scholarly journals Mining Public Metagenomes for Environmental Surveillance of Parasites: A Proof of Principle

2021 ◽  
Vol 12 ◽  
Author(s):  
Frits F. J. Franssen ◽  
Ingmar Janse ◽  
Dennis Janssen ◽  
Simone M. Caccio ◽  
Paolo Vatta ◽  
...  

Parasites often have complex developmental cycles that account for their presence in a variety of difficult-to-analyze matrices, including feces, water, soil, and food. Detection of parasites in these matrices still involves laborious methods. Untargeted sequencing of nucleic acids extracted from those matrices in metagenomic projects may represent an attractive alternative method for unbiased detection of these pathogens. Here, we show how publicly available metagenomic datasets can be mined to detect parasite specific sequences, and generate data useful for environmental surveillance. We use the protozoan parasite Cryptosporidium parvum as a test organism, and show that detection is influenced by the reference sequence chosen. Indeed, the use of the whole genome yields high sensitivity but low specificity, whereas specificity is improved through the use of signature sequences. In conclusion, querying metagenomic datasets for parasites is feasible and relevant, but requires optimization and validation. Nevertheless, this approach provides access to the large, and rapidly increasing, number of datasets from metagenomic and meta-transcriptomic studies, allowing unlocking hitherto idle signals of parasites in our environments.

1997 ◽  
Vol 35 (11-12) ◽  
pp. 363-368 ◽  
Author(s):  
T. R. Slifko ◽  
D. E. Friedman ◽  
J. B. Rose ◽  
S. J. Upton ◽  
W. Jakubowski

Cryptosporidium parvum is an infectious enteric protozoan parasite that causes waterborne disease, severe gastroenteritis and is associated with high mortality in immunocompromised individuals. Detection of oocysts in water is very difficult and current methodologies do not determine viability. This project has focused on low level detection of Cryptosporidium parvum in environmental samples using a unique cultural method. Previously, cell culture methods have been used to assess the developmental stages of Cryptosporidium; however, no cultural methods have been employed with environmental samples. The percentage of viable oocysts can be estimated by detecting intracellular developmental stages of the parasite using fluorescently labelled antibodies. Other methods are not capable of low level detection or high sensitivity. We are evaluating detection of single foci of infection, indicating that one of the four sporozoites released from the viable oocyst has infected a single cell.


2021 ◽  
Vol 937 (2) ◽  
pp. 022020
Author(s):  
Liang Chen ◽  
Zheng Huo ◽  
Xiafei Zhou ◽  
Baiqiang Niu ◽  
Wenting Zhang ◽  
...  

Abstract In this paper, we concentrate on the acute toxic effects of uranium on two aquatic organisms, Brachydanio rerio (B. rerio) and Cypridopsis vidua (C. vidua). We found that the toxicity of uranium on C. vidua was significantly greater than that of B. rerio. The results show that C. vidua has a higher sensitivity to uranium, even better than the commonly used test model organisms. In addition to its high sensitivity, C. vidua is a simple and cost-effective toxicological test organism. Therefore, C. vidua is an attractive alternative biological detection material. The acute toxicity results of the test are valuable for establishing water quality standards and protecting human health. At the same time, it enriches the relevant data of uranium on biological toxicity, provides clues for the study of the mechanism of toxicity, and deepens the understanding of the harm of uranium pollution to aquatic ecosystems.


