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Allergenicity to worldwide invasive grass Cortaderia selloana as environmental risk to public health

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Allergies to grass pollen affects about 20% of the population worldwide. In the last few decades, the South American grass Cortaderia selloana (CS, Pampas grass) has expanded worldwide in a variety of countries including the USA, Australia and Western Europe. In many of these locations, CS has strikingly spread and has now been classified an invasive species. Many pernicious consequences of CS have been reported for local biodiversity, landscape and structures. However, the effect on human health has not been studied. To investigate this issue, we have chosen a European region on the northern cost of Spain where CS spread is overwhelming, Cantabria. We obtained CS pollen extract and analysed the allergenic reaction of 98 patients that were allergic to pollen of local grasses. We determined the skin reaction and the presence of specific IgE antibodies (sIgE) to CS or to a typical autochthonous grass, Phleum pratense . We also compared the seasonal symptoms with reported grass pollen counts in the area. The results strongly suggest that CS can cause respiratory allergies at a similar extent to the local grasses. Given that CS pollinises later than the local grasses, this would extend the period of grass allergies in the region for about three months every year, as stated by most of the patients. This is the first study reported on the effects of the striking expansion of CS on human health. Considering the strong impact that respiratory allergies have on the population, our results suggest that CS can currently constitute a relevant environmental health issue.
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Allergenicity to worldwide
invasive grass Cortaderia selloana
as environmental risk to public
health
Fernando Rodríguez1*, Manuel Lombardero‑Vega2*, Lucía San Juan3, Leticia de las Vecillas1,
Sofía Alonso4, Eva Morchón1, Diego Liendo5, Marta Uranga6 & Alberto Gandarillas3,7*
Allergies to grass pollen aects about 20% of the population worldwide. In the last few decades, the
South American grass Cortaderia selloana (CS, Pampas grass) has expanded worldwide in a variety
of countries including the USA, Australia and Western Europe. In many of these locations, CS has
strikingly spread and has now been classied an invasive species. Many pernicious consequences of
CS have been reported for local biodiversity, landscape and structures. However, the eect on human
health has not been studied. To investigate this issue, we have chosen a European region on the
northern cost of Spain where CS spread is overwhelming, Cantabria. We obtained CS pollen extract
and analysed the allergenic reaction of 98 patients that were allergic to pollen of local grasses. We
determined the skin reaction and the presence of specic IgE antibodies (sIgE) to CS or to a typical
autochthonous grass, Phleum pratense. We also compared the seasonal symptoms with reported
grass pollen counts in the area. The results strongly suggest that CS can cause respiratory allergies at
a similar extent to the local grasses. Given that CS pollinises later than the local grasses, this would
extend the period of grass allergies in the region for about three months every year, as stated by most
of the patients. This is the rst study reported on the eects of the striking expansion of CS on human
health. Considering the strong impact that respiratory allergies have on the population, our results
suggest that CS can currently constitute a relevant environmental health issue.
Grass pollen is one of the main causes of respiratory allergies worldwide and the rst cause in North America and
Europe, with estimated 20% of the population aected1. Cortaderia selloana (CS) is a grass of the Poaceae family,
of the Danthonioideae subfamily, commonly known as Pampas grass and native to South America. However, in
the last few decades CS was introduced in a wide diversity of countries worldwide including the USA, Australia
and Western Europe2. In these locations, CS has strikingly spread, and it is classied as an invasive species.
Within Europe, France, Great Britain, Portugal and Spain are strongly colonised. e United States Department
of Agriculture, in a report of 2014, stated: ’Cortaderia selloana obtained a relatively high impact potential risk
score because it impacts natural, anthropogenic, and production systems’3. For this reason, it has been forbid-
den to commercialise, plant or maintain in a variety of countries. One of such countries is Spain, where CS has
intensively spread along the northern cost including the regions of Galicia, Asturias, Cantabria and the Basque
Country4,57. First report mentioning Cortaderia in Spain are from 1953 in Cantabria8.
e allergic incidence of CS is unknown. CS has been referred to as a danger to autochthonous species,
strongly aecting biodiversity and landscape. Moreover, it is sporadically mentioned in some venues and dis-
cussion groups as a danger to humans, because of material machinery damage and health, such as cuts due to
the sharp nature of its leaves, or allergic reactions in contact with the skin911. However, despite the striking
expansion of the grass in regions where it is not autochthonous, there are no studies on the impact on human
health so far reported worldwide.
