Application of a universal parasite diagnostic test to biological specimens collected from animals
- Lane, Meredith, Kashani, Mitra, Barratt, Joel, Qvarnstrom, Yvonne, Yabsley, Michael, Garrett, Kayla, Bradbury, Richard
- Authors: Lane, Meredith , Kashani, Mitra , Barratt, Joel , Qvarnstrom, Yvonne , Yabsley, Michael , Garrett, Kayla , Bradbury, Richard
- Date: 2023
- Type: Text , Journal article
- Relation: International Journal for Parasitology: Parasites and Wildlife Vol. 20, no. (2023), p. 20-30
- Full Text:
- Reviewed:
- Description: A previously described universal parasite diagnostic (nUPDx) based on PCR amplification of the 18S rDNA and deep-amplicon sequencing, can detect human blood parasites with a sensitivity comparable to real-time PCR. To date, the efficacy of this assay has only been assessed on human blood. This study assessed the utility of nUPDx for the detection of parasitic infections in animals using blood, tissues, and other biological sample types from mammals, birds, and reptiles, known to be infected with helminth, apicomplexan, or pentastomid parasites (confirmed by microscopy or PCR), as well as negative samples. nUPDx confirmed apicomplexan and/or nematode infections in 24 of 32 parasite-positive mammals, while also identifying several undetected coinfections. nUPDx detected infections in 6 of 13 positive bird and 1 of 2 positive reptile samples. When applied to 10 whole parasite specimens (worms and arthropods), nUPDx identified all to the genus or family level, and detected one incorrect identification made by morphology. Babesia sp. infections were detected in 5 of the 13 samples that were negative by other diagnostic approaches. While nUPDx did not detect PCR/microscopy-confirmed trichomonads or amoebae in cloacal swabs/tissue from 8 birds and 2 reptiles due to primer template mismatches, 4 previously undetected apicomplexans were detected in these samples. Future efforts to improve the utility of the assay should focus on validation against a larger panel of tissue types and animal species. Overall, nUPDx shows promise for use in both veterinary diagnostics and wildlife surveillance, especially because species-specific PCRs can miss unknown or unexpected pathogens. © 2022
- Authors: Lane, Meredith , Kashani, Mitra , Barratt, Joel , Qvarnstrom, Yvonne , Yabsley, Michael , Garrett, Kayla , Bradbury, Richard
- Date: 2023
- Type: Text , Journal article
- Relation: International Journal for Parasitology: Parasites and Wildlife Vol. 20, no. (2023), p. 20-30
- Full Text:
- Reviewed:
- Description: A previously described universal parasite diagnostic (nUPDx) based on PCR amplification of the 18S rDNA and deep-amplicon sequencing, can detect human blood parasites with a sensitivity comparable to real-time PCR. To date, the efficacy of this assay has only been assessed on human blood. This study assessed the utility of nUPDx for the detection of parasitic infections in animals using blood, tissues, and other biological sample types from mammals, birds, and reptiles, known to be infected with helminth, apicomplexan, or pentastomid parasites (confirmed by microscopy or PCR), as well as negative samples. nUPDx confirmed apicomplexan and/or nematode infections in 24 of 32 parasite-positive mammals, while also identifying several undetected coinfections. nUPDx detected infections in 6 of 13 positive bird and 1 of 2 positive reptile samples. When applied to 10 whole parasite specimens (worms and arthropods), nUPDx identified all to the genus or family level, and detected one incorrect identification made by morphology. Babesia sp. infections were detected in 5 of the 13 samples that were negative by other diagnostic approaches. While nUPDx did not detect PCR/microscopy-confirmed trichomonads or amoebae in cloacal swabs/tissue from 8 birds and 2 reptiles due to primer template mismatches, 4 previously undetected apicomplexans were detected in these samples. Future efforts to improve the utility of the assay should focus on validation against a larger panel of tissue types and animal species. Overall, nUPDx shows promise for use in both veterinary diagnostics and wildlife surveillance, especially because species-specific PCRs can miss unknown or unexpected pathogens. © 2022
Inactivating effects of common laboratory disinfectants, fixatives, and temperatures on the eggs of soil transmitted helminths
- Kines, Kristine, Fox, Mark, Ndubuisi, MacKevin, Verocai, Guilherme, Cama, Vitaliano, Bradbury, Richard
- Authors: Kines, Kristine , Fox, Mark , Ndubuisi, MacKevin , Verocai, Guilherme , Cama, Vitaliano , Bradbury, Richard
- Date: 2021
- Type: Text , Journal article
- Relation: Microbiology Spectrum Vol. 9, no. 3 (2021), p.
