Release and post-release monitoring of the biocontrol agent “Trichilogaster acaciaelongifoliae” for the control of the invasive “Acacia longifolia” in Portugal

Responsáveis pela equipa:
Elizabete Marchante, emarchante@uc.pt
Hélia Marchante, hmarchante@esac.pt
31-12-2020
Centro de Ecologia Funcional da Universidade de Coimbra
Escola Superior Agrária do Instituto Politécnico de Coimbra
Release and post-release monitoring of the biocontrol
agent “Trichilogaster acaciaelongifoliae” for the
control of the invasive “Acacia longifolia” in Portugal
FIFTH REPORT

Release and post-release monitoring of the biocontrol agent “Trichilogaster
acaciaelongifoliae” for the control of the invasive “Acacia longifolia” in Portugal
2020
1
Index
1. Preamble ............................................................................................................................................. 2
1. Release campaign in 2020................................................................................................................... 4
2. Post-release monitoring of the galls ................................................................................................... 7
3. Conclusions........................................................................................................................................ 14
4. References......................................................................................................................................... 15
Annex 1. FIRST REPORT – 2016.................................................................................................................. 17
Annex 2. SECOND REPORT – 2017............................................................................................................. 26
Annex 3. THIRD REPORT – 2018 ................................................................................................................ 35
Annex 4. FOURTH REPORT – 2019............................................................................................................. 50
Annex 5. FULL NAMES OF THE SITES REFERRED IN THE TEXT, FIGURES AND TABLES (2pages)................ 64
Annex 6. LIST OF NON-TARGET PLANTS MONITORED IN 2020 FOR GALLS OF TRICHILOGASTER
ACACIAELONGIFOLIAE. .............................................................................................................................. 66

2020
2
1. Preamble
This is the fifth annual report (previous reports available in Annex 1 to 4) about the release and post-release
monitoring of the biological control (hereafter biocontrol) agent Trichilogaster acaciaelongifoliae for the
control of the invasive plant Acacia longifolia (Figure 1) in Portugal.
Figure 1. Acacia longifolia, from left: plant, inflorescence, seeds in pods, and galls promoted by Trichilogaster acaciaelongifoliae.
Acacia longifolia is an Australian species invasive in Portugal (Presidência do Conselho de Ministros, 2019)
and one of the most widespread invasive plants in coastal areas of the mainland. It causes significant
adverse ecological impacts (López-Núñez et al., 2017; Marchante et al., 2008, 2015; Figure 2, bottom
images), which may persist over time and make restoration of invaded areas increasingly more difficult and
complex (Le Maitre et al., 2011; Marchante et al., 2009, 2011a). In addition, it reduces forest productivity,
mainly in littoral pine plantations, and increases fire hazard (Le Maitre et al., 2011), with consequent
negative socio-economic impacts. Control methods in use until recently (i.e., mechanical and/ or chemical
methods) are very expensive and often unsuccessful, due to the extensive, persistent and long-lived seed-
bank accumulated in the soil (Marchante et al., 2010), that promotes the quick reinvasion of cleared areas.
Figure 2. Coastal areas with native vegetation (top images) and areas densely invaded by Acacia longifolia (bottom images),
where the invasive species replaced the diverse native communities, composed by many shrubs and herbaceous species, by
almost monospecific woody stands.

2020
3
After more than 12 years of host speciﬁcity testing, risk assessment and permits (Jeger et al., 2016;
Marchante et al., 2011b; Shaw et al., 2016), in November 2015, the biocontrol agent Trichilogaster
acaciaelongifoliae (Australian bud-galling wasp) started to be released in pre-selected sites along the
Portuguese coast (Marchante et al., 2017, Annex 1). This biocontrol agent is a highly host-specific organism,
affecting almost exclusively A. longifolia and targets mainly seed reduction; it is a small Australian gall-
former (Hymenoptera: Pteromalidae, 3 mm), with an annual life cycle (in average), spending most of the
year inside the galls (Figure 3); emerging females search for flower (preferably, but also vegetative) buds,
oviposit the eggs, and die after 2 – 3 days (in average). As a result, galls are produced instead of flowers (or
new branches) and as such seed production is prevented (or vegetative growth reduced). In the short-term it
reduces the production and dispersal of seeds and in the longer-term a decrease of germination is expected
after control, fire or other disturbances, since the soil seed-bank receives less and less seeds each year; less
vegetative growth and physiological stress for the plants may also be expected, as they cannot cope with
heavy gall loads (Dennill, 1985).
This agent has been successfully used to control A. longifolia in South Africa for more than 30 years, where it
decreased seed production and vegetative growth (Dennill and Donnelly, 1991; Wilson et al., 2011). In
Portugal, in Spring-Summer 2016, 661
galls were detected in five2
of the release sites (Annex 1. FIRST REPORT
– 2016; Marchante et al., 2017), in 2017 around 1100 galls were detected in the same five sites (Annex 2.
SECOND REPORT – 2017)3
, during 2018 the monitoring effort detected almost 25000 galls in five sites (but
while four sites were the same as in previous years, one was new, since one of the sites with establishment
in 2016 burned in October 2017 and the population was lost, Annex 3. THIRD REPORT – 2018), and in 2019
establishment of T. acaciaelongifoliae was already observed in 334
sites along most of the Portuguese Coast,
from Riba de Âncora, in the north, to Faro, in the south (Annex 4. FOURTH REPORT – 2019).
Figure 3. The biocontrol agent Trichilogaster acaciaelongifoliae. From left: adult female; female ready to emerge from the gall and
pupae; and galls promoted by T. acaciaelongifoliae with emergence holes.
1
In the first report, 56 galls were mistakenly reported due to an error in the summing of galls (all galls were correctly
identified in each site of release - Table 1, Annex 1 - but the sum formula had an error).
2
In the first report (Annex 1), galls were reported in four sites, but later in the season, after report submission, one gall
was detected in Reserva Natural das Dunas de São Jacinto, raising the number of sites with establishment to five.
3
In the second report (Annex 2), galls were reported in five sites, but later on a few galls were detected in Coimbra, in a
new site (Patos), raising the number of sites with T. acaciaelongifoliae to six. However, one site burned in October 2017
(Perímetro Florestal das Dunas de Cantanhede) and as such the total number of sites with establishment was five.
4
In the 2019 report, 32 sites were reported with establishment, but a new site was detected in June 2020 most
probably resulting from the 2018 release campaign, raising the number of sites with establishment in 2019 to 33.

