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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

Photograph showing the primary collection area of resting mosquitoes in the toilet block at Elmley. Blood-fed Anopheles maculipennis s.l. mosquitoes were found resting directly on the walls and on or under the exposed wooden covering to the pipework
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Fig1: Photograph showing the primary collection area of resting mosquitoes in the toilet block at Elmley. Blood-fed Anopheles maculipennis s.l. mosquitoes were found resting directly on the walls and on or under the exposed wooden covering to the pipework

Mentions: An. maculipennis s.l. were collected over 15 visits from Elmley National Nature Reserve, Isle of Sheppey (51.377445, 0.784068), Kent, UK between June and September 2013. Elmley is a freshwater coastal marsh used to graze approximately 700 head of cattle. The collection site was within 200 meters of grazing cattle. The site is popular with birdwatchers all year-round owing to the abundance of local and seasonal migrant bird species that breed in the area. Mosquitoes were primarily collected using a mouth aspirator (John W Hock, Gainsville, Florida, USA) from inside the publically accessible toilet facilities where they were observed to be resting on walls and close to exposed sections of the wood enclosing the pipework (Fig. 1). Additional attempts to collect blood-feds from a similar area were made using four CDC resting traps [24] placed in close proximity (~25 m) to the toilet block and run overnight (~14 hours) for nine of the 15 nights. Finally any anophelines landing on and attempting to feed on the collector were captured where possible. Collected mosquitoes were placed into a cooler containing dry ice and transported to the laboratory. Blood-fed specimens were separated from non-blood-fed specimens on the same day as collection and stored at −20 °C until processing at the Animal and Plant Health Agency (APHA).Fig. 1


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)

Photograph showing the primary collection area of resting mosquitoes in the toilet block at Elmley. Blood-fed Anopheles maculipennis s.l. mosquitoes were found resting directly on the walls and on or under the exposed wooden covering to the pipework
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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

Fig1: Photograph showing the primary collection area of resting mosquitoes in the toilet block at Elmley. Blood-fed Anopheles maculipennis s.l. mosquitoes were found resting directly on the walls and on or under the exposed wooden covering to the pipework
Mentions: An. maculipennis s.l. were collected over 15 visits from Elmley National Nature Reserve, Isle of Sheppey (51.377445, 0.784068), Kent, UK between June and September 2013. Elmley is a freshwater coastal marsh used to graze approximately 700 head of cattle. The collection site was within 200 meters of grazing cattle. The site is popular with birdwatchers all year-round owing to the abundance of local and seasonal migrant bird species that breed in the area. Mosquitoes were primarily collected using a mouth aspirator (John W Hock, Gainsville, Florida, USA) from inside the publically accessible toilet facilities where they were observed to be resting on walls and close to exposed sections of the wood enclosing the pipework (Fig. 1). Additional attempts to collect blood-feds from a similar area were made using four CDC resting traps [24] placed in close proximity (~25 m) to the toilet block and run overnight (~14 hours) for nine of the 15 nights. Finally any anophelines landing on and attempting to feed on the collector were captured where possible. Collected mosquitoes were placed into a cooler containing dry ice and transported to the laboratory. Blood-fed specimens were separated from non-blood-fed specimens on the same day as collection and stored at −20 °C until processing at the Animal and Plant Health Agency (APHA).Fig. 1

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