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- Description: Soil-transmitted helminths (STH) are important and widespread intestinal pathogens of humans and animals. It is presently unknown which inactivating procedures may be universally effective for safe transport, preservation, and disinfection of STH-contaminated specimens, and this lack of knowledge may expose laboratory staff to higher risk of laboratory-acquired infections (LAI’s). There are limited data on the efficacy of commonly used disinfectants and fecal fixatives for inactivating the eggs of STH. This work tested five disinfectants for surface cleanup, four storage temperature conditions, and six transport/storage fixatives, to inactivate eggs of three species of STH of animal origin (Ascaris suum “roundworm,” Trichuris vulpis “whipworm” and Ancylostoma caninum “hookworm”) as surrogates for human STH. Among disinfectants, exposure to 10% povidone-iodine for $5 min inactivated 100% of the three species tested, while 5 min exposure to 95% ethanol inactivated T. vulpis and A. caninum eggs. All of the fixatives tested had inactivation effects on A. caninum hookworm eggs within 24 h of exposure, except potassium dichromate, which required 48 h. 95% ethanol for $48 h inactivated eggs from all three STH species. Freezing at #220°C for $24 h inactivated eggs of T. vulpis and A. caninum, but only freezing at 280°C for $24 h inactivated .99% eggs, including A. suum. This work provides an evidence base for health and safety guidelines and mitigation strategies for the handling, storage, and disposal of stool samples containing STH eggs in laboratory, health care, childcare, or veterinary settings. IMPORTANCE This study systematically evaluates common laboratory disinfectants and storage conditions for their effectiveness in inactivating the infective stages of soil-transmitted helminths (STH). Animal-infecting proxy species were chosen to represent three major groups of STH that infect humans: roundworms, whipworms, and hookworms. Previously published work in this area typically focuses on a particular inactivation method, either for a single STH species, or on a subset of closely related species. Because prediagnostic fecal specimens must be regarded as potentially infectious with a mix of species, such information may be of limited utility in a working laboratory. We provide a straightforward summary of storage and disinfection methods that can achieve complete inactivation across a range of STH species, which represents a significant advance for clinical, veterinary and research laboratory biosafety. Copyright © 2021 Kines et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.
- Authors: Kines, Kristine , Fox, Mark , Ndubuisi, MacKevin , Verocai, Guilherme , Cama, Vitaliano , Bradbury, Richard
- Date: 2021
- Type: Text , Journal article
- Relation: Microbiology Spectrum Vol. 9, no. 3 (2021), p.
- Full Text:
- Reviewed:
- Description: Soil-transmitted helminths (STH) are important and widespread intestinal pathogens of humans and animals. It is presently unknown which inactivating procedures may be universally effective for safe transport, preservation, and disinfection of STH-contaminated specimens, and this lack of knowledge may expose laboratory staff to higher risk of laboratory-acquired infections (LAI’s). There are limited data on the efficacy of commonly used disinfectants and fecal fixatives for inactivating the eggs of STH. This work tested five disinfectants for surface cleanup, four storage temperature conditions, and six transport/storage fixatives, to inactivate eggs of three species of STH of animal origin (Ascaris suum “roundworm,” Trichuris vulpis “whipworm” and Ancylostoma caninum “hookworm”) as surrogates for human STH. Among disinfectants, exposure to 10% povidone-iodine for $5 min inactivated 100% of the three species tested, while 5 min exposure to 95% ethanol inactivated T. vulpis and A. caninum eggs. All of the fixatives tested had inactivation effects on A. caninum hookworm eggs within 24 h of exposure, except potassium dichromate, which required 48 h. 95% ethanol for $48 h inactivated eggs from all three STH species. Freezing at #220°C for $24 h inactivated eggs of T. vulpis and A. caninum, but only freezing at 280°C for $24 h inactivated .99% eggs, including A. suum. This work provides an evidence base for health and safety guidelines and mitigation strategies for the handling, storage, and disposal of stool samples containing STH eggs in laboratory, health care, childcare, or veterinary settings. IMPORTANCE This study systematically evaluates common laboratory disinfectants and storage conditions for their effectiveness in inactivating the infective stages of soil-transmitted helminths (STH). Animal-infecting proxy species were chosen to represent three major groups of STH that infect humans: roundworms, whipworms, and hookworms. Previously published work in this area typically focuses on a particular inactivation method, either for a single STH species, or on a subset of closely related species. Because prediagnostic fecal specimens must be regarded as potentially infectious with a mix of species, such information may be of limited utility in a working laboratory. We provide a straightforward summary of storage and disinfection methods that can achieve complete inactivation across a range of STH species, which represents a significant advance for clinical, veterinary and research laboratory biosafety. Copyright © 2021 Kines et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.
