Comparative copro-diagnosis of Echinococcus multilocularis in experimentally infected foxes

Faecal samples from 15 foxes experimentally infected with Echinococcus multilocularis were examined until 90days post-infection (dpi) by microscopical identification of eggs isolated by flotation/sieving, by coproantigen-enzyme-linked immunosorbent assay (cELISA), by polymerase chain reaction (PCR) on DNA, respectively, isolated directly from the faecal samples (copro-DNA PCR) and from the eggs obtained by the flotation/sieving procedure (egg-DNA PCR). Based on egg counts, three periods of the infection were defined: pre-patent (2-29dpi), high patent (30-70dpi) and low patent periods (71-90dpi). Whereas all methods were highly sensitive with samples from the high patent period, cELISA was the most sensitive to detect pre-patent infections (63%). Samples from the low patent infections were positive in 77% by microscopy and in 80% by egg-DNA PCR, being significantly more sensitive than cELISA and copro-DNA PCR. The isolation of eggs from the faecal material proved to be more sensitive by the flotation/sieving procedure as compared to the classical concentration McMaster techniqu

Prevalence of molecular markers of anti-malarial drug resistance in Plasmodium vivax and Plasmodium falciparum in two districts of Nepal

<p>Abstract</p> <p>Background</p> <p>Sulphadoxine-pyrimethamine (SP) and chloroquine (CQ) have been used in treatment of falciparum and vivax malaria in Nepal. Recently, resistance to both drugs have necessitated a change towards artemisinin combination therapy (ACT) against <it>Plasmodium falciparum </it>in highly endemic areas. However, SP is still used against <it>P. falciparum </it>infections in low endemic areas while CQ is used in suspected cases in areas with lack of diagnostic facilities. This study examines the prevalence of molecular markers of CQ and SP resistance in <it>P. falciparum </it>and <it>Plasmodium vivax </it>to determine if high levels of <it>in vivo </it>resistance are reflected at molecular level as well.</p> <p>Methods</p> <p>Finger prick blood samples (n = 189) were collected from malaria positive patients from two high endemic districts and analysed for single nucleotide polymorphisms (SNPs) in the resistance related genes of <it>P. falciparum </it>and <it>P. vivax </it>for CQ (<it>Pfcrt, Pfmdr1, Pvmdr1</it>) and SP (<it>Pfdhfr, Pfdhps, Pvdhfr</it>), using various PCR-based methods.</p> <p>Results and discussion</p> <p>Positive <it>P. vivax </it>and <it>P. falciparum </it>infections were identified by PCR in 92 and 41 samples respectively. However, some of these were negative in subsequent PCRs. Based on a few <it>P. falciparum </it>samples, the molecular level of CQ resistance in <it>P. falciparum </it>was high since nearly all parasites had the <it>Pfcrt </it>mutant haplotypes CVIET (55%) or SVMNT (42%), though frequency of the <it>Pfmdr1 </it>wild type haplotype was relatively low (35%). Molecular level of SP resistance in <it>P. falciparum </it>was found to be high. The most prevalent <it>Pfdhfr </it>haplotype was double mutant CNRNI (91%), while frequency of <it>Pfdhps </it>double mutant SGEAA and AGEAA were 38% and 33% respectively. Combined, the frequency of quadruple mutations (CNRNI-SGEAA/AGEAA) was 63%. Based on <it>P. vivax </it>samples, low CQ and SP resistance were most likely due to low prevalence of <it>Pvmdr1 </it>Y976F mutation (5%) and absence of triple/quadruple mutations in <it>Pvdhfr</it>.</p> <p>Conclusions</p> <p>Based on the limited number of samples, prevalence of CQ and SP resistance at molecular levels in the population in the study area were determined as high in <it>P. falciparum </it>and low in <it>P. vivax</it>. Therefore, CQ could still be used in the treatment of <it>P. vivax </it>infections, but this remains to be tested <it>in vivo </it>while the change to ACT for <it>P. falciparum </it>seems justified.</p

