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Parasites & Vectors May 2018Paragonimus heterotremus is the main causative agent of paragonimiasis in Thailand. In Western blot diagnostic assays for paragonimiasis, the 35 kDa band present in...
BACKGROUND
Paragonimus heterotremus is the main causative agent of paragonimiasis in Thailand. In Western blot diagnostic assays for paragonimiasis, the 35 kDa band present in crude P. heterotremus somatic extracts represents one of the known diagnostic bands. This study aimed to use a P. heterotremus cDNA library to create a recombinant version of this antigen for use in immunodiagnosis of paragonimiasis.
METHODS
To accomplish this aim a cDNA expression library was constructed from adult worm mRNA and immuno-screened using antibodies from mice that had been immunized with the 35 kDa antigen. Screening resulted in the identification of an immunoreactive protein encoded by clone CE3, which contained an inserted sequence composed of 1292 base pairs. This clone was selected for use in the construction of a recombinant P. heterotremus protein because of its similarity to proactivator polypeptide. For recombinant protein expression, the CE3 gene sequence was inserted into the plasmid vector pRset and the resulting product had the expected molecular weight of 35 kDa. An IgG-ELISA based on the CE3 recombinant protein was evaluated by using sera from healthy individuals, from patients with paragonimiasis and other parasitic infections. This ELISA was performed by using human sera diluted at 1:2000, an optimized antigen concentration of 1 μg/ml, and anti-human IgG diluted at 1:4000.
RESULTS
The cut-off optical density value was set as the mean + 2 standard deviations (0.54), which resulted in the test having a sensitivity of 88.89% and a specificity of 95.51%. The recombinant antigen could react with antibodies from P. heterotremus, P. pseudoheterotremus and P. westermani infections. Cross-reactivity occurred with a few cases of Blastocystis hominis infection (2/3), Bancroftian filariasis (1/10), opisthorchiasis (3/10), strongyloidiasis (4/10) and neurocysticercosis (1/11).
CONCLUSIONS
Given the high test sensitivity and specificity, reflected in the low level of heterologous infection cross-reactivity (11/215 serum samples), observed in the IgG-ELISA, this 35 kDa antigen may be useful for the detection of paragonimiasis.
Topics: Animals; Antibodies, Helminth; Antigens, Helminth; Cross Reactions; Humans; Immunoglobulin G; Paragonimiasis; Paragonimus; Recombinant Proteins; Sensitivity and Specificity; Serologic Tests
PubMed: 29843786
DOI: 10.1186/s13071-018-2878-5 -
Mitochondrial DNA. Part B, Resources May 2018In this study, the complete mitochondrial genome of human lung fluke, was recovered through Illumina sequencing data. This complete mitochondrial genome of is...
In this study, the complete mitochondrial genome of human lung fluke, was recovered through Illumina sequencing data. This complete mitochondrial genome of is 13,927 bp in length and has a base composition of A (16.6%), T (41.8%), C (13.%), G (28.4%), demonstrating an obvious bias of high AT content (58.4%). The mitochondrial genome contains a typically conserved structure, encoding 12 protein-coding genes (PCGs), 22 transfer RNA genes (tRNA), 2 ribosomal RNA genes (12S rRNA and 16S rRNA) and a control region (D-loop region). All PCGs were located on the H-strand. ND4 gene and ND4L gene were overlapped by 39 bp. The nucleotide sequence of 12 PCGs of and other 10 parasite species were used for phylogenetic analysis. The result indicated a relative close relationship with species (AF219379.2).
PubMed: 33474240
DOI: 10.1080/23802359.2018.1462119 -
Open Forum Infectious Diseases 2017We report the case of a 15-year-old Burmese girl who presented with hemoptysis 3 years after immigrating to Australia with a background of previously treated pulmonary...