Medical parasitology taxonomy update, January 2018 to May 2020
- Mathison, Blaine, Bradbury, Richard, Pritt, Bobbi
- Authors: Mathison, Blaine , Bradbury, Richard , Pritt, Bobbi
- Date: 2021
- Type: Text , Journal article , Review
- Relation: Journal of Clinical Microbiology Vol. 59, no. 2 (2021), p.
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- Description: The taxonomy of parasites of medical and public health importance is rapidly evolving. This minireview provides an update of taxonomic revisions and additions in the field of medical parasitology from January 2018 to May 2020. Several established human parasites have been reassigned to different genera over the past 2 years, while a number of novel parasites of humans have been identified. A comprehensive summary of these changes is provided here, and Taenia suihominis is proposed as a replacement name for Taenia asiaticus Eom et al., which is a homonym of Taenia asiatica von Linstow. © 2021 American Society for Microbiology. All rights reserved. Erratum: Medical parasitology taxonomy update, January 2018 to May 2020 (Journal of Clinical Microbiology (2021) 59:2 (e01308-20) DOI: https://doi.org/10.1128/JCM.01308-20
- Authors: Mathison, Blaine , Bradbury, Richard , Pritt, Bobbi
- Date: 2021
- Type: Text , Journal article , Review
- Relation: Journal of Clinical Microbiology Vol. 59, no. 2 (2021), p.
- Full Text:
- Reviewed:
- Description: The taxonomy of parasites of medical and public health importance is rapidly evolving. This minireview provides an update of taxonomic revisions and additions in the field of medical parasitology from January 2018 to May 2020. Several established human parasites have been reassigned to different genera over the past 2 years, while a number of novel parasites of humans have been identified. A comprehensive summary of these changes is provided here, and Taenia suihominis is proposed as a replacement name for Taenia asiaticus Eom et al., which is a homonym of Taenia asiatica von Linstow. © 2021 American Society for Microbiology. All rights reserved. Erratum: Medical parasitology taxonomy update, January 2018 to May 2020 (Journal of Clinical Microbiology (2021) 59:2 (e01308-20) DOI: https://doi.org/10.1128/JCM.01308-20
Cloning, expression, purification and crystallographic studies of galectin-11 from domestic sheep (Ovis aries)
- Sakthivel, Dhanasekaran, Littler, Dene, Shahine, Adam, Troy, Sally, Johnson, Matthew, Rossjohn, Jamie, Piedrafita, David, Beddoe, Travis
- Authors: Sakthivel, Dhanasekaran , Littler, Dene , Shahine, Adam , Troy, Sally , Johnson, Matthew , Rossjohn, Jamie , Piedrafita, David , Beddoe, Travis
- Date: 2015
- Type: Text , Journal article
- Relation: Acta Crystallographica Section:F Structural Biology Communications Vol. 71, no. (2015), p. 993-997
- Full Text:
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- Description: Galectins are an evolutionarily conserved family of proteins that translate glycan recognition into cellular effects. Galectin-11 is a unique member of the galectin family that is only expressed in ruminants such as sheep, goat and cattle and that plays a critical role in several important biological processes, such as reproduction and parasite-mediated innate immune responses. Currently, these two areas are of major importance for the sustainability of ruminant livestock production. Despite the emerging biological significance of galectin-11, no structural information is available. It is expected that structural studies will unravel the functional mechanisms of galectin-11 activity. Here, the expression, purification and crystallization of the ruminant-specific galectin-11 from domestic sheep and the collection of X-ray data to 2.0 Å resolution are reported. © 2015.
- Authors: Sakthivel, Dhanasekaran , Littler, Dene , Shahine, Adam , Troy, Sally , Johnson, Matthew , Rossjohn, Jamie , Piedrafita, David , Beddoe, Travis
- Date: 2015
- Type: Text , Journal article
- Relation: Acta Crystallographica Section:F Structural Biology Communications Vol. 71, no. (2015), p. 993-997
- Full Text:
- Reviewed:
- Description: Galectins are an evolutionarily conserved family of proteins that translate glycan recognition into cellular effects. Galectin-11 is a unique member of the galectin family that is only expressed in ruminants such as sheep, goat and cattle and that plays a critical role in several important biological processes, such as reproduction and parasite-mediated innate immune responses. Currently, these two areas are of major importance for the sustainability of ruminant livestock production. Despite the emerging biological significance of galectin-11, no structural information is available. It is expected that structural studies will unravel the functional mechanisms of galectin-11 activity. Here, the expression, purification and crystallization of the ruminant-specific galectin-11 from domestic sheep and the collection of X-ray data to 2.0 Å resolution are reported. © 2015.
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