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Molecular species identification, host preference and detection of myxoma virus in the Anopheles maculipennis complex (Diptera: Culicidae) in southern England, UK.

Brugman VA, Hernández-Triana LM, Prosser SW, Weland C, Westcott DG, Fooks AR, Johnson N - Parasit Vectors (2015)

Bottom Line: Mosquitoes were then identified to species by sequencing a region of the internal transcribed spacer-2.A total of 94 blood-fed Anopheles maculipennis s.l. were collected, of which 43 (46%) provided positive blood-meal identification results.Seven mosquitoes were identified as Anopheles messeae, which had fed on cattle (n = 6, 86%) and dog (n = 1, 14%).

View Article: PubMed Central - PubMed

Affiliation: Vector-borne viral diseases programme, The Pirbright Institute, Ash Road, Woking, GU24 0NF, UK.

ABSTRACT

Background: Determining the host feeding patterns of mosquitoes by identifying the origin of their blood-meals is an important part of understanding the role of vector species in current and future disease transmission cycles. Collecting large numbers of blood-fed mosquitoes from the field is difficult, therefore it is important to maximise the information obtained from each specimen. This study aimed to use mosquito genome sequence to identify the species within Anopheles maculipennis sensu lato (An. maculipennis s.l.), identify the vertebrate hosts of field-caught blood-fed An. maculipennis s.l. , and to test for the presence of myxoma virus (Poxviridae, genus Leporipoxvirus) in specimens found to have fed on the European rabbit (Oryctolagus cuniculus).

Methods: Blood-fed An. maculipennis s.l. were collected from resting sites at Elmley Nature Reserve, Kent, between June and September 2013. Hosts that An. maculipennis s.l. had fed on were determined by a PCR-sequencing approach based on the partial amplification of the mitochondrial cytochrome C oxidase subunit I gene. Mosquitoes were then identified to species by sequencing a region of the internal transcribed spacer-2. DNA extracts from all mosquitoes identified as having fed on rabbits were subsequently screened using PCR for the presence of myxoma virus.

Results: A total of 94 blood-fed Anopheles maculipennis s.l. were collected, of which 43 (46%) provided positive blood-meal identification results. Thirty-six of these specimens were identified as Anopheles atroparvus, which had fed on rabbit (n = 33, 92%) and cattle (n = 3, 8%). Seven mosquitoes were identified as Anopheles messeae, which had fed on cattle (n = 6, 86%) and dog (n = 1, 14%). Of the 33 An. atroparvus that contained rabbit blood, nine (27%) were positive for myxoma virus.

Conclusions: Results demonstrate that a single DNA extract from a blood-fed mosquito can be successfully used for molecular identification of members of the An. maculipennis complex, blood-meal identification, and for the targeted detection of a myxoma virus. This study shows that An. atroparvus has a strong feeding preference for both healthy and myxoma-infected rabbits, providing evidence that this species may play a significant role in the transmission of myxomatosis among wild rabbit populations in the United Kingdom (UK).

No MeSH data available.


Related in: MedlinePlus

Gel image showing COI amplification products. The samples are PhiX174 DNA marker (M), negative control (1), mosquito blood-meal samples (2–4) and a positive control of DNA (5,6). The positive control was DNA extracted directly from horse blood
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Fig2: Gel image showing COI amplification products. The samples are PhiX174 DNA marker (M), negative control (1), mosquito blood-meal samples (2–4) and a positive control of DNA (5,6). The positive control was DNA extracted directly from horse blood

Mentions: In total, 94 blood-fed specimens belonging to the An. maculipennis s.l. were collected from the Elmley site over 15 collection days. The toilet block yielded the majority (n = 92), with only one blood-fed An. maculipennis s.l. collected in the CDC resting traps and one that alighted on the collector. Of the total blood-fed samples extracted, 43 (46 %) produced a 685 bp band when amplified with COI primers as illustrated in Fig. 2. An. maculipennis s.l. at Elmley fed on cow (Bos taurus), European rabbit (Oryctolagus cuniculus) and dog (Canis lupus familiaris) (Table 2).Fig. 2


Molecular species identification, host preference and detection of myxoma virus in the Anopheles maculipennis complex (Diptera: Culicidae) in southern England, UK.

