Journal of Pest Science (2022) 95:291–301

https://doi.org/10.1007/s10340-021-01390-3

ORIGINAL PAPER

Extended holding period and yeast hydrolysate in pre‑release diet

increase abundance of mature sterile Queensland fruit fly males
in the field
Md. Jamil Hossain Biswas1 · Bishwo Mainali1 · Jess R. Inskeep1,2 · Sushil K. Gaire1 · Dominic Cross1,3 ·
Lloyd D. Stringer4,5 · Phillip W. Taylor1 · Polychronis Rempoulakis1,6

Received: 22 December 2020 / Revised: 15 April 2021 / Accepted: 19 May 2021 / Published online: 30 May 2021
© The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature 2021

Abstract
Queensland fruit fly (Q-fly), Bactrocera tryoni (Froggatt), is a significant pest of horticultural crops in Australia. The sterile
insect technique (SIT) is currently employed to eradicate outbreaks in fruit fly free regions and may also be used to suppress
populations in endemic regions. For SIT to succeed, it is imperative that the released sterile males survive, disperse, attain
sexual maturity, and are sexually competitive against their wild rivals. Q-fly SIT programmes have conventionally held
adult flies for two to three days and have sometimes fed them only sugar before release, providing little time or nutrition for
development prior to release. We investigated whether a 5-d pre-release holding period and provision of yeast hydrolysate
(YH) together with sugar in the pre-release diet increase abundance of mature male Q-fly in the field. Indicating increased
survivorship and/or maturation, the combination of YH feeding and 5-d pre-release holding period resulted in 6–8 times
more recaptures of mature male flies in cuelure traps than was the case for flies released at 2 d with or without YH and for
flies released at 5 d without YH. Flies held for 5 d and fed YH were relatively more abundant than flies from other treatments
in traps close to the release point and were as abundant as other treatments in traps at the greatest assessed distances. These
findings strongly support a recommendation that sterile Q-flies be provided a pre-release diet of YH and sugar and be held
for 5-d post-eclosion before release.

Keywords Bactrocera tryoni · Q-fly · Sterile insect technique · Pre-release supplement · Sexual maturation

Key Messages

• Released sterile Q-flies held for 5 d and fed yeast hydro-

lysate (YH) were recaptured 6–8 times more than those
Communicated by Chris Cutler. held for 2 d with or without YH or for 5 d without YH.
• Higher recaptures indicate higher survival and/or sexual
* Md. Jamil Hossain Biswas
md-jamil-hossain.biswas@students.mq.edu.au maturation.
• Flies held for 5 d and fed YH tended to be captured close
1
Applied BioSciences, Macquarie University, Sydney, to the release point more than other treatments, but this
NSW 2109, Australia
was more than compensated by greater abundance.
2
Vector Control Branch, Hawaii Department of Health, • A 5-d pre-release holding period, with YH, is recom-
Kahului, HI 96732, USA
mended for operational Q-fly SIT programmes over the
3
The University of Sydney, Sydney, NSW, Australia historical 2–3-d standard.
4
The New Zealand Institute for Plant and Food Research Ltd., • Releasing 5-d-old YH-fed adults increases number of
Lincoln 7608, New Zealand sexually mature flies in the field, improving opportuni-
5
Better Border Biosecurity, Lincoln, New Zealand ties for SIT success.
6
New South Wales Department of Primary Industries,
Ourimbah, NSW 2258, Australia

