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Article ; Online: Comparative performance evaluation of four commercial multiplex real-time PCR assays for the detection of the diarrhoea-causing protozoa Cryptosporidium hominis/parvum, Giardia duodenalis and Entamoeba histolytica.

Silvia Paulos / José María Saugar / Aida de Lucio / Isabel Fuentes / María Mateo / David Carmena

PLoS ONE, Vol 14, Iss 4, p e

2019 Volume 0215068

Abstract: Background Multiplex molecular panels are relentlessly replacing conventional methods for the detection of enteric pathogens from stool samples in clinical and research laboratories. Here we evaluated four commercial multiplex real-time PCR assays for ... ...

Abstract	Background Multiplex molecular panels are relentlessly replacing conventional methods for the detection of enteric pathogens from stool samples in clinical and research laboratories. Here we evaluated four commercial multiplex real-time PCR assays for the detection of Cryptosporidium hominis/parvum, Giardia duodenalis and Entamoeba histolytica. Methods The diagnostic performance of the Gastroenteritis/Parasite Panel I (Diagenode), the RIDAGENE Parasitic Stool Panel (R-Biopharm), the Allplex Gastrointestinal Parasite Panel 4 (Seegene) and the FTD Stool Parasites (Fast Track) real-time PCR methods was assessed against a reference panel of 126 well-characterized DNA samples including Cryptosporidium hominis (n = 29), Cryptosporidium parvum (n = 3), Giardia duodenalis (n = 47), Entamoeba histolytica (n = 3), other parasite species (n = 20), and apparently healthy subjects (n = 24). Principal findings Obtained diagnostic sensitivities ranged from 53-88% for Cryptosporidium hominis/parvum, and from 68-100% for G. duodenalis. The R-Biopharm method achieved the best performance for the detection of Cryptosporidium hominis/parvum both in terms of diagnostic sensitivity (87.5%) and detection limit (a 100-fold increase compared to other tests). The Fast Track method was particularly suited for the detection of G. duodenalis, achieving a 100% sensitivity and a detection limit at least 10-fold superior. Detection of E. histolytica was similarly achieved by all compared methods except Diagenode. Conclusions Diagnostic performance varied largely depending on the method used and the targeted pathogen species. Factors including test sensitivity/specificity, cost, patient population surveyed, laboratory workflow, and diagnostic algorithm should be carefully considered when choosing the most appropriate multiplex PCR platform.
Keywords	Medicine ; R ; Science ; Q
Subject code	630
Language	English
Publishing date	2019-01-01T00:00:00Z
Publisher	Public Library of Science (PLoS)
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article: Occurrence and molecular epidemiology of Giardia duodenalis infection in dog populations in eastern Spain

Adell-Aledón, Manuel / Pamela C. Köster / Aida de Lucio / Paula Puente / Marta Hernández-de-Mingo / Paula Sánchez-Thevenet / María Auxiliadora Dea-Ayuela / David Carmena

BMC veterinary research. 2018 Dec., v. 14, no. 1

2018

Abstract: BACKGROUND: Giardia duodenalis is one of the most common enteric parasites in domestic animals including dogs. Young animals are more prone to the infection, with clinical manifestations ranging from asymptomatic to acute or chronic diarrhoea. Dogs are ... ...

