Tomberlin Et Al 2017
Tomberlin Et Al 2017
Tomberlin Et Al 2017
doi: 10.1093/aesa/saw086
Review
Special Collection: Filth Fly–Microbe Interactions
Abstract
Blow flies are commonly associated with decomposing material. In most cases, the larvae are found feeding on
decomposing vertebrate remains; however, some species have specialized to feed on living tissue or can sur-
vive on other alternate resources like feces. Because of their affiliation with such septic environments, these in-
sects have close associations with microbes. Historically, a tremendous amount of research focused on these
insects due to their veterinary importance. Within the past 40 yr, efforts have expanded this research to include
areas such as systems ecology, forensics, and even wound debridement (maggot) therapy. Initial research ef-
forts examining the relationship between microbes and these insects were hampered by the technology avail-
able. However, with the advent of high-throughput sequencing and modern molecular techniques, new ave-
nues of research examining these interactions have opened up. The purpose of this article is to highlight the
research exploring the interactions between microbes and blow flies with regards to blow fly biology, the appli-
cation of such information to benefit humanity, and potential future pathways of research.
Key words: interkingdom communication, veterinary entomology, medical entomology, trophic interaction, succession
Blow Fly Biology remains (Fig. 3; Hall 1948, Payne et al. 1968), while some are
known to cause myiasis (Broce 1985). The larvae pass through three
Blow flies (Diptera: Calliphoridae) (Figs. 1 and 2) are medium- to
stadia prior to pupating and emerging as adults. Total development
large-sized calyptrate flies, many of which are easily recognized for
time can vary depending on species and environmental conditions
their metallic blue or green coloration. They occur throughout the
encountered; in general, completion of development from egg to
year and have a global distribution except in areas of extreme cold.
adult takes between 8–14 d. Adult blow flies frequent carrion (Hall
This particular family of fly is of great medical, veterinary, and fo-
1948, Payne 1965, Pechal and Benbow 2016), wounds on verte-
rensic importance due to the resources they utilize and their associa-
brates (Sanford et al. 2014), feces (Mann et al. 2015, Brodie et al.
tion with people (Sanford et al. 2014), pets (Anderson and Huitson
2016), and even flowers (Fig. 4; Brodie et al. 2015). These resources
2004), livestock (Axtell 1986), poultry (Axtell and Arends 1990),
are used as sites for locating and securing mates, obtaining nutrition
and aquaculture (Fig. 3; Esser 1991, Aak et al. 2011). Globally, well
to meet the requirements of oogenesis, or supporting the develop-
over 1,000 species from 150 genera have been described (Shewell
ment of offspring. Because of the environment in which they inhabit,
1987). Of these, 54 species in 17 genera occur in North America
and their association with humans and other animals, a tremendous
(Whitworth 2006).
amount of research has been conducted to examine various aspects
The larvae of these flies are vermiform, and outside of the
of their biology, including:
Hawaiian genus Dyscritomyia, which larviposits (Wells et al. 2002),
adults deposit large numbers of eggs on a resource. In some cases, • Development (Byrd and Butler 1996, Byrd and Allen 2001,
the resulting larvae of these flies inhabit decomposing vertebrate Boatright and Tomberlin 2010, Owings et al. 2014)
C The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America.
V
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20 Annals of the Entomological Society of America, 2017, Vol. 110, No. 1
researchers began speculating that bacteria were regulating blow fly incubated with each of the Morganella–Proteus–Providencia spe-
attraction and colonization of sheep (Hobson 1935), but until re- cies and the combination of these four species. Significantly more
cently, technological barriers limited the ability of researchers to ex- oviposition occurred when a combination of Bacillus species was
plore these interactions and their relevance to the ecology and used than Proteus or a combination of Bacillus sp. and Proteus sp.
biology of this diverse group of flies. (Eddy et al. 1975). According to these authors, many of the results
DeVaney et al (1973) showed that bacteria-produced factors in were not consistent in their studies. For instance, peak attraction
bovine blood were highly attractive to primary screwworm (C. homi- of incubated blood was sometimes after 7 d of incubation and
nivorax). They determined cultures of several Gram-negative bacteria sometimes after 14 or 21 d. Similar inconsistent results were ob-
isolated from screwworm larval rearing media, homogenized third in- tained with homogenized screwworm larvae (Eddy et al. 1975).
