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The extracellular protozoan parasite Giardia duodenalis is a well-known and important causative agent of diarrhea on a global scale. Macrophage pyroptosis has been recognized as an important innate immune effector mechanism against intracellular pathogens. Yet, the effects of noninvasive Giardia infection on macrophage pyroptosis and the associated molecular triggers and regulators remain poorly defined. Here we initially observed that NLRP3 inflammasome-mediated pyroptosis was activated in Giardia -treated macrophages, and inhibition of ROS, NLRP3, or caspase-1 could block GSDMD cleavage, IL-1β, IL-18 and LDH release, and the cell viability reduction. We also confirmed that Giardia -induced NLRP3 inflammasome activation was involved in its K63 deubiquitination. Thus, six candidate deubiquitinases were screened, among which A20 was identified as an effective regulator. We then screened TLRs on macrophage membranes and found that upon stimulation TLR4 was tightly correlated to ROS enhancement, A20-mediated NLRP3 deubiquitination, and pyroptotic signaling. In addition, several Giardia -secreted proteins were predicted as trigger factors via secretome analysis, of which peptidyl-prolyl cis-trans isomerase B (PPIB) independently induced macrophage pyroptosis. This was similar to the findings from the trophozoite treatment, and also led to the TLR4-mediated activation of NLRP3 through K63 deubiquitination by A20. Collectively, the results of this study have significant implications for expanding our understanding of host defense mechanisms after infection with G. duodenalis.

Introduction An association of Helicobacter pylori and common protozoal parasites in patients with abdominal discomfort and chronic diarrhoea is unclear and may be pathological. Materials and methods One hundred and sixty-one patients with diarrhoea were compared to 114 age and sex matched controls. Stool samples were examined by microscopy and DNA extracted for PCR with specific primers for H. pylori and protozoal parasites Blastocystis sp., Entamoeba sp. (Entamoeba histolytica, Entamoeba dispar and Entamoeba moshkovskii) and Giardia duodenalis (G. duodenalis). Results There was a marked difference in the presence of parasites between patients and controls: no parasite 42/75%, one parasite 42/15%, two or more parasites 16/10%, respectively (p < 0.001). Patients with diarrhoea were more likely to be infected with Blastocystis sp (p < 0.001), E. histolytica (p = 0.027) and E moshkovskii (p = 0.003). There was no difference in the frequency of H. pylori (p = 0.528), G duodenalis (p = 0.697) or E dispar (p = 0.425). Thirty-three patients and 27 controls had H. pylori infection. Of these, 22 patients and 6 controls were infected with Blastocystis sp (p = 0.001), 6 patients and no controls were infected with E. histolytica (p = 0.02), whilst 7 patents and 9 controls were infected with E dispar (p = 0.292). Conclusion In this population, diarrhoea is linked to infection with Blastocystis sp, E. histolytica and E moshkoviskii. In H. pylori infection, diarrhoea is linked to Blastocystis sp and E. histolytica infection. These associations may be linked pathogenically.

Bovine neonatal diarrhea is common due low immunity in newborn calves, poor management (or absence) of sanitary barriers, and other factors. Newborn calves with diarrhea in the first days of life suffer failure to thrive and may die if left untreated. The aim of this study was to evaluate whether prophylactic administration of a homeopathic product (Dia 100 ® ) can control bovine neonatal diarrhea in calves born on a farm with substantial sanitary challenges. We counted total bacteria and protozoan parasites in fecal samples. We measured serum glucose, total protein, globulin, albumin, cholesterol and triglycerides on days 1, 7 and 14 of life. Twenty newborn calves were maintained in individual stalls, and were divided in two groups: ten untreated animals (control) and ten animals treated with Dia 100 ® . Fecal consistency was evaluated daily. We diagnosed diarrhea in five animals in the treated group, and in all animals from the control group. Infections with Escherichia coli and Giardia duodenalis were identified as the responsible organisms. The E. coli count was low in the treatment group on day 7 of life compared with the control group. Antibiotics were given to eight animals in the control group, and to two animals in the treatment group. On day of life 7, serum levels of total protein and globulins were higher in the control group, but were lower on day 14. Serum levels of glucose and triglycerides were greater in treated animals on days 7 and 14, suggesting that the homeopathic product contributes to improvement of intestinal health and absorption and nutrients. We conclude that Dia 100 ® controls diarrhea with 50% of efficacy, and reduces antibiotic utilization., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Giardia duodenalis is a zoonotic protozoan parasite with public health importance worldwide. While articles about animal model infectivity have been published for G. duodenalis, the studies have used diverse protocols and parameters to evaluate the infectivity of this protozoan parasite. Hence, the objectives of this study were to (1) conduct a meta-analysis of published literature for cyst shedding and diarrhea outcomes in animal models and (2) develop recommendations to help standardize experimental dose response studies. Results showed that, for the outcome of cyst shedding in faeces, the covariates of infective stage (cyst versus trophozoite), Giardia dose, and the interactions between doses and infective stage, as well as dose and species of experimental host, were all significant (P value ≤ 0.05). This study suggests inoculation of the experimental host with cysts rather than trophozoites and administration of higher doses of Giardia will most likely result in cyst shedding. Based on the results of this meta-analysis, the infective stage (cyst versus trophozoite), parasite dose, and the interactions between dose and infective stage, as well as dose and species of experimental host, should be considered when designing experimental dose response studies that will assist in the study of zoonotic neglected tropical diseases globally.

