Wolbachia Biocontrol Strategies for Arboviral Diseases and the Potential Influence of Resident Wolbachia Strains in Mosquitoes
Curr Trop Med Rep
Wolbachia Biocontrol Strategies for Arboviral Diseases and the Potential Influence of Resident Wolbachia Strains in Mosquitoes
Claire L. Jeffries 0 1
Thomas Walker 0 1
0 Department of Disease Control, Faculty of Infectious Diseases, London School of Hygiene and Tropical Medicine , Keppel Street, London WC1E 7HT , UK
1 Claire L. Jeffries
2 Thomas Walker
Arboviruses transmitted by mosquitoes are a major cause of human disease worldwide. The absence of vaccines and effective vector control strategies has resulted in the need for novel mosquito control strategies. The endosymbiotic bacterium Wolbachia has been proposed to form the basis for an effective mosquito biocontrol strategy. Resident strains of Wolbachia inhibit viral replication in Drosophila fruit flies and induce a reproductive phenotype known as cytoplasmic incompatibility that allows rapid invasion of insect populations. Transinfection of Wolbachia strains into the principle mosquito vector of dengue virus, Stegomyia aegypti, has resulted in dengue-refractory mosquito lines with minimal effects on mosquito fitness. Wolbachia strains have now been established in wild St. aegypti populations through open releases in dengue-endemic countries. In this review, we outline the current state of Wolbachia-based biocontrol strategies for dengue and discuss the potential impact of resident Wolbachia strains for additional target mosquito species that transmit arboviruses.
Mosquitoes; Arboviruses; Mosquito biocontrol; Wolbachia bacteria
Arboviruses that cause human disease are predominantly
transmitted by mosquitoes. Although there are more than 80
different arboviruses, most human cases result from infection
with dengue virus (DENV) and other closely related
Flaviviruses. The genus Flavivirus also includes West Nile
virus (WNV), Yellow fever virus (YFV), Zika virus (ZIKV)
and Japanese encephalitis virus (JEV). These medically
important arboviruses are transmitted by several species of
Culicine mosquitoes (Table 1). It is estimated that 40 % of
the world’s population live in areas at risk for DENV infection
in more than 100 countries. Global DENV infections range
from 100–390 million per year, with 100 million symptomatic
infections leading to 12,500 deaths per year [
]. Dengue is an
epidemic disease that occurs in tropical areas of Southeast
Asia and South America and has a significant impact on
developing countries [
]. Rare cases have been documented in
the USA and southern Europe, and dengue is ‘re-emerging’
mostly due to the expansion of the geographical range of the
principal mosquito vector, Stegomyia (St.) (Aedes) aegypti,
through globalization and climate change [
]. There are
currently no effective vaccines for DENV, so supportive
treatment is the only method to reduce morbidity or prevent
mortality for infected patients. Prevention of DENV transmission
relies on mosquito vector control but this has not seen much
success in recent years. Attempts to reduce the larval breeding
of St. aegypti, predominantly an urban species, over a large
geographical area have been difficult to achieve. There is also
a delay between implementing larval control and achieving an
impact on the adult mosquito population already transmitting
disease. When dengue epidemics occur, the usual response is
to use outdoor space spraying of insecticides. Although this
targets the adult mosquito, insecticide resistance has been
problematic in many countries. The options for developing
DENV dengue virus, YFV yellow fever virus, CHIKV chikungunya virus,
ZIKV zika virus, WNV West Nile virus, EEEV Eastern equine
encephalitis virus, JEV Japanese encephalitis virus, SLEV St Louis encephalitis
virus, RVFV Rift Valley fever virus, WEEV Western equine encephalitis
virus, SINDV Sindbis virus, MVEV Murray Valley encephalitis virus,
VEEV Venezuelan equine encephalitis virus, RRV Ross River virus,
CEV California encephalitis virus, LCV La Crosse virus
new mosquito control strategies can be classified as either
suppression (reduce or eliminate the wild mosquito
population) or replacement with mosquitoes that are unable to
transmit disease. A recent novel approach is the use of
endosymbiotic Wolbachia bacteria to prevent DENV from replicating
within the mosquito. In recent years, Wolbachia-based
biocontrol has emerged as a very promising method that is
environmentally friendly, safe to humans and potentially
cost effective [
]. The ‘eliminate dengue’ project (www.
eliminatedengue.com) has shown that Wolbachia can
prevent DENV transmission in mosquitoes without
significant fitness costs.