2020
4
1. Release campaign in 2020
As in previous campaigns, some release and monitoring was done by a team from Centre for Functional
Ecology, which includes members of Escola Superior Agrária, Instituto Politécnico de Coimbra (ESAC) and
Universidade de Coimbra (UC), but in 2020 galls were also released and monitored by technicians from the
Instituto da Conservação da Natureza e das Florestas (ICNF), municipalities, NGOs and forest companies in a
strategic collaboration to involve other stakeholders. In what concerns ESAC and UC, work occurred in the
context of two ongoing research projects (INVADER-IV - PTDC/AAGREC/4896/2014, funded by the
Portuguese Foundation for Science and Technology, and GANHA - POSEUR-03-2215-FC-000052, funded by
Portugal 2020 through Programa Operacional Sustentabilidade e Eficiência no Uso de Recursos, POSEUR).
Both projects were complete during 2020 and, as such, funding for the next campaigns is not yet secured.
Efforts are being made to ensure these, including the involvement of technicians from different entities
and citizen science, but the monitoring effort may have to be limited and reduced due to lack of funding.
In 2020, T. acaciaelongifoliae was released on 18 sites, five of which had releases from previous years, but
had undetected or limited establishment (Table I). This biocontrol project has been disseminated in scientific
and awareness activities over the years and several site owners / managers, when becoming aware of it,
showed interest in releasing galls on their sites to increase the speed of establishment and therefore galls
were released on such sites in 2020; with the number of sites with establishment in previous years, it was
expected that T. acaciaelongifoliae would eventually colonize all these sites in the coming years, but this way
the rate of establishment was accelerated. Site owners / managers have been instructed to register release
locations so that we can monitor the establishment and spread of the biocontrol agent, but the number of
galls or wasps has not always been reported (Table I, 2020 section). Other releases assisted by humans
without our knowledge may have happened, since the galls are conspicuous and visible and people can
collect and release them in other locations. Table I includes all the release campaigns from 2015 to 2020.
Table I. Number of female wasps* of Trichilogaster acaciaelongifoliae released in the six release campaigns (2015-2020) in
Portugal. In bold, sites with more than one release. *In 2019 & 2020, most releases were made by technicians from ICNF, forestry
companies or municipalities and on some sites galls were left in place instead of emerged wasps, so it was not possible to know
the exact number of wasps released; in 2020, on a few sites, no information was reported on the number of galls left on the site.
CODE SITE
#
COORDINATES PROJECT YEAR WASPS OBJECTIVE
$
ESP Esposende 41.508999,-8.784351 INVADER-B 2015 30 Monitoring
SJD Dunas de São Jacinto 40.672717,-8.740729 INVADER-B 2015 88 Monitoring
TOC Tocha 40.34837,-8.81704 INVADER-B 2015 105 Monitoring
QUD Dunas de Quiaios 40.22476,-8.88622 INVADER-B 2015 80 Monitoring
EST Coimbra (ESAC) 40.216722,-8.450210 INVADER-B 2015 9 Monitoring
COI1 Coimbra (Patos) 40.209237,-8.401237 INVADER-B 2015 39 Monitoring
SBV Serra da Boa Viagem 40.20037,-8.88969 INVADER-B 2015 65 Monitoring
COI Coimbra (Pólo II) 40.18588,-8.41358 INVADER-B 2015 44 Monitoring
SPM São Pedro de Moel 39.75711,-9.02338 INVADER-B 2015 74 Monitoring
TOTAL 2015* 534
ESP Esposende 41.508999,-8.784351 INVADER-IV 2016 139 Monitoring
SPI Espinho 40.923320,-8.658262 INVADER-IV 2016 135 Biocontrol
SJD Dunas de São Jacinto 40.672717,-8.740729 INVADER-IV 2016 74 Monitoring
EIX Eixo 40.620281,-8.568342 INVADER-IV 2016 54 Biocontrol
MIR Mira 40.394569,-8.789050 INVADER-IV 2016 45 Biocontrol
TOC1 Tocha N 40.320083,-8.845127 INVADER-IV 2016 129 Monitoring
LAG Lagoa da Vela 40.269481,-8.799287 INVADER-IV 2016 70 Biocontrol

2020
5
Table I (cont). Number of female wasps of Trichilogaster acaciaelongifoliae released during the six release campaigns in Portugal.
CODE SITE
#
$
QUP Quiaios N 40.241645,-8.854573 INVADER-IV 2016 75 Monitoring
EST Coimbra (ESAC) 40.216722,-8.450210 INVADER-IV 2016 83 Founder pop.
COI1 Coimbra (Patos) 40.209237,-8.401237 INVADER-IV 2016 38 Biocontrol
COV Covões 40.194632,-8.466302 INVADER-IV 2016 37 Biocontrol
ANO Anobra 40.161626,-8.510096 INVADER-IV 2016 21 Biocontrol
HEL Condeixa 40.113693,-8.513775 INVADER-IV 2016 5 Biocontrol
LAV Lavos 40.097376,-8.856496 INVADER-IV 2016 66 Biocontrol
LEI Leirosa 40.076190,-8.865008 INVADER-IV 2016 77 Biocontrol
MUR Mata do Urso 39.983285,-8.914007 INVADER-IV 2016 89 Biocontrol
PED Pedrogão 39.937940,-8.927712 INVADER-IV 2016 45 Biocontrol
VLE Vieira de Leiria 39.865863,-8.971235 INVADER-IV 2016 76 Monitoring
PAT Paredes da Vitoria (Pataias) 39.707963,-9.048827 INVADER-IV 2016 138 Biocontrol
TOTAL 2016** 1396
ESP Esposende 41.508999,-8.784351 INVADER-IV 2017 39 Monitoring
SEI Seixo 40.497200,-8.754436 INVADER-IV 2017 46 Monitoring
EST Coimbra (ESAC) 40.216722,-8.450210 INVADER-IV 2017 32 Founder pop.
COV Covões
##
40.194632,-8.466302 INVADER-IV 2017 69 Biocontrol
ALH Alhadas (Cabecinho) 40.174834,-8.787518 INVADER-IV 2017 6 Biocontrol
FIG2 Figueira da Foz (McDonald´s) 40.166223,-8.853586 INVADER-IV 2017 6 Biocontrol
ANO Anobra 40.161626,-8.510096 INVADER-IV 2017 6 Biocontrol
ANO1 Anobra1
##
40.160576,-8.498345 INVADER-IV 2017 20 Biocontrol
FIG1 Figueira da Foz (Rotunda LIDL) 40.157502,-8.849582 INVADER-IV 2017 22 Biocontrol
VIV Vila Verde 40.154741,-8.795803 INVADER-IV 2017 16 Biocontrol
FIG Figueira da Foz 40.148682,-8.836959 INVADER-IV 2017 48 Biocontrol
MOR Morraceira 40.139481,-8.844876 INVADER-IV 2017 8 Biocontrol
GAL Gala, Parque de Merendas 40.123295,-8.859909 INVADER-IV 2017 10 Biocontrol
HEL Condeixa 40.113693,-8.513775 INVADER-IV 2017 2 Biocontrol
PAT Paredes da Vitoria (Pataias) 39.707963,-9.048827 INVADER-IV 2017 21 Biocontrol
LSA Lagoas de Santo André 37.991658,-8.854934 INVADER-IV 2017 46 Monitoring
FAR Faro 37.028391,-8.005642 INVADER-IV 2017 28 Monitoring
TOTAL 2017*** 425
RBA Riba de Âncora (Baldios de Riba de Âncora) 41.808889,-8.796121 OTHERS 2018 48 Biocontrol
PEN Quinta Pentieiros 41.775557,-8.657753 GANHA 2018 25 Biocontrol
ANH Anha 41.674618,-8.801127 GANHA 2018 9 Biocontrol
ANT Antas 41.61092,-8.80828 GANHA 2018 54 Biocontrol
ESP Esposende 41.508999,-8.784351 GANHA 2018 83 Monitoring
BEZ Barrinha de Esmoriz 40.963981,-8.647378 GANHA 2018 45 Biocontrol
EIX2 Eixo (Quinta de S. Francisco) 40.616127,-8.567075 OTHERS 2018 18 Biocontrol
IP3 Acesso IP3 40.586589,-8.015036 OTHERS 2018 3 Biocontrol
VAG Dunas Vagos (GANHA) 40.535932,-8.74104 GANHA 2018 146 Monitoring
BZM Belazaima do Chão 40.527697,-8.330153 OTHERS 2018 20 Biocontrol
VNR Vila Nova da Rainha 40.47055,-8.09458 OTHERS 2018 3 Biocontrol
POC1 Pocariça (Cantanhede1) 40.382712,-8.574368 OTHERS 2018 39 Biocontrol
POC2 Pocariça (Cantanhede2) 40.37305,-8.579074 OTHERS 2018 21 Biocontrol
TOC2 Tocha (Caniceira) 40.3476,-8.77368 GANHA 2018 79 Biocontrol
TOC4 Charco Berlengas 40.326566,-8.782187 GANHA 2018 4 Biocontrol

