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Evidence for a role of the host-specific flea (Paraceras melis) in the transmission of Trypanosoma (Megatrypanum) pestanai to the European badger.

Lizundia R, Newman C, Buesching CD, Ngugi D, Blake D, Sin YW, Macdonald DW, Wilson A, McKeever D - PLoS ONE (2011)

Bottom Line: High levels of genotypic polymorphism were observed between the isolates.Wet smears and Giemsa-stained preparations from dissected fleas revealed large numbers of trypanosome-like forms in the hindgut, some of which were undergoing binary fission.We conclude that P. melis is the primary vector of T. pestanai in European badgers.

View Article: PubMed Central - PubMed

Affiliation: Royal Veterinary College, University of London, Hatfield, United Kingdom.

ABSTRACT
We investigated the epidemiology of Trypanosoma pestanai infection in European badgers (Meles meles) from Wytham Woods (Oxfordshire, UK) to determine prevalence rates and to identify the arthropod vector responsible for transmission. A total of 245 badger blood samples was collected during September and November 2009 and examined by PCR using primers derived from the 18S rRNA of T. pestanai. The parasite was detected in blood from 31% of individuals tested. T. pestanai was isolated from primary cultures of Wytham badger peripheral blood mononuclear cells and propagated continually in vitro. This population was compared with cultures of two geographically distinct isolates of the parasite by amplified fragment length polymorphism (AFLP) and PCR analysis of 18S rDNA and ITS1 sequences. High levels of genotypic polymorphism were observed between the isolates. PCR analysis of badger fleas (Paraceras melis) collected from infected individuals at Wytham indicated the presence of T. pestanai and this was confirmed by examination of dissected specimens. Wet smears and Giemsa-stained preparations from dissected fleas revealed large numbers of trypanosome-like forms in the hindgut, some of which were undergoing binary fission. We conclude that P. melis is the primary vector of T. pestanai in European badgers.

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Related in: MedlinePlus

Comparison by PCR of 3 geographically distinct isolates of T. pestanai.DNA extracted from different T. pestanai isolates in axenic culture (1. East Anglia isolate; 2. Oxford isolate; 3. France isolate) was analysed by PCR using primers derived from the 18S rRNA (A. TPEF1/B1 primers; B. TPEF2/B2 primers) and ITS1 sequences of T. pestanai. (C. KIN1/KIN2 primers).
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pone-0016977-g003: Comparison by PCR of 3 geographically distinct isolates of T. pestanai.DNA extracted from different T. pestanai isolates in axenic culture (1. East Anglia isolate; 2. Oxford isolate; 3. France isolate) was analysed by PCR using primers derived from the 18S rRNA (A. TPEF1/B1 primers; B. TPEF2/B2 primers) and ITS1 sequences of T. pestanai. (C. KIN1/KIN2 primers).

Mentions: Total DNA extracted from cultures of three geographically distinct T. pestanai isolates was analysed by PCR (Fig. 3). PCR analysis using primers derived from the 18S rRNA of T. pestanai resulted in amplification of an identical band from all three isolates when using TPEF1/ TPEB1 (Fig. 3A) and TPEF2/TPEB2 (Fig. 3B) primers. However, PCR analysis using primers specific for the ITS1 sequence (KIN1, KIN2) revealed different size bands (Fig. 3C). More detailed genetic characterization using AFLP revealed clear genetic polymorphism between all three isolates (Figure S1). Four selective AFLP primer combinations were used, yielding 56 markers, 41 of which were polymorphic for one or more isolates. The Jaccard index of similarity ranged from 34 to 64%, indicating elevated levels of genetic heterogeneity among the isolates. The highest coefficient of similarity was found between the France and East Anglia isolates (64%), followed by the Oxford and East Anglia isolates (48%) and by the Oxford and France isolates (34%).


Evidence for a role of the host-specific flea (Paraceras melis) in the transmission of Trypanosoma (Megatrypanum) pestanai to the European badger.

Lizundia R, Newman C, Buesching CD, Ngugi D, Blake D, Sin YW, Macdonald DW, Wilson A, McKeever D - PLoS ONE (2011)

Comparison by PCR of 3 geographically distinct isolates of T. pestanai.DNA extracted from different T. pestanai isolates in axenic culture (1. East Anglia isolate; 2. Oxford isolate; 3. France isolate) was analysed by PCR using primers derived from the 18S rRNA (A. TPEF1/B1 primers; B. TPEF2/B2 primers) and ITS1 sequences of T. pestanai. (C. KIN1/KIN2 primers).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3038870&req=5

pone-0016977-g003: Comparison by PCR of 3 geographically distinct isolates of T. pestanai.DNA extracted from different T. pestanai isolates in axenic culture (1. East Anglia isolate; 2. Oxford isolate; 3. France isolate) was analysed by PCR using primers derived from the 18S rRNA (A. TPEF1/B1 primers; B. TPEF2/B2 primers) and ITS1 sequences of T. pestanai. (C. KIN1/KIN2 primers).
Mentions: Total DNA extracted from cultures of three geographically distinct T. pestanai isolates was analysed by PCR (Fig. 3). PCR analysis using primers derived from the 18S rRNA of T. pestanai resulted in amplification of an identical band from all three isolates when using TPEF1/ TPEB1 (Fig. 3A) and TPEF2/TPEB2 (Fig. 3B) primers. However, PCR analysis using primers specific for the ITS1 sequence (KIN1, KIN2) revealed different size bands (Fig. 3C). More detailed genetic characterization using AFLP revealed clear genetic polymorphism between all three isolates (Figure S1). Four selective AFLP primer combinations were used, yielding 56 markers, 41 of which were polymorphic for one or more isolates. The Jaccard index of similarity ranged from 34 to 64%, indicating elevated levels of genetic heterogeneity among the isolates. The highest coefficient of similarity was found between the France and East Anglia isolates (64%), followed by the Oxford and East Anglia isolates (48%) and by the Oxford and France isolates (34%).

Bottom Line: High levels of genotypic polymorphism were observed between the isolates.Wet smears and Giemsa-stained preparations from dissected fleas revealed large numbers of trypanosome-like forms in the hindgut, some of which were undergoing binary fission.We conclude that P. melis is the primary vector of T. pestanai in European badgers.

View Article: PubMed Central - PubMed

Affiliation: Royal Veterinary College, University of London, Hatfield, United Kingdom.

ABSTRACT
We investigated the epidemiology of Trypanosoma pestanai infection in European badgers (Meles meles) from Wytham Woods (Oxfordshire, UK) to determine prevalence rates and to identify the arthropod vector responsible for transmission. A total of 245 badger blood samples was collected during September and November 2009 and examined by PCR using primers derived from the 18S rRNA of T. pestanai. The parasite was detected in blood from 31% of individuals tested. T. pestanai was isolated from primary cultures of Wytham badger peripheral blood mononuclear cells and propagated continually in vitro. This population was compared with cultures of two geographically distinct isolates of the parasite by amplified fragment length polymorphism (AFLP) and PCR analysis of 18S rDNA and ITS1 sequences. High levels of genotypic polymorphism were observed between the isolates. PCR analysis of badger fleas (Paraceras melis) collected from infected individuals at Wytham indicated the presence of T. pestanai and this was confirmed by examination of dissected specimens. Wet smears and Giemsa-stained preparations from dissected fleas revealed large numbers of trypanosome-like forms in the hindgut, some of which were undergoing binary fission. We conclude that P. melis is the primary vector of T. pestanai in European badgers.

Show MeSH
Related in: MedlinePlus