Abstract	BACKGROUND: Giardia duodenalis is one of the most common enteric parasites in domestic animals including dogs. Young animals are more prone to the infection, with clinical manifestations ranging from asymptomatic to acute or chronic diarrhoea. Dogs are primarily infected by canine-specific (C-D) assemblages of G. duodenalis. However, zoonotic assemblages A and B have been increasingly documented in canine isolates, raising the question of whether and to which extent dogs can act as natural reservoirs of human giardiosis. METHODS: In this cross-sectional epidemiological survey we assessed the molecular diversity of G. duodenalis in dogs in the province of Castellón, Eastern Spain. A total of 348 individual faecal samples from sheltered (n = 218), breeding (n = 24), hunting (n = 68), shepherd (n = 24), and pet (n = 14) dogs were collected between 2014 and 2016. Detection of G. duodenalis cysts in faecal material was carried out by direct fluorescence microscopy as a screening test, whereas a qPCR targeting the small subunit ribosomal RNA gene of the parasite was subsequently used as a confirmatory method. RESULTS: Giardia duodenalis was detected in 36.5% (95% CI: 31.6–41.7%) of dogs. No significant differences in prevalence rates could be demonstrated among dogs according to their sex and geographical origin, but breeding (45.8%; 95% CI: 27.9–64.9%) and sheltered (40.4%; 95% CI: 34.1–47.0%) dogs harboured significantly higher proportions of G. duodenalis. Multi-locus sequence-based genotyping of the glutamate dehydrogenase and β-giardin genes of G. duodenalis allowed the characterization of 35 canine isolates that were unambiguously assigned to assemblages A (14.3%), B (22.9%), C (5.7%), and D (37.1%). A number of inter-assemblage mixed infections including A + B (11.4%), A + D (2.9%), and A + B + D (5.7%) were also identified. CONCLUSIONS: Data presented here are strongly indicative of high infection pressures in kennelled animals. Zoonotic sub-assemblages AII, BIII, and BIV were responsible for a considerable proportion of the G. duodenalis infections detected, but very few of the genotypes identified have been previously documented in Spanish human populations. Although possible, zoonotic transmission between dogs and humans seems an infrequent event in this Spanish region.
Keywords	Giardia lamblia ; breeding ; diarrhea ; dog diseases ; dogs ; epidemiological studies ; feces ; fluorescence microscopy ; genes ; genotype ; genotyping ; giardiasis ; giardin protein ; glutamate dehydrogenase ; human population ; humans ; mixed infection ; molecular epidemiology ; parasites ; pets ; provenance ; quantitative polymerase chain reaction ; ribosomal RNA ; veterinary medicine ; young animals ; Spain
Language	English
Dates of publication	2018-12
Size	p. 26.
Publishing place	BioMed Central
Document type	Article
ISSN	1746-6148
DOI	10.1186/s12917-018-1353-z
Database	NAL-Catalogue (AGRICOLA)

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Article: Occurrence and subtype distribution of Blastocystis sp. in humans, dogs and cats sharing household in northern Spain and assessment of zoonotic transmission risk

Paulos, Silvia / Pamela C. Köster / Aida de Lucio / Marta Hernández‐de‐Mingo / Guillermo A. Cardona / Juan C. Fernández‐Crespo / Christen R. Stensvold / David Carmena

Zoonoses and public health. 2018 Dec., v. 65, no. 8

2018

Abstract: Blastocystis sp. is probably the most common enteric parasite in humans globally. Although the role of Blastocystis in human disease is still controversial, epidemiological and experimental evidence suggests that pathogenicity may be associated with ... ...

Abstract	Blastocystis sp. is probably the most common enteric parasite in humans globally. Although the role of Blastocystis in human disease is still controversial, epidemiological and experimental evidence suggests that pathogenicity may be associated with certain subtypes of the protist. Since the life cycle of Blastocystis is maintained through still elusive pathways, companion animals have attracted the attention of researchers as potential reservoirs of human infections. In order to evaluate the risk of zoonotic transmission of Blastocystis, we investigated the occurrence and molecular diversity of this microorganism in human, canine and feline populations sharing temporal and spatial settings in the province of Álava, northern Spain. A total of 268 (including 179 human, 55 canine and 34 feline) faecal specimens were obtained from 63 family households during February–December 2014. Detection of Blastocystis was achieved by PCR amplification and sequencing of small subunit rRNA genes. Blastocystis was found in 35.2% (95% CI: 0.29%–0.42%) of the human stool samples analysed, but not in any of the canine or feline faecal specimens investigated. Out of the 63 PCR‐positive human samples, 84.1% (53/63) were successfully subtyped, allowing the identification of the subtypes ST2 (62.3%), ST3 (17.0%), ST1 (13.2%) and ST4 (7.5%). No mixed subtype infections were identified. Blastocystis carriage was independent of the gender and region of origin of the affected individuals, but children in the age groups of >5‒10 years and >10‒15 years were significantly more affected by the protist. None of the risk factors considered (water‐use practices, contact with livestock, contact with individual undergoing diarrhoeal episodes) were associated with increased prevalence of Blastocystis. Our data demonstrate that pet dogs and cats play a negligible role as natural reservoirs of human Blastocystis infection in this geographic region, although the applicability of these results should be corroborated in future molecular epidemiological studies.
Keywords	Blastocystis ; cats ; children ; dogs ; epidemiological studies ; feces ; gender ; genes ; households ; human diseases ; humans ; livestock ; parasites ; pathogenicity ; pets ; polymerase chain reaction ; protists ; provenance ; ribosomal RNA ; risk factors ; Spain
Language	English
Dates of publication	2018-12
Size	p. 993-1002.
Publishing place	John Wiley & Sons, Ltd
Document type	Article
Note	JOURNAL ARTICLE
ZDB-ID	2271118-1
ISSN	1863-2378 ; 1863-1959
ISSN (online)	1863-2378
ISSN	1863-1959
DOI	10.1111/zph.12522
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Occurrence and molecular epidemiology of Giardia duodenalis infection in dog populations in eastern Spain