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Introduction
The sterile insect technique (SIT) involves mass produc-
tion and release of large numbers of reproductively sterile
insects to suppress reproduction in pest populations and is
widely used to control major tephritid fruit fly pests (Kni-
pling 1955, 1959; Krafsur 1998; Enkerlin 2005; Shelly and
McInnis 2015). There are many aspects of SIT operations,
such as quality of larval or adult diets, holding conditions,
and handling that can affect key quality measures such as
body mass, emergence, survival, flight ability, and mat-
ing ability (Shelly and Kennelly 2003; Dyck et al. 2005;
Chang et al. 2006). In many fruit fly SIT programmes,
pupae are produced and irradiated at a mass-rearing facil-
ity and are then delivered to a rear-out facility in proximity
to the release area, where the adults emerge and are held
at high density until release (Pereira et al. 2021). The pre-
release holding period may be kept short for logistical rea-
sons such as the cost of material and labour and to avoid
the need for larger infrastructure to handle overlapping
cohorts, as well as to reduce mortality of the flies that can
arise from stress when kept at high density. On the other
hand, releasing insects before they reach sexual matura-
tion can lead to reduced performance in the field (McInnis
et al. 1996; Lance et al. 2000; Lux et al. 2002). In SIT, it
is imperative that the quality and number of mature sterile
males are maintained at high levels in order to out-number
and out-compete wild males to impede reproduction of
wild females (Shelly et al. 2007). Decisions of how long
to hold sterile males before release are hence driven by the
competing demands of logistics, holding costs, pre-release
mortality, and post-release performance.
The Queensland fruit fly (Q-fly) Bactrocera tryoni
(Froggatt) (Diptera: Tephritidae), is Australia’s most eco-
nomically damaging fruit fly pest of horticulture, caus-
ing more than $300 M/yr in losses (Hancock et al. 2000;
Dominiak et al. 2003a; Clarke et al. 2011; Hort Innovation
2016). The success of SIT for Q-fly control depends on
the abundance, distribution, and performance of sexually
mature males in the field (Meats and Edgerton 2008). In
SIT programmes, Q-flies have typically been held for two
to three days before release and in some cases have only
been provided sugar and water for sustenance (Dominiak
et al. 2003a; Meats et al. 2006; Weldon and Meats 2007;
Reynolds et al. 2014). Adult Q-flies require adequate nutri-
tion, particularly a protein source, in order to complete
sexual development (Vijaysegaran et al. 2002; Prabhu
et al. 2008; Weldon et al. 2008; Pérez-Staples et al. 2009,
2011; Weldon and Taylor 2011; Fanson and Taylor 2012).
As a result, flies released in SIT programmes without pre-
release access to a protein source need to acquire protein
as well as other nutrients in the field to sustain sexual
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Journal of Pest Science (2022) 95:291–301
development (Gilchrist and Meats 2012). Laboratory and
field cage studies have found that sexual development and
mating competitiveness of male Q-fly can be sustained by
providing yeast hydrolysate (YH) as a protein source con-
tinuously for 24 or 48 h (Weldon et al. 2008; Pérez-Staples
et al. 2007, 2009; Reynolds et al. 2014). Yeast hydrolysate
is an important component of adult diet for mass-reared
fruit flies, providing a rich source of amino acids, carbohy-
drates, sterols, vitamins, and minerals (Barry et al. 2007;
Chang 2009). There is also evidence that pre-release YH
feeding can improve post-release performance of sterile
Q-fly in open field settings. In releases carried out when
flies were 2—3 days post-eclosion, Reynolds et al. (2014)
found that cuelure traps recaptured more flies from a group
that was provided sugar and YH as a pre-release diet than
from a group that had been provided only sugar. Because
only sexually mature males are attracted to cuelure traps
(Weldon et al. 2008), the findings of Reynolds et al. (2014)