2020
6
Table I (cont). Number of female wasps of Trichilogaster acaciaelongifoliae released during the six release campaigns in Portugal.
CODE SITE
#
$
TOC3 Cruzamento Tocha 40.3254768,-8.8131098 GANHA 2018 9 Biocontrol
COV Covões 40.194632,-8.466302 INVADER-IV 2018 2 Biocontrol
FIG2 Figueira da Foz (McDonald´s) 40.166223,-8.853586 INVADER-IV 2018 8 Biocontrol
FIG1 Figueira da Foz (Rotunda LIDL) 40.157502,-8.849582 INVADER-IV 2018 16 Biocontrol
FIG Figueira da Foz 40.148682,-8.836959 INVADER-IV 2018 12 Biocontrol
SOU Soure 40.099786,-8.619687 OTHERS 2018 45 Biocontrol
PAT Paredes da Vitoria (Pataias) 39.707963,-9.048827 GANHA 2018 77 Biocontrol
STC Praia do navio, Santa Cruz 39.144362,-9.371916 GANHA 2018 13 Biocontrol
CAR Carapinheira, Mafra 38.935492,-9.276104 OTHERS 2018 34 Biocontrol
CAP Praias da Costa da Caparica 38.599935,-9.20743 GANHA 2018 48 Biocontrol
ARR Estrada Setúbal - praias 38.496597,-8.930805 GANHA 2018 12 Biocontrol
SAD Estrada Tróia - Comporta 38.426573,-8.824844 GANHA 2018 46 Biocontrol
BRJ Brejinhos 38.031267,-8.808383 GANHA 2018 43 Biocontrol
PQS Pesqueiro Sancha 38.02586,-8.82275 GANHA 2018 52 Biocontrol
SIN2 Monte Feio - Sines 2 37.996261, -8.842971 GANHA 2018 20 Biocontrol
LSA Lagoas de Santo André 37.991658,-8.854934 GANHA 2018 103 Monitoring
SIN1 Monte Feio - Sines 1 37.981616, -8.845499 GANHA 2018 16 Biocontrol
FAR Faro 37.028391,-8.005642 GANHA 2018 109 Monitoring
TOTAL 2018**** 1262
ALHQ Alhadas (areeiro) 40.2021111, -8.7884639 OTHERS 2019 50 Biocontrol
VNM Vila Nova de Mil Fontes(Praia das Furnas) 37.714934, -8.7845 GANHA 2019 24 Biocontrol
TOTAL 2019***** 74
RBA Riba de Âncora (Baldios de Riba de Âncora) 41.808889,-8.796121 OTHERS 2020 548 Biocontrol
AFI Afife 41.789493,-8.869711 OTHERS 2020 310 Biocontrol
CARR Carreço 41.754159,-8.87547 OTHERS 2020 300 Biocontrol
VIC Viana do Castelo 41.702256,-8.837369 OTHERS 2020 225 Biocontrol
ESP Esposende 41.508999,-8.784351 OTHERS 2020 505 Biocontrol
ESP 2 Esposende 2 41.499313,-8.775334 OTHERS 2020 540 Biocontrol
BZM Belazaima do Chão 40.527697,-8.330153 OTHERS 2020 ? Biocontrol
VIG Vigía 40.516265,-8.725941 OTHERS 2020 200 Biocontrol
ARE Praia do Areão 40.491751,-8.788856 OTHERS 2020 35 Biocontrol
CAL Calvão 40.470903,-8.685942 OTHERS 2020 ? Biocontrol
LOB Covão do Lobo 40.447997,-8.646407 OTHERS 2020 35 Biocontrol
COV Covões 40.194632,-8.466302 OTHERS 2020 150 Biocontrol
PAT 2 Pataias 2 39.662347,-9.002107 OTHERS 2020 375 Biocontrol
FAL Falca 39.640147,-9.069502 OTHERS 2020 ? Biocontrol
MMA Mata da Machada 38.619271,-9.038819 OTHERS 2020 ? Biocontrol
EVR Évora 38.533685,-8.031304 OTHERS 2020 7 Biocontrol
BRE Brescos 38.105037,-8.77737 OTHERS 2020 468 Biocontrol
BRJ Brejinhos 38.031267,-8.808383 OTHERS 2020 1655 Biocontrol
TOTAL 2020****** 5353
#
for simplification, site names were shortened, for full name check Annex 5;
##
many trees were cut after release; $ "Monitoring"-
sites monitored for establishment and effects of the BCA; "Biocontrol"- sites monitored only for establishment of the BCA; "Founder
pop."- greenhouse where a “founder population” was initially maintained; *releases from 20/11 to 07/12/2015, with South African
galls; **releases from 12/11 to 13/12/2016, with South African galls; ***releases from 06/12/2017 to 08/01/2018, with South Afri-
can galls; ****releases from 14/06 to 20/07/2018, with Portuguese galls; ***** release at 28/06 and 15/08/2019, with Portuguese
galls; galls were left in field instead of wasps, we assumed 1 wasp emerged/ gall; ****** In 2020, number of wasps are underesti-
mated, because galls were released instead of emerged wasps; sites with "?" - no information about the number of wasps released.

2020
7
2. Post-release monitoring of the galls
The majority of release sites were monitored in 2020 (Figure 4, Table II). As in previous years, because the
number of sites to be monitored is currently too high to screen them all in detail, some sites were selected
to more detailed monitoring (sites signalled as “Monitoring” in Table I), while others were monitored with a
less demanding protocol whose only aim was to detect establishment of T. acaciaelongifoliae, recording
presence or absence of galls in the plants where galls were released or nearby (sites signalled as “Biocontrol”
in Table I). Depending on the size of the galls, their detectability, and proximity to Coimbra, for some sites
more than one monitoring was performed; when this happened, the maximum number of galls recorded
was used for Table II (also for other years); additionally, in 2020, in some sites, more than one distinct
generation of galls was observed after the “normal” season, which is most frequently in the end of
Spring/early Summer (May – June; the others were one in August/September - Summer - and another in
November - Autumn) and in such cases the sum of galls of the different generations is shown in Table II.
Because release sites are now widespread along the Portuguese coast and funding is limited, several sites
were monitored with the help of technicians from ICNF, municipalities, NGOs and forest companies in a
highly positive collaborative effort.
Figure 4. Field sites where the biocontrol agent Trichilogaster acaciaelongifoliae was released and monitored along the
Portuguese coast during the first (2015, black dots in the legend), second (2016, dark red dots), third (2017, green dots), fourth
(2018, blue dots), fifth (2019, brown dots), and sixth (2020, orange dots) release campaigns. Sites where galls were detected
(establishment of the biocontrol agent confirmed) are marked with symbols in full, and sites with no establishment confirmed are
open; for 2020 releases information about establishment it is not yet possible for most of the sites. Source: López-Núñez et al.,
n.d. and updated for 2020.