OPEN
1Allergy Service, Hospital Universitario Marqués de Valdecilla (HUM), Valdecilla 25, 39008 Santander, Spain. 2CMC
R&D Department, ALK-Abelló S.A., Miguel Fleta 19, 28037 Madrid, Spain. 3Institute for Research Marqués de
Valdecilla, Ave Herrera Oria SN, 39011 Santander, Spain. 4Allergy Service, Sierrallana Hospital, Torrelavega,
Spain. 5Department of Plant Biology and Ecology, University of the Basque Country (UPV/EHU), Bilbao,
Spain. 6Asociación Cantabria Sin Plumeros, Liaño, Spain. 7INSERM, Occitanie, Montpellier, France. *email:
fernando.rodriguezf@scsalud.es; Manuel.LombarderoVega@alk.net; agandarillas@idival.org
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CS has strongly colonised extensive areas of Cantabria, a typical Northern Spanish region (Fig.1A,B) of about
500,000 inhabitants, most signicantly during 1990–2008, a period of intensive road and house building6,12. CS
has mainly invaded the coast but it also has reached the inland mountains (Fig.1B–F). e plant has spread
by human activities. It is used in motorways to retain the road slope soil and as a natural barrier13,14 and it is
transported with construction aggregates and gravel from stone quarries. erefore, it is consistently found next
to roads, new buildings or small paths covered with gravel and is abundant around stone quarries (Fig.1C–F).
Plans for limiting and eradicating the growth of this invasive plant have been debated in the local parliament
due to pressure of ecologist organisations although only limited programmes were implemented. Currently,
the European Union is funding a regional network for ghting the inland expansion of the grass and diusing
Figure1. Cortaderia selloana (CS) has strongly invaded northern Spain. (A) Current spread of CS worldwide
(yellow/orange spots). From GBIF.org, GBIF Home Page. Available from: https:// www. gbif. org/ speci es/ 27045
19. (B) Le: location of Cantabria region in Spain (le). Right: current spread of CS in the region is striking, not
only on the coast but also inland (blue line). Source: LIFE Stop Cortaderia, http:// stopc ortad eria. org/ langu age/
en/ early warni ngnet work/. (CF) Representative photographs of the overwhelming presence of CS in Cantabria
region, on the northern coast of Spain. CS has spread near the coast, next to motorways and new house
buildings (C) but is also notorious inland, next to newly constructed areas (D) and even in discrete locations at
the mountains, where gravel has been used on small paths (E). Stone quarries where the gravel is transported
from, are frequently surrounded by CS (F).
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the rapidly increasing problem among the European society (3.5 million euros for 2018–202215). However, the
presence of CS on the northern cost of Spain is still overwhelming.
Cantabria is a good paradigmatic territory to investigate the allergenic eects of CS on the human population.
In Cantabria, autochthonous grass pollens peak from April to July1,16,17, when they cause a concomitant peak of
hay fever. It is estimated that the percent of the population suering from grass-associated hay fever in Cantabria
is about 19% of patients diagnosed of rhinoconjunctivitis and 14% of asthmatic patients18. In contrast to the
autochthonous grasses, CS in the North of Spain ourishes from mid August to October 5. Grass pollens of the
Pooideae subfamily, the main grasses found in temperate climates of the North Hemisphere, contain proteins
with similarities in their antigens19,20. We questioned whether patients allergic to the autochthonous grass pol-
len (Phleum pratense, Phl, as representative species) might also be allergic to CS pollen. Allergenic molecules
of groups 1 and 5 (Phl p1 and Phl p5) are main antigens inducing allergies due to their high capacity to bind to
immunoglobulin IgE of the human immune system. To investigate this issue, we analysed the skin reaction to
Phl and CS extracts of 98 patients of Cantabria that were allergic to local grass pollen. In addition, we determined
the presence of specic IgE antibodies (sIgE) to Phl and CS pollen extracts and to the single allergens Phl p 1,
Phl p 5, Phl p 7 and Phl p 12 in blood serum. We also compared the seasonal symptoms with reported grass
pollen counts. e results very strongly suggest that CS is a signicant cause of respiratory allergies, at a similar
extent as the local grass. is might thus extend the period of respiratory allergies in the region for more than
three months every year. is is the rst study reported on the eects of the striking expansion of CS on human
health and it has implications in all the regions of the world where CS has become a widespread invasive grass.