Natural Wolbachia Infections in Mosquitoes
Wolbachia is a genus of obligate intracellular alphaproteobacteria
within the family Rickettsiaceae [
]. Wolbachia was initially
found in Culex pipiens mosquitoes in 1924 [
]. Since then, many
different strains of Wolbachia have been discovered in a wide
range of invertebrate hosts, including many filarial nematode
and arthropod species. A recent study estimated that
Wolbachia naturally infects more than 65 % of arthropod
]. Natural Wolbachia infections are present in some
mosquito species that transmit human pathogens including
Culex (Cx.) quinquefasciatus and Stegomyia albopicta
(Aedes albopictus) (Table 1). Additional Wolbachia strains
have been discovered in minor mosquito vectors of YFV
including Aedes (Ae.) bromeliae [
] and Ae. fluviatilis which
is infected with the wFlu strain [
]. Wolbachia infections are
also present in wild mosquito populations of Coquilletidia
] and Mansonia titillans [
]. However, natural
infections are absent from most Anopheles spp. (that transmit
malaria), St. aegypti and Cx. tritaeniorhynchus (the major
vector of JEV). Wolbachia strains are currently divided into
eight (A–H) supergroups according to sequence information
]. Wolbachia strains have the ability to manipulate host
reproduction in order to enhance their own reproduction and
transmission through an insect population. Wolbachia strains
can induce various phenotypic effects in insects including
parthenogenesis, feminization, male killing and cytoplasmic
In mosquitoes, Wolbachia strains can induce cytoplasmic
incompatibility (CI), which results in the generation of unviable
offspring when an uninfected female mates with a
Wolbachiainfected male [
]. In contrast, Wolbachia-infected females can
produce viable progeny when they mate with both infected
and uninfected males, resulting in a reproductive advantage
over uninfected females. The CI phenotype allows the
maternally transmitted Wolbachia to efficiently invade host
populations without being infectious or moving horizontally between
CI can also be the result when mating occurs between
mosquitoes with two different, incompatible Wolbachia strains.
This CI can occur bidirectionally if crosses of both males
and females with each different Wolbachia strain are
incompatible, resulting in embryonic death in both crosses.
However, some crosses between mosquitoes with differing
strains can instead lead to unidirectional CI, where a cross of
males with strain ‘x’ and females with strain ‘y’ results in CI,
whereas the reciprocal cross (males with strain y and females
with strain x) allows the production of viable offspring. In this
case, mosquitoes with Wolbachia strain x would have a
reproductive advantage over those with strain y and strain x and
would be predicted to successfully spread through the
population, Bsweeping over^ strain y [
Transinfection of Wolbachia Into Mosquitoes
The various phenotypic effects of Wolbachia strains on natural
insect hosts have led to a range of ideas on how this could be
applied for mosquito-borne disease vector control. One
potential strategy was investigated in the 1960s using repeated
releases of Wolbachia-infected male Culex mosquitoes to
suppress wild populations using CI [
]. A later discovery of the
wMelPop strain of Wolbachia in Drosophila melanogaster
fruit flies, which dramatically lowered the lifespan of its host
], led to the potential use of ‘life-shortening’ strains to
manipulate the population age structure of important mosquito
vectors. Recently, Wolbachia strains from Drosophila fruit
flies were found to protect their native hosts against infection
by pathogenic RNA viruses [
]. Wolbachia strains are
associated with a significant reduction in the viral density of
a range of viruses in flies, which delays insect mortality .
The use of Wolbachia for mosquito biocontrol first required
the stable infection (transinfection) of target mosquito species.
The first target species selected was St. aegypti, the principle
vector of DENV. The wAlbB strain of Wolbachia was
successfully established in St. aegypti using embryo cytoplasm transfer
from closely related St. albopicta mosquitoes [
]. St. aegypti
lines were later stably transinfected with wMelPop and wMel
strains from the native host Drosophila melanogaster [
These transinfected Wolbachia strains significantly reduced the
vector competence of St. aegypti for DENV in laboratory
]. High levels of Wolbachia bacteria infected
the tissues that play a crucial role in DENV replication within
mosquitoes. The presence of infectious DENV in saliva was
completely inhibited by Wolbachia .