2020
8
In 2020, there is establishment of T. acaciaelongifoliae confirmed along most of the Portuguese Coast and
some more inland sites, from north to south of the country, in a total of 40 sites (Figure 4, Table II), many of
them being the first or second year of establishment: more than 67000 galls were counted, but this is clearly
an underestimation as an intensive search was not possible in most places (check notes in Table II). Figure 4
and Table II summarize the information not only of the sixth release and fifth monitoring campaigns (in
2020), but include also the previous campaigns in order to give a complete picture or this biocontrol project.
Like in 2019, it is noteworthy that while in the previous years the number of first generation galls/site was
always low (at most in the order of tens), in 2020 in several sites this number was much higher, sometimes
several hundred, e.g., Pataias 2 and Falca (Table II). This difference is most probably because the wasps
released until 2017 (establishment until 2018) were of South African origin (and so the life cycle was not
synchronized with the season and phenology of A. longifolia in Portugal), while since 2018 the releases were
done with wasps originating from Portuguese populations, and hence more synchronized with the phenolo-
gy of the host-plant (A. longifolia) and also with more adequate meteorological conditions. In addition, when
wasp spread naturally by their own means or when galls are released instead of wasps, wasps have a longer
lifespan as they emerge in the field instead of emerging in the laboratory and being transported a few hours
or days later - since they are short-lived (on average 2-3 days as adults), this can make some difference.
Despite T. acaciaelongifoliae has apparently synchronized its life cycle with the Northern hemisphere, in
2020 (as before) in a few places mature galls were detected (including insects emerging) outside the
expected main season, i.e., not synchronized with the majority of the other galls in the end of Spring/early
Summer (May – June; which seems to be the “normal” season) and so it was not possible to be certain of the
number of generations per year; nevertheless, galls resulting from the 2015 releases were possibly the fifth
generation. In fact, in 2020 these galls were in higher numbers with apparently a second (even third)
generation observed in August/ September, and in November in some sites, e.g., in Pólo II from University of
Coimbra, Paredes da Vitoria (Pataias), Mata Nacional das Dunas de Quiaios (Dunas Quiaios), Mata Nacional
de Leiria (São Pedro de Moel) and Covões (and less so in Reserva Natural das Dunas de S. Jacinto and Vagos).
Nevertheless, as the population growth is expected to be exponential, these numbers of galls observed
outside the May - June season may be an “illusion” and be in fact “low” numbers when compared with the
next season; we expect to confirm this in 2021 or in the following years.
Table II. Number of galls of Trichilogaster acaciaelongifoliae detected during post-release monitoring campaigns (2016 - 2020),
including the one in 2020, to which this reports refers specifically. Sites are ordered first from the year of establishment and,
secondly, from north to south. Except for 2020, only sites and years where galls were detected are shown. For 2019 and 2020, for
sites that have establishment for more than three years and in sites where releases were done with galls of Portuguese
populations the numbers shown in Table II should be interpreted as merely indicative and are certainly underestimated because
either the T. acaciaelongifoliae spread further away from the release trees and galls were not detected or the numbers were too
high, making it impossible to have an accurate count.
CODE SITE
MONITORING
YEAR
NUMBER of
DETECTED
GALLS
(max
$
)
COORDINATES
SJD Reserva Natural das Dunas de S. Jacinto 2016 1 40.672717,-8.740729
TOC Perímetro Florestal das Dunas de Cantanhede (Tocha) 2016 38 40.34837,-8.81704
TOC Perímetro Florestal das Dunas de Cantanhede (Tocha) 2017 29 40.34837,-8.81704
TOC Perímetro Florestal das Dunas de Cantanhede (Tocha)##
2018 0 40.34837,-8.81704

2020
9
Table II (cont.). Number of galls of Trichilogaster acaciaelongifoliae detected during all post-release monitoring campaigns.
CODE SITE
MONITORING
YEAR
NUMBER of
DETECTED
GALLS
(max
$
)
COORDINATES
QUD Mata Nacional das Dunas de Quiaios (Dunas Quiaios) 2016* 9 40.22476,-8.88622
QUD Mata Nacional das Dunas de Quiaios (Dunas Quiaios) 2017 73 40.22476,-8.88622
COI1 Coimbra (Patos) 2016 0 40.209237,-8.401237
COI1 Coimbra (Patos) 2017*** 21 40.209237,-8.401237
COI Coimbra (Pólo II from University of Coimbra) 2016 9 40.18588,-8.41358
COI Coimbra (Pólo II from University of Coimbra) 2017** 413 40.18588,-8.41358
SPM Mata Nacional de Leiria (São Pedro de Moel) 2016 9 39.75711,-9.02338
SPM Mata Nacional de Leiria (São Pedro de Moel) 2018**** 16415 39.75711,-9.02338
RBA Riba de Âncora (Baldios de Riba de Âncora) 2019 153 41.808889,-8.796121
RBA Riba de Âncora (Baldios de Riba de Âncora) 2020 tbc 41.808889,-8.796121
PEN Quinta Pentieiros 2019 26 41.775557,-8.657753
PEN Quinta Pentieiros 2020 tbc 41.775557,-8.657753
ANT Antas 2019 51 41.61092, -8.80828
ANT Antas 2020 tbc 41.61092, -8.80828
ESP Esposende 2019 2546 41.508999,-8.784351
ESP Esposende 2020 1165 41.508999,-8.784351
BEZ Barrinha de Esmoriz (BEZ) 2019 77 40.963981,-8.647378
BEZ Barrinha de Esmoriz (BEZ) 2020 2100 40.963981,-8.647378
EIX2 Eixo (Quinta de S. Francisco) 2019 107 40.616127,-8.567075
EIX2 Eixo (Quinta de S. Francisco) 2020 tbc 40.616127,-8.567075
VAG Dunas Vagos (GANHA) 2019 733 40.535932,-8.74104
VAG Dunas Vagos (GANHA) 2020 ? 40.535932,-8.74104
BZM Belazaima do Chão 2019 3 40.527697,-8.330153
BZM Belazaima do Chão 2020 0 40.527697,-8330153
POC1 Pocariça (Cantanhede1) 2019 37 40.382712,-8.574368
POC1 Pocariça (Cantanhede1) 2020 tbc 40.382712,-8.574368
POC2 Pocariça (Cantanhede2) 2019 39 40.37305,-8.579074
POC2 Pocariça (Cantanhede2) 2020 tbc 40.37305,-8.579074
TOC2 Tocha (Caniceira) 2019 11 40.3476,-8.77368
TOC2 Tocha (Caniceira) 2020 tbc 40.3476,-8.77368
TOC4 Charco Berlengas 2019 3 40.326566,-8.782187
TOC4 CharcoBerlengas 2020 tbc 40.326566,-8.782187
COV Covões 2019 3 40.194632,-8.466302
COV Covões 2020 3015 40.194632,-8.466302
FIG2 Figueira da Foz (McDonald´s) 2019 31 40.166223,-8853586
FIG2 Figueira da Foz (McDonald´s) 2020 tbc 40.166223,-8853586