Introduction: Acute diarrhea continues to be a major cause of morbidity and mortality in children from developing countries. Determination of the frequency of diarrhea in an area, along with the proportion of disease caused by specific enteric agents of different origins, is considered the first step in controlling diarrheal diseases., Methodology: From 2005 to 2007, a hospital-based surveillance was conducted in two locations in Egypt to determine the causes of acute diarrhea in children younger than 5-years seeking treatment. Five additional enteric viral and parasitic pathogens were tested using commercially-available enzyme immunoassays (EIA) to re-evaluate the prevalence of diarrheal pathogens in undiagnosed cases., Results: Adenovirus, astrovirus, norovirus and G. lamblia were detected as the sole pathogen in 2% (n=34), 3% (n=56), 9% (n=191) and 7% (n=146) of the cases, respectively. E. histolytica was never detected as the sole pathogen. The percentage of diarrheal cases with a known cause increased significantly, from 48% (n=1,006) to 74% (n=1,568) (P < 0.0001)., Conclusion: In our study, the incorporation of immunoassays yielded useful data in identifying pathogens in previously pathogen-negative diarrhea cases.

Wastewater reuse contributes to closing the nutrient recycling loop as a sustainable way of managing water resources. Bangkok has over a thousand man-made drainage and irrigation canals for such purposes. Its use for agricultural and recreational purposes has a long tradition in rural and peri-urban areas. However, the continuation of these practices is increasingly questioned since potential health risks are an issue if such practices are not appropriately managed. The microbial and chemical quality of canal water has considerably deteriorated over the last decade, mainly because of discharged, untreated domestic and industrial wastewater. It is important to understand the health risks of wastewater reuse and identify risky behaviors from the most highly exposed actors promote the safe use of wastewater. This study assessed diarrhea infection risks caused by the use of and contact with wastewater in Klong Luang municipality, a peri-urban setting in Northern Bangkok, using quantitative microbial risk assessment. Wastewater samples were collected from canals, sewers at household level, and vegetables grown in the canals for consumption. Samples were also collected from irrigation water from the agricultural fields. Two protozoa, Giardia lamblia and Entamoeba histolytica, were quantified and analyzed by real-time PCR, exposure assessment was conducted, and finally, the risk of infection due to contact with wastewater in different scenarios was calculated. The results showed that canal water and vegetables were heavily contaminated with G. lamblia and E. histolytica. Infection risk was high in tested scenarios and largely exceeded the acceptable risk given by WHO guidelines.

Giardiasis is a significant cause of diarrheal disease and associated morbidity in children and adults worldwide. In addition to diarrhea, it can also lead to malnutrition and cognitive deficits in children from developing countries. Giardia duodenalis is considered to be a species complex of several assemblages, of which assemblage A and B are predominantly associated with human infections. Assemblage type has been associated with risk of occurrence of symptoms and duration of illness. Hence genotyping of giardial isolates may help understand better the epidemiology and transmission ecology of the disease in a particular setting or area. In India, prevalence rates of Giardia infection in patients with diarrhea range from 0.4% to 70%, and asymptomatic cyst passage has been found to be as high as 50% in rural southern India. In this review, the global distribution of giardial assemblage, zoonotic transmission and the association of assemblage with disease have been discussed, followed by epidemiology of giardiasis in India.

