Current Understanding
Japanese encephalitis virus (JEV) is an important cause of viral encephalitis in Asia,
with an estimated 67,900 cases annually [1]. Mosquito-borne zoonoses, including JEV,
present some of the most complex disease systems, often involving multiple mosquito
and vertebrate species.
The first investigations of JEV transmission ecology were undertaken in the 1950s
in Saitama Prefecture, Japan (Fig 1A) [2–10]. As a result of these studies, Culex
tritaeniorhynchus was implicated as the primary vector and pigs as the amplifying
hosts, with a minor role described for ardeid birds [10]. Scherer et al. [3] justified
the intensive investigation of pigs and birds in Japan by emphasizing that, within
this context, only these animals and wild rodents underwent population turnover high
enough to provide the continuous supply of susceptible individuals necessary to explain
the occurrence of annual epidemics. Research was focused on these species, in preference
to other animals, including cattle, whose total and susceptible populations were smaller.
Among potential bird hosts, ardeid birds in particular were studied because they possessed
anti-JEV antibodies, were numerous, colonial, could be caught in large numbers, and
were large enough to withstand repeated bleedings adequate for testing. Their selection
was not meant to imply that other birds were not potentially important in JEV ecology
[3].
10.1371/journal.pntd.0004074.g001
Fig 1
Study locations in Japan (A) and Bangladesh (B) where host community composition has
been estimated (Fig 2).
The transmission cycle proposed from this initial research in Japan arose from careful
study of the transmission context in that location, at that time. Vertebrate population
density (Fig 2), life span, and JEV viremia were considered when implicating primary
hosts [6–8]. Baited mosquito traps were used to determine numbers of mosquitoes attracted
to a variety of bird species, pigs, and humans [5]. The relative abundance of each
mosquito species caught in baited traps and their JEV infection status were compared
when implicating vectors in transmission [4]. Cx. tritaeniorhynchus was found to be
most abundant in traps baited with hosts able to produce JEV viremia [2,4], providing
circumstantial evidence for this species’ role in transmission, which was strengthened
by laboratory experiments demonstrating this mosquito’s competence for JEV replication
and transmission [2].
10.1371/journal.pntd.0004074.g002
Fig 2
Comparison of JEV transmission contexts between Saitama Prefecture, Japan (A), and
three districts of Bangladesh (Rajshahi, Naogaon, and Chapai Nawabganj) (B), with
respect to host community composition.
Arrows represent hypothesized transmission of JEV between hosts and mosquitoes. Each
square represents approximately 10,000 animals; smaller squares for pigs in Bangladesh
and ardeid birds in Japan represent proportionately smaller numbers. Saitama Prefecture
covers approximately 3,800 km2 and the three districts of Bangladesh approximately
7,500 km2. Data for Bangladesh were, therefore, scaled so that densities between the
two regions are comparable. Saitama Prefecture census data for cattle and pigs were
taken from [8], field estimates of ardeid birds from [7]. Bangladesh data for pigs
were from [20], and cattle, pigeons, ducks, and chickens from Bangladesh Yearbook
of Agricultural Statistics [19]. As ducks and chickens were reported together, an
approximate ratio of 1:5 was calculated from FAOSTAT for 2012 [21]. Cattle and bird
absolute numbers and density in Bangladesh are much higher than was observed in Japan.
Given the context in Bangladesh, it is not currently fully understood how JEV transmission
is maintained, and we propose that domesticated birds may play an important role.
As highlighted by numerous review articles, the initial investigations in Japan have
formed the basis for describing the JEV transmission cycle, primarily involving Cx.
tritaeniorhynchus, pigs, and, to a lesser extent, ardeid birds [11–15].
Considering Transmission Context
The Cx. tritaeniorhynchus–pig transmission cycle first described in Japan occurred
in a context where pigs were intensively farmed and were the most numerous of possible,
competent, vertebrate hosts. Yet not all regions of Asia experiencing Japanese encephalitis
(JE) outbreaks reflect this scenario.
JE cases do occur in the absence of intensive pig farming and where pig density is
low relative to other livestock, including in regions of Bangladesh and India [1].
Unlike Japan, in Bangladesh, Islam is the largest religion. As a consequence, pig
farming can be associated with social stigma in this region, thus restricting its
growth as an industry [16].