Brugman VA, Hernández-Triana LM, Prosser SW, Weland C, Westcott DG, Fooks AR, Johnson N - Parasit Vectors (2015)

Gel image showing COI amplification products. The samples are PhiX174 DNA marker (M), negative control (1), mosquito blood-meal samples (2–4) and a positive control of DNA (5,6). The positive control was DNA extracted directly from horse blood
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4536751&req=5

Fig2: Gel image showing COI amplification products. The samples are PhiX174 DNA marker (M), negative control (1), mosquito blood-meal samples (2–4) and a positive control of DNA (5,6). The positive control was DNA extracted directly from horse blood
Mentions: In total, 94 blood-fed specimens belonging to the An. maculipennis s.l. were collected from the Elmley site over 15 collection days. The toilet block yielded the majority (n = 92), with only one blood-fed An. maculipennis s.l. collected in the CDC resting traps and one that alighted on the collector. Of the total blood-fed samples extracted, 43 (46 %) produced a 685 bp band when amplified with COI primers as illustrated in Fig. 2. An. maculipennis s.l. at Elmley fed on cow (Bos taurus), European rabbit (Oryctolagus cuniculus) and dog (Canis lupus familiaris) (Table 2).Fig. 2

Bottom Line: Mosquitoes were then identified to species by sequencing a region of the internal transcribed spacer-2.A total of 94 blood-fed Anopheles maculipennis s.l. were collected, of which 43 (46%) provided positive blood-meal identification results.Seven mosquitoes were identified as Anopheles messeae, which had fed on cattle (n = 6, 86%) and dog (n = 1, 14%).

View Article: PubMed Central - PubMed

Affiliation: Vector-borne viral diseases programme, The Pirbright Institute, Ash Road, Woking, GU24 0NF, UK.

ABSTRACT

Background: Determining the host feeding patterns of mosquitoes by identifying the origin of their blood-meals is an important part of understanding the role of vector species in current and future disease transmission cycles. Collecting large numbers of blood-fed mosquitoes from the field is difficult, therefore it is important to maximise the information obtained from each specimen. This study aimed to use mosquito genome sequence to identify the species within Anopheles maculipennis sensu lato (An. maculipennis s.l.), identify the vertebrate hosts of field-caught blood-fed An. maculipennis s.l. , and to test for the presence of myxoma virus (Poxviridae, genus Leporipoxvirus) in specimens found to have fed on the European rabbit (Oryctolagus cuniculus).

Methods: Blood-fed An. maculipennis s.l. were collected from resting sites at Elmley Nature Reserve, Kent, between June and September 2013. Hosts that An. maculipennis s.l. had fed on were determined by a PCR-sequencing approach based on the partial amplification of the mitochondrial cytochrome C oxidase subunit I gene. Mosquitoes were then identified to species by sequencing a region of the internal transcribed spacer-2. DNA extracts from all mosquitoes identified as having fed on rabbits were subsequently screened using PCR for the presence of myxoma virus.

Results: A total of 94 blood-fed Anopheles maculipennis s.l. were collected, of which 43 (46%) provided positive blood-meal identification results. Thirty-six of these specimens were identified as Anopheles atroparvus, which had fed on rabbit (n = 33, 92%) and cattle (n = 3, 8%). Seven mosquitoes were identified as Anopheles messeae, which had fed on cattle (n = 6, 86%) and dog (n = 1, 14%). Of the 33 An. atroparvus that contained rabbit blood, nine (27%) were positive for myxoma virus.

Conclusions: Results demonstrate that a single DNA extract from a blood-fed mosquito can be successfully used for molecular identification of members of the An. maculipennis complex, blood-meal identification, and for the targeted detection of a myxoma virus. This study shows that An. atroparvus has a strong feeding preference for both healthy and myxoma-infected rabbits, providing evidence that this species may play a significant role in the transmission of myxomatosis among wild rabbit populations in the United Kingdom (UK).

No MeSH data available.


Related in: MedlinePlus