reflect effects of YH on survivorship and/or maturation.
Regardless of how the effect is derived, the results still
indicate more sexually mature sterile males in the field.
Laboratory, field cage, and open field studies are quite
consistent in supporting the provision of YH to Q-flies
released in SIT programmes at 2–3 days of age. In one cau-
tionary exception, pre-release provisioning with YH has
been found to render Q-flies more vulnerable to starvation
if they are unable to acquire sufficient further nutrition after
release (Taylor et al. 2013a). This is likely because providing
YH in the diet places the flies on a developmental trajectory
that imposes high nutritional demands and exposes them
to risk of starvation if they are insufficiently advanced in
development when food shortages are encountered. Perhaps
higher survivorship might be achieved if flies are held for
longer and released when in a more mature state (Kaspi and
Yuval 2000; Yuval et al. 2002, 2007; Maor et al. 2004).
Pre-release holding period in fruit fly SIT varies amongst
programmes and species, such as a minimum of 4 d for
Ceratitis capitata, 5 to 7 d for Anastrepha ludens and 5 d
for A. obliqua and Zeugodacus cucurbitae (Enkerlin 2007;
Shelly et al. 2007; McInnis et al. 2007, 2013). Meats et al.
(2003) considered effects of pre-release period and condi-
tions in Q-fly SIT and reported very poor recapture rates
of flies that were released after a week of access to sugar
and YH compared with flies released as teneral adults that
had access only to sugar. However, the authors concede that
poor recapture rates could have reflected stressful pre-release
conditions that reduced the quality of released mature flies.
Despite weaknesses in Meats et al. (2003), the consensus
view has been that a 2–3-day pre-release holding is preferred
over longer pre-release holding periods for Q-fly, and there
has been no subsequent investigation. In the present study,
we revisit the question of pre-release holding period for
Q-fly SIT and also consider the effects of diet. Sterile Q-flies
Journal of Pest Science (2022) 95:291–301
were released at two or five days of age after receiving a
pre-release diet of sugar only (YH-deprived) or sugar plus
YH (YH-fed), and the field abundance and flight distance of
mature males were compared using cuelure traps. We also
consider the potential influence of weather and landscape
features on the spatial distribution of the flies based on the
proportion of Q-flies released and recaptured.
Materials and methods
Insects
Q-fly were obtained as eggs from a colony that was reared
on a gel-based larval diet (Moadeli et al. 2017) in con-
trolled environment conditions (25 ± 0.5 °C, 65 ± 5% RH)
at Macquarie University, New South Wales, Australia.
The adult Q-fly colony was fed a 3:1 mixture of sugar and
YH (MP Biomedicals, Aurora, OH, USA) in a mesh cage
(40 × 40 × 110 cm). Tap water was continuously available
through a soaked sponge. Three parental generations from
the laboratory colony were used to collect eggs to conduct
five release-recapture experiments; G23 for the first and sec-
ond field release; G24 for the third and fourth release; and
G25 for the fifth release. To collect eggs, we placed three
oviposition devices inside the colony cages from 9 am to
4 pm. Oviposition devices consisted of white 250-mL plastic
bottles with numerous puncture holes and containing ~ 2 mL
of water to generate high humidity to keep the eggs moist.
The eggs were collected from the inner surface of the ovi-
position devices by rinsing with fresh tap water. Using a
pipette, the collected eggs were then seeded (~3500 eggs)
onto gel larval diet (150 mL) in a plastic tray for rearing fol-
lowing a standard protocol (Moadeli et al. 2017). When the
larvae reached the third instar, we placed the plastic tray con-
taining the developing larvae onto a 0.5-cm deep layer of fine
vermiculite (Ausperl, Orica Australia, Banksmeadow, NSW,
Australia) inside a clear plastic bin (67.5 × 50 × 43 cm). The
larvae exited the larval tray and landed in the vermiculite