stars of C. hominivorax, and infested wounds on sheep produced at- Most likely, these variations were caused by un-even presence of
tractive odors. These results suggested bacterial contamination of bacteria in the test materials used.
oviposition substrates was a prerequisite for attraction and oviposi- The above work was followed by Hammack et al. (1987) using a
tion by gravid flies (DeVaney et al. 1973, Eddy et al. 1975). These steam distillate of culture medium inoculated with P. rettgeri to as-
workers showed that the source of odors in incubated bovine blood sess the attraction of C. hominivorax adults. Results showed that fe-
that was attractive to gravid flies was bacteria (and compounds pro- males with previtellogenic ovaries and males were not attracted.
duced by them). Olfactometer tests showed that the blood containing Gravid mated females of 10–12 d old were most strongly attracted.
Providencia (previously Proteus) rettgeri was the most attractive. These results confirmed that P. rettgeri produces an attractant for
Comparatively, Morganella (previously Proteus) morganii was some- screwworm flies, and that this attractant lures females rather than
what more attractive than Proteus vulgaris and significantly more so males, and older females than younger ones, and more mated than
than Proteus mirabilis. They also compared the attraction of these virgin females (Hammack et al. 1987). Later, Hammack (1991) ex-
bacteria species alone with the four species in combination, and amined factors affecting oviposition by C. hominivorax using host-
found that the combination was considerably more attractive than P. originated fluids in laboratory bioassays. She reported that fresh
mirabilis, M. morganii, or P. vulgaris alone, but there was no signifi- blood (with no attractive odor) was as attractive for oviposition as
cant difference between this combination and P. rettgeri (Eddy et al the other attractive fluids tested including fluids from screwworm-
1975). They also tested a combination of 12 Bacillus species against infested wounds and cultures of P. rettgeri. However, she noted that
the combination of four species mentioned above. Results from these oviposition varied depending on the substrates to which the blood
studies showed that the Bacillus combination trapped only 3% while was applied, suggesting that an interaction exists between olfactory
the Morganella–Proteus–Providencia combination trapped 34% of cues and tactile stimuli to bring about oviposition (Hammack
the flies. Eventually, the cultures of P. rettgeri were found to be 1991). More recent work indicates color could also play an im-
the most attractive (Eddy et al. 1975). portant role in blow fly foraging behavior (Brodie et al. 2014,
Screwworm flies normally deposit more egg masses when they Brodie et al. 2015).
have a suitable substrate such as horse meat, liver, bovine blood, Chaudhury et al. (2002) continued the above research on screw-
etc. Eddy et al. (1975) compared oviposition on substrates worms using eight species of bacteria that were isolated from
22 Annals of the Entomological Society of America, 2017, Vol. 110, No. 1
screwworm-infested animal wounds. The species of bacteria were: (Chaudhury et al. 2016). These tests showed that significantly more
Enterobacter cloacae, E. sakazakii, Klebsiella oxytoca, P. mirabilis, flies landed on substrates containing P. mirabilis than on substrates
P. vulgaris, P. rettgeri, P. stuartii, and Serratia liquefaciens. Both fer- with other species of bacteria. Substrates treated with K. oxytoca at-
tile and sterile (irradiated) male and female C. hominivorax were tracted the least flies. Substrates containing bacteria incubated for
tested in a cage bioassay system for assessing attraction and oviposi- 72 h attracted significantly more flies than those incubated for 24,
tion using bovine blood inoculated with all eight species of bacteria 48, or 96h period. In 3-h duration oviposition tests, substrates with
and incubated for varying time periods. Substrates incubated for P. rettgeri attracted significantly more flies to oviposit than the other
48–72 h attracted more (>50% of flies were attracted to the re- four species. The most oviposition events were recorded from sub-
source) 7-d-old fertile females than did the substrates incubated for strates treated with all five species of bacteria. At least 72 h of incu-
24 and 96 h (<25% attraction). Significantly more fertile females bation seems to be required to obtain the most active volatiles.