Successful Wolbachia-based biocontrol would also require
invasion of wild mosquito populations. Wolbachia-infected
females must vertically transmit the bacteria to their progeny
at a high frequency. All three transinfected Wolbachia strains
(wAlbB, wMel and wMelPop) show maternal transmission
rates close to 100 % and induce high levels of CI in St. aegypti
]. The ability of transinfected Wolbachia to successfully
invade wild St. aegypti mosquito populations will depend on a
balance between negative selection imposed by fitness costs of
the bacteria on the mosquito and positive selection associated
with CI induction. The wMelPop strain results in significant
fitness costs (including impacts on adult longevity and fecundity)
so was considered inappropriate for initial test releases into wild
The Release of Wolbachia-Infected Mosquitoes
The invasive potential of Wolbachia strains was first tested in
a semi-field facility that simulated the natural habitat of St.
aegypti in north Queensland, Australia [
]. Successful trials
led to mosquitoes infected with the wMel strain being released
into the wild through open releases in two locations near
Cairns in north Queensland, Australia [
]. The wMel
strain successfully invaded the two natural populations,
infecting nearly 100 % of the local population within a
few months following releases. Prior to the release of
Wolbachia-infected mosquitoes in Australia, extensive
engagement with the communities in the release areas took
place to determine the attitudes and levels of knowledge
about dengue and mosquitoes. The success of these initial
trials has led to further releases in DENV endemic countries
such as Indonesia, Vietnam and Brazil (www.eliminatedengue.
com). Continued success of a release program will require
maintenance of an inhibitory effect on DENV replication in
wild Wolbachia-infected St. aegypti populations. Vector
competence experiments carried out with field wMel-infected
mosquitoes, collected 1 year following field release, indicated
insignificant DENV replication and dissemination [
Currently, work is being undertaken to determine the
optimal Wolbachia strain for applied use that can balance
the effects on pathogen transmission and fitness costs to
the mosquito. Recent mathematical models of DENV
transmission incorporating the dynamics of viral infection
in humans and mosquitoes predict that wMel would reduce
the basic reproduction number, R0, of DENV transmission
by approximately 70 % [
]. At the current time, it
remains unclear what effect Wolbachia will have on
DENV transmission and dengue epidemiology in the
field. A cluster-randomized trial is premature because the
choice of Wolbachia strain for release and deployment
strategies is still being optimized [
Potential Use of Wolbachia for Additional Arboviral
Although Wolbachia-infected St. aegypti were originally
generated for biocontrol of dengue, they are likely to have
the added benefit of reducing transmission of additional
arboviruses vectored by this mosquito species, including
chikungunya virus (CHIKV) [
] and YFV [
potentially ZIKV. Like St. aegypti, mosquito species that are
the principle vector of arboviruses and contain no natural
Wolbachia infections are likely to represent the most feasible
transinfection targets. JEV is predominantly transmitted by
Cx. tritaeniorhynchus mosquitoes, and a Wolbachia-based
biocontrol strategy has the potential to reduce transmission if
stably infected lines can be generated [
]. JEV is part of the
same genus as DENV (Flavivirus), so Wolbachia strains
would likely provide similar inhibitory effects on
transinfected mosquitoes. Drosophila Wolbachia strains
grow to high densities in their native and transinfected
hosts and provide strong inhibition of both insect viruses
in Drosophila [
] and DENV in mosquitoes [
transinfection of Drosophila Wolbachia strains in Cx.