where they pupated. Pupae were removed from vermicu-
lite by using a fine mesh sieve and were transferred to 5-L
clear plastic cages (23 × 16 × 16 cm) that had a mesh-covered
opening (14 × 10 cm) for ventilation. The pupae were kept
under standard conditions (25 ± 0.5 °C, 65 ± 5% RH). Two
days prior to eclosion, 300 mL of pupae (~12,000 pupae)
for each treatment was placed in hypoxic conditions over-
night inside the sealed zip-lock plastic bag (230 × 320 mm,
Shantou Zhiyuan Commodity Co. Ltd) and then irradiated at
65 Gy using a C ­ o60 gamma irradiation source at Macquarie
University (dose rate 9 Gy/min). The irradiated pupae were
marked using four fluorescent dyes—stellar green 8, astral
pink 1, comet blue 60, and lunar yellow 27 (Swada, 7 Stan-
ley Street, Stalybridge, Cheshire, SK15 1SS, England)—at
293
a rate of 2 g of dye per L of pupae (Meats and Edgerton
2008; Akter et al. 2020) according to pre-release holding
period and pre-release diet treatment. Dye colour alternated
among treatments between releases to ensure that any dye
effects were dispersed across treatments rather than incur-
ring systematic bias. Approximately 60 mL of marked pupae
from each treatment group (~2400 pupae) was transferred
to an open 90-mm-diameter Petri dish and then placed in
separate fine mesh cages (47.5 × 47.5 × 47.5 cm, Megaview
BugDorm-4F4545).
Quality control of dyed sterile flies
Fly quality for each release was assessed using a flight abil-
ity test (25 ± 0.5 °C with 65 ± 5% RH) following standard
procedures (Collins et al. 2008; FAO/IAEA/USDA 2014;
Moadeli et al. 2017). Three replicates were carried out for
each treatment for each release. One hundred pupae were
placed in 5.5-cm plastic Petri dish lids which were then cen-
tred on 90-mm Petri dishes that were lined with black filter
paper. The Petri dishes containing pupae were transferred to
mesh cage (32.5 × 32.5 × 32.5 cm, BugDorm 43030F, Meg-
aview Science, Taichung, Taiwan). A black acrylic ‘flight
tube’ (10-cm tall and 8.3 cm inner diameter with 3-mm-thick
walls) was placed over the Petri dish. The inner surface of
the flight tube was coated with a thin layer of unscented
talcum powder to avoid the flies escaping from the flight
tubes by walking. A second ‘fly-back tube’, also coated
with talcum powder, was positioned ~ 5 cm from the flight
tube. The fly-back tube enables estimation of the number of
flies that escaped from the flight tube and later returned and
died inside. Each mesh cage with prepared tubes and pupae
was positioned beneath 20-W fluorescent tubes, generating
∼1250 lx at the top of the flight tubes and ∼900 lx at the
base. No food and water were provided to the flies during
flight ability assays. Flies that escaped from the flight tubes
were collected from the mesh cages every second day. All
remaining flies and pupae were collected seven days after
the first flies eclosed.
The flies were categorized as (1) fliers (the number of flies
were collected from the mesh cage plus fly-back), (2) fully
emerged (adults that entirely emerged from the puparium),
(3) not emerged (flies within an unopened puparium), (4)
partially emerged (adult body partially stuck in puparium),
(5) fly-back (the number of flies inside the fly-back tube plus
the same number of morphologically normal flies inside the
flight tube), (6) non-fliers (morphologically normal flies that
were collected from the flight tube minus the number of flies
inside the fly-back tube), and (7) deformed (fully emerged
flies with morphological abnormalities, such as twisted or
deformed wings). The following parameters were calculated
to assess the quality of flies released in the field:
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(1) Emergence rate: percentage of fully emerged adults
calculated, as (N pupae – [N not emerged + N partially
emerged]/N pupae) × 100.
(2) Percentage of fliers: percentage of fully emerged flies
that can fly, calculated as (N pupae – [N not emerged + N
partially emerged + N deformed + N non-f liers]/N
pupae) × 100.
Release site and trapping network
The field study was performed at the experimental field sta-