were attracted to these substrates than sterile females (<20% attrac- These results suggest that C. macellaria uses similar chemical cues as
tion) of the same age group. Males of all tested age groups were C. hominivorax from bacteria volatiles as an oviposition attractant,
unresponsive (<1% attraction). Oviposition tests lasting for 1 h re- supporting the generalizability of at least some blow fly interactions
sulted in significantly more oviposition in treated substrates com- with microbes.
pared to untreated control. Results indicate volatiles from five Interestingly, several of the key microbial players in the
individual species of bacteria (K. oxytoca, P. mirabilis, P. vulgaris, Cochliomyia studies also appear in the Lucilia system where myiasis
P. rettgeri, and P. stuartii) were responsible for attracting more fe- manifests in association with animal feces in wool. The larvae of
males resulting in more oviposition than volatiles from the remain- sheep blow flies L. cuprina and L. sericata are the primary cause of
ing three species (E. cloacae, E. sakazakii, and S. liquefaciens) sheep myiasis in Australia, Europe, and New Zealand. The re-
(Chaudhury et al. 2010). Volatiles from the same five species sponses of these two species are similar and attraction to host and
were also tested in a two-choice bioassay to study landing response subsequent oviposition appear to involve volatile chemicals resulting
and oviposition of the secondary screwworm, C. macellaria from bacterial decomposition. Emmens and Murray (1982) selected
Annals of the Entomological Society of America, 2017, Vol. 110, No. 1 23
four species of bacteria isolated from the fleece of Merino sheep, sp. to similar classes of molecules. In some instances, these bacterial
namely, Pseudomonas aeruginosa, Bacillus subtilis, E. cloacae, and cues/signals are clearly important to numerous organisms (see Davis
P. mirabilis for their study. Extracts from cultures of these bacteria et al. 2013), indicating that the blow fly system is a useful model for
were incorporated into nutrient agar and exposed to females of dissecting insect and animal interactions with microbes.
L. cuprina in cages. Odors from the cultures of bacteria attracted At the time the aforementioned studies were conducted, the eco-
L. cuprina females to oviposit but the attraction was not consistent logical relevance of the VOCs was not known. However, within the
for all the cultures. The order of the bacterial species with respect past decade it was determined that these volatiles are associated
to decreasing overall response was P. mirabilis with highest number with bacterial activity, specifically bacterial communication and
of eggs laid, followed by E. cloacae, B. subtilis and least being decision-making (i.e., quorum sensing (Lee et al. 2007)). More re-
P. aeruginosa (Emmens and Murray 1982). In another experiment, cent research is beginning to work toward bridging the nutritional
extracts from unsterile sheep fleeces treated with these four species ecology of bacteria with blow fly behavioral ecology, as many of
of bacteria singly stimulated oviposition of L. cuprina equally dur- these volatiles are by-products of the break-down of essential amino
ing 24-h period; however, with increasing length of incubation, the acids (Liu et al. 2016). Specific examples of VOCs include dimethyl
cultures of P. mirabilis, E. cloacae, and B. subtilis became contami- disulfide, which is produced by the breakdown of methionine
nated with increasing numbers of P. aeruginosa resulting in greater (Hayward et al. 1977), is a recognized by-product of vertebrate car-
responses of the flies (Emmens and Murray 1983). The maximum rion decomposition (Vass et al. 2002, Paczkowski et al. 2015), and a
number of eggs was deposited over 4-d-old cultures. This response regulator of blow fly attraction to such resources (Urech et al.
was significantly associated with the presence of P. mirabilis and 2004). Other by-products include isobutyl amine and phenylacetic
E. cloacae but not B. subtilis. The relatively lower oviposition re- acid, which are produced by the degradation of valine (Richardson
sponses to P. aeruginosa in pure culture as seen in these experi- 1966) and phenylalanine (Erdmann and Khalil 1986), respectively.