Considering the implications that respiratory allergies have on health, not only by the direct eects but also
by allowing opportunist infections, our results suggest that CS can constitute a signicant public health issue.
is risk must be added to the ecological impact, in order to encourage eorts for eradicating CS from invaded,
non-autochthonous regions.
Materials and methods
Setting. is study was conducted in Cantabria, a region of the North coast of Spain.
Design and patients. A cross-sectional study with prospective data collection was performed at the
Allergy Services of the Marqués de Valdecilla University Hospital in Santander and the Sierrallana Hospital in
Torrelavega (Cantabria, Spain).
98 patients diagnosed of rhinoconjunctivitis, asthma or both, caused by sensitization to grass pollen, were
included in a sequential way from October 2015 to March 2016.
Written informed consent was obtained from all patients before entering the study. e study met the prin-
ciples of the 1975 Helsinki declaration and was reviewed and approved by the local Research Committee of
Cantabria (CEIC reference number 2015.207).
A serum sample was obtained from each patient and stored at –20°C until used.
Pollen extract preparation. All methods were performed in accordance with the relevant guidelines and
regulations.
Cortaderia selloana (CS) pollen was obtained commercially (Iber-Polen, Jaén, Spain) and then extracted at a
1:10 (w/v) ratio in PBS pH 6.5 with magnetic stirring for 90min. at 5°C. e soluble fraction was separated by
centrifugation. Aer dialysis against PBS, the extract was ltered through 0, 22µm lters. Protein content was
determined by Bradford method (BioRad, Hercules, CA, USA). Two dierent batches were obtained (07 and
09) with consistent results.
Part of the extract was adjusted to 0.25mg protein/ml and formulated in PBS with 50% glycerol, phenol 0.51%
(SPT buer). e remaining extract was stored in aliquots at −20°C.
Phleum pratense (Phl) pollen extract was made as described for CS. e origin of the pollen in this case was
ALK Source Materials, Post Falls, Idaho, USA.
e protein proles of the CS or the Phl extracts were determined by polyacrylamide electrophoresis in the
presence of sodium dodecyl sulphate (SDS-PAGE) under reducing conditions (Invitrogen-Novex tricine gels
10–20% acrylamide, Fisher Scientic, SL, Madrid Spain).
Skin prick test. Patients were skin prick tested (SPT) with a commercial extract (ALK-Abelló, S.A. Madrid,
Spain) of Phl and the CS extract. Histamine dihydrochloride solution (10mg/ml) and SPT buer were used as
positive and negative control(no reaction), respectively.
e SPT wheal areas were measured by planimetry. A cut-o area of 7 mm2 (about 3mm average diameter)or
higher was considered a positive test result(histamine).
e CS extract was tested in 10 control subjects, that were not sensitised to grass pollen, with negative
result(no reaction).
IgE assays. Serum samples were tested for IgE antibodies against Phleum pratense (Phl) pollen extract and
the allergens Phl p 1, Phl p 5, Phl p 7 (polcalcin) and Phl p 12 (prolin) (ImmunoCap FEIA, ermo Fisher
Scientic, Barcelona, Spain).
In addition, specic IgE against Phl and CS pollen extracts was determined by RAST (Radio Allergo Sorb-
ent Test). Paper discs were activated with CNBr and sensitised with the pollen extracts as described by Ceska
etal.21. Phl and CS discs were incubated overnight with 50 µL of the patient’s serum and aer washing (0.1%
Tween-20 in PBS), with approximately 100,000cpm of the iodine 125–labeled anti-IgE mAb HE-2 for 3h as
described22. Finally, the discs were washed, and their radioactivity was determined in a gamma counter. sIgE
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values in kilounits per litre were determined by interpolating in a standard curve built up with Lolium perenne
sensitised discs and 4 dilutions of a serum pool from patients with grass allergy, which was previously calibrated
in arbitrary kU/l.
A cut-o value of 0.35 kU/l was considered positive for both ImmunoCap and RAST. ere was a very sig-
nicant correlation between the sIgE against Phl determined by both methods (r Spearman = 0.8874, p < 0.0001).