2020
10
CODE SITE
MONITORING
YEAR
NUMBER of
DETECTED
GALLS
(max
$
)
COORDINATES
FIG1 Figueira da Foz (Rotunda LIDL) 2019 81 40.157502,-8849582
FIG1 Figueira da Foz (Rotunda LIDL) 2020 tbc 40.157502,-8849582
FIG Figueira da Foz 2019 5 40.148682,-8836959
FIG Figueira da Foz 2020 tbc 40.148682,-8836959
SOU Soure (RAIZ - Pai Daniel) 2019 35 40.099786,-8.619687
SOU Soure (RAIZ - Pai Daniel) 2020 ? 40.099786,-8.619687
PAT Paredes da Vitoria (Pataias) 2019 67 39.707963,-9.048827
PAT Paredes da Vitoria (Pataias) 2020 3992 39.707963,-9.048827
STC praia do navio, Santa Cruz 2019 5 39.144362,-9.371916
STC praia do navio, Santa Cruz 2020 tbc 39.144362,-9.371916
CAR Carapinheira, Mafra 2019 25 38.935492,-9.276104
CAR Carapinheira, Mafra 2020 ? 38.935492,-9.276104
CAP Praias da Costa da Caparica 2019 77 38.599935,-9.20743
CAP Praias da Costa da Caparica 2020 tbc 38.599935,-9.20743
SAD estrada Tróia - Comporta (RN Estuario Sado) 2019 23 38.426573,-8824844
SAD estrada Tróia - Comporta (RN Estuario Sado) 2020 ? 38.426573,-8.824844
BRJ Brejinhos 2019 321 38.031267,-8.808383
BRJ Brejinhos 2020 14777 38.031267,-8.808383
PQS Pesqueiro Sancha 2019 11 38.02586,-8.82275
PQS Pesqueiro Sancha 2020 tbc 38.02586,-8.82275
SIN2 Monte Feio - Sines 2 2019 2 37.996261,-8.842971
SIN2 Monte Feio - Sines 2 2020 ? 37.996261,-8.842971
LSA Lagoas de Santo André 2019 818 37.991658,-8.854934
LSA Lagoas de Santo André 2020 20 37.991658,-8.854934
SIN1 Monte Feio - Sines 1 2019 6 37.981616,-8.845499
SIN1 Monte Feio - Sines 1 2020 ? 37.981616,-8.845499
FAR Faro 2019 1117 37.028391,-8.005642
FAR Faro 2020 1597 37.028391,-8.005642
AFI Afife 2020 tbc 41.789493,-8.869711
CAR Carreço 2020 tbc 41.754159,-8.87547
VIC Viana do Castelo 2020 tbc 41.702256,-8.837369
ESP2 Esposende 2 2020 tbc 41.499313,-8.775334
VIG Vigía 2020 tbc 40.516265,-8.725941
ARE Praia do Areão 2020 tbc 40.491751,-8.788856
CAL Calvão 2020 tbc 40.470903,-8.685942
LOB Covão do Lobo 2020 tbc 40.447997,-8.646407
ALHQ Alhadas (areeiro) 2020 8 40.20211,-8.78846
HEL Condeixa 2020 23 40.113693,-8.513775
PAT 2 Pataias 2 2020 1446 39.662347,-9.002107
FAL Falca 2020 683 39.640147,-9.069502
MMA Mata da Machada 2020 tbc 38.619271,-9.038819
EVR Évora 2020 tbc 38.533685,-8.031304
BRE Brescos 2020 205 38.105037,-8.77737
VNM Vila Nova de Mil Fontes (Praia das Furnas) 2020 4 37.714934,-8.7845

2020
11
CODE SITE
MONITORING
YEAR
NUMBER of
DETECTED
GALLS
(max
$
)
COORDINATES
TOTAL 2016 66
TOTAL 2017 1124
TOTAL 2018 24793
TOTAL 2019 11957
TOTAL 2020 67245
$
For some sites, more than one monitoring/year was performed; when this happened, the maximum number of galls
recorded was used for this table;
##
The area in Tocha burned in October 2017 and the population was lost; *In the
end of 2017, 16 dried galls were detected that had not previously been detected; **413 corresponds to the
maximum number of galls detected in 3 monitoring events during 2017 (1st 413; 2nd 304; 3rd 363); ***These 21
galls were detected only after conclusion of the second report; **** 5690 galls were counted and the rest were
estimated from observations; In 2020, sites with "tbc" (to be confirmed) means that are still to be monitored; "?" -
sites where gall formation is confirmed, but where they have not been counted.
Despite the exact counting of galls is now almost impossible, in most sites where establishment occurred in
previous years it was clear an exponential growth on the number of the galls each year. This was expected
since each female wasp can oviposit on average around 300 eggs, and therefore when populations are
established and (more) synchronized with the seasons and phenology of A. longifolia in the Northern
Hemisphere, population growth can increase quite fast. In addition to the increase in the number of sites,
the size of the areas/ range of dispersal is also increasing, with T. acaciaelongifoliae spreading into new
areas, sometimes several hundred meters or even a few kilometres (at least up to 7 km) from the focal trees
where wasps were initially released. Therefore, in 2020 we started a citizen science project, asking citizens to
report galls of T. acaciaelongifoliae in order to increase detectability in places further away from the release
sites. Although this was the first year, a few citizens reported galls in sites where we were not aware of
establishment.
In what concerns the monitoring and counting of galls, we need to highlight a few points: 1) although
establishment of T. acaciaelongifoliae is clearly confirmed and galls increasing in space and number, gall
counting is becoming increasingly more difficult and less accurate because i) the biocontrol agent is
spreading farther and farther from the release trees, sometimes a few kilometres, decreasing the chances of
detectability, ii) the number of galls is too high to allow an absolute or accurate counting, iii) the number of
sites is very high and the workload for detailed monitoring is unaffordable, even more now that we have no
dedicated funding; iv) in some sites and years, several generations of galls are observed and it is difficult to
delimit each one in time; and v) some sites are separated by only a few kilometres and after a certain time
the populations will most likely come together and it will no longer be possible to distinguish different
populations; 2) the counts reflect a limited period of time and there are certainly galls not detected because
they were not visible when monitoring was done, i.e., it must be kept in mind that galls may not yet be
detected and establishment may be confirmed later on; and 3) sites that have establishment for more than
three years and in sites where releases were done with galls of Portuguese populations (releases in 2018 and
after that) the numbers shown in Table II should be interpreted as merely indicative and are certainly
underestimated because either the T. acaciaelongifoliae spread further away from the release trees and was
not detected or the numbers were too high, making it impossible to have an accurate count.