Perhaps the increased survival rates seen with mixed bacterial en- associated with blow fly larvae and the larvae themselves has been
vironments are because different bacteria are more advantageous conducted (Cazander et al. 2009a, Cazander et al. 2009b, Cazander
at different stages of growth dependent on the metabolites they et al. 2010, Barnes and Gennard 2011, Cazander et al. 2012, Barnes
produce and the nutritional needs of the arthropod at that stage. and Gennard 2013, Cazander et al. 2013). Two of these antimicro-
Zurek et al. (2000) collected third-instars of house fly larvae from bials, phenyl acid acid (noted above) and phenylacetaldehyde, were
two common sources found at animal production facilities. They identified from P. mirabilis isolated from larvae of C. hominivorax
found that Bacillus coagulans, Bacillus sp., Clavibacter michiga- (Erdmann and Khalil 1986). Genetic and microscopic evidence indi-
nese, Corynebacterium aquaticum, Lactococcus garviae, cating that P. mirabilis can reside in L. sericata salivary glands
Microbacterium esteraromaticum, Microbacterium lacticum, (Singh et al. 2015, Blenkiron et al. 2015), which are relatively mi-
Microbacterium liquefaciens, Ochrobacter anthropic, crobe free, supports the concept that these two species coevolved to
Sphingobacterium spiritivorum, Sphinomonas capsulataa, kill other microbes. However, it should be noted that while many
Staphylococcus epidermidis, Staphylococcus lentus, Streptococcus san- bacteria are suppressed by ES, some were able to survive pupation
guis, Xanthobacter flavus, and Yersinia pseudotuberculosis, all sup- of C. macellaria, indicating some level of "resistance" to being di-
ported larval growth to some extent to the pupal stage, whereas gested by the larvae (Ahmad et al. 2006). This ability to persist in
Corynebacterium seminale, Gordona amarae, Microbacterium barkei, the alimentary canal of the insect through development is cause for
Morganella morganii, Providencia rettgeri, Providencia stuartii, and concern as these flies could serve as mechanical vectors for these path-
Serratia marcescens did not. Streptococcus sanguis and ogens, as well as creating an environment in which antibiotic resis-
Sphingobacterium spiritivorum supported larval development through tance is developed and amplified in some circumstances (Wei et al.
eclosion as adult flies, whereas Bacillus sp. and Staphylococcus epider- 2014a,b), but also provides further support for the concept that cer-
midis did not. tain flies and microbes act as mutualists to compete against others.
(Caporaso et al. 2010), mothur (Schloss et al. 2009), RDPipeline Enterobacteriaceae. It can be an opportunistic pathogen living in
(Cole et al. 2014)). Metagenomic sequencing approaches provide in- soil, water, and sewage, causing diarrhea with fever and tachycar-
formation not only on microbial community structure but also on dia, and leading to low blood pressure. The other Proteobacterium,
microbial function, and avoid PCR-based biases associated with Ignatzchineria was first isolated as the dominant species in the ante-
marker gene approaches (Lee et al. 2012). Recovery of rare taxa re- rior portion of the digestive tract in larval Wohlfahrtia magnifica
quires high sequencing depth in metagenomic sequencing Schiner (Diptera: Sarcophagidae) flesh flies (Toth et al. 2006, Gupta
approaches, which ultimately increases sequencing cost, and compli- et al. 2011). This species has been associated with myiasis, but was
cates already complicated metagenomic data analysis pipelines not generally associated with severe human disease until reports in
(Sharpton 2014). One method for balancing the advantages of both which it was isolated in cases of bacteremia and urinary tract infec-
methods is to use single-molecule information to extrapolate the tion (Maurin et al. 2007, Roudiere et al. 2007, Barker et al. 2014,
metagenomic gene content by using prior knowledge of sequenced Le Brun et al. 2015). Neither the mechanism nor the epidemiology
bacterial genomes (Langille et al. 2013). Given the importance of of an Ignatzchineria infection has been defined and the cases were
blow fly interactions with microbes, we predict many more studies all associated with a corresponding fly larvae infestation where
of microbial communities in blow flies in the coming years. Ignatzchineria was isolated concurrent with other bacteria, such as
Enterococcus and Providencia (Le Brun et al. 2015). Lactobacillus
is a Firmicute that is a member of the lactic acid bacterial group.