tritaeniorhynchus and the release of stably infected mosquitoes
which enable the Wolbachia strain to invade wild
populations, with strong viral interference characteristics, would be
likely to significantly reduce JEV transmission [
Resident Wolbachia Strains in Mosquitoes and Effects on Arbovirus Replication
Some major mosquito vectors of arboviral diseases harbor
natural Wolbachia infections. Both DENV and CHIKV
are also transmitted by St. albopicta, which contain two
resident strains of Wolbachia; wAlbA and wAlbB [
The principle vectors of WNV in the USA, Cx. pipiens
and Cx. quinquefasciatus, contain a natural Wolbachia
strain, wPip. Minor differences in vector competence
may be due to the presence of these resident Wolbachia
strains. For example, Cx. quinquefasciatus, infected with
the wPip strain of Wolbachia, is generally less susceptible
to WNV than Cx. tarsalis , which is not infected with
Wolbachia. However, resident Wolbachia infections in
mosquitoes do not impact arboviral transmission to the
same extent as transinfected Drosophila Wolbachia strains,
as reviewed in [
]. One mechanism suggested for this
difference is that the newly transinfected strains trigger
an immune response in their new invertebrate hosts, which
also has antiviral effects, whereas native Wolbachia strains
have been present in the host long enough that the host
no longer generates such an immune response. However,
the complexity in the interaction between Wolbachia, the
insect host and arboviruses remains unclear .
Although the mechanism of how Wolbachia inhibits
arboviruses is not fully known, Wolbachia density is
correlated with viral interference in both native Drosophila
 and transinfected St. aegypti [
]. In general, resident
Wolbachia strains in wild populations do not appear to
grow to such high densities as transinfected strains and
therefore have less impact on arboviral vector competence.
For example, a high-density wPip strain in a laboratory
colonized line of Cx. quinquefasciatus was observed to
show resistance to WNV, compared to a wPip cleared line
. However, lower density wPip infections found in
field-collected Cx. quinquefasciatus and Cx. pipiens
mosquitoes do not appear to be capable of inhibiting WNV
infection and transmission . Recent studies have
shown that the removal of resident Wolbachia strains from
St. albopicta, followed by transinfection of the Drosophila
wMel strain, results in strong inhibition of both DENV
and CHIKV [35, 36]. Therefore, the transinfection of mosquito
species with Drosophila Wolbachia strains would likely
provide strong inhibitory effects on arbovirus transmission.
Resident Wolbachia Strains and Mosquito Population
The potential impact of resident Wolbachia strains on the
ability of introduced transinfected strains must be considered if
species such as Cx. quinquefasciatus and St. albopicta are
considered targets for Wolbachia biocontrol. The first point
to consider is whether the resident strain actually inhibits the
ability to form stable transinfected lines with Drosophila
Wolbachia strains. Wolbachia bacteria are maternally
inherited so are found at high densities in the reproductive
tissues (ovaries for females). As Wolbachia transinfection
involves the injection of preblastoderm embryos for the
infection of the pole cells (that form the germline), the presence of
high-density resident Wolbachia strains is likely to decrease
the chances of success. However, an artificial triple Wolbachia
infection in St. albopicta was successfully created with the
wRi strain of Wolbachia from Drosophila simulans and
yielded a new pattern of cytoplasmic incompatibility .
This generation of a ‘superinfected’ line with a resident strain
and a transinfected strain has not been possible so far in Cx.
quinquefasciatus (despite significant efforts).
There may be species-specific differences between
Wo l b a c hi a - m o s q u i t o h o s t i n t er a c t i o n s t h a t i m p a c t
transinfection success. A stable, Wolbachia superinfected line
with new transinfected strains (wMel and wAlbB) was
recently generated in St. aegypti (Joubert et al. unpublished data).
The transinfection options for target mosquito species such as
Cx. quinquefasciatus that contains a single resident strain
(wPip) include removal of the resident strain through
antibiotic treatment or attempting to create a resident strain/
transinfected strain superinfection. If a single infection
containing a novel transinfected Wolbachia strain (eg. wMel)
was generated, the crossing patterns induced between infected
mosquitoes could still result in the invasion of the transinfected
strain. Matings involving two different Wolbachia strains in
Cx. quinquefasciatus would result in bidirectional CI 
and two strains cannot stably co-exist in a given mosquito
population. The strain that reaches the highest local frequency
would likely reach fixation given that females infected with
this strain would be at a reproductive advantage (more males
to mate with that are compatible). However, if significant
fitness costs are associated with a transinfected Wolbachia strain
this would prevent the invasion of the population and
replacement of the resident wPip strain.
The potential role of Wolbachia strains in the speciation
and genetic evolution of mosquito populations over time
through unidirectional or bidirectional CI patterns between
different native or introduced strains should also be
considered, in addition, to the resultant phenotypic effects .