RAST inhibition assay. Paper discs were sensitised as above in the IgE assays section and then incubated
with 50 µL of a serum pool from all patients combined. 50 µL of (inhibitory) CS extract solution (in serial dilu-
tions) were added onto the paper discs and incubated overnight at room temperature. All other incubations were
performed as indicated above in the IgE assays section. e % of inhibition was determined for each extract
dilution by radioactive counts (cpm) and calculated by means of the following equation:
Cpmx corresponds to the mean radioactivity of the discs incubated with inhibitor at a given X dilution.
cpm100% corresponds to the blank control samples of the assay (no serum pool added). cpm0% corresponds to
the signal obtained with no inhibitor extract added.
Results
To investigate whether patients allergic to the local pollen react to CS pollen, we chose a cohort of 98 patients
from Cantabria. Table1 shows the demographic and clinical characteristics of the patients. All of them were
diagnosed with rhinitis during the spring season and grass pollen sensitisation. In addition to nasal symptoms,
98% had also associated conjunctivitis, 31.6% suered from asthma and 8.2% from urticaria. Only 12.2% had
food allergies and 2 out of 98 drug allergies. 53.06% of the patients underwent grass pollen immunotherapy.
76.5% of the patients referred living in areas with high presence of CS. 78.6% of patients presented a worsening
of their pollen allergic symptoms from August to November (“delayed reactivation”). In addition, 56.12% of the
cohort were polysensitised including other pollens such as Plantago spp. (18/98), trees (9/98), Parietaria spp.
(6/98), animal dander (11/98) or house dust mites (38/98).
CS pollen extract is not commercially available to run skin prick tests or sIgE determination. erefore, we
isolated and prepared a CS pollen extract by a standard extraction protocol used for pollens (see Materials and
Methods). e yield protein/pollen was about 50mg/g, a typical concentration obtained for other grass pollens
(our own unpublished data). Grass-specic ELISA assays showed that the CS extract did not contain group 5
antigen, as expected for a non-Pooideae subfamily grass (< 0.3µg group 5/mL23). e prole of the protein extract
by SDS-PAGE shows a group of 25–37 kD bands with the mobility of the grass group 1 allergens and it might
correspond to the homologous CS group 1 (arrow, Fig.2A;19,24).
Isolated CS pollen extract was used on cutaneous tests on the patient cohort, in parallel with Phl pollen
extract, as a representative of the local autochthonous grass pollens. All 98 patients gave a positive response by
skin prick test to Phl pollen extract and 89% of the patients were also positive to CS pollen extract (Table1).
Moreover, there was a signicant correlation between the area of the papule to Phl and to CS (rPearson = 0.2558,
p = 0.01; Fig.2B). As a control, 10 patients negative for skin reaction to Phl were found also negative for CS
extract. ese results show a strong coincidence in the cutaneous reaction to CS and to the local grass. To further
study the interspecies cross reaction of the patient sera, we run by RAST (radio allergo sorbent test) inhibition
assays. As shown in Fig.2C, Phl extract signicantly competed with CS extract to bind the serum sIgE from the
patients.
Supplementary TableI displays the results of sIgE masurement. We determined sIgE to Phl and to the aller-
gens Phl p 1, Phl p 5, Phl p 7 and Phl p 12 by ImmunoCap (ermo Fisher) and to CS by RAST. All patients had
serum sIgE to Phl by both ImmunoCap and RAST, in agreement with the skin prick test results. We determined
the correlation between both techniques in detecting the sIgE for Ph. e relation was rSpearman = 0.8874,
p < 0.0001. Values obtained by RAST were below those obtained by ImmunoCAP (factor = 0.36) and the linear
range for RAST (0.17–27) was shorter than for ImmunoCAP (0.35–100). Nevertheless, the correlation between
both techniques was good, indicating that the sIgE data obtained by ImmunoCap can be compared with the
sIgE data obtained by RAST (Supplementary Fig.1). All patients but seven contained sIgE specic to CS extract.
Interestingly, within the seven patients with a negative sIgE test to CS, 5 displayed a negative skin response to CS
and the other 2 displayed a weal smaller than 14 mm2. erefore, there was a strong correlation between the skin
response and the sIgE to CS in serum (Fishers exact test, p < 0.0001; Supplementary TableII).