These bacteria constitute a major portion of the microbiota of the
Bacteria Associated With Blow Flies gut and other body sites. They produce alcohol and lactic acid from
Blow flies and other filth flies (e.g., Sarcophagidae and Muscidae) sugars which lowers pH and that controlled fermentation is ex-
use decaying organic matter for nutrition and larval habitat and ploited by industry in the production of items such as yogurts, beer,
among a variety of arthropods, including some blow flies, is In an interesting study by Pechal (2012), the microbiome associated
Wolbachia (Stouthamer et al. 1999, Baudry et al. 2003, Mingchay with decomposing swine carcasses that were accessible to flies or in
et al. 2014). Studies suggest that a benefit of vertically-transmitted which the flies were excluded was characterized over five days. They
infection is the prevention of more virulent infection, by horizontally demonstrated significant changes in the bacterial community during
transmitted organisms (Lively et al. 2005). Wolbachia also has the decomposition between insect access and exclusion carcasses. When
capability to cause reproductive isolation between infected and flies were not present, Proteobacteria was a dominant taxon
uninfected flies, although no such evidence for Wolbachia-induced throughout the 5-d sampling period, whereas relative abundance of
reproductive isolation has been documented in blow flies (Baudry Firmicutes decreased as decomposition progressed. However, when
et al. 2003, Mingchay et al. 2014). flies were present, the opposite occurred, as Proteobacteria de-
Horizontal transmission of pathogenic bacteria by the exterior creased over time and Firmicutes became the dominant taxon by the
surfaces and mouthparts, of flies, along with internal transfer via fe- fifth day of decomposition. At the genus level, Psychrobacter and
ces and vomit has long been known to occur in many fly species Moraxella were dominant for both exclusion and access carcasses.
(Förster et al. 2007, 2009; Pava-Ripoll et al. 2012). Blaak et al. But other bacterial succession patterns differed as decomposition
(2014) demonstrated the horizontal acquisition of Escherichia coli progressed. Without insect access, Aeromonas and Shewanellaceae
with extended-spectrum antibiotic resistance to most beta-lactam were detected only on the first day, Peptostretpococcus was detected
antibiotics by blow flies at a poultry facility. Horizontal transmis- only on the fifth day and Proteus transitioned to the dominant taxon
sion of various bacteria in blow flies was recently assessed in a meta- by the third and fifth day. When insects were present, Providencia
genomic study of L. sericata and L. cuprina, that characterized and was dominant on the first and third day, but Bacillales was domi-
compared the bacteria on adult flies and the fresh liver before they nant by the fifth day. Proteus and Corynebacterium were present on
oviposited on it, followed by the resulting larvae and the aged liver the third day of decomposition, but by the fifth day, Psychrobacillus
associated communities of bacteria, there can be additional valuable features of that strain. First, there are two high quality reference ge-
information gleaned from the whole genomes of specific species and nomes for the species and the blow fly derived strain is much more sim-
strains isolated from blow flies and blow fly resources. One area ilar to the BB2000 strain used to study self-recognition during
where this endeavor is important relates to bacterial community swarming of P. mirabilis (Gibbs et al. 2008, Cardarelli et al. 2015)
studies. For example, algorithms can be used to predict community than is was to the original reference genome obtained from a clinical
function from the community structure information provided by the strain of the species. Second, it is clear that the fly-derived strain con-
16S rDNA gene (Langille et al. 2013). This is done by extrapolating tains lineage specific insertions and deletions. It remains to be seen if
from the presence of specific taxa (via 16S sequences) to the gene there are genes relevant to fly interactions in those regions of the ge-
compositions of entire genomes and communities using microbial nome, but such a hypothesis will be important to test. It is clear that P.
genome databases to provide a prediction of metabolic potential es- mirabilis and other bacteria often have a core genome (composed of se-
timated from the types and predicted functions of genes in the se- quences shared by all members) as well as numerous auxiliary compo-
quenced genomes associated with 16S sequences. This capability nents (composed of sequences not shared by all members of the species
enables researchers to extend their interpretations of 16S data to the and sometimes unique to one strain; Collins and Higgs 2012).