tion (~ 200 ha) of New South Wales (NSW) Department
of Primary Industries at Somersby (~ 70 km North of Syd-
ney, 33°22′10.7"S 151°18′12.4"E), NSW, Australia. This
site contained diverse habitats and landscape features (e.g.
orchards, abandoned orchards, open fields, bush, ponds). We
established a coaxial network of traps around the centre of
the field, containing 12 cuelure-baited traps (BioTrap V2 X,
containing 2 g of cuelure, an attractant specific to mature
males, plus Malathion as a toxicant) (BioTrap Australia Pty
Ltd). The effective sampling area for cuelure traps has been
estimated at 1.5 hectares (Kean 2015), so we positioned
traps to avoid overlapping effective sampling area. We esti-
mated that a maximum distance of 370 m was adequate to
provide information for the movement of the flies in the
landscape, as per Meats and Edgerton (2008). We used the
circular arrangement to recapture flies from all cardinal
points around the centre of release in order to gain insights
about the distance and orientation of dispersal.
Release and recapture
Twenty fine mesh cages (47.5 × 47.5 × 47.5 cm, Megaview
BugDorm-4F4545) were prepared to rear the adult Q-flies
for release. Ten cages were assigned to the flies held for
two days and ten cages to the flies held for five days. Out
of the ten cages for each pre-release holding period, five
cages were YH-fed and five were YH-deprived. All cages
were kept in a controlled environment room at Mac-
quarie University (25 ± 0.5 °C, 65 ± 5% RH and 11:1:11:1
light:dusk:dark:dawn photoperiod).
The effect of holding period and pre-release diet on abun-
dance and dispersal of Q-fly was assessed by five replicate
release-recapture trials between October 2017 and January
2018. Over the five releases, ~ 24,000 flies per treatment
were released from their maintenance cages at a central
release point. Recapture of flies was performed three, seven,
ten, and 14 days after each release. Trapped flies were col-
lected using 1-mL plastic vials labelled with the date of col-
lection and trap number. The recaptured flies were stored at
− 20 °C until examining the colour of dye in the ptilinum
under a dissecting microscope (Leica S6E, Germany) using
an ultraviolet (UV) LED light source in a dark room. The
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Journal of Pest Science (2022) 95:291–301
flies were classified as either wild, with absence of any of
the dye colours, or sterile with one of the colours used for
dyeing based on the treatment groups.
Effect of landscape on dispersal
Since the experimental location contained different habitats
such as forest, grassland, kiwi, and citrus orchards, effect
of those landscape features on the dispersal of Q-flies was
estimated based on the number of flies recaptured by the
traps at different habitats.
Statistical analysis
Data for flight ability quality control assays were subjected
to analysis of variance (ANOVA) using GLM procedure
in SAS. Effect of fluorescent dyes on emergence and flight
ability of the flies over the five releases was considered as
a fixed effect. Sex ratio of the flies over the releases was
analysed by running PROC FREQ to perform Chi-square
test. For other analyses, data that did not meet the normal-
ity assumption (from D’Agostino and Pearson test) or equal
variance assumption (from Bartlett’s test), were square root
transformed prior to analyses (Zar 1996). Total seasonal
recapture of the YH-deprived and YH-fed Q-flies held either
for 2 or 5 d was analysed by one-way ANOVA using GLM
procedure in SAS. For each treatment, the mean number
of sterile flies recaptured in the 12 cuelure traps for every
release was considered to analyse their field prevalence by
repeated-measures ANOVA using GLM procedure in SAS
with the assumption that the errors have a Gaussian distribu-
tion over the five releases which were considered as fixed
effects. Tukey’s HSD test was used as a post hoc analysis to
separate means (SAS Institute Inc., 2000).
We performed linear regression in Microsoft Excel
using trap distance as the continuous predictor variable and