Pathogens ◽  
2021 ◽  
Vol 10 (5) ◽  
pp. 523
Author(s):  
Marianne Lebbad ◽  
Jadwiga Winiecka-Krusnell ◽  
Christen Rune Stensvold ◽  
Jessica Beser

The intestinal protozoan parasite Cryptosporidium is an important cause of diarrheal disease worldwide. The aim of this study was to expand the knowledge on the molecular epidemiology of human cryptosporidiosis in Sweden to better understand transmission patterns and potential zoonotic sources. Cryptosporidium-positive fecal samples were collected between January 2013 and December 2014 from 12 regional clinical microbiology laboratories in Sweden. Species and subtype determination was achieved using small subunit ribosomal RNA and 60 kDa glycoprotein gene analysis. Samples were available for 398 patients, of whom 250 (63%) and 138 (35%) had acquired the infection in Sweden and abroad, respectively. Species identification was successful for 95% (379/398) of the samples, revealing 12 species/genotypes: Cryptosporidium parvum (n = 299), C. hominis (n = 49), C. meleagridis (n = 8), C. cuniculus (n = 5), Cryptosporidium chipmunk genotype I (n = 5), C. felis (n = 4), C. erinacei (n = 2), C. ubiquitum (n = 2), and one each of C. suis, C. viatorum, C. ditrichi, and Cryptosporidium horse genotype. One patient was co-infected with C. parvum and C. hominis. Subtyping was successful for all species/genotypes, except for C. ditrichi, and revealed large diversity, with 29 subtype families (including 4 novel ones: C. parvum IIr, IIs, IIt, and Cryptosporidium horse genotype VIc) and 81 different subtypes. The most common subtype families were IIa (n = 164) and IId (n = 118) for C. parvum and Ib (n = 26) and Ia (n = 12) for C. hominis. Infections caused by the zoonotic C. parvum subtype families IIa and IId dominated both in patients infected in Sweden and abroad, while most C. hominis cases were travel-related. Infections caused by non-hominis and non-parvum species were quite common (8%) and equally represented in cases infected in Sweden and abroad.


Author(s):  
Fatemeh Rahmani ◽  
Yahya Ehteshaminia ◽  
Hamid Mohammadi ◽  
Seif Ali Mahdavi

Introduction: Trichomoniasis is the most common non-viral sexually transmitted infection in the world, caused by the protozoan parasite Trichomonas vaginalis, which infects the urogenital tract of men and women. Approximately, 250 million new cases of Trichomonas vaginalis Infection are reported worldwide each year. Trichomoniasis is also considered an important HIV co-infection. The infection is often asymptomatic but can be accompanied by symptoms such as severe inflammation, itching and irritation, foamy discharge, and malodorous smell mucus, but the signs and symptoms of the disease are not sufficient for specific diagnosis. Material and Methods: In this study, the websites of PubMed, Google Scholar, SID, and Margiran were searched and related articles were reviewed. Results: Only screening and the use of highly sensitive and specific diagnostic methods can identify asymptomatic individuals. Today, the most common way to diagnose the infection is to use wet slide, Pap smear and culture methods that do not have high sensitivity and specificity. Also, due to the increase in infection and its complications, finding an efficient, rapid, and easy test to detect the parasite and differentiate Trichomoniasis vaginitis from other sexually transmitted diseases is considered important and necessary. Conclusion: Nowadays, there are several diagnostic methods that differentiate trichomoniasis infection from other sexually transmitted infections with high accuracy and sensitivity. Of course, existing diagnostic methods mostly use women's urine and vaginal samples for diagnosis, and methods that specifically diagnose the infection in men are more limited.


2015 ◽  
Vol 71 (11) ◽  
pp. 2236-2247 ◽  
Author(s):  
Cristiana M. Sousa ◽  
Philippe Carpentier ◽  
Pedro M. Matias ◽  
Fabrizio Testa ◽  
Filipa Pinho ◽  
...  