Manuel Adell-Aledón / Pamela C. Köster / Aida de Lucio / Paula Puente / Marta Hernández-de-Mingo / Paula Sánchez-Thevenet / María Auxiliadora Dea-Ayuela / David Carmena

BMC Veterinary Research, Vol 14, Iss 1, Pp 1-

2018 Volume 11

Abstract: Abstract Background Giardia duodenalis is one of the most common enteric parasites in domestic animals including dogs. Young animals are more prone to the infection, with clinical manifestations ranging from asymptomatic to acute or chronic diarrhoea. ... ...

Abstract	Abstract Background Giardia duodenalis is one of the most common enteric parasites in domestic animals including dogs. Young animals are more prone to the infection, with clinical manifestations ranging from asymptomatic to acute or chronic diarrhoea. Dogs are primarily infected by canine-specific (C-D) assemblages of G. duodenalis. However, zoonotic assemblages A and B have been increasingly documented in canine isolates, raising the question of whether and to which extent dogs can act as natural reservoirs of human giardiosis. Methods In this cross-sectional epidemiological survey we assessed the molecular diversity of G. duodenalis in dogs in the province of Castellón, Eastern Spain. A total of 348 individual faecal samples from sheltered (n = 218), breeding (n = 24), hunting (n = 68), shepherd (n = 24), and pet (n = 14) dogs were collected between 2014 and 2016. Detection of G. duodenalis cysts in faecal material was carried out by direct fluorescence microscopy as a screening test, whereas a qPCR targeting the small subunit ribosomal RNA gene of the parasite was subsequently used as a confirmatory method. Results Giardia duodenalis was detected in 36.5% (95% CI: 31.6–41.7%) of dogs. No significant differences in prevalence rates could be demonstrated among dogs according to their sex and geographical origin, but breeding (45.8%; 95% CI: 27.9–64.9%) and sheltered (40.4%; 95% CI: 34.1–47.0%) dogs harboured significantly higher proportions of G. duodenalis. Multi-locus sequence-based genotyping of the glutamate dehydrogenase and β-giardin genes of G. duodenalis allowed the characterization of 35 canine isolates that were unambiguously assigned to assemblages A (14.3%), B (22.9%), C (5.7%), and D (37.1%). A number of inter-assemblage mixed infections including A + B (11.4%), A + D (2.9%), and A + B + D (5.7%) were also identified. Conclusions Data presented here are strongly indicative of high infection pressures in kennelled animals. Zoonotic sub-assemblages AII, BIII, and BIV were responsible for a ...
Keywords	Giardia duodenalis ; Protozoa ; Dogs ; Molecular epidemiology ; Castellón ; Spain ; Veterinary medicine ; SF600-1100
Subject code	630
Language	English
Publishing date	2018-01-01T00:00:00Z
Publisher	BMC
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Molecular Genotyping of Giardia duodenalis Isolates from Symptomatic Individuals Attending Two Major Public Hospitals in Madrid, Spain.