trap catch as the response variable to analyse the relation-
ship between total number of flies recaptured over the five
releases and distance from the release point. The regres-
sion slopes (between treatments) were then compared (Zar
2010). Means ± SE are presented. The figures for fly dis-
tribution among all treatments were generated in Origin V
2019 (9.60) software.
Results
Flight ability
We did not find any differences between the fluorescent dyes
in either emergence rate (F3,8 = 1.72, P = 0.239) or percent-
age of fliers (F3,8 = 1.05, P = 0.423). The overall emergence
rate over the release period was 83.52 ± 0.04 (SE) %, and
Journal of Pest Science (2022) 95:291–301
percentage of fliers was 70.20 ± 0.06 (SE) %. There were
significant differences among the releases in emergence rate
(F4,32 = 15.59, P < 0.0001), but not in percentage of fliers
(F4,32 = 0.19, P = 0.1661).
Field abundance
There was no difference in the sex ratio of the flies by release
(χ2 = 2.49, df = 4, P < 0.649), and overall, the sex ratio was
49.98 male: 50.02 female. Over the release period, total
recaptures were higher for flies that were YH-fed and held
for 5d (10.20%) than for flies that were YH-deprived and
held for 5d (2.46%), YH-fed and held for 2 d (2.16%), and
YH-deprived and held for 2 d (1.25%). Repeated-measures
ANOVA revealed a significant interaction between release
and pre-release diet in number of flies recaptured (between-
subject effect: Diet F3,44 = 9.10, P < 0.0001, within-sub-
ject effect: Release F4,176 = 20.75, P < 0.0001, Interaction
F12,176 = 2.19, P = 0.0231). For each release, the number of
Fig. 1  Number of male Q-flies recaptured in the 12 cuelure traps
(mean ± SE) over the five releases separated as YH-fed and YH-
deprived flies that had been held for 2 or 5 d prior to release. Col-
umns with the same letter are not significantly different (Tukey test
α = 0.05)
Fig. 2  The relationship between
total number of flies recaptured
over the five releases and flight
distance for each of the pre-
release treatment groups. For
each treatment group a linear
regression was plotted with trap
distance as the continuous pre-
dictor variable and number of
flies recaptured as the response
variable
295
recaptured YH-fed flies held for 5 d was higher than for
all other groups, although the extent of this difference var-
ied across the releases. For example, the YH-fed flies held
for 5 d were recaptured approximately 16, 8, 20, 6, and 4
times more than the YH-deprived flies held for 2d in the five
releases, respectively. On average, the number of recaptured
YH-fed flies held for 5 d was ~ 8.0-fold more than for YH-
deprived flies held for 2 d (Fig. 1).
Flight distance
The relationship between recapture of male Q-flies and
flight distance was significant for all treatments (5 d
YH-deprived: F 1,10 = 24.73, R 2 = 0.71, P = 0.0006; 5 d
YH-fed: F 1,10 = 29.86, R 2 = 0.75, P = 0.0003; 2 d YH-
deprived: F1,10 = 16.85, R2 = 0.63, P = 0.0021; 2 d YH-
fed: F1,10 = 22.66, R2 = 0.69, P = 0.0007). For all treatment
groups, recapture rates decreased as the trap distance from
the release point increased. However, there was a significant
difference in the slope coefficient between the treatments
(t = 2.2064, df = 21, P = 0.0386). The differences in slope
coefficients arise from a much steeper decline in the recap-
ture rate of YH-fed flies held for 5 days with distance from
the release point, which is driven mainly by a far greater
recapture rate of this treatment group in traps close to the
release point (Fig. 2).
The temporal and spatial distribution of sterile
Q‑flies
The spatial distribution and fly catch over time in the experi-
mental field are represented in heatmaps (Fig. 3). Most of the
flies that were held for 5 d (YH-fed and YH-derived) were
recaptured in the same area, just north of the release point
(Fig. 3A, B). The same trend was found for the flies that had
been held for 2 d (YH-fed and YH-derived) (Fig. 3C, D).
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296 Journal of Pest Science (2022) 95:291–301