Background: Asymptomatic carriage of Giardia intestinalis is highly prevalent among children in developing countries, and evidence regarding its role as a diarrhea-causing agent in these settings is controversial. Impaired linear growth and cognition have been associated with giardiasis, presumably mediated by malabsorption of nutrients. In a prospective cohort study, we aim to compare diarrhea rates in pre-school children with and without Giardia infection. Because the study was conducted in the context of an intervention trial assessing the effects of multi-nutrients on morbidity, we also assessed how supplementation influenced the relationship between Giardia and diarrhoea rates, and to what extent Giardia modifies the intervention effect on nutritional status., Methods and Findings: Data were collected in the context of a randomized placebo-controlled efficacy trial with 2×2 factorial design assessing the effects of zinc and/or multi-micronutrients on morbidity (n=612; height-for-age z-score <-1.5 SD). Outcomes measures were episodes of diarrhea (any reported, or with ≥3 stools in the last 24 h) and fever without localizing signs, as detected with health-facility based surveillance. Giardia was detected in stool by enzyme-linked immunosorbent assay. Among children who did not receive multi-nutrients, asymptomatic Giardia infection at baseline was associated with a substantial reduction in the rate of diarrhea (HR 0.32; 0.15-0.66) and fever without localizing signs (HR 0.56; 0.36-0.87), whereas no such effect was observed among children who received multi-nutrients (p-values for interaction 0.03 for both outcomes). This interaction was independent of age, HAZ-scores and distance to the research dispensary. There was no evidence that Giardia modified the intervention effect on nutritional status., Conclusion: Although causality of the Giardia-associated reduction in morbidity cannot be established, multi-nutrient supplementation results in a loss of this protection and thus seems to influence the proliferation or virulence of Giardia or associated intestinal pathogens.

Giardia lamblia is the most prevalent human intestinal protozoan worldwide, but only a minority of infections result in diarrhea. We tested here whether the 2 major G. lamblia genotypes, assemblages A and B, differ in their propensity to cause disease. To determine whether an association exists between infection with assemblage A or B and diarrhea, 2534 Bangladeshi patients were enrolled in a case-control study. A total of 322 Giardia infections were identified and assayed for genotype by real-time polymerase chain reaction. Higher odds ratios for diarrhea were observed for assemblage A and A2 infections, whereas higher parasite DNA loads and a higher overall prevalence were observed for assemblage B infections. Our findings indicate that genotypic differences in virulence and fecundity may help to explain why not every Giardia infection results in disease, but they need to be confirmed in other urban populations of the developing world.

Objective: To evaluate the pathogenicity of Dientamoeba fragilis by comparing it with Giardia lamblia and to investigate the effect of a single dose of secnidazole in dientamoebiasis., Methods: Stool samples of 400 patients, admitted to the Department of Parasitology, Celal Bayar University, were examined by direct wet mount, formalin-ethyl acetate concentration and trichrome staining methods on three consecutive days. All cases positive for D. fragilis were treated with a single dose of secnidazole, 30 mg/kg for children, and 2 g for adults. On the seventh and fourteenth days, at the end of the treatment, stool samples were examined by the same methods, and clinical symptoms were again evaluated., Results: D. fragilis and G. lamblia were detected in 35 (8.8%) and 34 (8.5%) cases, respectively. The most frequent symptoms were found to be abdominal pain and diarrhea in both infections. D. fragilis was eradicated in 34 (97.1%) patients with a single dose of secnidazole, but a second dose was necessary in one patient. Clinical symptoms disappeared in 27 (77.1%) and decreased in eight (22.9%) cases, after eradication., Conclusions: These data suggested that D. fragilis is as prevalent and pathogenic as G. lamblia, and secnidazole seems to be highly effective in achieving parasitologic and clinical cure. To our knowledge, this is the first report of secnidazole being used in the treatment of dientamoebiasis.