Putative biomarkers of environmental enteric disease fail to correlate in a cross-sectional study in two study sites in Sub-Saharan Africa

Environmental enteric dysfunction (EED) is an elusive, inflammatory syndrome of the small intestine thought to be associated with enterocyte loss and gut leakiness and lead to stunted child growth. To date, the gold standard for diagnosis is small intestine biopsy followed by histology. Several putative biomarkers for EED have been proposed and are widely used in the field. Here, we assessed in a cross-sectional study of children aged 2-5 years for a large set of biomarkers including markers of protein exudation (duodenal and fecal alpha-1-antitrypsin (AAT)), inflammation (duodenal and fecal calprotectin, duodenal, fecal and blood immunoglobulins, blood cytokines, C-reactive protein (CRP)), gut permeability (endocab, lactulose-mannitol ratio), enterocyte mass (citrulline) and general nutritional status (branched-chain amino acids (BCAA), insulin-like growth factor) in a group of 804 children in two Sub-Saharan countries. We correlated these markers with each other and with anemia in stunted and non-stunted children. AAT and calprotectin, CRP and citrulline and citrulline and BCAA correlated with each other. Furthermore, BCAA, citrulline, ferritin, fecal calprotectin and CRP levels were correlated with hemoglobin levels. Our results show that while several of the biomarkers are associated with anemia, there is little correlation between the different biomarkers. Better biomarkers and a better definition of EED are thus urgently needed

A stochastic model of Echinococcus multilocularis transmission in Hokkaido, Japan, focusing on the infection process

Echinococcus multilocularis causes human alveolar echinococcus. In Japan, high prevalence of E. multilocularis among the fox population has been reported throughout Hokkaido. Accordingly, control measures, such as fox hunting and the distribution of bait containing Praziquantel, have been conducted. This study developed a transmission model for individuals in the fox population and included a stochastic infection process to assess the prevalence of E. multilocularis. To make our model realistic, we used the worm burden for each individual in the fox population. We assumed that the worm burden depends on the number of protoscoleces in a predated vole and the number of infection experiences. We carried out stochastic simulations with 1,000 trials for the situations of Koshimizu and Sapporo, Hokkaido, Japan. The distribution of the worm burden among foxes obtained using the model agreed with dissection data. The simulation indicates that a careful choice of season is necessary for an effective distribution of Praziquantel-containing bait. A stochastic model for E. multilocularis, which can assess the range of the prevalence in the fox population, would be helpful in analyzing their complex life-cycle and also in designing control strategies.</p

Stunted children display ectopic small intestinal colonization by oral bacteria, which cause lipid malabsorption in experimental models

Environmental enteric dysfunction (EED) is an inflammatory syndrome postulated to contribute to stunted child growth and to be associated with intestinal dysbiosis and nutrient malabsorption. However, the small intestinal contributions to EED remain poorly understood. This study aimed to assess changes in the proximal and distal intestinal microbiota in the context of stunting and EED and to test for a causal role of these bacterial isolates in the underlying pathophysiology. We performed a cross-sectional study in two African countries recruiting roughly 1,000 children aged 2 to 5 years and assessed the microbiota in the stomach, duodenum, and feces. Upper gastrointestinal samples were obtained from stunted children and stratified according to stunting severity. Fecal samples were collected. We then investigated the role of clinical isolates in EED pathophysiology using tissue culture and animal models. We find that small intestinal bacterial overgrowth (SIBO) is extremely common (>80%) in stunted children. SIBO is frequently characterized by an overgrowth of oral bacteria, leading to increased permeability and inflammation and to replacement of classical small intestinal strains. These duodenal bacterial isolates decrease lipid absorption in both cultured enterocytes and mice, providing a mechanism by which they may exacerbate EED and stunting. Further, we find a specific fecal signature associated with the EED markers fecal calprotectin and alpha-antitrypsin. Our study shows a causal implication of ectopic colonization of oral bacterial isolated from the small intestine in nutrient malabsorption and gut leakiness in vitro. These findings have important therapeutic implications for modulating the microbiota through microbiota-targeted interventions