2020
12
In three of the sites selected for “Monitoring” where galls were detected since 2016 and hence the number
of galls is in the order of thousands (not possible to count anymore; Mata Nacional das Dunas de Quiaios,
Reserva Natural das Dunas de São Jacinto and Mata Nacional de Leiria), the effects of T. acaciaelongifoliae
on A. longifolia were evaluated, analysing different parameters at vegetative (number of secondary branches
and phyllodes, and total branch length) and reproductive (number of pods and seeds) level (Figure 5). In
each site, 12 plants with galls were selected and similar number with no galls (the latter ones working as
Controls); in plants with galls, five branches with galls and five without galls were selected; in plants without
galls, five branches were selected, totalizing 60 branches for each type of evaluation. The same branches
were marked and measured in two consecutive years; these measures were done from 2018 to 2020,
comparing branches from 2018 to 2019 and from 2019 to 2020, i.e., two periods of two consecutive years.
Figure 5. (a) Impacts of Trichilogaster acaciaelongifoliae on reproductive (number of seeds and pods) and vegetative (number of
secondary branches and phyllodes, and total branch length) output of Acacia longifolia. The impacts are depicted as mean
percentage ratio between the periods 2018-19 and 2019-20, considering average values of the three sites evaluated: São Jacinto
dunes, Quiaos and São Pedro de Moel. Error bars show the standard error. Letters above bars show results of Tukey post-hoc test.
(b) Net percentage impact (i.e., the percentage ratio between the average of galled and un-galled branches in galled trees versus
un-galled branches in un-galled trees) of T. acaciaelongifoliae on reproductive and vegetative output of A. longifolia. Source:
(López-Núñez et al., n.d.)

2020
13
Although it is important to perform the evaluation of the effects of T. acaciaelongifoliae from the beginning
of the biocontrol establishment, it must be kept in mind that the number of plants with galls and the
quantity of galls in each plant is still small and sparse compared to the numbers of available floral and
vegetative buds of A. longifolia in very extensive invaded areas, and consequently the effects detected are
expected to be still indicative at this stage. Nevertheless, this monitoring showed that T. acaciaelongifoliae is
promoting already clearly detectable effects in the monitored trees (Figure 5): it was possible to observe a
significant reduction in the number of secondary branches (-33%), pods (-84%) and seeds (-95%) in the
branches of trees with galls, independently of having or not galls in the measured branch (Figure 5). This
suggests that the effects of T. acaciaelongifoliae are not limited to branches with galls, but are reflected at
the tree level. This could be expected, since the formation of galls represents an energy expenditure for the
entire plant and not only for the branches where galls develop. These measured effects are noticeable when
looking at plants more broadly, with some A. longifolia almost not producing flowers in 2020.
As in previous years, amongst all the galls observed, some exhibited parts of A. longifolia phyllodes (revea-
ling that they were probably originated from oviposition in vegetative buds, Figure 6, bottom right), but
most galls were more probably originated from flowers buds (Figure 6, bottom left), confirming the better
synchrony between the early stages of flower bud development and emergency of T. acaciaelongifoliae
wasps. A few wasps were observed flying and ovipositing in the field, with most galls revealing one or more
emergency holes (depending on the number of chambers) indicating that insects of T. acaciaelongifoliae
must have emerged.
Figure 6. Galls of Trichilogaster acaciaelongifoliae detected during post-release monitoring campaigns in 2020. Top, branches
heavily colonized. Bottom, from left: flower and vegetative galls.

2020
14
As said above, and like in South Africa, even if there is a peak for emergency, considerable variation was
observed in the phenology of galls and sites suggesting that wasps do not emerge all at the same time and
probably are not taking the same time to complete the life cycle, raising some doubt about the number of
generations per year. Nevertheless and despite the variation, in Portugal, T. acaciaelongifoliae seems to be
approaching to take around one year to complete the life cycle, with wasps more synchronized with the
conditions (climate and phenology of A. longifolia) of the northern hemisphere, as observed in the southern
hemisphere.
Like in previous years, observations to detect galls of T. acaciaelongifoliae were also done in many non-
target plants (please check list of species in Annex 6), located in the areas surrounding A. longifolia trees
where there is establishment of the agent. Despite intensive search no (zero) galls of T. acaciaelongifoliae
were observed in any non-target species. These observations included Cytisus striatus (giesta-das-serras) and
Acacia retinodes, the first because in quarantine tests eggs were detected in the buds of this species, and the
second because it has commercial value in Italy and was later tested. Nevertheless, T. acaciaelongifoliae was
not able to complete the cycle in any of them, with no galls observed in quarantine (Marchante et al. 2011b,
2017) or in the field.
3. Conclusions
Overall, there is establishment of T. acaciaelongifoliae along most of the Portuguese Coast and some more
inland sites, from Riba de Âncora, in the north, to Faro, in the south of the country, in a total of 40 sites.
With increasing number of sites and galls and dispersal to larger areas it is becoming more and more difficult
to detect and count the galls, but there is clearly a very accentuated increase in the establishment of the
biocontrol agent, including both an increase in the number of galls and colonized plants and also in the size
of colonized areas. Most galls in sites with establishment since 2015 correspond probably to fifth generation
of galls, although synchronization of T. acaciaelongifoliae with Northern hemisphere conditions may be not
complete and galls are observable in different months, and as such we cannot be certain about the number
of generations. Nevertheless, independently of the number of generations, T. acaciaelongifoliae is
establishing and spreading by its own means in Portugal, and at some sites galls were observed several
kilometres (up to 7km) from the establishment sites. Although it may be expected that this spread happens
naturally, given the rate of spread of T. acaciaelongifoliae observed in South Africa of up to 20 km in two
years (Dennill and Donnelly, 1991), it is not possible anymore to be certain about the original populations or
if dispersal was done by natural means or aided by any human activity (e.g., transport of A. longifolia wood
including galled branches; or unknown citizens collecting galls and releasing them after).
It is noteworthy the high number of first and second generation galls/site in some sites with recent
establishment; this is probably due to the releases since 2018 being made with wasps originating from
Portuguese populations, and hence more synchronized not only with the phenology of the host-plant
(A. longifolia), but also with more adequate meteorological conditions.
Although the number of plants with galls and the quantity of galls in each plant is still very small and sparse
compared to the numbers of available floral and vegetative buds of A. longifolia in the very extensive
invaded areas, the monitoring of effects shows that T. acaciaelongifoliae is reducing the number of
secondary branches, seeds and pods in colonized plants, reducing both reproductive and vegetative growth
of A. longifolia and consequently its invasive potential.