We report the case of a 15-year-old Burmese girl who presented with hemoptysis 3 years after immigrating to Australia with a background of previously treated pulmonary tuberculosis at 6 years of age. Cavitation in the right upper lobe had originally been identified on her baseline chest radiograph following arrival to Australia; extensive investigations were conducted thereafter to exclude causes of cavitary lung disease; these were negative. was finally diagnosed on serological grounds 3 years after this child's original presentation, with subsequent identification of in sputum and in stool. Clinicians should be alert to the possibility of Paragonimiasis in children who have traveled to or originate from endemic countries who present with a clinically compatible illness. Treatment is simple and effective. Failure to consider this pathogen early may result in unnecessary investigative workup and delayed diagnosis.
PubMed: 29308402
DOI: 10.1093/ofid/ofx224 -
PloS One 2017MF6p/FhHDM-1 is a small cationic heme-binding protein which is recognized by the monoclonal antibody (mAb) MF6, and abundantly present in parenchymal cells and secreted...
MF6p/FhHDM-1 is a small cationic heme-binding protein which is recognized by the monoclonal antibody (mAb) MF6, and abundantly present in parenchymal cells and secreted antigens of Fasciola hepatica. Orthologs of this protein (MF6p/HDMs) also exist in other causal agents of important foodborne trematodiasis, such as Clonorchis sinensis, Opisthorchis viverrini and Paragonimus westermani. Considering that MF6p/FhHDM-1 is relevant for heme homeostasis in Fasciola and was reported to have immunomodulatory properties, this protein is expected to be a useful target for vaccination. Thus, in this study we mapped the epitope recognized by mAb MF6 and evaluated its antigenicity in sheep. The sequence of the MF6p/FhHDM-1 ortholog from F. gigantica (MF6p/FgHDM-1) was also reported. By means of ELISA inhibitions with overlapping synthetic peptides, we determined that the epitope recognized by mAb MF6 is located within the C-terminal moiety of MF6p/FhHDM-1, which is the most conserved region of MF6p/HDMs. By immunoblotting analysis of parasite extracts and ELISA inhibitions with synthetic peptides we also determined that mAb MF6 reacted with the same intensity with F. hepatica and F. gigantica, and in decreasing order of intensity with C. sinensis, O.viverrini and P. westermani orthologs. On the contrary, mAb MF6 showed no reactivity against Dicrocoelium dendriticum and Schistosoma mansoni. The study of the recognition of peptides covering different regions of MF6p/FhHDM-1 by sera from immunized sheep revealed that the C-terminal moiety is the most antigenic, thus being of potential interest for vaccination. We also demonstrated that the production of antibodies to MF6p/FhHDM-1 in sheep infected by F. hepatica occurs relatively early and follows the same pattern as those produced against L-cathepsins.
Topics: Animals; Antibodies, Helminth; Antibodies, Monoclonal; Antigens, Helminth; Carrier Proteins; Dendrites; Enzyme-Linked Immunosorbent Assay; Epitope Mapping; Epitopes; Fasciola hepatica; Fascioliasis; Heme; Heme-Binding Proteins; Hemeproteins; Protein Conformation; Sheep; Vaccination
PubMed: 29161330
DOI: 10.1371/journal.pone.0188520 -
BMC Pediatrics Nov 2017Paragonimiasis, particularly hepatic paragonimiasis (HP), is a type of zoonotic parasitic disease rarely encountered in infants. There have been only a few reports of...
BACKGROUND
Paragonimiasis, particularly hepatic paragonimiasis (HP), is a type of zoonotic parasitic disease rarely encountered in infants. There have been only a few reports of HP, and no case of HP has been reported in an infant.
CASE PRESENTATION
A 15-month-old girl presented with persistent mild fever with a duration of 1 month, hepatomegaly, and low-density lesions in the right hepatic lobe on abdominal ultrasound and computer tomography. Pathological examination and serum antibody detection were performed to verify HP. The diagnosis of HP was established based on findings of Charcot-Leyden crystals on liver lesion biopsy and antibodies against paragonimus westermani detected by enzyme-linked immunosorbent assay. After initiation of praziquantel (75 mg/kg/day for 3 days), all clinical findings promptly improved and the patient was discharged.
CONCLUSION
It is very important to consider paragonimiasis in the clinical examination of infants from an area with paragonimiasis epidemic presenting with fever, hepatomegaly, low-density lesions in the liver.