The genetic basis of apparently idiopathic ventricular fibrillation:A retrospective overview

Aims: During the diagnostic work-up of patients with idiopathic ventricular fibrillation (VF), next-generation sequencing panels can be considered to identify genotypes associated with arrhythmias. However, consensus for gene panel testing is still lacking, and variants of uncertain significance (VUS) are often identified. The aim of this study was to evaluate genetic testing and its results in idiopathic VF patients. Methods and results: We investigated 419 patients with available medical records from the Dutch Idiopathic VF Registry. Genetic testing was performed in 379 (91%) patients [median age at event 39 years (27-51), 60% male]. Single-gene testing was performed in 87 patients (23%) and was initiated more often in patients with idiopathic VF before 2010. Panel testing was performed in 292 patients (77%). The majority of causal (likely) pathogenic variants (LP/P, n = 56, 15%) entailed the DPP6 risk haplotype (n = 39, 70%). Moreover, 10 LP/P variants were found in cardiomyopathy genes (FLNC, MYL2, MYH7, PLN (two), TTN (four), RBM20), and 7 LP/P variants were identified in genes associated with cardiac arrhythmias (KCNQ1, SCN5A (2), RYR2 (four)). For eight patients (2%), identification of an LP/P variant resulted in a change of diagnosis. In 113 patients (30%), a VUS was identified. Broad panel testing resulted in a higher incidence of VUS in comparison to single-gene testing (38% vs. 3%, P &lt; 0.001). Conclusion: Almost all patients from the registry underwent, albeit not broad, genetic testing. The genetic yield of causal LP/P variants in idiopathic VF patients is 5%, increasing to 15% when including DPP6. In specific cases, the LP/P variant is the underlying diagnosis. A gene panel specifically for idiopathic VF patients is proposed.</p

Mitral Annular Disjunction in Idiopathic Ventricular Fibrillation Patients: Just a Bystander or a Potential Cause?

AIMS: Previously, we demonstrated that inferolateral mitral annular disjunction (MAD) is more prevalent in patients with idiopathic ventricular fibrillation (IVF) than in healthy controls. In the present study, we advanced the insights into the prevalence and ventricular arrhythmogenicity by inferolateral MAD in an even larger IVF cohort. METHODS AND RESULTS: This retrospective multicentre study included 185 IVF patients (median age 39 [27, 52] years, 40% female). Cardiac magnetic resonance images were analysed for mitral valve and annular abnormalities and late gadolinium enhancement. Clinical characteristics were compared between patients with and without MAD. MAD in any of the 4 locations was present in 112 (61%) IVF patients and inferolateral MAD was identified in 24 (13%) IVF patients. Mitral valve prolapse (MVP) was found in 13 (7%) IVF patients. MVP was more prevalent in patients with inferolateral MAD compared with patients without inferolateral MAD(42% vs. 2%, p < 0.001). Proarrhythmic characteristics in terms of a high burden of premature ventricular complexes (PVC) and non-sustained ventricular tachycardia (VT) were more prevalent in patients with inferolateral MAD compared to patients without inferolateral MAD (67% vs. 23%, p < 0.001 and 63% vs 41%, p = 0.046, respectively). Appropriate implantable cardioverter defibrillator therapy during follow-up was comparable for IVF patients with or without inferolateral MAD (13% vs. 18%, p = 0.579). CONCLUSION: A high prevalence of inferolateral MAD and MVP is a consistent finding in this large IVF cohort. The presence of inferolateral MAD is associated with a higher PVC burden and non-sustained VTs. Further research is needed to explain this potential interplay