performed through direct examination and Ritchie concentration technique and Giardia duodenalis antigen detection by rapid immunochromatographic test. Modified Ziehl-Neelsen staining was used for coccidia detection. Results: The global prevalence of pathogenic intestinal parasites was 31.6%. G. duodenalis (23.9%) was by far the most prevalent parasite followed by Strongyloides stercoralis (4.1%) and Cryptosporidium sp. (3.4%). Intestinal parasites were more frequent in older children (p = 0.005; aOR = 1.025). Conclusions: This work is one of the few published studies reporting intestinal parasites infection in hospitalized children. The percentage of children affected with G. duodenalis is higher than found in other studies in the African region. This highlights the need of particular attention being given to this intestinal protozoan and its resistance to water treatment, as well as to environmental health and personal hygiene.

Determining the optimal number of stool samples for accurate parasite detection is crucial in healthcare. However, previous studies have reported conflicting results. This study investigated factors associated with the detection of pathogenic intestinal parasites in additional stool microscopy examinations. This retrospective cross-sectional study at a tertiary care hospital outpatient clinic included patients who submitted three stool samples between 2012 and 2021 and had at least one pathogenic intestinal parasite-positive stool sample. Diagnostic yields for one, two, and three stool specimens were the primary outcome. The secondary outcome was the identification of factors associated with the time to the first positive stool microscopy result, which was analysed using ordinal logistic regression. Among the 103 infected patients, compared with the infection detection rate in the first specimen, the rate increased with the second specimen and further increased with the third specimen, achieving a cumulative detection rate of 100%. Some parasites, such as hookworms, were easily detected in the first sample. However, more than half the patients infected with Trichuris trichiura and all patients infected with Isospora belli were missed if only one stool specimen was collected. Immunocompetent hosts were significantly more likely to have pathogenic intestinal parasites detected in later stool specimens (adjusted ordinal odds ratio = 3.94 [95% confidence interval: 1.34–14.05]). Collecting multiple stool specimens was associated with a higher likelihood of detecting pathogenic intestinal parasites, particularly in immunocompetent patients. Clinicians may request a second or even a third sample if the first is negative.

Intestinal helminthic parasites pose a serious threat to public health. The prevalence of helminth contamination in fresh vegetables collected from nine farms in Chachoengsao Province, Thailand was investigated. Sedimentation technique was used to screen 359 fresh vegetable samples for helminths following washing with a detergent solution. The overall 38% prevalence of helminth contamination in the study highlighted the widespread nature of this issue. Of the nine vegetables, 63% of lettuce had the highest proportion of contamination, followed by 51% for coriander, 44% for celery, 41% for sweet basil, 37% for hoary basil, 34% for spring onion, 32% for kale, 24% for water spinach, and 15% for yearlong beans. Six genera of helminths were identified, present in both egg and larval stages, namely, Ancylostomatoidea (hookworm), Ascaris spp, Strongyloides spp, Taenia spp, Trichuris spp, and Toxocara spp. Of note, hookworms were the most prevalent helminth in egg (8%) and larval (15%) stages. Contamination by the helminths significantly differed among the nine vegetables (p-value = 0.0003, using the Chi-square test). The high prevalence of helminth contamination in fresh vegetables obtained from local farms in this province raises concerns about potential transmission to humans. This finding underscores the critical importance of stringent hygiene practices, such as thorough washing, disinfection and proper cooking, to ensure vegetables are devoid of parasitic and pathogenic contaminants. Education and communication of these measures will assist public awareness of proper food handling practices to ensure vegetables are safe for human consumption.

Background A close connection between a protozoan parasite and the balance of the other gut microbes of the host has been demonstrated. The calves may be naturally co-infected with many parasites, and the co-effects of parasites on other intestinal microbes of calves remain unclear. This study aims to preliminarily reveal the relationship between intestinal parasites and other intestinal microbes in calves. Methods Fecal samples were collected from four calves with bloody diarrhea, four calves with watery diarrhea, and seven normal calves, and the microbial flora of the samples were analyzed by whole-genome sequencing. Protozoal parasites were detected in the metagenome sequences and identified using polymerase chain reaction (PCR). Results Cryptosporidium , Eimeria , Giardia , Blastocystis , and Entamoeba were detected by metagenomic analysis, and the identified species were Giardia duodenalis assemblage E, Cryptosporidium bovis , Cryptosporidium ryanae , Eimeria bovis , Eimeria subspherica , Entamoeba bovis , and Blastocystis ST2 and ST10. Metagenomic analysis showed that the intestinal microbes of calves with diarrhea were disordered, especially in calves with bloody diarrhea. Furthermore, different parasites show distinct relationships with the intestinal microecology. Cryptosporidium , Eimeria , and Giardia were negatively correlated with various intestinal bacteria but positively correlated with some fungi. However, Blastocystis and Entamoeba were positively associated with other gut microbes. Twenty-seven biomarkers not only were significantly enriched in bloody diarrhea, watery diarrhea, and normal calves but were also associated with Eimeria , Cryptosporidium , and Giardia . Only Eimeria showed a distinct relationship with seven genera of bacteria, which were significantly enriched in the healthy calves. All 18 genera of fungi were positively correlated with Cryptosporidium , Eimeria , and Giardia , which were also significantly enriched in calves with bloody diarrhea. Functional genes related to parasites and diseases were found mainly in fungi. Conclusions This study revealed the relationship between intestinal protozoan parasites and the other calf gut microbiome. Different intestinal protozoan parasites have diametrically opposite effects on other gut microecology, which not only affects bacteria in the gut, but also is significantly related to fungi and archaea. Graphical Abstract