Topics: Female; Humans; Infant; Liver Diseases, Parasitic; Paragonimiasis
PubMed: 29141594
DOI: 10.1186/s12887-017-0942-5 -
The Korean Journal of Parasitology Jun 2017During civil engineering construction near Sejong-ro, Jongro-ku, Seoul, cultural sites were found that are thought to have been built in the 15th century. This area was...
During civil engineering construction near Sejong-ro, Jongro-ku, Seoul, cultural sites were found that are thought to have been built in the 15th century. This area was home to many different people as well as the leaders of the Yi dynasty. To gain further insight into the life styles of the inhabitants of the old capital, soil samples were collected from various areas such as toilets, water foundations, and drainage ways. Parasite eggs were examined by microscopy after 5 g soil samples were rehydrated in 0.5% trisodium phosphate solution. A total of 662 parasite eggs from 7 species were found. Species with the highest number of eggs found were (n=483), followed by (138), (21), (8), (6), (4), and (2). These findings indirectly indicate the food habits of the people in Yi dynasty.
Topics: Animals; Archaeology; Ascaris lumbricoides; Clonorchis sinensis; Fasciola hepatica; Feeding Behavior; Heterophyidae; History, 15th Century; Humans; Life Style; Paragonimus westermani; Parasite Egg Count; Parasitology; Republic of Korea; Soil; Trichuris
PubMed: 28719964
DOI: 10.3347/kjp.2017.55.3.357 -
The Korean Journal of Parasitology Jun 2017An epidemiological study was performed to know the recent infection status of metacercariae (PwMc) in freshwater crayfish, , from 2 streams in Jeollanam-do, Republic of...
An epidemiological study was performed to know the recent infection status of metacercariae (PwMc) in freshwater crayfish, , from 2 streams in Jeollanam-do, Republic of Korea. Crayfish were collected from creeks in Bogil-do (Island), Wando-gun, and in a creek near Daeheung Temple in Haenam-gun. The infection rate of crayfish with PwMc in Bogil-do was 89.8%, and the metacercarial burden was 37 PwMc per the infected crayfish. Crayfish in a creek near Daeheung Temple were larger and twice heavier than those in Bogil-do. Of them, 96.5% were infected with PwMc. An average of 140 metacercariae was found in the infected crayfish, almost quadruple to those of Bogil-do. There was a strong correlation between the number of PwMc and body weight of the crayfish. These results suggest that metacercariae are still prevalent in crayfish of the 2 regions in Jeollanam-do, Korea.
Topics: Animals; Astacoidea; Body Weight; Fresh Water; Incidence; Metacercariae; Paragonimiasis; Paragonimus westermani; Republic of Korea
PubMed: 28719962
DOI: 10.3347/kjp.2017.55.3.347 -
The Korean Journal of Parasitology Jun 2017Paragonimiasis is a parasitic disease caused by species. The primary site of infection is the lung, and extrapulmonary involvement is also reported. When infected with...
Paragonimiasis is a parasitic disease caused by species. The primary site of infection is the lung, and extrapulmonary involvement is also reported. When infected with , which is the dominant species in Korea, the central nervous system is frequently involved along with the liver, intestine, peritoneal cavity, retroperitoneum, and abdominal wall. Ectopic paragonimiasis raises diagnostic challenge since it is uncommon and may be confused with malignancy or other inflammatory diseases. Here, we report an ectopic paragonimiasis case initially presented with recurrent abdominal pain. The patient developed abdominal pain 3 times for the previous 3 years and the computed tomography (CT) of the abdomen revealed fluid collection with wall enhancement. Recurrent diverticulitis was initially suspected and part of the ascending colon was resected. However, the specimen showed intact colon wall without evidence of diverticulitis and multiple parasite eggs and granulomas were found instead. The size of about 70 μm, the presence of an operculum and relatively thick egg shell suggested eggs of species. With appropriate exposure history and a positive antibody test, the definitive diagnosis was made as peritoneal paragonimiasis.