Aida de Lucio / Rocío Martínez-Ruiz / Francisco J Merino / Begoña Bailo / María Aguilera / Isabel Fuentes / David Carmena

PLoS ONE, Vol 10, Iss 12, p e

2015 Volume 0143981

Abstract: The flagellate protozoan Giardia duodenalis is an enteric parasite causing human giardiasis, a major gastrointestinal disease of global distribution affecting both developing and industrialised countries. In Spain, sporadic cases of giardiasis have been ... ...

Abstract	The flagellate protozoan Giardia duodenalis is an enteric parasite causing human giardiasis, a major gastrointestinal disease of global distribution affecting both developing and industrialised countries. In Spain, sporadic cases of giardiasis have been regularly identified, particularly in pediatric and immigrant populations. However, there is limited information on the genetic variability of circulating G. duodenalis isolates in the country.In this longitudinal molecular epidemiological study we report the diversity and frequency of the G. duodenalis assemblages and sub-assemblages identified in 199 stool samples collected from 184 individual with symptoms compatible with giardiasis presenting to two major public hospitals in Madrid for the period December 2013-January 2015. G. duodenalis cysts were initially detected by conventional microscopy and/or immunochomatography on stool samples. Confirmation of the infection was performed by direct immunofluorescence and real-time PCR methods. G. duodenalis assemblages and sub-assemblages were determined by multi-locus genotyping of the glutamate dehydrogenase (GDH) and β-giardin (BG) genes of the parasite. Sociodemographic and clinical features of patients infected with G. duodenalis were also analysed.Of 188 confirmed positive samples from 178 giardiasis cases a total of 124 G. duodenalis isolates were successfully typed at the GDH and/or the BG loci, revealing the presence of sub-assemblages BIV (62.1%), AII (15.3%), BIII (4.0%), AI (0.8%), and AIII (0.8%). Additionally, 6.5% of the isolates were only characterised at the assemblage level, being all of them assigned to assemblage B. Discordant genotype results AII/AIII or BIII/BIV were also observed in 10.5% of DNA isolates. A large number of multi-locus genotypes were identified in G. duodenalis assemblage B, but not assemblage A, isolates at both the GDH and BG loci, confirming the high degree of genetic variability observed in other molecular surveys. BIV was the most prevalent genetic variant of G. duodenalis found in individuals with symptomatic giardiasis in the population under study.Human giardiasis is an ongoing public health problem in Spain affecting primarily young children under four years of age but also individuals of all age groups. Our typing and sub-typing results demonstrate that assemblage B is the most prevalent G. duodenalis assemblage circulating in patients with clinical giardiasis in Central Spain. Our analyses also revealed a large genetic variability in assemblage B (but not assemblage A) isolates of the parasite, corroborating the information obtained in similar studies in other geographical regions. We believe that molecular data presented here provide epidemiological evidence at the population level in support of the existence of genetic exchange within assemblages of G. duodenalis.
Keywords	Medicine ; R ; Science ; Q
Language	English
Publisher	Public Library of Science (PLoS)
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Molecular diversity and frequency of the diarrheagenic enteric protozoan Giardia duodenalis and Cryptosporidium spp. in a hospital setting in Northern Spain.

José Manuel Azcona-Gutiérrez / Aida de Lucio / Marta Hernández-de-Mingo / Concepción García-García / Luis Miguel Soria-Blanco / Lucía Morales / María Aguilera / Isabel Fuentes / David Carmena

PLoS ONE, Vol 12, Iss 6, p e

2017 Volume 0178575

Abstract: Human giardiosis and cryptosporidiosis are caused by the enteric protozoan parasites Giardia duodenalis and Cryptosporidium spp. Both pathogens are major contributors to the global burden of diarrhoeal disease, affecting primarily children and ... ...