We measured the presence of sIgE to the individual allergens Phl p 1, Phl p 5, Phl p 7 and Phl p 12 in the sera
from the patients (Supplementary TableI). For the pan-allergens Phl p 7 (polcalcin) and Phl p 12 (prolin), only
27 patients (27.5%) had sIgE to any of them. Consequently, the patient sensitisation to these allergens cannot
explain the high cross-sensitisation to Phl and CS in this group of patients. e prevalence of sIgE to Phl p 1 was
very high (98%) and only two patients (# 45 and 83) were negative for IgE to Phl p 1. Consistently, these patients
also displayed a negative skin response to CS extract. e prevalence of Phl p 5 was lower but still important
(72%). Twenty-seven patients of the cohort displayed no IgE to Phl p 5 in serum. However, of these, only ve
patients were negative for skin response to the CS extract. ere was a signicant linear regression between the
sIgE to the whole Phl extract and the sIgE to Phl p 1 (Fig.3A) or Phl p 5 (Fig.3B). From the slope of the regression
line, we can conclude that every allergen accounts for about 50% of the total IgE response to the whole extract,
being the IgE-response to Phl p 1 slightly higher. e reaction to Phl p 1 plus Phl p 5 is similar to the reaction to
whole Phl extract (Fig.3C), strongly suggesting that groups 1 and 5 are the main allergens of Phl and they account
for most of the IgE to the whole Phl extract. ere is a signicant correlation between thesIgE to CS extract and
thesIgE to Phl whole extract, toPhl p1 or toPhl p5 (Table2). e correlation is stronger for the whole extract or
100
×
1

cpmx cpm100%
/
cpm0% cpm100%

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Patient Age (years) SexaYears living in
Cantabria ExposurebClinical symptomscMonths with symptoms Other sensitisationsd
Cutaneous reactione
C. selloana P. pr at en s e
E0416001 50 M 10 (*) RC May–Oct HDM/plantago 47 59
E0416002 38 F 38 (*) RCA Mar–Oct 40 24
E0416003 23 M 23 (–) RCAU Mar–Sep HDM/dog 30 21
E0416004 44 10 (*) RCA Apr–Sep HDM 47 45
E0416005 46 M 46 (*) RC Mar–Oct HDM/platanus 10 22
E0416006 49 M 20 (*) RC Jun–Oct – 67 34
E0416007 41 F 41 (–) RCA Mar–Aug Dog 32 113
E0416008 27 F 27 (*) RC May–Aug HDM 31 65
E0416009 27 F 27 (*) RC Mar–Oct 49 76
E0416010 55 M 55 (*) RC Apr/Oct – 30 54
E0416011 50 F 20 (*) RC Jul–Oct – 35 38
E0416012 36 M 36 (*) RC May–Oct – 16 33
E0416013 20 M 20 (*) RC Apr–Aug HDM 39 94
E0416014 42 M 37 (*) RC Mar–Sep – 19 92
E0416015 39 M 12 (*) RCA May–Oct HDM 18 25
E0416016 45 M 45 (*) RC May–Sep – 21 20
E0416017 52 M 31 (*) RCA Mar–Jul 63 92
E0416018 45 M 40 (*) RC May–Sep – 42 48
E0416019 34 M 34 (*) RC May–Oct – 67 78
E0416020 30 M 25 (*) RC Feb–Nov Cat 29 48
E0416021 44 M 21 (*) RCA May–Set 111 162
E0416022 38 M 38 (*) RC Apr–Aug HDM 737
E0416023 43 M 7 (–) RC Feb–Nov Plantago 14 23
E0416024 50 M 50 (*) RC May–Set – 6(N) 39
E0416025 33 F 33 (*) RC Apr–Oct 49 28
E0416026 29 F 29 (*) RC Apr–Set – 37 77
E0416027 48 M 48 (*) RC May–Oct HDM 68 35
E0416028 41 M 4 (*) RC Apr–Jun – 86 48
E0416029 42 M 14 (*) RCU Apr–Sep HDM 21 48
E0416030 29 M 29 (*) RC May–Sep HDM 48 50
E0416031 42 M 42 (*) RCA Mar–Aug HDM 36 48
E0416032 48 M 15 (–) RC Mar–Aug 0 49 69
E0416033 25 M 25 (–) RC May–Sep HDM 34 169
E0416034 39 F 15 (*) RC Apr–Jul 6(N) 83
E0416035 53 F 17 (*) RC Apr–Nov 27 38