Topics: Abdominal Abscess; Abdominal Pain; Animals; Colonic Diseases; Diagnosis, Differential; Diverticulitis, Colonic; Humans; Intestinal Diseases, Parasitic; Male; Middle Aged; Paragonimiasis; Paragonimus westermani; Parasite Egg Count; Recurrence
PubMed: 28719956
DOI: 10.3347/kjp.2017.55.3.313 -
The Journal of Veterinary Medical... Aug 2017Infection of boar-hunting dogs with Paragonimus westermani was investigated in Western Japan. Blood and rectal feces were collected from 441 dogs in the three districts...
Infection of boar-hunting dogs with Paragonimus westermani was investigated in Western Japan. Blood and rectal feces were collected from 441 dogs in the three districts (205 in Kinki, 131 in Chugoku and 105 in Shikoku District). In a screening ELISA for serum antibody against P. westermani antigen, 195 dogs (44.2%) showed positive reaction. In the 195 dogs, 8 dogs were found excreting P. westermani eggs after molecular analysis of fecal eggs, and additional 7 were identified serologically for the parasite infection because of their stronger reactivity against P. westermani antigen than against antigens of other species of Paragonimus. A spatial analysis showed that all of the P. westermani infections were found in Kinki and Chugoku Districts. In this area, dogs' experience of being fed with raw boar meat showed high odds ratio (3.35) to the sero-positivity in the screening ELISA, and the frequency of such experiences was significantly higher in sero-positive dogs. While clear relationship was not obtained between predation of boars by dogs during hunting and their sero-positivity. Therefore, it is suggested that human activity of feeding with wild boar meat is the risk factor for P. westermani infection in boar-hunting dogs. Considering that hunting dogs could play as a major definitive host and maintain the present distribution of P. westermani in Western Japan, control measures for the infection in hunting dogs, such as prohibition of raw meat feeding and regular deworming, should be undertaken.
Topics: Animal Feed; Animals; Dog Diseases; Dogs; Enzyme-Linked Immunosorbent Assay; Japan; Meat; Paragonimiasis; Paragonimus; Sus scrofa; Swine; Swine Diseases
PubMed: 28717056
DOI: 10.1292/jvms.17-0149 -
The Journal of Biological Chemistry May 2017MF6p/FhHDM-1 is a small protein secreted by the parasitic flatworm (trematode) that belongs to a broad family of heme-binding proteins (MF6p/helminth defense molecules...
MF6p/FhHDM-1 is a small protein secreted by the parasitic flatworm (trematode) that belongs to a broad family of heme-binding proteins (MF6p/helminth defense molecules (HDMs)). MF6p/HDMs are of interest for understanding heme homeostasis in trematodes and as potential targets for the development of new flukicides. Moreover, interest in these molecules has also increased because of their immunomodulatory properties. Here we have extended our previous findings on the mechanism of MF6p/HDM-heme interactions and mapped the protein regions required for heme binding and for other biological functions. Our data revealed that MF6p/FhHDM-1 forms high-molecular-weight complexes when associated with heme and that these complexes are reorganized by a stacking procedure to form fibril-like and granular nanostructures. Furthermore, we showed that MF6p/FhHDM-1 is a transitory heme-binding protein as protein·heme complexes can be disrupted by contact with an apoprotein ( apomyoglobin) with higher affinity for heme. We also demonstrated that (i) the heme-binding region is located in the MF6p/FhHDM-1 C-terminal moiety, which also inhibits the peroxidase-like activity of heme, and (ii) MF6p/HDMs from other trematodes, such as and , also bind heme. Finally, we observed that the N-terminal, but not the C-terminal, moiety of MF6p/HDMs has a predicted structural analogy with cell-penetrating peptides and that both the entire protein and the peptide corresponding to the N-terminal moiety of MF6p/FhHDM-1 interact with cell membranes in hemin-preconditioned erythrocytes. Our findings suggest that MF6p/HDMs can transport heme in trematodes and thereby shield the parasite from the harmful effects of heme.
Topics: Animals; Carrier Proteins; Cattle; Fasciola hepatica; Helminth Proteins; Heme; Opisthorchis; Paragonimus westermani; Protein Domains
PubMed: 28348084
DOI: 10.1074/jbc.M116.771675