Other forms of reproductive interference can also affect
mosquito vector population dynamics and could therefore
potentially impact on the spread of Wolbachia. For example,
satyrization is a form of reproductive interference whereby
males of one species mate with females of another, producing
no viable offspring but leading to the mated female no longer
being receptive to further insemination for the remaining life
of that mosquito [40, 41]. This phenomenon is believed to
result from male accessory gland products and has been linked
to population displacement in certain areas of St. aegypti by St.
albopicta . As St. albopicta has a resident Wolbachia
strain and St. aegypti does not, if CI of Wolbachia in St.
albopicta is not itself involved in this mechanism, satyrization
could potentially assist in or lead to the amplification of the
driving force of Wolbachia into certain populations in a
population replacement strategy.
In addition to the presence of resident Wolbachia strains,
the presence of other microorganisms within the mosquito
population is another important factor to consider, which
could affect the introduction of any transinfected Wolbachia
strain into that population. For example, the bacteria Asaia is
stably associated with many mosquito species  and has
been found to compete with Wolbachia within a host,
seemingly preventing or reducing Wolbachia establishment. It has
also been suggested that the presence of Asaia in certain
mosquito species could be a reason for the absence of native
Wolbachia strains in such species .
Wolbachia-based biocontrol strategies are likely to provide an
environmentally benign and effective long-term control
option for arboviral diseases such as dengue. Invasion and
maintenance of transinfected Wolbachia strains in natural mosquito
populations is likely to be cost effective given this should be
self-sustaining through the CI phenotype and maternal
transmission. Although significant advancements have been made
in the implementation of Wolbachia for dengue biocontrol, the
optimal Wolbachia strain (or combination of strains) to
balance the inhibitory effects of DENV and fitness costs to the
mosquito is still to be determined. A major area of research
that needs addressing is how Wolbachia inhibits arboviruses
in mosquitoes as further knowledge would allow more
accurate predictions of the evolutionary consequences of strong
selection pressure on DENV. This would likely provide better
estimates of the effect Wolbachia would have on dengue
transmission. Wolbachia’s broad protective inhibition of human
arboviruses suggests the potential of this strategy to reduce
the transmission of other arboviral diseases. The presence of
resident Wolbachia strains in some mosquito vectors must be
considered if transinfection (with subsequent favourable
phenotypic effects) is to be successfully implemented. If
promising initial laboratory research and field trials can translate into
effective biocontrol programs, Wolbachia may provide a
novel method to control additional mosquito-borne diseases.
Compliance with Ethical Standards
Conflict of Interest The authors declare that they have no competing
Human and Animal Rights and Informed Consent This article does
not contain any studies with human or animal subjects performed by any
of the authors.
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Papers of particular interest, published recently, have been
1. Bhatt S , Gething PW , Brady OJ , Messina JP , Farlow AW , Moyes CL , et al. The global distribution and burden of dengue . Nature . 2013 ; 496 : 504 - 7 . A recent review of worldwide dengue transmission and resulting disease .
2. Guzman MG , Halstead SB , Artsob H , Buchy P , Farrar J , Gubler DJ , et al. Dengue: a continuing global threat . Nat Rev Microbiol . 2010 ; 8 : S7 - 16 .
3. Kilpatrick AM , Randolph SE . Drivers, dynamics, and control of emerging vector-borne zoonotic diseases . Lancet . 2012 ; 380 : 1946 - 55 .
4. Iturbe-Ormaetxe I , Walker T , ONeill SL . Wolbachia and the biological control of mosquito-borne disease . EMBO Rep . 2011 ; 12 : 508 - 18 .
5. Werren JH , Baldo L , Clark ME . Wolbachia: master manipulators of invertebrate biology . Nat Rev Microbiol . 2008 ; 6 : 741 - 51 .
6. Hertig M , Wolbach SB . Studies on rickettsia-like micro-organisms in insects . J Med Res . 1924 ; 44 : 329 - 374 .7.
7. Hilgenboecker K , Hammerstein P , Schlattmann P , Telschow A , Werren JH . How many species are infected with Wolbachia?-a statistical analysis of current data . FEMS Microbiol Lett . 2008 ; 281 : 215 - 20 .
8. Osei-Poku J , Han C , Mbogo CM , Jiggins FM . Identification of Wolbachia strains in mosquito disease vectors . PLoS ONE . 2012 ; 7 , e49922 .