E0416036 48 M 6 (*) RC Apr–Oct HDM 61 33
E0416037 63 F 63 (*) RC Apr–Jul 27 33
E0416038 58 M 58 (*) RC May–Aug – 23 22
E0416039 39 M 39 (*) RC May–Aug HDM 50 114
E0416040 40 F 40 (*) RC May–Oct HDM 18 37
E0416041 31 F 31 (*) RC May–Aug – 1(N) 26
E0416042 29 F 29 (*) RCAU May–Jul 13 31
E0416043 32 F 18 (*) RC Jul–Sep HDM 24 171
E0416044 42 F 8 (*) RCA Apr–Aug Parietaria 17 22
E0416045 42 M 2 (*) RC Jul–Sep – 1(N) 37
E0416046 22 F 22 (*) RCA Mar–Aug HDM/parietaria 79 64
E0416047 34 M 34 (*) RCA May–Oct HDM/cat 1(N) 18
E0416048 39 M 39 (*) RCA May–Jul 41 42
E0416049 41 M 15 (*) RC Apr–Oct HDM 57 44
E0416050 28 F 27 (*) RC Apr–Nov HDM 15 23
E0416051 30 M 30 (*) RC Mar–May HDM/plantago/cupresa-
ceous/parietaria 32 128
E0416052 63 M 63 (*) RC May–Oct HDM 40 37
E0416053 22 M 20 (*) RC Apr–Aug Dog 21 75
E0416054 32 F 32 (*) RC May–Oct HDM 22 67
Continued
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Table 1. Demographic and clinical data and SPT results. SPT Skin prick test. a F female, M male. b Exposure
(*) means that the patient lives in an area in which C. selloana plants have been identied. c A athma, C
conjunctivitis, R rhinitis, U urticaria. d HDM House Dust Mites. e Wheal area (mm2). Negative reaction to C.
selloana is highlighted in italic numbers (N). Numbers in bold indicate positive reaction (>6).
Patient Age (years) SexaYears living in
Cantabria ExposurebClinical symptomscMonths with symptoms Other sensitisationsd
Cutaneous reactione
C. selloana P. pr at en s e
E0416055 41 F 7 (*) RCA May–Sep HDM/parietaria/
plantago 15 41
E0416056 44 F 44 (*) RC Apr–Nov HDM/horse/dog/cat 34 79
E0416057 23 F 23 (–) RCA May–Sep HDM 13 49
E0416058 41 F 41 (*) RC Apr–Sep – 38 36
E0416059 31 F 31 (*) RCAU Feb–Nov Cat/dog/plantago/HDM 815
E0416060 41 M 36 (*) RC May–Sep – 37 45
E0416061 29 F 29 (*) RC Apr–Sep – 29 68
E0416062 44 M 43 (–) RCA May–Jul Cat 535
E0416063 50 F 50 (–) RC Jun–Nov HDM 11 21
E0416064 26 F 1.5 (*) RC Mar–Oct 17 46
E0416065 69 M 69 (–) RCA May–Nov 5(N) 66
E0416066 39 F 31 (*) RCA Apr–Aug 27 47
E0416067 40 M 40 (*) RCA May–Nov Plantago 10 14
E0416068 26 F 26 (*) RC May–Sep Plantago 34 39
E0416069 67 M 67 (–) RC May–Sep – 12 42
E0416070 70 F 70 (*) RC May–Nov HDM 48 46
E0416071 32 F 32 (*) RC Apr–Oct HDM 20 20
E0416072 30 F 30 (*) RC Apr–Jul 130 34
E0416073 18 F 18 (–) R May–Jun HDM/plantago 13 26
E0416074 50 M 24 (*) RC May–Oct – 54 53
E0416075 35 M 35 (–) RC Apr–Aug 75 77
E0416076 23 M 23 (*) RCA Apr–Aug HDM/plantago 71 96
E0416077 38 F 38 (*) RCA May–Oct HDM 38 22
E0416078 34 F 34 (–) RC Apr–Oct Parietaria 37 57
E0416079 23 M 23 (*) RC Apr–Oct HDM 1(N) 24
E0416080 36 M 36 (*) RCA Apr–Sep 37 36
E0416081 32 F 32 (*) RCA Apr–Jul HDM/parietaria 28 78
E0416082 36 F 9 (–) RC May–Jun – 18 28
E0416083 31 M 31 (–) RC Apr–Jul 329
E0416084 23 M 23 (*) RCA May–Jul 27 56
E0416085 39 F 39 (*) RC May–Aug HDM 67 55
E0416086 29 F 10 (–) RCAU May–Sep Platanus/cupresaceous/
plantago 11 39
E0416087 18 F 12 (*) RCA Apr–Sep Plantago 16 96
E0416088 46 M 15 (–) RCA Mar–Sep HDM/cat/dog/horse/
platanus/cupresaceous/
plantago 32 32
E0416089 30 F 30 (–) RCAU Mar–Jul Platanus/plantago 33 138
E0416090 23 M 23 (–) RCU Mar–Oct 18 52
E0416091 39 F 39 (*) RCA Mar–Nov HDM/plantago 17 80
E0416092 20 M 20 (*) RCU Apr–Jul 107 65
E0416093 63 F 63 (–) RC Apr–Aug 20 23