expected functionality of the community, facilitating the ability to Evolutionary comparisons, such as those described below, of fly-
ask questions like, does the community have the ability to catabolize derived and non-fly-derived bacteria will help to determine if core ge-
a specific amino acid or produce a specific antibiotic? Such capabil- nome components, auxiliary components, or both components of bac-
ity with blow fly bacteria communities may allow us to expand our terial genomes are important to fly–bacterial interactions.
understanding of the specific biological processes involved in fly–mi-
crobe interactions. However, the quality of a result from any appli-
cation that relies on a database will depend greatly on the content of
Fig. 5. Comparisons of phylogenies of host flies and their associated bacteria can inform our understanding of the habitats in which flies live. Phylogenies show
the evolutionary relationships of nine fly species and bacterial samples taken from each of the nine flies. On the left, the phylogenies of the flies and bacteria are
in perfect congruence, demonstrating that the bacteria have evolved within each species and without horizontal transmission between species. On the right, the
phylogenies of the flies and bacteria are discordant, suggesting that fly habitat is a better predictor of bacterial association than evolutionary ancestry. Three can-
didate habitat categories are defined for each fly species based on the evolutionary relationships of their associated bacteria.
28 Annals of the Entomological Society of America, 2017, Vol. 110, No. 1
life history (Fig. 5). For example, the gut microbial communities in allow for the identification of specific mutations that promote colo-
mammals and insects are associated with the host diet, and not nec- nization, and the targeting of the products of those genes could re-
essarily congruent with the phylogenetic relationships of the hosts duce the ability of pathogenic bacteria to be vectored by blow flies.
(Muegge et al. 2011, Yun et al. 2014). As described above, the pre- In addition, host-specific adaptation should result in an evolved bac-
ferred substrate for oviposition differs across blow fly species, and teria strain that affects fly physiology differently than the ancestral
we therefore expect blow fly species that oviposit in the same sub- strain. Aside from immune-related genes (see below), little is known
strate to have associated microbial communities that resemble each about specific fly traits that regulate bacterial proliferation. It would
other. In addition, the microbial communities associated with blow thus be informative of fly traits involved in bacterial interactions to
fly species whose larvae are blood-feeding parasites of birds compare the phenotypes of blow flies fed ancestral and derived
(Protocalliphora sp.) should differ from blow flies whose larvae de- strains of host-evolved bacteria.
velop in carrion or flesh (Sabrosky et al. 1989).
Comparisons of host-associated bacteria across blow fly species
have tremendous potential to resolve cryptic variation and similari- Comparative Genomics of Blow Flies to Study
ties in life history strategies across species. While the larval habitats Ecological Interactions Between Bacteria and
of blow flies are easily characterized, the breadth of adult diet Hosts
choice and greater adult mobility can expose the flies to bacteria
Genome sequences of blow flies will also improve our understanding
whose origin is more difficult to determine. For example, a female
of interactions with bacteria and the specific aspects of fly physiol-
bias in visits to oviposition sites could result in sexually dimorphic
ogy responsible for regulating bacteria. For example, comparisons
microbial communities within blow fly adults (Mohr and
of gene content and sequence evolution in the genomes of related fly
Tomberlin 2014), which may translate into variation in the bacte-
demonstrated that the expression of genes encoding AMPs is up- and Firmicutes (30–70%; dominated by Enterococaceae,
regulated upon exposure to bacteria, but sequenced genomes now Lachnospiraceae, and Ruminococaceae)) and less so by
allow for the identification of novel recognition and effector compo- Proteobacteria (5–25%; dominated by Enterobacteriaceae) and
nents of the immune system and other genes involved in the physio- Actinobacteria (<10% dominated by Bifidobacteriaceae).