9. Baton LA , Pacidonio EC , Goncalves DS , Moreira LA . wFlu: characterization and evaluation of a native Wolbachia from the mosquito Aedes fluviatilis as a potential vector control agent . PLoS ONE . 2013 ; 8 , e59619 .
10. Andrews ES , Xu G , Rich SM . Microbial communities within fieldcollected Culiseta melanura and Coquillettidia perturbans . Med Vet Entomol. 2014 ; 28 : 125 - 32 .
11. de Oliveira CD , Goncalves DS , Baton LA , Shimabukuro PH , Carvalho FD , and Moreira LA . Broader prevalence of Wolbachia in insects including potential human disease vectors . Bull Entomol Res . 2015 : 1 - 11 .
12. Sinkins SP , Braig HR , O'Neill SL . Wolbachia superinfections and the expression of cytoplasmic incompatibility . Proc Biol Sci . 1995 ; 261 : 325 - 30 .
13. Laven H . Eradication of Culex pipiens fatigans through cytoplasmic incompatibility . Nature . 1967 ; 216 : 383 - 4 .
14. Min KT , Benzer S . Wolbachia, normally a symbiont of Drosophila, can be virulent, causing degeneration and early death . Proc Natl Acad Sci U S A . 1997 ; 94 : 10792 - 6 .
15. Teixeira L , Ferreira A , Ashburner M. The bacterial symbiont Wolbachia induces resistance to RNA viral infections in Drosophila melanogaster . PLoS Biol . 2008 ; 6 , e2 .
16. Hedges LM , Brownlie JC , O'Neill SL , Johnson KN . Wolbachia and virus protection in insects . Science . 2008 ; 322 : 702 .
17. Osborne SE , Leong YS , O'Neill SL , Johnson KN . Variation in antiviral protection mediated by different Wolbachia strains in Drosophila simulans . PLoS Pathog . 2009 ; 5 , e1000656 .
18. Xi Z , Khoo CC , Dobson SL . Wolbachia establishment and invasion in an Aedes aegypti laboratory population . Science . 2005 ; 310 : 326 - 8 .
19. McMeniman CJ , Lane RV , Cass BN , Fong AW , Sidhu M , Wang YF , et al. Stable introduction of a life-shortening Wolbachia infection into the mosquito Aedes aegypti . Science . 2009 ; 323 : 141 - 4 .
20. Walker T , Johnson PH , Moreira LA , Iturbe-Ormaetxe I , Frentiu FD , McMeniman CJ , et al. The wMel Wolbachia strain blocks dengue and invades caged Aedes aegypti populations . Nature . 2011 ; 476 : 450 - 3 .
21. Moreira LA , Iturbe-Ormaetxe I , Jeffery JA , Lu G , Pyke AT , Hedges LM , et al. A Wolbachia symbiont in Aedes aegypti limits infection with dengue, Chikungunya, and Plasmodium . Cell . 2009 ; 139 : 1268 - 78 .
22. Marinotti O , de Brito M , Moreira CK . Apyrase and alpha-glucosidase in the salivary glands of Aedes albopictus . Comp Biochem Physiol B: Biochem Mol Biol . 1996 ; 113 : 675 - 9 .
23. Hoffmann AA , Montgomery BL , Popovici J , Iturbe-Ormaetxe I , Johnson PH , Muzzi F , et al. Successful establishment of Wolbachia in Aedes populations to suppress dengue transmission . Nature . 2011 ; 476 : 454 - 7 .
24. Frentiu FD , Zakir T , Walker T , Popovici J , Pyke AT , van den Hurk A , et al. Limited dengue virus replication in field-collected Aedes aegypti mosquitoes infected with Wolbachia . PLoS Negl Trop Dis . 2014 ; 8 , e2688. Research showing strong dengue inhibition in wild Wolbachia-infected Stegomyia aegypti mosquitoes .
25. Ferguson NM , Kien DT , Clapham H , Aguas R , Trung VT , Chau TN , et al. Modeling the impact on virus transmission of Wolbachiamediated blocking of dengue virus infection of Aedes aegypti . Sci Transl Med . 2015 ; 7 : 279ra - 37 .