Pig density relative to cattle density is particularly important to consider in JEV
transmission ecology. Cattle are unable to produce viremia sufficient to infect mosquitoes
under experimental conditions and, thus, are a “dead end” for JEV [17]. During outbreaks
of JE in the 1950s in Saitama Prefecture, Japan (Fig 1A), there was a high pig population
turnover, with approximately 100,000 pigs slaughtered annually, and pig densities
were reported to be ten times higher than cattle (Fig 2A) [8]. In contrast, in some
JE-endemic regions of India, cattle can outnumber pigs by up to 20:1 [18]. In three
JE-endemic districts of Rajshahi Division, Bangladesh (Fig 1B), which, together, cover
an area almost twice the size of Saitama, the pig population is estimated to be 11,000
and the cattle population over 1 million—140 cattle for every pig (Fig 2B) [19,20].
When given a choice between feeding on a cow or a pig under experimental conditions,
42% of 496 Cx. tritaeniorhynchus fed on the cow and 5% on the pig [22]. Blood feeding
of natural populations of Cx. tritaeniorhynchus in India has been observed to be between
85% and 98% on cattle and less than 10% on pigs [18,23,24]. This compares with 36%
on cattle and 55% on pigs in Japan [25]. These differences are likely due to differences
in the availability of the respective hosts. Theoretical models of vector-borne pathogen
transmission [26] demonstrate that the rate of pathogen spread is particularly sensitive
to the proportion of vector bloodmeals taken from competent versus dead-end hosts.
This is because the proportion of bloodmeals taken on each host species influences
both mosquito-to-host and host-to-mosquito transmission rates, forming a squared term
in an equation for the basic reproduction number of a vector-borne pathogen. If mammalophilic
vectors such as Cx. tritaeniorhynchus are more likely to feed on cattle than pigs,
transmission intensity may decrease if cattle density substantially exceeds pig density
[27]. While it is possible there are sufficient mosquitoes per host in tropical regions
for pigs to maintain transmission irrespective of the proportion of bites on pigs,
the size of the reservoir community required for JEV amplification to levels that
are a risk to human populations is unknown. Are pig population densities in regions
of India and Bangladesh sufficient for maintaining JEV transmission? Our understanding
of the drivers of JEV transmission in regions that differ in transmission context
from Japan is currently deficient.
In light of the potential expansion of JEV to new geographic regions that support
a range of livestock and agricultural practices (http://faostat.fao.org/) [14,28,29],
it is important that the transmission cycle be reconsidered for regions of Asia where
the transmission context may differ substantially from that first described in Japan
(Fig 2).
Reassessing the JEV Transmission Cycle
During entomological investigations in ten randomly selected villages in a JE-endemic
region of Bangladesh [30], birds—including chickens, ducks, and pigeons—were observed
to be the most abundant domestic animals, often comprising between 50% and 100% of
household animal communities. These birds are also reported to be the most numerous
domestic animals by census data for the three districts where surveyed villages are
located [19].
In addition to density, the amount of virus present in host blood after a bite by
an infectious mosquito is also an important parameter in determining the extent to
which a host may contribute to transmission [31]. Viremia profiles of pigeons, ducks,
chickens, and pigs have, to our knowledge, yet to be compared with respect to the
probability of mosquito infection; however, experimental infection studies for these
animals are available [2,8,32–35]. Whilst the amount and strain of JEV administered
likely differ between studies, the amount of virus detected in an individual host
on any day post-infection was similar between species. Pig viremia has been recorded
to vary between 0.4 and 3.3 log10 lethal dose (LD) 50 / 0.03 ml, compared with 0.2
to 1.7 for pigeons, 0.5 to 3.4 for chickens, and 0.6 to 4.5 for ducks [2,8,32–34].
Although ducks and chickens are, therefore, likely to produce JEV viremia sufficient
to infect mosquitoes [2,8, 31,32–35], the role of domesticated birds in JEV transmission
remains unknown. The involvement of ducks in JEV transmission, in particular, was
suggested as a possibility in Borneo, but their contribution to transmission there
also remains to be quantified [36]. Quantifying the relative contributions of pigs
and domesticated birds to JEV transmission is essential for understanding JEV ecology
in regions where the pig population density is relatively low compared with the domesticated
bird population density (Fig 2B). We propose that several competing hypotheses should
be evaluated: (i) pigs contribute more than domesticated birds to JEV transmission;
(ii) domesticated birds contribute more than pigs to JEV transmission; (iii) the relative
contributions of domesticated birds and pigs varies in space and time. There are,
however, currently insufficient data to fully assess these hypotheses.