logical response to bacteria exposure through genome-wide analyses Interestingly, the absence of these gut microbes resulted in a thinner
of gene expression (Joyner et al. 2013, Nayduch et al. 2013, Poppel layer of olfactory cilia, a reduced epithelial cell turnover rate and a
et al. 2015, Sackton et al. 2016). For example, 47 expressed AMPs reduced expression of many genes associated with the olfactory sig-
were identified in a high throughput RNA sequencing experiment in nal transduction pathway. Thus, the microbial community structure
which L. sericata larvae were infected with P. aeruginosa and S. au- may indeed influence the odor sensory capabilities of its host. Flies
reus (Poppel et al. 2015). The activities of 23 of these AMPs were use the olfactory sensilla and receptors on their antenna and maxil-
tested against a panel of Gram-positive and -negative bacteria, and lary palps to smell odors and locate resources (Shanbhag et al. 1999,
their effects varied across AMP–bacteria combinations and de- Wasserman and Itagaki 2003). We know that the area around
pended on which other AMPs were present in the treatment (Poppel mouthparts can be heavily contaminated with bacteria (Barro et al.
et al. 2015). Additional experiments could determine which AMPs 2006). Could the bacteria present around the antennae and fly
(or combinations of AMPs) are best suited to be used as antimicro- mouthparts, as well as their gut microbiome, affect development of
bial agents for wound therapy or provide targets for pest control odor perception organs in the fly? Could raising flies under certain
measures. lab conditions or on specific resources, which affect the community
The power of genomic approaches to identify previously unchar- structure of the gut microbiome, change adult olfactory capabilities
acterized components of the blow fly response to bacteria can be in- and ultimately the behavior of the fly being investigated? What im-
creased by comparing multiple species or different strains within a pact would this have on the olfactory capabilities of wild flies that
enzymes that cleave other proteins. Thus, while mRNA studies can mRNA and proteomic levels described above. However, metabolic
provide valuable information regarding interactions between bacte- shifts can also be studied. It is possible to assay basic metabolic re-
ria and hosts, there are also levels of regulation independent of RNA sponses of flies and bacteria with any number of commercially avail-
that are equally valuable. As the genomes of blow flies and their mi- able panels and tests, such as Biolog plates (Garland and Mills 1991,
crobial associates are produced, these applications will become Garland 1997, Smalla et al. 1998, Choi and Dobbs 1999, Dobranic
more feasible—opening up new areas of inquiry. and Zak 1999, Classen et al. 2003, Bochner et al. 2011), which can
be used to evaluate metabolic profiles of both bacteria and eukary-
otes. Given the connections between bacterial signaling molecules
and essential amino acids, the metabolic profiles of both bacteria
Metabolic Studies and flies will be useful in dissecting their interactions. Some work
Metabolic consequences of insect–microbe interactions are important has already been done with this tool in carrion ecology (Pechal et al.
for understanding the mechanisms underpinning them. As an exam- 2013), using plates that included molecules found to be of impor-
ple, this review has demonstrated the importance of indole to a num- tance in previous blow fly studies (Ma et al. 2012, Tomberlin et al.
ber of blow fly–microbe interactions. This amino acid-derived 2012). However, individual blow flies and bacteria are small com-
molecule is also important to bacterial signaling and mammalian reg- ponents of that complex system, making it difficult to assign meta-
ulation of inflammation. In this era where we are learning through bolic responses to specific bacteria. Further, and more specific,
studies of bacteria communities that bacteria impact and are associ- dissection of blow fly interactions with bacteria using these and sim-
ated with a wide array of eukaryotic phenotypes, it will be necessary ilar tools will be useful in determining and differentiating among hy-
to step beyond bacterial community structures to the mechanisms un- potheses regarding the mechanisms regulating them.
derpinning the importance of certain community members on eukary- In addition, mass spectrometry (MS) and other chemical analysis
rapidly and cheaply has opened the door to developing a more com- Akhtar, M., H. Hirt, and L. Zurek. 2009. Horizontal transfer of the tetracycline
prehensive genetic, physiological, and metabolic framework for the resistance gene tetM mediated by pCF10 among Enterococcus faecalis in the
types of studies described above that surpass the community sam- house fly (Musca domestica L.) alimentary canal. Microb. Ecol. 58: 509–518.
Aleklett, K., M. Hart, and A. Shade. 2014. The microbial ecology of flowers:
pling information provided by the small number of 16S rDNA se-
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Anderson, G. S., and N. R. Huitson. 2004. Myiasis in pet animals in British
advances can be determined through genomic studies of both the mi-
Columbia: The potential of forensic entomology for determining duration
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