Abstract	Human giardiosis and cryptosporidiosis are caused by the enteric protozoan parasites Giardia duodenalis and Cryptosporidium spp. Both pathogens are major contributors to the global burden of diarrhoeal disease, affecting primarily children and immunodebilitated individuals in resource-poor settings. Giardiosis and cryptosporidiosis also represent an important, often underestimate, public health threat in developed countries. In Spain only limited information is currently available on the epidemiology of these infections. Molecular data on the diversity, frequency, geographical distribution, and seasonality of G. duodenalis assemblages/sub-assemblages and Cryptosporidium species/sub-genotypes are particularly scarce.A longitudinal molecular epidemiological survey was conducted between July 2015 to September 2016 in patients referred to or attended at the Hospital San Pedro (La Rioja, Northern Spain) that tested positive for G. duodenalis (N = 106) or Cryptosporidium spp. (N = 103) by direct microscopy and/or a rapid lateral flow immunochromatographic assay. G. duodenalis infections were subsequently confirmed by real-time PCR and positive isolates assessed by multi-locus sequence genotyping of the glutamate dehydrogenase and β-giardin genes of the parasite. Cryptosporidium species and sub-genotypes were investigated at the 60 kDa glycoprotein or the small subunit ribosomal RNA genes of the parasite. Sociodemographic and clinical parameters of infected patients were also gathered and analysed.Out of 90 G. duodenalis-positive isolates by real-time PCR a total of 16 isolates were successfully typed. AII (44%, 7/16) was the most prevalent sub-assemblage found, followed by BIV (31%, 5/16) and BIII (19%, 3/16). A discordant genotype result AII/AIII was identified in an additional (6%, 1/16) isolate. No mixed infections A+B were detected. Similarly, a total of 81 Cryptosporidium spp. isolates were successfully typed, revealing the presence of C. hominis (81%, 66/81) and C. parvum (19%, 15/81). Obtained GP60 sequences ...
Keywords	Medicine ; R ; Science ; Q
Subject code	610
Language	English
Publishing date	2017-01-01T00:00:00Z
Publisher	Public Library of Science (PLoS)
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article: Detection and molecular diversity of Giardia duodenalis and Cryptosporidium spp. in sheltered dogs and cats in Northern Spain

Gil, Horacio / Aida de Lucio / Begoña Bailo / David Carmena / Guillermo A. Cardona / José A. Fernández-Basterra / Juan Aramburu-Aguirre / Lourdes Cano / Marta Hernández de Mingo / Nuria López-Molina

Infection, genetics, and evolution. 2017 June, v. 50

2017

Abstract: Domestic dogs and cats may act as natural reservoirs of a large number of zoonotic pathogens, including the enteric parasites Giardia duodenalis and Cryptosporidium spp., the most relevant protozoan species causing gastrointestinal disease worldwide. A ... ...