The objective of this study conducted in 75 herds was to investigate the presence and significance of Criptosporidium parvum and Giardia intestinalis in Swedish dairy calves in comparison with rotavirus, coronavirus and Escherichia coli K99+. The farmers were asked to collect faecal samples from each heifer calf that had diarrhoea between birth and 90 days of age, and also from a healthy calf of the same age. In total, 270 samples were collected and analysed. C. parvum, either alone or together with G. intestinalis and/or rotavirus, was detected in 16 (11%) and 6 (5%) of the samples from diarrhoeic and healthy calves, respectively. Even though a higher proportion of diarrhoeic calves shed C. parvum, the difference between the groups was not statistically significant (p = 0.067), possibly due to the low number of positive samples. G. intestinalis was found in 42 (29%) of the diarrhoea samples and in 29 (23%) of the samples from healthy calves. Rotavirus and coronavirus were demonstrated in 24% and 3% of the diarrhoea samples, respectively, whereas E. coli K99+ was only found in samples from 2 healthy calves. C. parvum and G. intestinalis were found in samples from calves 7 to 84 days of age and during all seasons. The results confirm that C. parvum is present in Swedish dairy herds and might have clinical significance. G. intestinalis was the most common agent found but the importance of this parasite remains unclear. Both parasites have suggested zoonotic potential and thus warrant further attention. In addition, rotavirus is a major pathogen in neonatal enteritis in Sweden, whereas coronavirus and E. coli K99+ seem to be of less importance.

Cryptosporidium parvum and Giardia lamblia are important agents of waterborne diarrheal illness worldwide. While giardiasis is routinely diagnosed in Russia with a chemical staining technique, data on the prevalence of cryptosporidiosis are scarce. Monitoring of the respective parasites in water supplies in Russia is very limited. A health survey conducted in the city of Cherepovets and three other cities in the European part of Russia using enzyme-linked immunosorbent assays (ELISA) demonstrated that 6.9% of diarrheal patients tested had C. parvum antigens in their fecal samples; 9.4% had G. lamblia antigens. A survey of occurrence of these parasites in water supplies in Cherepovets and seven other cities demonstrated that source and finished water samples from several of these cities were contaminated with either C. parvum oocysts or G. lamblia cysts. The surveys were not designed to assess associations between presence or concentrations of C. parvum and G. lamblia in water and related gastrointestinal diseases in exposed populations. Rather, the goals were to demonstrate the presence of disinfection-resistant protozoan parasites in untreated and treated waters, and the importance of these pathogens as causative agents of diarrheal illnesses in a number of Russian cities.

Infectious diarrhoea is one of the most common illness affecting mankind. Recent advancement have led to detailed understanding of causative agents and the pathogenesis of the infections. Fluid and electrolyte replacement remains the most important aspect of treatment. The role of antimicrobial agents is discussed.

Giardia has been frequently implicated as a causative agent for acute as well as chronic diarrheal diseases in children. The present study was aimed at exploring the determinants of manifestations of Giardiasis in childhood, in relation to various host and parasite related factors. A total of 200 children with acute (100), chronic (50) or without (50) diarrhea in last 15 days were recruited for the study and evaluated with regards to nutritional status, serum immunoglobulins, secretory IgA levels, presence of Giardia in stool/duodenal aspirate/duodenal biopsy specimen and for associated infections. Lysates from acute giardiasis cases were further studied for zymodeme (banding) pattern. After correlation of all investigations, humoral immune defect in the host was found to be the major determinant of whether the Giardial infestation would be symptomatic or not, while associated bacterial infections and zymodeme patterns were not found to be important in determining the pathogenicity or presentation of giardiasis.

The transmucosal fluxes of Na+ and Cl- were studied in Giardia lamblia infected mice in the presence or absence of phorbol-12-myristate-13-acetate (PMA), the activator of protein kinase C (PKC) or 1-(5-isoquinolinylsulphonyl)-2-methylpiperazine (H-7), the inhibitor of PKC or Ca(2+)-calmodulin. There was net secretion of Na+ and Cl- in infected animals, while in control animals there was net absorption of these ions. The addition of ionophore or PMA resulted in net secretion of Na+ and Cl- in the control group while in the infected group there was no change in the fluxes of these ions. The selective potent inhibitor of protein kinase C, H-7, reversed the secretion of Na+ and Cl- in infected group to absorption. The addition of PMA and Ca(2+)-ionophore together in the infected group had a partial additive effect. This study suggests that G. lamblia induced fluid secretion involves protein kinase C and further protein kinase C acts in synergism with calcium.