Efforts to accurately quantify the contribution of different hosts and vectors to
JEV transmission are hindered by the need to simultaneously assess multiple parameters
[26]. These parameters include population density of multiple species, mosquito species’
blood feeding habits, and the ability of species to become infected and subsequently
transmit JEV. As applied to the study of West Nile virus in the USA [37], the use
of mathematical models parameterized with data from entomological and host-based studies
would be useful in quantifying the relative roles of potential species in JEV transmission,
but this approach has not, thus far, been applied to the JEV system, in part due to
inadequate data.
Estimation of the parameters necessary for implicating host and vector species may
be affected by method bias (for example, mosquito collection methods that favor one
species over another) [37], and parameter estimates may differ across scales, space,
and time due to ecological heterogeneity (Table 1). These factors are important to
consider, as bias and heterogeneity may influence parameters for each species under
consideration in different ways that would need to be accounted for when using mathematical
models (Table 1). In addition, many mosquito species can become infected and, subsequently,
transmit JEV. Further investigations—including bloodmeal analyses, use of mosquito
sampling methods that focus collections on competent rather than dead-end host species
present in an area, and JEV competence experiments—would improve our understanding
of the host and vector species driving JEV transmission.
10.1371/journal.pntd.0004074.t001
Table 1
Summary of potential sources of bias and heterogeneity that may influence estimation
of parameters used to implicate host and vector species in Japanese encephalitis virus
transmission.
Parameter
Data source
Bias/ heterogeneity
Potential implications
Recommendation
Mosquito species relative abundance
Often estimated from sampling near large domestic animals, particularly cattle, at
dusk [30].
Over-representation of dusk-biting and/or mammalophilic species, including the Cx.
vishnui subgroup (Cx. tritaeniorhynchus, Cx. vishnui, Cx. pseudovishnui) in the studied
mosquito community relative to other species and under-representation of day-biting
and/or ornithophilic species [30].
May reinforce current theory of a Cx. tritaeniorhynchus–pig cycle, creating a barrier
to recognition of alternative transmission cycles.
Use a combination of methods. These may include: collections focused near hosts known
to produce JEV viremia both during the day and in the evening; collections of resting
mosquitoes away from potential host animals, indoors as well as outdoors.
Host and mosquito species competence (ability to become infected and subsequently
transmit a pathogen)
Estimated from experimental laboratory transmission experiments.
Usually taken to be two constant parameters that are not influenced by environmental
factors. Mosquito competence is, however, affected by host viremia (aspect of host
competence), and this relationship may be temperature-dependent [38].
Assuming constant host-to-mosquito and mosquito-to-host transmission probabilities
may lead to failure to account for regional differences in host and vector species
competence due to environmental conditions.
Experimental infections should be conducted to quantify how the probability of mosquito
midgut and salivary gland infection varies with dose and temperature. Such experiments
will give insight into the relationships between environmental factors and transmission
probabilities.
Mosquito species’ host-feeding patterns
Usually averaged over a region including multiple villages [18,23,39].
May not account for poor mixing between host species and vectors across spatial scales
[40].
May overestimate the proportion of bloodmeals taken on dead-end rather than competent
species in an area, resulting in failure to understand how transmission is maintained.
Identification of the scale at which host community composition varies. Quantification
of the proportion of bloodmeals on each host species at this scale (for example, at
the household rather than village level).
Implications for Control
Quantifying the relative contributions of species involved in JEV transmission, and
the role of birds in particular, would improve assessments of both the potential for
JEV to spread to new geographic regions [14,28,29] and the potential impact of particular
farming systems, including duck farming in rice paddies [41].
Japanese encephalitis is a vaccine-preventable disease and has been successfully controlled
by national human immunization programs in Japan, Taiwan, China, and Korea [1]; however,
the disease is still a major public health problem in many regions of Asia, including
Bangladesh and India [1]. The cost of national immunization programs and the logistics
of vaccinating all individuals in at-risk areas currently restrict use in some JE-endemic
regions [42]. Furthermore, as human infection does not contribute to transmission
and the human vaccine does not reduce transmission of JEV in the reservoir community,
no herd immunity is generated, and vaccination has to be sustained indefinitely. Implicating
host and vector species would improve understanding of transmission risk in space
and time, and could, therefore, inform targeted vaccination efforts toward those at
highest risk.