Abstract	Domestic dogs and cats may act as natural reservoirs of a large number of zoonotic pathogens, including the enteric parasites Giardia duodenalis and Cryptosporidium spp., the most relevant protozoan species causing gastrointestinal disease worldwide. A cross-sectional epidemiological study aiming to assess the prevalence and molecular diversity of G. duodenalis and Cryptosporidium spp. was conducted in an animal rescue centre in the province of Álava (Northern Spain). A total of 194 and 65 faecal dropping samples from individual dogs and cats, respectively, were collected between November 2013 and June 2016. G. duodenalis cysts and Cryptosporidium spp. oocysts were detected by direct fluorescence microscopy and PCR-based methods targeting the small subunit ribosomal RNA gene of these parasites.Overall, G. duodenalis and Cryptosporidium spp. were detected in 33% (63/194) and 4.1% (8/194) of dogs, and 9.2% (6/65) and 4.6% (3/65) of cats, respectively. G. duodenalis and Cryptosporidium co-infections were observed in 1.5% (3/194) of dogs, but not in cats. No significant differences in infection rates could be demonstrated among dogs or cats according to their sex, age group, status, or geographical origin. Multi-locus sequence-based genotyping of the glutamate dehydrogenase and β-giardin genes of G. duodenalis allowed the characterization of 19 canine isolates that were unambiguously assigned to sub-assemblages AII (n=7), BIII (n=1), and BIV (n=7), and assemblages C (n=3) and D (n=1). Two feline isolates were genotyped as assemblages A and F, respectively. No mixed assemblage or sub-assemblage infections were identified. C. canis (n=5) and C. hominis (n=1) were the Cryptosporidium species found in dogs, whereas C. felis (n=1) was identified in cats. The finding of G. duodenalis sub-assemblages AII, BIII, and BIV circulating in dogs (but not cats) may have zoonotic potential, although most of the AII and BIV isolates sub-genotyped corresponded to genetic variants not previously found in Spanish human populations. Dogs may also act as novel suitable hosts for C. hominis. We recommend to considerer companion animals as sentinel surveillance system for zoonotic giardiasis and cryptosporidiosis in order to minimize the risk of spreading of these parasitic diseases among the human population.
Keywords	cats ; cryptosporidiosis ; Cryptosporidium ; digestive system diseases ; dogs ; epidemiological studies ; fluorescence microscopy ; genes ; genetic variation ; genotyping ; Giardia lamblia ; giardiasis ; giardin protein ; glutamate dehydrogenase ; hosts ; human population ; mixed infection ; monitoring ; oocysts ; parasites ; pathogens ; pets ; polymerase chain reaction ; provenance ; ribosomal RNA ; risk ; zoonoses ; Spain
Language	English
Dates of publication	2017-06
Size	p. 62-69.
Publishing place	Elsevier B.V.
Document type	Article
ZDB-ID	2037068-4
ISSN	1567-1348
ISSN	1567-1348
DOI	10.1016/j.meegid.2017.02.013
Database	NAL-Catalogue (AGRICOLA)

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Article: Evaluation of five commercial methods for the extraction and purification of DNA from human faecal samples for downstream molecular detection of the enteric protozoan parasites Cryptosporidium spp., Giardia duodenalis, and Entamoeba spp

Paulos, Silvia / Aida de Lucio / BegoÃ±a Bailo / David Carmena / Guillermo A. Cardona / Isabel Fuentes / JosÃ© M. Saugar / MarÃa Mateo / Marta HernÃ¡ndez-de Mingo / Marta Mateo

Journal of microbiological methods. 2016 Aug., v. 127

2016

Abstract: High quality, pure DNA is required for ensuring reliable and reproducible results in molecular diagnosis applications. A number of in-house and commercial methods are available for the extraction and purification of genomic DNA from faecal material, each ...

Abstract	High quality, pure DNA is required for ensuring reliable and reproducible results in molecular diagnosis applications. A number of in-house and commercial methods are available for the extraction and purification of genomic DNA from faecal material, each one offering a specific combination of performance, cost-effectiveness, and easiness of use that should be conveniently evaluated in function of the pathogen of interest. In this comparative study the marketed kits QIAamp DNA stool mini (Qiagen), SpeedTools DNA extraction (Biotools), DNAExtract-VK (Vacunek), PowerFecal DNA isolation (MoBio), and Wizard magnetic DNA purification system (Promega Corporation) were assessed for their efficacy in obtaining DNA of the most relevant enteric protozoan parasites associated to gastrointestinal disease globally. A panel of 113 stool specimens of clinically confirmed patients with cryptosporidiosis (n=29), giardiasis (n=47) and amoebiasis by Entamoeba histolytica (n=3) or E. dispar (n=10) and apparently healthy subjects (n=24) were used for this purpose. Stool samples were aliquoted in five sub-samples and individually processed by each extraction method evaluated. Purified DNA samples were subsequently tested in PCR-based assays routinely used in our laboratory. The five compared methods yielded amplifiable amounts of DNA of the pathogens tested, although performance differences were observed among them depending on the parasite and the infection burden. Methods combining chemical, enzymatic and/or mechanical lysis procedures at temperatures of at least 56Â°C were proven more efficient for the release of DNA from Cryptosporidium oocysts.
Keywords	amebiasis ; cost effectiveness ; cryptosporidiosis ; Cryptosporidium ; digestive system diseases ; DNA ; Entamoeba histolytica ; feces ; Giardia lamblia ; giardiasis ; humans ; oocysts ; parasites ; pathogens ; patients ; polymerase chain reaction ; temperature
Language	English
Dates of publication	2016-08
Size	p. 68-73.
Publishing place	Elsevier B.V.
Document type	Article
ZDB-ID	604916-3
ISSN	1872-8359 ; 0167-7012
ISSN (online)	1872-8359
ISSN	0167-7012
DOI	10.1016/j.mimet.2016.05.020
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Prevalence and Genetic Diversity of Giardia duodenalis and Cryptosporidium spp. among School Children in a Rural Area of the Amhara Region, North-West Ethiopia.