Superoxide reductase (SOR), which is commonly found in prokaryotic organisms, affords protection from oxidative stress by reducing the superoxide anion to hydrogen peroxide. The reaction is catalyzed at the iron centre, which is highly conserved among the prokaryotic SORs structurally characterized to date. Reported here is the first structure of an SOR from a eukaryotic organism, the protozoan parasiteGiardia intestinalis(GiSOR), which was solved at 2.0 Å resolution. By collecting several diffraction data sets at 100 K from the same flash-cooled protein crystal using synchrotron X-ray radiation, photoreduction of the iron centre was observed. Reduction was monitored using an online UV–visible microspectrophotometer, following the decay of the 647 nm absorption band characteristic of the iron site in the glutamate-bound, oxidized state. Similarly to other 1Fe-SORs structurally characterized to date, the enzyme displays a tetrameric quaternary-structure arrangement. As a distinctive feature, the N-terminal loop of the protein, containing the characteristic EKHxP motif, revealed an unusually high flexibility regardless of the iron redox state. At variance with previous evidence collected by X-ray crystallography and Fourier transform infrared spectroscopy of prokaryotic SORs, iron reduction did not lead to dissociation of glutamate from the catalytic metal or other structural changes; however, the glutamate ligand underwent X-ray-induced chemical changes, revealing high sensitivity of theGiSOR active site to X-ray radiation damage.


Pathogens ◽  
2021 ◽  
Vol 11 (1) ◽  
pp. 21
Author(s):  
Jiawen Nie ◽  
Jigang Yin ◽  
Dongqiang Wang ◽  
Chenchen Wang ◽  
Guan Zhu

Phosphoglucomutase 1 (PGM1) catalyzes the conversion between glucose-1-phosphate and glucose-6-phosphate in the glycolysis/glucogenesis pathway. PGM1s are typically cytosolic enzymes in organisms lacking chloroplasts. However, the protozoan Cryptosporidium parasites possess two tandemly duplicated PGM1 genes evolved by a gene duplication after their split from other apicomplexans. Moreover, the downstream PGM1 isoform contains an N-terminal signal peptide, predicting a non-cytosolic location. Here we expressed recombinant proteins of the two PGM1 isoforms from the zoonotic Cryptosporidium parvum, namely CpPGM1A and CpPGM1B, and confirmed their enzyme activity. Both isoforms followed Michaelis–Menten kinetics towards glucose-1-phosphate (Km = 0.17 and 0.13 mM, Vmax = 7.30 and 2.76 μmol/min/mg, respectively). CpPGM1A and CpPGM1B genes were expressed in oocysts, sporozoites and intracellular parasites at a similar pattern of expression, however CpPGM1A was expressed at much higher levels than CpPGM1B. Immunofluorescence assay showed that CpPGM1A was present in the cytosol of sporozoites, however this was enriched towards the plasma membranes in the intracellular parasites; whereas CpPGM1B was mainly present under sporozoite pellicle, although relocated to the parasitophorous vacuole membrane in the intracellular development. These observations indicated that CpPGM1A played a house-keeping function, while CpPGM1B played a different biological role that remains to be defined by future investigations.


2004 ◽  
Vol 72 (5) ◽  
pp. 2772-2779 ◽  
Author(s):  
Tarek K. Zaalouk ◽  
Mona Bajaj-Elliott ◽  
John T. George ◽  
Vincent McDonald

ABSTRACT Invasion of enterocytes by pathogenic microbes evokes both innate and adaptive immune responses, and microbial pathogens have developed strategies to overcome the initial host immune defense. β-Defensins are potentially important endogenous antibiotic-like effectors of innate immunity expressed by intestinal epithelia. In this study, the interplay between the enteric protozoan parasite Cryptosporidium parvum and host epithelial β-defensin expression was investigated. Using human and murine models of infection, we demonstrated that C. parvum infection differentially regulates β-defensin gene expression. Downregulation of murine β-defensin-1 mRNA and protein was observed in both in vitro and in vivo models of infection. Infection of the human colonic HT29 cell line with the parasite resulted in differential effects on various members of the defensin gene family. Partial reduction in human β-defensin-1 (hBD-1), induction of hBD-2, and no effect on hBD-3 gene expression was observed. Recombinant hBD-1 and hBD-2 peptides exhibited significant antimicrobial activity against C. parvum sporozoites in vitro. These findings demonstrate that C. parvum infection of enterocytes may affect the expression of various defensins in different ways and suggest that the overall outcome of the effect of antimicrobial peptides on early survival of the parasite may be complex.


