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Detection of Brucella abortus DNA and RNA in different stages of development of the sucking louse Haematopinus tuberculatus

BACKGROUND: Brucellosis is considered the world’s most widespread zoonotic infection. It causes abortion and sterility in livestock leading to serious economic losses and has even more serious medical impact in humans, since it can be a trigger to more than 500,000 infections per year worldwide. The...

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Autores principales: Neglia, Gianluca, Veneziano, Vincenzo, De Carlo, Esterina, Galiero, Giorgio, Borriello, Giorgia, Francillo, Matteo, Campanile, Giuseppe, Zicarelli, Luigi, Manna, Laura
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4220825/
https://www.ncbi.nlm.nih.gov/pubmed/24289112
http://dx.doi.org/10.1186/1746-6148-9-236
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author Neglia, Gianluca
Veneziano, Vincenzo
De Carlo, Esterina
Galiero, Giorgio
Borriello, Giorgia
Francillo, Matteo
Campanile, Giuseppe
Zicarelli, Luigi
Manna, Laura
author_facet Neglia, Gianluca
Veneziano, Vincenzo
De Carlo, Esterina
Galiero, Giorgio
Borriello, Giorgia
Francillo, Matteo
Campanile, Giuseppe
Zicarelli, Luigi
Manna, Laura
author_sort Neglia, Gianluca
collection PubMed
description BACKGROUND: Brucellosis is considered the world’s most widespread zoonotic infection. It causes abortion and sterility in livestock leading to serious economic losses and has even more serious medical impact in humans, since it can be a trigger to more than 500,000 infections per year worldwide. The aim of this study was to evaluate the role of Haematopinus tuberculatus, a louse that can parasitize several ruminants, as a new host of brucellosis. Louse specimens were collected from seropositive and seronegative water buffaloes and divided in 3 developmental stages: adults, nymphs and nits. All samples were separately screened for Brucella spp. DNA and RNA detection by Real Time PCR. In particular, primers and probes potentially targeting the 16S rRNA and the Brucella Cell Surface 31 kDalton Protein (bcsp31) genes were used for Real Time PCR and buffalo β actin was used as a housekeeping gene to quantify host DNA in the sample. A known amount of B. abortus purified DNA was utilized for standard curve preparation and the target DNA amount was divided by the housekeeping gene amount to obtain a normalized target value. A further molecular characterization was performed for Brucella strain typing and genotyping by the Bruce-ladder, AMOS-PCR and MLVA assays. Data were statistically analysed by ANOVA. RESULTS: Brucella abortus DNA and RNA were detected in all developmental stages of the louse, suggesting the presence of viable bacteria. Data obtained by MLVA characterization support this finding, since the strains present in animals and the relative parasites were not always identical, suggesting bacterial replication. Furthermore, the detection of Brucella DNA and RNA in nits samples demonstrate, for the first time, a trans-ovarial transmission of the bacterium into the louse. CONCLUSIONS: These findings identified H. tuberculatus as a new host of brucellosis. Further studies are needed to establish the role of this louse in the epidemiology of the disease, such as vector or reservoir.
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spelling pubmed-42208252014-11-06 Detection of Brucella abortus DNA and RNA in different stages of development of the sucking louse Haematopinus tuberculatus Neglia, Gianluca Veneziano, Vincenzo De Carlo, Esterina Galiero, Giorgio Borriello, Giorgia Francillo, Matteo Campanile, Giuseppe Zicarelli, Luigi Manna, Laura BMC Vet Res Research Article BACKGROUND: Brucellosis is considered the world’s most widespread zoonotic infection. It causes abortion and sterility in livestock leading to serious economic losses and has even more serious medical impact in humans, since it can be a trigger to more than 500,000 infections per year worldwide. The aim of this study was to evaluate the role of Haematopinus tuberculatus, a louse that can parasitize several ruminants, as a new host of brucellosis. Louse specimens were collected from seropositive and seronegative water buffaloes and divided in 3 developmental stages: adults, nymphs and nits. All samples were separately screened for Brucella spp. DNA and RNA detection by Real Time PCR. In particular, primers and probes potentially targeting the 16S rRNA and the Brucella Cell Surface 31 kDalton Protein (bcsp31) genes were used for Real Time PCR and buffalo β actin was used as a housekeeping gene to quantify host DNA in the sample. A known amount of B. abortus purified DNA was utilized for standard curve preparation and the target DNA amount was divided by the housekeeping gene amount to obtain a normalized target value. A further molecular characterization was performed for Brucella strain typing and genotyping by the Bruce-ladder, AMOS-PCR and MLVA assays. Data were statistically analysed by ANOVA. RESULTS: Brucella abortus DNA and RNA were detected in all developmental stages of the louse, suggesting the presence of viable bacteria. Data obtained by MLVA characterization support this finding, since the strains present in animals and the relative parasites were not always identical, suggesting bacterial replication. Furthermore, the detection of Brucella DNA and RNA in nits samples demonstrate, for the first time, a trans-ovarial transmission of the bacterium into the louse. CONCLUSIONS: These findings identified H. tuberculatus as a new host of brucellosis. Further studies are needed to establish the role of this louse in the epidemiology of the disease, such as vector or reservoir. BioMed Central 2013-12-01 /pmc/articles/PMC4220825/ /pubmed/24289112 http://dx.doi.org/10.1186/1746-6148-9-236 Text en Copyright © 2013 Neglia et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Neglia, Gianluca
Veneziano, Vincenzo
De Carlo, Esterina
Galiero, Giorgio
Borriello, Giorgia
Francillo, Matteo
Campanile, Giuseppe
Zicarelli, Luigi
Manna, Laura
Detection of Brucella abortus DNA and RNA in different stages of development of the sucking louse Haematopinus tuberculatus
title Detection of Brucella abortus DNA and RNA in different stages of development of the sucking louse Haematopinus tuberculatus
title_full Detection of Brucella abortus DNA and RNA in different stages of development of the sucking louse Haematopinus tuberculatus
title_fullStr Detection of Brucella abortus DNA and RNA in different stages of development of the sucking louse Haematopinus tuberculatus
title_full_unstemmed Detection of Brucella abortus DNA and RNA in different stages of development of the sucking louse Haematopinus tuberculatus
title_short Detection of Brucella abortus DNA and RNA in different stages of development of the sucking louse Haematopinus tuberculatus
title_sort detection of brucella abortus dna and rna in different stages of development of the sucking louse haematopinus tuberculatus
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4220825/
https://www.ncbi.nlm.nih.gov/pubmed/24289112
http://dx.doi.org/10.1186/1746-6148-9-236
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