Background Spain’s recent abrupt rise in numbers of registered pet dogs and cats has intensified the need for proper animal health care, as 60% of infectious diseases are zoonotic. While pathogen detection has improved through advances in molecular techniques, pet owners often fail to adhere to veterinary guidelines, increasing infection risks. Among the diagnostic tools available, faecal analysis plays a key role in detecting zoonotic parasites such as Giardia duodenalis , Cryptosporidium spp. and Toxocara spp. This study was designed to assess intestinal parasite prevalence in dogs and cats along with epidemiological trends. Methods Between 2013 and 2023, a total of 15,899 faecal samples from dogs and cats submitted to a reference laboratory of parasitology in Madrid (Spain) were analysed using Mini-FLOTAC ® , merthiolate–iodine–formalin (MIF), Baermann–Wetzel (for lungworms and S. stercoralis when indicated) and direct immunofluorescence assays, with molecular confirmation by polymerase chain reaction (PCR) when required. Epidemiological variables were statistically analysed. Results Overall, 26% of dogs and 21.4% of cats tested positive for at least one parasite. Protozoan infections were more prevalent overall, particularly G. duodenalis in dogs (16.0%) and Cystoisospora spp. in cats (7.8%). In contrast, helminth infections such as T. cati (7.6%) were more frequent in cats than in dogs. Dogs were more commonly infected by protozoa than helminths, while cats showed a more balanced distribution between both groups. Co-infections occurred in 13.6% of positive samples, with G. duodenalis being frequently involved. Puppies and kittens were more susceptible to infection, likely owing to an immature immune system. In animals from shelters, infection rates were higher than in owned pets. Seasonal variations were clearly observed, such that G. duodenalis peaked in winter and helminths in autumn. Over time, the prevalence of G. duodenalis increased, while that of Cystoisospora spp. declined. Conclusions These findings highlight the importance of parasite control for purposes of both animal and public health, and emphasize a need for regular faecal testing, deworming and improving owner awareness of parasites. To minimize zoonotic risks and improve pet health management, we would recommend standardizing diagnostic procedures and designing suitable veterinary interventions. Graphical Abstract

Background Intestinal parasitic infections (IPIs) are still among the major public health issues in developing countries. Assessing the prevalence of IPIs and potential risk factors in different localities is essential to enhance control strategies. To date, no prevalence assessment study was conducted in Debre Berhan town. Therefore, the aim of this study was to assess the prevalence of IPIs and associated habit and culture-related risk factors among primary schoolchildren in Debre Berhan town, Northeast Ethiopia. Method School based cross-sectional study was conducted from April to June 2017. A total of 645 children aged 6–15 years were selected from six primary schools in Debre Berhan town via a multistage random sampling technique. A structured questionnaire was used to collect data about sociodemographic and potential risk factor variables. Fresh stool samples were collected from each child and examined using direct smear and formal-ether concentration technique. Result Among the 645 children participated in the study, 341 (52.9%) were infected by one or more intestinal parasites. Helminths (33.8%) were more prevalent than protozoa (20%). Double parasitic infection rate was 0.9%. The predominant parasites were Ascaris lumbricoides (22.6%), Entamoeba histolytica/dispar/moshkovskii (18.1%) and Hymenolepis nana (5.7%). Multivariable logistic regression analysis showed that age of child (6–9 years), family size (above 5), mother’s illiteracy and primary education, father’s illiteracy, urban-farmer father, manual-worker father, not washing hands before eating, unclean fingers, open defecation site (ODS) near residence, latrine type, cultural response to dropped food (cleaning and eating; ‘kiss and replace’), habit of playing with waste water, habit of playing with soil, habit of sucking fingers and habit of eating when playing were significantly associated with IPIs ( p < 0.05). Likewise, age (6–9 years), mother’s illiteracy, urban-farmer father, not washing hands before eating, ODS near residence, tradition of cleaning and eating dropped food, habit of playing with soil, sucking fingers and eating when playing were identified as significant risk factors of A. lumbricoides infection. Conclusion High prevalence of IPIs among the study participants demands improvement of environmental sanitation, personal hygiene, and health education regarding the potential habit and culture-related risk factors.