2020
15
While the team from the Centre for Functional Ecology (both from ESAC and UC) is committed to apply for
new funding, it is somewhat worrying that both projects under which release and monitoring campaigns
were implemented (INVADER-IV and GANHA) are now finished and funding to continue these campaigns is
not yet secured. Nevertheless, monitoring of establishment of T. acaciaelongifoliae and its effects on
A. longifolia will continue, even if more limited, with the collaboration of technicians / managers of release
site and of the citizen science project initiated in 2020.
4. References
Dennill, G.B., 1985. The effect of the gall wasp Trichilogaster acaciaelongifoliae (Hymenoptera: Pteromalidae) on
reproductive potential and vegetativa growth of the Acacia longifolia. Agric. Ecosyst. Environ. 14, 53–61.
Dennill, G.B., Donnelly, D., 1991. Biological control of Acacia longifolia and related weed species (Fabaceae) in South
Africa. Agric. Ecosyst. Environ. 37, 115–135.
Jeger, M.J., Pautasso, M., Stancanelli, G., Vos, S., 2016. The EFSA assessment of Trichilogaster acaciaelongifoliae as
biocontrol agent of the invasive alien plant Acacia longifolia: a new area of activity for the EFSA Plant Health
Panel? EPPO Bull. 46, 270–274. https://doi.org/10.1111/epp.12306
Le Maitre, D.C., Gaertner, M., Marchante, E., Ens, E.J., Holmes, P.M., Pauchard, A., O’Farrell, P.J., Rogers, A.M.,
Blanchard, R., Blignaut, J., Richardson, D.M., 2011. Impacts of invasive Australian acacias: Implications for
management and restoration. Divers. Distrib. 17, 1015–1029. https://doi.org/10.1111/j.1472-4642.2011.00816.x
López-Núñez, F.A., Heleno, R.H., Ribeiro, S., Marchante, H., Marchante, E., 2017. Four-trophic level food webs reveal
the cascading impacts of an invasive plant targeted for biocontrol. Ecology 98, 782–793.
https://doi.org/10.1002/ecy.1701
López-Núñez, F.A., Marchante, E., Heleno, R., Duarte, L., Palhas, J., Impson, F.A.C., Freitas, H., Marchante, E., n.d.
Establishment, spread and early impacts of the first biocontrol agent against an invasive plant in continental
Europe. Submitt. to J. Appl. Ecol.
Marchante, E., Kjøller, A., Struwe, S., Freitas, H., 2009. Soil recovery after removal of the N2-fixing invasive Acacia
longifolia: Consequences for ecosystem restoration. Biol. Invasions 11, 813–823. https://doi.org/10.1007/s10530-
008-9295-1
Marchante, E., Kjøller, A., Struwe, S., Freitas, H., 2008. Short and long-term impacts of Acacia longifolia invasion on the
belowground processes of a Mediterranean coastal dune ecosystem. Appl. Soil Ecol. 40, 210–217.
https://doi.org/10.1016/j.apsoil.2008.04.004
Marchante, H., Freitas, H., Hoffmann, J.H., 2011a. Post-clearing recovery of coastal dunes invaded by Acacia longifolia:
Is duration of invasion relevant for management success? J. Appl. Ecol. 48, 1295–1304.
https://doi.org/10.1111/j.1365-2664.2011.02020.x
Marchante, H., Freitas, H., Hoffmann, J.H., 2011b. Assessing the suitability and safety of a well-known bud-galling wasp,
Trichilogaster acaciaelongifoliae, for biological control of Acacia longifolia in Portugal. Biol. Control 56, 193–201.
https://doi.org/10.1016/j.biocontrol.2010.11.001
Marchante, H., Freitas, H., Hoffmann, J.H., 2010. Seed ecology of an invasive alien species, Acacia longifolia (Fabaceae),
in Portuguese dune ecosystems. Am. J. Bot. 97, 1780–1790. https://doi.org/10.3732/ajb.1000091
Marchante, H., López-núñez, F.A., Freitas, H., Hoffmann, J.H.J.H., Impson, F., Marchante, E., 2017. First report of the
establishment of the biocontrol agent Trichilogaster acaciaelongifoliae for control of invasive Acacia longifolia in
Portugal. EPPO Bull. 47, 274–278. https://doi.org/10.1111/epp.12373

2020
16
Marchante, H., Marchante, E., Freitas, H., Hoffmann, J.H., 2015. Temporal changes in the impacts on plant communities
of an invasive alien tree, Acacia longifolia. Plant Ecol. 216, 1481–1498. https://doi.org/10.1007/s11258-015-0530-
4
Presidência do Conselho de Ministros, 2019. Decreto-Lei n
o
92/2019, 10 de julho de 2019, Diário da República, 1.
a
série
— N.
o
130.
Shaw, R., Schaffner, U., Marchante, E., 2016. The regulation of biological control of weeds in Europe – an evolving
landscape. EPPO Bull. 46, 254–258. https://doi.org/10.1111/epp.12308
Wilson, J.R.U., Gairifo, C., Gibson, M.R., Arianoutsou, M., Bakar, B., Baret, S., Celesti-Grapow, L., Ditomaso, J.M., Dufour-
Dror, J.M., Kueffer, C., Kull, C.A., Hoffmann, J.H., Impson, F.A.C., Loope, L., Marchante, E., Marchante, H., Moore,
J.L., Murphy, D.J., Tassin, J., Witt, A., Zenni, R.D., Richardson, D.M., 2011. Risk assessment, eradication, and
biological control: Global efforts to limit Australian acacia invasions. Divers. Distrib. 17, 1030–1046.
https://doi.org/10.1111/j.1472-4642.2011.00815.x

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Annex 1. FIRST REPORT – 2016

2020
18

2020
19

2020
20

2020
21

2020
22

2020
23

2020
24

2020
25

2020
26
Annex 2. SECOND REPORT – 2017

2020
27

2020
28

2020
29

2020
30

2020
31

2020
32

2020
33

2020
34

2020
35
Annex 3. THIRD REPORT – 2018

2020
36

2020
37

2020
38

2020
39

2020
40

2020
41

2020
42

2020
43

2020
44

2020
45

2020
46

2020
47

2020
48

2020
49

2020
50
Annex 4. FOURTH REPORT – 2019

2020
51

2020
52

2020
53

2020
54

2020
55

2020
56

2020
57

2020
58

2020
59

2020
60

2020
61

2020
62

2020
63

2020
64
Annex 5. FULL NAMES OF THE SITES REFERRED IN THE TEXT, FIGURES AND TABLES (2pages)
CODE SITE SHORT NAME SITE FULL NAME COORDINATES
IP3 Acesso IP3 Acesso IP3, S. Miguel do Outeiro, Viseu 40.586589,-8.015036
AFI Afife Afife, Viana do Castelo 41.789493,-8.869711
ALHQ Alhadas (areeiro) Alhadas (areeiro) 40.2021111, -8.7884639
ALH Alhadas (Cabecinho) Alhadas (Cabecinho), Figueira da Foz 40.174834,-8.787518
ANH Anha Anha, Viana do Castelo 41.674618,-8.801127
ANO Anobra Anobra, Condeixa-a-Nova 40.161626,-8.510096
ANT Antas Parque Natural do Litoral Norte (Antas) 41.61092,-8.80828
BEZ Barrinha de Esmoriz Barrinha de Esmoriz, Esmoriz 40.963981,-8.647378
BZM Belazaima do Chão Belazaima do Chão, Águeda 40.527697,-8.330153
BRJ Brejinhos Brejinhos, Vila Nova de Santo André 38.031267,-8.808383
BRE Brescos Brescos, Santiago do Cacém 38.105037,-8.77737
CAL Calvão Calvão, Vagos 40.470903,-8.685942
CAR Carapinheira, Mafra Carapinheira, Mafra 38.935492,-9.276104
CARR Carreço Carreço, Viana do Castelo 41.754159,-8.87547
TOC4 Charco Berlengas Charco Berlengas 40.32656634,-8.7821874
EST Coimbra (ESAC) Coimbra (ESAC, greenhouse) 40.216722,-8.450210
COI1 Coimbra (Patos) Coimbra (Patos) 40.209237,-8.401237
COI Coimbra (Pólo II) Coimbra (Pólo II from University of Coimbra) 40.18588,-8.41358
HEL Condeixa Condeixa-a-Nova 40.113693,-8.513775
LOB Covão do Lobo Covão do Lobo, Vagos 40.447997,-8.646407
COV Covões Covões, Coimbra 40.194632,-8.466302
TOC3 Cruzamento Tocha
Perímetro Florestal das Dunas de Cantanhede
(cruzamento Tocha)
40.3254768,-8.8131098
QUD Dunas de Quiaios Mata Nacional das Dunas de Quiaios (Dunas Quiaios) 40.22476,-8.88622
SJD Dunas de São Jacinto Reserva Natural das Dunas de S. Jacinto 40.672717,-8.740729
VAG Dunas Vagos (GANHA)
Perímetro Florestal das Dunas de Vagos (Dunas
Vagos)
40.535932,-8.74104
EIX Eixo Eixo, Aveiro 40.620281,-8.568342
EIX2 Eixo (Quinta de S. Francisco) Eixo (Quinta de S. Francisco), Aveiro 40.616127,-8.567075
SPI Espinho
Perímetro Florestal Dunas de Ovar (Maceda,
Espinho)
40.923320,-8.658262
ESP Esposende Parque Natural do Litoral Norte (Esposende) 41.508999,-8.784351
ESP 2 Esposende 2 Esposende 2 41.499313,-8.775334
ARR Estrada Setúbal - praias Parque Natural da Arrábida 38.496597,-8.930805
SAD Estrada Tróia - Comporta
Reserva Natural do Estuário do Sado (Estrada Tróia -
Comporta)
38.426573,-8.824844
EVR Évora Valverde, Évora 38.533685,-8.031304
FAL Falca Praia da Falca, Alcobaça 39.640147,-9.069502
FAR Faro Parque Natural da Ria Formosa (Ludo, Faro) 37.028391,-8.005642
FIG Figueira da Foz Figueira da Foz 40.148682,-8.836959
FIG2 Figueira da Foz (McDonald´s) Figueira da Foz (McDonald´s) 40.166223,-8.853586
FIG1
Figueira da Foz (Rotunda
LIDL)
Figueira da Foz (Rotunda LIDL) 40.157502,-8.849582
GAL Gala, Parque de Merendas Gala, Parque de Merendas, Figueira da Foz 40.123295,-8.859909
LAG Lagoa da Vela Mata Nacional das Dunas de Quiaios (Lagoa da Vela) 40.269481,-8.799287