E0416094 23 F 23 (–) RC Mar–Sep Plantago/platanus 529
E0416095 44 F 26 (–) RC Apr–Jul Platanus/plantago/
cupresaceous 34 39
E0416096 43 F 43 (–) R May–Jun 1(N) 89
E0416097 37 F 37 (*) RC May–Oct – 80 27
E0416098 30 M 30 (*) RCA Mar–Jul Dog/cupresaceous/
plantago 20 56
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for Phl p 1 (r = 0.75) than for Phl p 5 (r = 0.55). ese results suggest that the common reaction observed in the
patients to pollen extract of Phl and CS might reside in the antigenic Group 1 that is ubiquitous in all grasses25.
We analysed the measured grass pollen concentration along the year in the region. e regional agency
Health Department of the nearby Basque Country detected a spring main peak of grass pollen around May
and a second, August-to-October peak, in the air of Bilbao, a city 50km o Cantabria with a similar climate
and density of CS. It is interesting that most patients (78.6%) in the study mentioned a second allergic reaction
around September–October (Table1, Supplementary Fig.2). is indicates a timely correlation between grass
pollen and the referred allergic symptoms by the patients.
Discussion
We could not nd in the literatureany report on the impact of CS on human health. is is somehow surpris-
ing and highlights the need of studies on the issue, considering the widespread presence of this invasive plant
worldwide15. Concerns about the consequences of CS expansion are evident among professionals regarding the
impact of CS in ecology, industry or health912. Our study addresses for the rst time the potential allergenic
eects of CS pollen. Given the wide impact of grass allergy in the population, this constitutes a public health issue.
We here present several lines of evidence strongly suggesting that patients allergic to pollen of northern Span-
ish autochthonous grasses, such as Phl, are also allergic to pollen of CS: (i) 89% of the patients allergic to Phl
were sensitised to CS, as evident both by skin reaction and by sIgE in serum; (ii) the timely coincidence along
the year of allergy symptoms reported by patients, grass pollen counts and ourishing of CS; (iii) the presence
in CS of a protein band with a mobility compatible with grass allergenic group 1 and the strong prevalence of
this group in the sIgE to Phl.
050100 15
0200
0
50
100
150
SPT Phleum
(mm
2
)
SPT Cortaderia
(mm
2
)
BA
C
CS PhL
12M
250
150
100
75
50
37
25
20
15
10
43 kD
G1[ ]G1,5
Figure2. Cortaderia selloana (CS) pollen shares antigens and inmmunogenicity with authoctonous grass
Phleum pratense (Phl). (A) SDSPAGE prole of CS (lanes 1, 2) or Phl pollen extract (lanes 3,4) extract. Lane
1 and 2 corresponds to 20µl and 40µl of CS pollen extract, respectively (see also Supplementary Fig.3)
representative of two independent batches. Lane 3 and 4 correspond to 20µl from two dierent batches of Phl
pollen extract. M: the molecular weight markers. Brackets indicate the position of the allergenic groups (G)
according to the documented apparent molecular weights. (B) Correlation between SPT result for PhL and
CS Pearson r: 0.2558; R2: 0.06543; p value : 0.0110 (twotailed). (C) IgE Crossreactivity of CS and Phl pollen
antigens as measured by radioallergosorbent (RAST) inhibition assays. Note that if there were no crossreaction
the Phl plot should be at to cero (broken line).