Fig. 3  The mean proportion of

YH-fed Q-flies held for 5 days
(A), YH-deprived Q-flies held
for 5 days (B), YH-fed Q-flies
held for 2 days (C), and YH-
deprived Q-flies held for 2 days
(D) that were recaptured 0–3,
4–7, 8–10, and 11–14 days
after release (colouring based
on proportion catch over all
traps). The smoother the colours
through space the more equal
the proportions. The warmer the
colour the greater the propor-
tion of flies caught in that time
period. Black x indicates cue-
lure trap position; the red cross
indicates the central release
point. The numbers on Eastings
(X-axis) and Northings (Y-axis)
are spatial measurements in
metres instead of degrees as in
latitude and longitude measure-
ments

Effects of landscape on dispersal the release point, and habitat (Fig. 4). The charts illustrate
that recapture rates were highest in traps close to the release
To display the number of recaptured Q-flies a spider chart point, with YH-fed flies held for 5-d dominating abundance.
was plotted according to treatment group, trap distance from Regardless of the feeding regime and holding period, the

13
Journal of Pest Science (2022) 95:291–301 297

Fig. 3  (continued)

released Q-flies appeared to aggregate in the citrus orchard Discussion

more than in other habitats.
The present study demonstrates a substantial positive
effect of longer pre-release holding period and the provi-
sion of YH on the abundance of sexually mature sterile
male Q-flies in the field. The recapture rate of the YH-
fed Q-flies held for 5 d was eight times more than that of