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FULL NAMES OF THE SITES REFERRED IN THE TEXT, FIGURES AND TABLES (cont., 2/2pages)
CODE SITE SHORT NAME SITE FULL NAME COORDINATES
LSA Lagoas de Santo André
Reserva Natural das Lagoas de Santo André e da
Sancha
37.991658,-8.854934
LAV Lavos Mata Nacional das Dunas da Costa de Lavos (Lavos) 40.097376,-8.856496
LEI Leirosa Mata Nacional das Dunas da Leirosa (Leirosa) 40.076190,-8.865008
MMA Mata da Machada Mata Nacional da Machada 38.619271,-9.038819
MUR Mata do Urso Mata Nacional do Urso 39.983285,-8.914007
MIR Mira Perímetro Florestal Dunas e Pinhais de Mira (Mira) 40.394569,-8.789050
SIN1 Monte Feio - Sines 1 Monte Feio - Sines 1 37.981616, -8.845499
SIN2 Monte Feio - Sines 2 Monte Feio - Sines 2 37.996261, -8.842971
MOR Morraceira Morraceira, Figueira da Foz 40.139481,-8.844876
PAT Paredes da Vitoria (Pataias) Paredes da Vitoria (Pataias) 39.707963,-9.048827
PAT 2 Pataias 2 39.662347,-9.002107
PED Pedrogão Mata Nacional do Pedrogão 39.937940,-8.927712
PQS Pesqueiro Sancha
Reserva Natural das Lagoas de Santo André e da
Sancha (Pesqueiro Sancha)
38.02586,-8.82275
POC1 Pocariça (Cantanhede1)
Pocariça, propriedade do Sr. Mário Mendes
(Cantanhede)
40.382712,-8.574368
POC2 Pocariça (Cantanhede2) Pocariça, vizinho Sr. Mário Mendes (Cantanhede) 40.37305,-8.579074
ARE Praia do Areão Praia do Areão, Vagos 40.491751,-8.788856
STC Praia do navio, Santa Cruz Praia do navio, Santa Cruz 39.144362,-9.371916
CAP Praias da Costa da Caparica
Paisagem Protegida da Arriba Fóssil da Costa da
Caparica (margem)
38.599935,-9.20743
QUP Quiaios N Mata Nacional das Dunas de Quiaios (Pinhal Quiaios) 40.241645,-8.854573
PEN Quinta Pentieiros Quinta Pentieiros, Viana do Castelo 41.775557,-8.657753
RBA
Riba de Âncora (Baldios de
Riba de Âncora)
Perímetro Florestal de Serras de Vieira e Monte
Crasto (por confirmar) - Riba de Âncora (Baldios de
Riba de Âncora)
41.808889,-8.796121
SPM São Pedro de Moel Mata Nacional de Leiria (São Pedro de Moel) 39.75711,-9.02338
SEI Seixo
Perímetro Florestal Dunas e Pinhais de Mira (Seixo,
Vagos)
40.497200, -8.754436
SBV Serra da Boa Viagem
Mata Nacional do Prazo de Santa Marinha (Serra
Boa Viagem)
40.20037,-8.88969
SOU Soure Soure 40.099786,-8.619687
TOC Tocha
(Tocha)
40.34837,-8.81704
TOC2 Tocha (Caniceira) Caniceira, Tocha 40.3476,-8.77368
TOC1 Tocha N
(Tocha Norte)
40.320083,-8.845127
VIC Viana do Castelo Viana do Castelo 41.702256,-8.837369
VLE Vieira de Leiria Mata Nacional de Leiria (Vieira de Leiria) 39.865863,-8.971235
VIG Vigía Vigía, Vagos 40.516265,-8.725941
VNR Vila Nova da Rainha Vila Nova da Rainha, Tondela 40.47055,-8.09458
VNF Vila Nova de Mil Fontes Vila Nova de Mil Fontes (Praia das Furnas) 37.714934, -8.7845
VIV Vila Verde Vila Verde, Figueira da Foz 40.154741,-8.795803

2020
66
Annex 6. LIST OF NON-TARGET PLANTS MONITORED IN 2020 FOR GALLS OF
TRICHILOGASTER ACACIAELONGIFOLIAE.
Non-target species Number of plants observed Number of biocontrol galls
Acacia dealbata 7 0
Acacia mearnsii 1 0
Acacia melanoxylon 27 0
Acacia retinodes 4 0
Acacia saligna 5 0
Arbutus unedo 3 0
Cistus psilosepalus 1 0
Cistus salviifolius 17 0
Cistus sp. 5 0
Corema album 18 0
Cytisus sp. (C. striatus or C. grandiflorus) 19 0
Erica cinerea 1 0
Erica sp. 1 0
Fraxinus angustifolia 1 0
Juniperus turbinata 3 0
Myrica faya 14 0
Other species 1 0
Quercus faginea 2 0
Quercus robur 1 0
Quercus suber 2 0
Salix atrocinerea 1 0
Stauracanthus genistoides 3 0
Ulex sp. 23 0
Total 160 0

Release and post-release monitoring of the biocontrol agent “Trichilogaster acaciaelongifoliae” for the control of the invasive “Acacia longifolia” in Portugal

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Release and post-release monitoring of the biocontrol agent “Trichilogaster acaciaelongifoliae” for the control of the invasive “Acacia longifolia” in Portugal