Development of a real-time PCR to detect Demodex canis DNA in different tissue samples

Ravera, Iván; Altet, Laura; Francino, Olga; Bardagí, Mar; Sánchez, Armand; Ferrer, L.

doi:10.1007/s00436-010-2062-0

Cited by 38 publications

(41 citation statements)

References 9 publications

(8 reference statements)

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“…However, in spite of the high prevalence of canine demodicosis, limited papers show molecular data on Demodex mites from dogs (Toops et al 2010;Ravera et al 2011;. Moreover, the newly observed species of Demodex from dogs, D. injai and D. sp.…”

Section: Introductionmentioning

confidence: 93%

Molecular study on three morphotypes of Demodex mites (Acarina: Demodicidae) from dogs

et al. 2012

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Canine demodicosis is a severe and highly prevalent dermatologic disease in dogs. Pet dogs can be affected by three recognized Demodex species that can produce clinical effects. In this paper, three morphological types of Demodex mites have been isolated from Spanish dogs. A complete morphobiometrical study of each one has been carried out. Morphological and biometrical studies revealed three closely related populations with some distinctive characteristics and could be identified as Demodex canis, Demodex injai, and Demodex sp. "cornei." Furthermore, one population of D. canis from China, different populations of Demodex folliculorum from human skin (Spain and China), D. folliculorum from human eyelashes (Spain), and Demodex brevis from human skin (China) were considered to find out the level of variation between different species and geographical origin. The aim of the present study is to assess the usefulness of mitochondrial DNA molecular markers in establishing phylogenetic relationships and resolve taxonomic questions in Demodex mites. Molecular studies based on the amplification and sequencing of the 16S rDNA and cytochrome oxidase I mitochondrial genes did not show clear differences between the three morphotypes considered. Furthermore, phylogenetic relationships in Demodex mites were analyzed. The resulting phylogenetic trees show that Demodex species from dogs were gathered together, and populations of D. folliculorum from humans appear together in a different branch; however, D. brevis from humans seemed to be more distant. Our results show that cytochrome oxidase I region is a useful tool to solve different taxonomic questions at the species and population level and to infer phylogenetic relationships in Demodex species. However, 16S mitochondrial rDNA seems a good marker for comparisons at an interspecies level, but not at a population level in this group of mites. Furthermore, from genetic distance and divergence data, we would suggest that D. canis, D. injai, and Demodex sp. cornei are polymorphisms of the same species.

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Section: Introductionmentioning

confidence: 93%

Molecular study on three morphotypes of Demodex mites (Acarina: Demodicidae) from dogs

et al. 2012

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“…The failure of the culture of Demodex in vitro directly hampers the confirmation of its pathogenicity by Koch's postulates. In recent years, the usage of molecular biology technique in the classification and identification of Demodex population lays an experimental foundation for both the DNA barcode study here and the pathogenicity study of Demodex population in the future (Ravera et al 2011;de Rojas Fig. 2 Frequency distribution diagram of intra-and inter-specific mitochondrial cox1 fragment divergences of the four Demodex species Zhao et al 2012cZhao et al , 2013bSilbermayr et al 2013).…”

Section: Discussionmentioning

confidence: 99%

Molecular identification and phylogenetic study of Demodex caprae

Zhao

Juan

et al. 2014

Parasitol Res

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The DNA barcode has been widely used in species identification and phylogenetic analysis since 2003, but there have been no reports in Demodex. In this study, to obtain an appropriate DNA barcode for Demodex, molecular identification of Demodex caprae based on mitochondrial cox1 was conducted. Firstly, individual adults and eggs of D. caprae were obtained for genomic DNA (gDNA) extraction; Secondly, mitochondrial cox1 fragment was amplified, cloned, and sequenced; Thirdly, cox1 fragments of D. caprae were aligned with those of other Demodex retrieved from GenBank; Finally, the intra- and inter-specific divergences were computed and the phylogenetic trees were reconstructed to analyze phylogenetic relationship in Demodex. Results obtained from seven 429-bp fragments of D. caprae showed that sequence identities were above 99.1% among three adults and four eggs. The intraspecific divergences in D. caprae, Demodex folliculorum, Demodex brevis, and Demodex canis were 0.0-0.9, 0.5-0.9, 0.0-0.2, and 0.0-0.5%, respectively, while the interspecific divergences between D. caprae and D. folliculorum, D. canis, and D. brevis were 20.3-20.9, 21.8-23.0, and 25.0-25.3, respectively. The interspecific divergences were 10 times higher than intraspecific ones, indicating considerable barcoding gap. Furthermore, the phylogenetic trees showed that four Demodex species gathered separately, representing independent species; and Demodex folliculorum gathered with canine Demodex, D. caprae, and D. brevis in sequence. In conclusion, the selected 429-bp mitochondrial cox1 gene is an appropriate DNA barcode for molecular classification, identification, and phylogenetic analysis of Demodex. D. caprae is an independent species and D. folliculorum is closer to D. canis than to D. caprae or D. brevis.

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“…The development of molecular biology techniques has made studies of molecular markers and Demodex species identification at the molecular level feasible. However, our database searches found only a few reports of effective molecular identification of Demodex (Zhao and Cheng, 2009;Toops et al, 2010;Ravera et al, 2011;de Rojas et al, 2012a;Zhao and Wu, 2012a;Zhao et al, 2009b;2012c).…”

Section: Introductionmentioning

confidence: 99%

Discrimination between Demodex folliculorum (Acari: Demodicidae) isolates from China and Spain based on mitochondrial cox1 sequences

Zhao

et al. 2013

J. Zhejiang Univ. Sci. B

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For a long time, classification of Demodex mites has been based mainly on their hosts and phenotypic characteristics. A new subspecies of Demodex folliculorum has been proposed, but not confirmed. Here, cox1 partial sequences of nine isolates of three Demodex species from two geographical sources (China and Spain) were studied to conduct molecular identification of D. folliculorum. Sequencing showed that the mitochondrial cox1 fragments of five D. folliculorum isolates from the facial skin of Chinese individuals were 429 bp long and that their sequence identity was 97.4%. The average sequence divergence was 1.24% among the five Chinese isolates, 0.94% between the two geographical isolate groups (China (5) and Spain (1)), and 2.15% between the two facial tissue sources (facial skin (6) and eyelids (1)). The genetic distance and rate of third-position nucleotide transition/transversion were 0.0125, 2.7 (3/1) among the five Chinese isolates, 0.0094, 3.1 (3/1) between the two geographical isolate groups, and 0.0217, 4.4 (3/1) between the two facial tissue sources. Phylogenetic trees showed that D. folliculorum from the two geographical isolate groups did not form sister clades, while those from different facial tissue sources did. According to the molecular characteristics, it appears that subspecies differentiation might not have occurred and that D. folliculorum isolates from the two geographical sources are of the same population. However, population differentiation might be occurring between isolates from facial skin and eyelids.

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Development of a real-time PCR to detect Demodex canis DNA in different tissue samples

Cited by 38 publications

References 9 publications

Molecular study on three morphotypes of Demodex mites (Acarina: Demodicidae) from dogs

Molecular study on three morphotypes of Demodex mites (Acarina: Demodicidae) from dogs

Molecular identification and phylogenetic study of Demodex caprae

Discrimination between Demodex folliculorum (Acari: Demodicidae) isolates from China and Spain based on mitochondrial cox1 sequences

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