Aida de Lucio / Aranzazu Amor-Aramendía / Begoña Bailo / José M Saugar / Melaku Anegagrie / Ana Arroyo / Beatriz López-Quintana / Derjew Zewdie / Zimmam Ayehubizu / Endalew Yizengaw / Bayeh Abera / Mulat Yimer / Wondemagen Mulu / Tadesse Hailu / Zaida Herrador / Isabel Fuentes / David Carmena

PLoS ONE, Vol 11, Iss 7, p e

2016 Volume 0159992

Abstract: Giardia duodenalis and Cryptosporidium spp. are enteric protozoan causing gastrointestinal illness in humans and animals. Giardiasis and cryptosporidiosis are not formally considered as neglected tropical diseases, but belong to the group of poverty- ... ...

Abstract	Giardia duodenalis and Cryptosporidium spp. are enteric protozoan causing gastrointestinal illness in humans and animals. Giardiasis and cryptosporidiosis are not formally considered as neglected tropical diseases, but belong to the group of poverty-related infectious diseases that impair the development and socio-economic potential of infected individuals in developing countries.We report here the prevalence and genetic diversity of G. duodenalis and Cryptosporidium spp. in children attending rural primary schools in the Bahir Dar district of the Amhara Region, Ethiopia. Stool samples were collected from 393 children and analysed by molecular methods. G. duodenalis was detected by real-time PCR, and the assemblages and sub-assemblages were determined by multilocus sequence-based genotyping of the glutamate dehydrogenase and β-giardin genes of the parasite. Detection and identification of Cryptosporidium species was carried out by sequencing of a partial fragment of the small-subunit ribosomal RNA gene.The PCR-based prevalences of G. duodenalis and Cryptosporidium spp. were 55.0% (216/393) and 4.6% (18/393), respectively. A total of 78 G. duodenalis isolates were successfully characterized, revealing the presence of sub-assemblages AII (10.3%), BIII (28.2%), and BIV (32.0%). Discordant typing results AII/AIII and BIII/BIV were identified in 7.7% and 15.4% of the isolates, respectively. An additional five (6.4%) isolates were assigned to assemblage B. No mixed infections of assemblages A+B were found. Extensive genetic variation at the nucleotide level was observed within assemblage B (but no within assemblage A), resulting in the identification of a large number of sub-types. Cryptosporidium diversity was demonstrated by the occurrence of C. hominis, C. parvum, and C. viatorum in the population under study.Our data suggest an epidemiological scenario with an elevated transmission intensity of a wide range of G. duodenalis genetic variants. Importantly, the elevated degree of genetic diversity observed within assemblage B is consistent with the occurrence of intra-assemblage recombination in G. duodenalis.
Keywords	Medicine ; R ; Science ; Q
Subject code	580
Language	English
Publisher	Public Library of Science (PLoS)
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article: Occurrence and molecular genotyping of Giardia duodenalis and Cryptosporidium spp. in wild mesocarnivores in Spain

Mateo, Marta / Aida de Lucio / Alberto Espí / Ana Balseiro / David Carmena / Guillermo A. Cardona / José Francisco Lima Barbero / José L. Fernández-García / Lucía Morales / Marta Barral / Marta Hernández de Mingo / Miguel Ángel Habela / Pamela C. Köster / Rafael Calero Bernal

Veterinary parasitology. 2017 Feb. 15, v. 235

2017

Abstract: There is a surprisingly scarce amount of epidemiological and molecular data on the prevalence, frequency, and diversity of the intestinal protozoan parasites Giardia duodenalis and Cryptosporidium spp. in wildlife in general and mesocarnivore species in ... ...