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Fig. 4  Spider chart displaying spatial distribution of YH-fed and YH-
deprived sterile male Q-flies held for either 2 or 5 d before release
depicted by trap distance and habitat
the YH-deprived flies held for 2 d and at least four times
greater than other treatments. Several previous studies
have demonstrated the effectiveness of YH in sustaining
sexual maturation in Q-flies (Meats et al. 2004; Prabhu
et al. 2008; Pérez-Staples et al. 2009, 2011; Weldon and
Taylor 2011; Collins et al. 2014). Recapture rates of
released sterile Q-flies tend to be variable and low (Meats
et al. 2003, 2006), with this likely reflecting poor survival
as well as poor rates of sexual maturation due to failure
of the flies to secure essential nutrients in the field (Reyn-
olds et al. 2012). We report here an overall recapture rate
of more than 10% for flies that were held for five days
and provided YH. The dominance of YH-fed Q-flies held
for 5 d in the recapture data likely reflects earlier sexual
maturity of these flies and increased rates of survival until
sexual maturity and beyond. Sterile Q-flies commonly start
mating at about six days after eclosion, peaking about ten
days after eclosion (Pérez-Staples et al. 2007).
Once adult flies are released into nature, they require
nutrients including carbohydrates, amino acids, vitamins,
and minerals for sustenance (Fletcher 1987; Drew and Yuval
2000; for a review see Taylor et al. 2013b). Bacteria and
fungi are the key components of adult fruit fly diet (Hen-
drichs et al. 1993), including Q-fly (Drew et al. 1983; Drew
and Lloyd 1987; Majumder et al. 2019, 2020). Like some
other fruit flies (Hendrichs et al. 1993; Manrakhan and Lux
2006), both wild and domesticated Q-flies feed on bat guano,
bird faeces, and pollen in the laboratory, but these dietary
components have not been found to be sufficient to sustain
sexual maturation of Q-fly (Weldon and Taylor 2011).
Flies held for 2–3 days before release are still far from
mature and are susceptible to predators and adverse weather
conditions through the maturation phase. As a consequence,
few sterile males released at such young age may attain sex-
ual maturity, resulting in a lower than required overflooding
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Journal of Pest Science (2022) 95:291–301
ratio of sterile to wild flies. This can greatly reduce preva-
lence of mating between sterile males and wild females and
hence reduces the efficacy of SIT. One common response
to low abundance of sterile males in the field is to increase
release rates to maintain the minimum required overflood-
ing ratio. However, by simply extending the pre-release
holding period by several days and providing the flies with
YH, substantial improvements may be made in the effective
overflooding ratio for a given number of released flies. By
extending the pre-release holding period and providing YH
the number of mature sterile flies can be at least quadrupled
without any increase in production or release costs, and this
increased performance of SIT will easily mitigate any addi-
tional expenses incurred in the rear-out centres.
Field studies of Q-fly have reported recapture rates to
be affected by a wide diversity of factors (Dominiak et al.
2003a, b, 2011; Weldon and Meats 2007; Worsley et al.
2008; Reynolds and Orchard 2015). The maximum trap
distance in this study was 370 m from the release point,
and Q-flies were found to disperse up to that distance. In
the field, we also observed that the sterile flies were recap-
tured in higher numbers in traps that were associated with
host plants than in the traps that were in non-host habitats.
It is likely that released Q-flies that entered host habitats,
by attraction or by chance, tended to remain in those habi-
tats because of the availability of shelter, food, and other
resources. Increased and long-distance travel of Q-fly has
been linked to favourable habitats; flies tend to accumulate in
resource-rich habitat and avoid or move away from resource
poor habitats such as areas with limited food, water, and
shelter (Fletcher 1973, 1974; Dalby-Ball and Meats 2000;
Edge et al. 2001; Meats and Edgerton 2008; see review from
Dominiak 2012). Meats (1996) suggested 400 m as an effec-
tive spacing interval for static release points based on the
known dispersal characteristics of Q-fly and predicted that
a release programme at one-km intervals would be ineffec-
tive owing to incomplete coverage. In accord with previous
studies of Meats (1998), we found a decrease in the propor-
tion of Q-flies recaptured with increased trap distance from
point of release (see Gavriel et al. (2012) for similar findings
in medfly).
While we conducted static releases from a single point,
this is not the case in many operational releases in which the
flies are released from an aircraft (‘aerial release’) or from
a moving vehicle (‘roving ground release’) (Dominiak et al.
2000; Reynolds and Orchard 2015). By increasing the ini-
tial dispersion of flies through the release zone, both aerial
releases and roving ground releases are expected to be much
less vulnerable to factors affecting dispersal of released ster-
ile flies than is the case for static ground releases. While
there was a tendency for YH-fed flies that were held for
5 days to be recaptured closer to the release point than was
the case for other treatments, the vastly greater numbers of
Journal of Pest Science (2022) 95:291–301
flies from this treatment group recaptured mean that sexually
mature males from this treatment were far more abundant
than for other treatments across the trapping grid. While
the tendency of YH-fed flies that were held for 5 days to
be trapped relatively more often close to the release point
may indicate reduced dispersal tendency, this finding instead
likely reflects earlier maturation of this group such that they
are more often captured at early stages of dispersal when still
close to the release point.
Conclusion
The results of the present study indicate that abundance
of sexually mature sterile Q-fly males can be significantly
increased in SIT programmes by extending the pre-release
holding period from the historical standard of two to three
days to five days and by providing the flies a pre-release
diet that includes YH. The increased abundance of cuelure-
responsive male Q-flies indicates increases in the number
that survive until sexual maturity, and hence increases in the
numbers available in the field to compete with wild males
for mating with wild females. While high rates of matura-
tion are important, the sexually mature males still need to be
competitive with wild males for mating with wild females.
Further studies are now needed to investigate the effect of an
extended pre-release holding period combined with dietary
YH on Q-fly mating compatibility with wild females and
competitiveness with wild males.
Author contributions
Md Jamil Hossain Biswas (MJHB) conceived and designed
the work, conducted the experiments, analysed the data, and
wrote the manuscript. Bishwo Mainali (BM) and Polychro-
nis Rempoulakis (PR) designed the work, conducted the
experiments, analysed the data. Jess R. Inskeep (JRI), Sushil
Gaire (SG), and Dominic Cross (DC) conducted the experi-
ments. Lloyd D. Stringer (LDS) analysed the data. Phillip
W. Taylor (PWT) completed data curation. All authors con-
tributed to the revising of the manuscript.
Acknowledgements Project Raising Q-fly Sterile Insect Technique
to World Standard (HG14033) is funded by the Hort Frontiers Fruit
Fly Fund, part of the Hort Frontiers strategic partnership initiative
developed by Hort Innovation, with co-investment from Macquarie
University (NSW, Australia) and contributions from the Australian
Government. MJHB is funded by the Macquarie University Research
Excellence (MQRTP) scholarship from Macquarie University. Addi-
tional support came from the Australian Research Council Industrial
Transformation Training Centre (ITTC) for Fruit Fly Biosecurity Inno-
vation (Project IC50100026), funded by the Australian Government.
299
Declarations
Competing interest There was no conflict of interest regarding the
preparation and submission of this manuscript.
Ethical approval All applicable international, national, and/or institu-
tional guidelines for the care and use of animals were followed.
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