Growth and development of Gnathostoma spinigerum (Nematoda: Gnathostomatidae) larvae in Mesocyclops aspericornis (Cyclopoida: Cyclopidae)

<p>Abstract</p> <p>Background</p> <p><it>Gnathostoma spinigerum </it>larva is pathogenic, causing gnathostomiasis in humans and certain animals, and is prevalent mainly in Asia. Growth and development of <it>Gnathostoma spinigerum </it>larvae in the cyclopoid copepod <it>Mesocyclops aspericornis</it>, the first intermediate host, were examined.</p> <p>Results</p> <p>When newly hatched, ensheathed second-stage larvae (L2) were ingested by <it>M. aspericornis</it>, they immediately appeared exsheathed in the stomach of <it>M. aspericornis</it>. They then penetrated the stomach wall and entered the body cavity, where they immediately metamorphosed to a stunted form with the body length/width ratio equal to the early third-stage larvae (EL3) up to 2 h after being ingested. During metamorphosis, the anterior spine-like structure of L2 transformed into unequal transparent lips. The larvae moulted into EL3 in the body cavity of the copepod at around day 5-7 post-infection. Minute cuticular striations were seen on the whole body, with prominent single-pointed spines on the anterior part of the body. The head bulb had four rows of hooklets and two lateral trilobed lips. The size of EL3 in copepods continuously increased towards day 12 and showed a negative correlation to their density per copepod (R = -0.881, <it>P </it>< 0.05 for body length, and R = -0.906, <it>P </it>< 0.05 for body width).</p> <p>Conclusions</p> <p>The results revealed for the first time that <it>M. aspericornis</it>, one of the most abundant freshwater copepods in Thailand, is a suitable first intermediate host for <it>G. spinigerum</it>. High susceptibility of <it>M. aspericornis </it>suggests its importance for the maintenance of the life cycle of <it>G. spinigerum </it>in Thailand.</p

Development of a PCR assay and pyrosequencing for identification of important human fish-borne trematodes and its potential use for detection in fecal specimens

BACKGROUND: Small liver and minute intestinal flukes are highly prevalent in Southeast Asia. Definitive diagnosis of parasite infection is usually achieved parasitologically by finding the fluke eggs in feces. However, their eggs are difficult to differentiate morphologically in fecal samples, even for experienced technicians. The present study developed a PCR assay coupled with DNA pyrosequencing for identification of the fish-borne trematodes (FBT), Opisthorchis viverrini, Clonorchis sinensis, Haplorchis taichui, H. pumilio and Stellantchasmus falcatus, and to evaluate potential detection in fecal specimens, and identification and differentiation of cercarial and metacercarial stages. METHODS: Primers targeting the partial 28S large subunit ribosomal RNA gene were designed and about 46–47 nucleotides were selected as the target region for species identification by a PCR assay coupled with a pyrosequencing technique. RESULTS: The nucleotide variations at 24 positions, which is sufficient for the identification of the five species of FBT were selected. The method could identify O. viverrini and C. sinensis eggs in feces, cercarial and metacercarial stages of O. viverrini, and metacercarial stage of H. pumilio and H. taichui. The detection limit was as little as a single O. viverrini or C. sinensis egg artificially inoculated in 100 mg of non-infected fecal sample (equivalent to 10 eggs per gram), indicating highly sensitivity. The method was found to be superior to the traditional microscopy method and was more rapid than Sanger DNA sequencing. CONCLUSIONS: DNA pyrosequencing-based identification is a valuable tool for differentiating O. viverrini and other Opisthorchis-like eggs, and can be applied to epidemiological studies and for molecular taxonomic investigation of FBT in endemic areas

Data

The data for the project "Automatic detection of Opisthorchis viverrini egg in stool examination using convolutional-based neural networks.""1 original dataset" folder, consisting of original microscopic images containing artifacts (class 0) and O.viverrini eggs (class 1) in preparation for the training and validation step."2 training dataset" folder, consisting of microscopic images containing artifacts (class 0) and O.viverrini eggs (class 1), was augmented using image rotation, filtering, adding noise, and sharpening techniques. This augmentation increased the image dataset from 1 time (100 images/class) to 36 times (3600 images/class) in preparation for the training and validation step."3 testing dataset" folder, consisting of 148 microscopic images for the testing process."4 result_heatmap" folder; our patch search algorithm detects eggs and generates a heat mapping image."5 result_detection" folder; an object detection method was proposed using a patch search algorithm to detect the O.viverrini egg and its location.</p

Codes

The code for the project "Automatic detection of Opisthorchis viverrini egg in stool examination using convolutional-based neural networks.""ImageAugmentation.ipynb" file is a Python code for image augmentation in our work."meinlabproj_02_modeltrainingvalidation_rev.ipynb" file is a Python code for the model construction and validation and then export to the h5 file."ObjectDetection_Rev.ipynb" is a Python code for importing testing images and h5 files for patch searching and creating a bounding box around the O. viverrini egg.</p