Abstract	There is a surprisingly scarce amount of epidemiological and molecular data on the prevalence, frequency, and diversity of the intestinal protozoan parasites Giardia duodenalis and Cryptosporidium spp. in wildlife in general and mesocarnivore species in particular. Consequently, the extent of the cyst/oocyst environmental contamination attributable to these wild host species and their potential implications for public veterinary health remain largely unknown. In this molecular epidemiological survey a total of 193 individual faecal samples from badgers (Meles meles, n=70), ferrets (Mustela putorius furo, n=2), genets (Genetta genetta, n=6), Iberian lynxes (Lynx pardinus, n=6), beech martens (Martes foina, n=8), mongooses (Herpestes ichneumon, n=2), otters (Lutra lutra, n=2), polecats (Mustela putorius, n=2), red foxes (Vulpes vulpes, n=87), wildcats (Felis silvestris, n=2), and wolves (Canis lupus, n=6) were obtained from road-killed, hunted, and accidentally found carcasses, and from camera-trap surveys or animals entering rescue shelters, during the period December 2003–April 2016. Investigated specimens were collected in five Spanish autonomous regions including Andalusia (n=1), Asturias (n=69), Basque Country (n=49), Castile-La Mancha (n=38), and Extremadura (n=36). The presence of cysts/oocysts was confirmed by PCR-based methods targeting the small subunit (ssu) ribosomal RNA gene of these parasite species. Genotyping of the obtained isolates were attempted at appropriate markers including the glutamate dehydrogenase (G. duodenalis) and the 60-kDa glycoprotein (C. parvum and C. ubiquitum) loci. Overall, G. duodenalis was detected in 8% (7/87) of red foxes, a single beech marten, and a single wolf, respectively. Cryptosporidium was identified in 3% (2/70) of badgers, 8% (7/87) of red foxes, a single genet, and a single mongoose, respectively. None of the nine G. duodenalis isolates generated could be genotyped at the assemblage/sub-assemblage level. Out of the nine Cryptosporidium isolates successfully characterized, three were identified as C. canis (one in a mongoose and two in red foxes), and three as C. parvum (one in a badger and three in red foxes). The remaining three isolates were assigned to C. felis (in a red fox), C. hominis (in a badger), and C. ubiquitum (in a red fox), respectively. Two additional Cryptosporidium isolates infecting a badger and a genet, respectively, were untypable. The red fox was confirmed as a suitable host of potentially zoonotic Cryptosporidium species, mainly C. parvum and C. ubiquitum. The high mobility and wide home range of red foxes, together with their increasing presence in urban and peri-urban settings, may led to the overlapping of sylvatic and domestic cycles of the parasite, and consequently, to an increased risk of cryptosporidiosis in production animals and humans. The detection of C. hominis oocysts in a badger raises the question of whether this finding represents a true infection or a sporadic event of mechanical passage of C. hominis oocyst of anthroponotic origin.
Keywords	badgers ; cameras ; Canis lupus ; carnivores ; cryptosporidiosis ; Cryptosporidium ; epidemiological studies ; Fagus ; feces ; Felis ; ferrets ; genes ; genotyping ; Giardia lamblia ; glutamate dehydrogenase ; glycoproteins ; Herpestes ; home range ; hosts ; humans ; loci ; Lutra lutra ; Lynx pardinus ; Martes foina ; Meles meles ; Mustela putorius ; oocysts ; parasites ; pollution ; polymerase chain reaction ; ribosomal RNA ; risk ; Vulpes vulpes ; wildlife ; wolves ; Spain
Language	English
Dates of publication	2017-0215
Size	p. 86-93.
Publishing place	Elsevier B.V.
Document type	Article
ZDB-ID	196831-2
ISSN	1873-2550 ; 0304-4017
ISSN (online)	1873-2550
ISSN	0304-4017
DOI	10.1016/j.vetpar.2017.01.016
Database	NAL-Catalogue (AGRICOLA)

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