26. Lambrechts L , Ferguson NM , Harris E , Holmes EC , McGraw EA , O'Neill SL , Ooi EE , Ritchie SA , Ryan PA , Scott TW , Simmons CP , and Weaver SC. Assessing the epidemiological effect of Wolbachia for dengue control . Lancet Infect Dis . 2015 . Paper highlights a practical approach for dengue reduction through field release of Wolbachia-infected mosquitoes .
27. van den Hurk AF , Hall-Mendelin S , Pyke AT , Frentiu FD , McElroy K , Day A , et al. Impact of Wolbachia on infection with chikungunya and yellow fever viruses in the mosquito vector Aedes aegypti . PLoS Negl Trop Dis . 2012 ; 6 , e1892 .
28. Jeffries CL , Walker T. The potential use of Wolbachia-based mosquito biocontrol strategies for Japanese encephalitis . PLoS Negl Trop Dis . 2015 ; 9 , e0003576 .
29. Sinkins SP , Braig HR , O'Neill SL . Wolbachia pipientis: bacterial density and unidirectional cytoplasmic incompatibility between 30.
infected populations of Aedes albopictus . Exp Parasitol . 1995 ; 81 : 284 - 91 .
Zug R , Hammerstein P. Bad guys turned nice? A critical assessment of Wolbachia mutualisms in arthropod hosts . Biol Rev Camb Philos Soc . 2015 ; 90 : 89 - 111 .
Appl Environ Microbiol . 2012 ; 78 : 6922 - 9 .
Glaser RL , Meola MA . The native Wolbachia endosymbionts of Drosophila melanogaster and Culex quinquefasciatus increase host resistance to West Nile virus infection . PLoS ONE . 2010 ; 5 , e11977 .
Micieli MV , Glaser RL . Somatic Wolbachia (Rickettsiales: Rickettsiaceae) levels in Culex quinquefasciatus and Culex pipiens (Diptera: Culicidae) and resistance to West Nile virus infection . J Med Entomol . 2014 ; 51 : 189 - 99 .
Blagrove MS , Arias-Goeta C , Di Genua C , Failloux AB , Sinkins SP . A Wolbachia wMel transinfection in Aedes albopictus is not detrimental to host fitness and inhibits Chikungunya virus . PLoS Negl Trop Dis . 2013 ; 7 , e2152 .
Fu Y , Gavotte L , Mercer DR , Dobson SL . Artificial triple Wolbachia infection in Aedes albopictus yields a new pattern of unidirectional cytoplasmic incompatibility . Appl Environ Microbiol . 2010 ; 76 : 5887 - 91 .
Sinkins SP , Walker T , Lynd AR , Steven AR , Makepeace BL , Godfray HC , et al. Wolbachia variability and host effects on crossing type in Culex mosquitoes . Nature . 2005 ; 436 : 257 - 60 .
Atyame CM , Labbe P , Rousset F , Beji M , Makoundou P , Duron O , et al. Stable coexistence of incompatible Wolbachia along a narrow contact zone in mosquito field populations . Mol Ecol . 2015 ; 24 : 508 - 21 .
Craig Jr GB . Mosquitoes: female monogamy induced by male accessory gland substance . Science . 1967 ; 156 : 1499 - 501 .
Tripet F , Lounibos LP , Robbins D , Moran J , Nishimura N , Blosser EM . Competitive reduction by satyrization? Evidence for interspecific mating in nature and asymmetric reproductive competition between invasive mosquito vectors . Am J Trop Med Hyg . 2011 ; 85 : 265 - 70 .
Bargielowski IE , Lounibos LP , Carrasquilla MC . Evolution of resistance to satyrization through reproductive character displacement in populations of invasive dengue vectors . Proc Natl Acad Sci U S A . 2013 ; 110 : 2888 - 92 .
Favia G , Ricci I , Damiani C , Raddadi N , Crotti E , Marzorati M , et al. Bacteria of the genus Asaia stably associate with Anopheles stephensi, an Asian malarial mosquito vector . Proc Natl Acad Sci U S A . 2007 ; 104 : 9047 - 51 .
Rossi P , Ricci I , Cappelli A , Damiani C , Ulissi U , Mancini MV , et al. Mutual exclusion of Asaia and Wolbachia in the reproductive organs of mosquito vectors . Parasit Vectors . 2015 ; 8 : 278 .