Effectiveness of Strongyloides Recombinant IgG Immunoreactive Antigen in Detecting IgG and IgG4 Subclass Antibodies for Diagnosis of Human Strongyloidiasis Using Rapid Immunochromatographic Tests

Human strongyloidiasis is an important soil-transmitted helminthiasis that affects millions worldwide and can develop into fatal systemic strongyloidiasis in immunosuppressed patients. We have developed two new rapid and simple-to-use immunochromatographic test (ICT) kits for rapid serodiagnosis that support stool examination for clinical diagnosis. Strongyloides stercoralis recombinant IgG immunoreactive antigen (GenBank: AAB97359.1; rSsIR-based ICT kit) was used for detection of IgG and IgG4 antibodies. The diagnostic efficacy of both kits was evaluated using human serum samples from strongyloidiasis patients, healthy individuals, and those with other parasitosis. At a prevalence of infection of 36.4%, the diagnostic sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of the rSsIR-based IgG ICT kit were 91.7%, 83.8%, 76.4%, 94.6%, and 86.7%, respectively, and those of the rSsIR-based IgG4 ICT kit were 78.3%, 84.8%, 74.6%, 87.3%, and 82.4% respectively. The concordance between the two kits was 89.7%. The recombinant antigen can be produced to an unlimited extent and the kits can be used as point-of-care diagnostic tools and in large-scale surveys in endemic areas throughout tropical regions without necessitating additional facilities or ancillary supplies

Current high prevalences of Strongyloides stercoralis and Opisthorchis viverrini infections in rural communities in northeast Thailand and associated risk factors

Abstract Background Two important helminths, Strongyloides stercoralis (an intestinal roundworm) and Opisthorchis viverrini (a liver fluke), are endemic in northeast Thailand. There have been national campaigns in place aimed at the control and eradication of soil-transmitted helminthiasis and opisthorchiasis in Thailand for several decades. However, these helminths still exist and raise concerns regarding public health. This study aimed to evaluate the current prevalence of S. stercoralis and O. viverrini infections in rural communities in northeast Thailand. The data from this study will be useful to improve strategies for future helminth prevention and control. Methods A cross-sectional study was conducted from December 2016 to June 2017 in Mueang Khon Kaen district in Khon Kaen, Thailand. The participants were selected using a simple random sampling method. Demographic data were collected using a questionnaire. Stool samples were collected and processed using agar plate culture to determine the presence of S. stercoralis infection and an in-house formalin-ethyl acetate concentration technique to determine the presence of O. viverrini and other intestinal parasite infections (IPIs). Results In total, 602 persons were enrolled. However, only 526 were analyzed for S. stercoralis and 387 for O. viverrini risk factors. The overall prevalence of S. stercoralis infection was 23.0% (95% confidence interval [95%CI]: 19.4 to 26.6). The prevalence of O. viverrini infection and IPIs other than S. stercoralis was 20.4% (95%CI: 16.5 to 24.8). The prevalence of O. viverrini infection was 19.4% (95%CI: 15.6 to 23.7). Male sex was significantly associated with S. stercoralis infection [Adjusted Odds Ratio (aOR) 4.0; 95%CI: 2.5 to 6.2; P-value < 0.001]. Males were significantly more likely to be infected with O. viverrini and other IPIs (aOR 4.1; 95%CI: 2.3 to 7.2, P-value < 0.001). Conclusions This study demonstrated that the updated prevalence of intestinal parasite infections is still high in rural communities in northeast Thailand, especially that of strongyloidiasis and opisthorchiasis

Microbiome Composition and Microbial Community Structure in Mosquito Vectors <i>Aedes aegypti</i> and <i>Aedes albopictus</i> in Northeastern Thailand, a Dengue-Endemic Area

Bacterial content in mosquito larvae and adults is altered by dynamic interactions during life and varies substantially in variety and composition depending on mosquito biology and ecology. This study aimed to identify the microbiota in Aedes aegypti and Aedes albopictus and in water from their breeding sites in northeastern Thailand, a dengue-endemic area. Bacterial diversity in field-collected aquatic larvae and subsequently emerged adults of both species from several locations were examined. The microbiota was characterized based on analysis of DNA sequences from the V3-V4 region of the 16S rRNA gene and exhibited changes during development, from the mosquito larval stage to the adult stage. Aedes aegypti contained a significantly higher number of bacterial genera than did Ae. albopictus, except for the genus Wolbachia, which was present at significantly higher frequencies in male Ae. albopictus (p < 0.05). Our findings also indicate likely transstadial transmission from larva to adult and give better understanding of the microbial diversity in these mosquitoes, informing future control programs against mosquito-borne diseases