Parasitology ◽  
2008 ◽  
Vol 136 (2) ◽  
pp. 149-157 ◽  
Author(s):  
B.-K. NA ◽  
J.-M. KANG ◽  
H.-I. CHEUN ◽  
S.-H. CHO ◽  
S.-U. MOON ◽  
...  

SUMMARYCryptosporidium parvum is an intracellular protozoan parasite that causes cryptosporidiosis in mammals including humans. In the current study, the gene encoding the cysteine protease of C. parvum (cryptopain-1) was identified and the biochemical properties of the recombinant enzyme were characterized. Cryptopain-1 shared common structural properties with cathepsin L-like papain family enzymes, but lacked a typical signal peptide sequence and contained a possible transmembrane domain near the amino terminus and a unique insert in the front of the mature domain. The recombinant cryptopain-1 expressed in Escherichia coli and refolded to the active form showed typical biochemical properties of cathepsin L-like enzymes. The folding determinant of cryptopain-1 was characterized through multiple constructs with or without different lengths of the pro-domain of the enzyme expressed in E. coli and assessment of their refolding abilities. All constructs, except one that did not contain the full-length mature domain, successfully refolded into the active enzymes, suggesting that cryptopain-1 did not require the pro-domain for folding. Western blot analysis showed that cryptopain-1 was expressed in the sporozoites and the enzyme preferentially degraded proteins, including collagen and fibronectin, but not globular proteins. This suggested a probable role for cryptopain-1 in host cell invasion and/or egression by the parasite.


2015 ◽  
Vol 82 (2) ◽  
pp. 431-437 ◽  
Author(s):  
Pamela Cameron ◽  
Birgit K. Gaiser ◽  
Bidha Bhandari ◽  
Paul M. Bartley ◽  
Frank Katzer ◽  
...  

ABSTRACTOocysts of the waterborne protozoan parasiteCryptosporidium parvumare highly resistant to chlorine disinfection. We show here that both silver nanoparticles (AgNPs) and silver ions significantly decrease oocyst viability, in a dose-dependent manner, between concentrations of 0.005 and 500 μg/ml, as assessed by an excystation assay and the shell/sporozoite ratio. For percent excystation, the results are statistically significant for 500 μg/ml of AgNPs, with reductions from 83% for the control to 33% with AgNPs. For Ag ions, the results were statistically significant at 500 and 5,000 μg/ml, but the percent excystation values were reduced only to 66 and 62%, respectively, from 86% for the control. The sporozoite/shell ratio was affected to a greater extent following AgNP exposure, presumably because sporozoites are destroyed by interaction with NPs. We also demonstrated via hyperspectral imaging that there is a dual mode of interaction, with Ag ions entering the oocyst and destroying the sporozoites while AgNPs interact with the cell wall and, at high concentrations, are able to fully break the oocyst wall.


2017 ◽  
Vol 73 (7) ◽  
pp. 387-394
Author(s):  
Agnieszka Kaupke ◽  
Artur Rzeżutka

Cryptosporidium parvum is a zoonotic protozoan parasite occurring in a wide range of hosts. Invasions caused by this parasite have been reported in humans and in many animal species including birds. Despite its worldwide prevalence, infections have usually generated considerable losses in the livestock industry, mostly affecting calves, lambs and goat kids. It has previously been shown that ruminants are a major reservoir of zoonotic Cryptosporidium parvum and contact with an infected animal can lead to human infection. The application of molecular methods for parasitological diagnostics has increased our knowledge on the parasite hosts and its prevalence in humans and animals. They also confirmed their usefulness during epidemiological investigations and in surveillance studies of human and animal cryptosporidiosis. In this review the current state of knowledge concerning the importance of Cryptosporidium parvum invasions in farm and wild animals was presented.


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