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020406080100
0
20
40
60
80
100
sIgE Phleum, kU/L
sIgE Phl p 1, kU/L
A
02040608
01
00
0
20
40
60
80
100
sIgE Phleum, kU/L
sIgE Phl p 5, kU/L
B
020406080
100
0
20
40
60
80
100
sIgE Phleum, kU/L
sIgE Phl p 1 + 5, kU/L
C
Figure3. Linear regression of sIgE to Phleum pratense whole extract (ImmunoCap) versus sIgE to Phl p 1 (A),
sIgE to Phl p 5 (B) and sIgE to Phl p 1 + Phl p 5 (C).
Table 2. Correlation between sIgE to C. selloana and to Phl p 1 sIgE, to Phl p 5 sIgE and to P. pratense sIgE.
Phl p: Phleum antigen group.
sIgECS (Ku/L) vs. sIgE Phl p 1 (kU/L) sIgECS (Ku/L) vs. sIgE Phl p 5 (kU/L) sIgECS (Ku/L) vs. sIgE Phl (kU/L)
ImmunoCap
r Spearman 0.755 0.552 0.7476
95% condence interval 0.6513 to 0.8311 0.3920 to 0.6795 0.6400 to 0.8264
P (twotailed) < 0.0001 < 0.0001 < 0.0001
Signicant? (alpha = 0.05) Ye s Ye s Yes
Number of XY pairs 98 98 96
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e high cross-sensitisation to Phl and CS pollen in this cohort of patients is not explained by a possible
reaction to pan-allergens, such as prolin (Phl p 12) and polcalcin (Phl p 7), since only 27.5% of the patients
contained serum sIgE against them. Group 1 is a major grass allergen ubiquitous in all grasses in contrast to
group 5 which is absent in non-Pooideae grasses as is the case of CS. e data presented in this study strongly
suggests that grass group 1 might be the culprit of the observed cross-sensitisation between autochthonous
grasses (in this study, Phl) and CS.
Autochthonous grasses in Northern Spain ourish from April to July1,16,17,26, while CS ourishes from August
to October5,7. From a clinical point of view, most patients (78.6%) referred a late allergic symptoms reactivation
around September–October coincident with a second, August-to-October, peak of grass pollen counts in the
air. At present, there are no commercial extracts of CS for immunotherapy. However, the overall improvement
of symptoms usually reported by allergic patients that were treated with conventional grass immunotherapy,
during both pollination peaks, suggests that they might have been protected also to CS pollen. is in addition
holds clinical interest to those regions where CS is autochthonous and possibly allergenic.
e implications of the results into public health-related issues are many and diverse. First, the results encour-
age the international community to run allergenic tests to CS and to biochemically characterise the reaction
to CS. Second, the results suggest that CS might lengthen the grass allergy season in territories where CS has
expanded, by causing a second later peak, additional to the peak due to the autochthonous grasses. To note,
commercially available grass immunotherapy might be benecial to patients allergic to CS worldwide. ird,
given that CS is banned in many countries and states, since it is considered aninvasive species3, a demonstrated
impact on human health would encourage policy makers to run programmes for eradicating this plant in non-
autochthonous areas. e results provide an example of the global eects that alien invasive species can have
on human health.
Received: 5 August 2021; Accepted: 25 November 2021
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Acknowledgements
We thank Daniel Liébana Uranga (Cantabria, Spain) for permission to include authorship and data obtained
by Marta Uranga, sadly deceased before preparation of the manuscript. We thank alia Burn (England, UK)
for manuscript text revision and Life Stop Cortaderia (Cantabria, Spain) for image in Fig.2B-right. is work
received nancial support from ALK-Abelló S.A (Madrid, Spain). is article is dedicated to the memory of
Marta Uranga.
Author contributions
F.R.: conceptualization, data curation, investigation, methodology, project administration, supervision, valida-
tion, writing—review & editing. M.L.V.: conceptualization, data curation, formal analysis, investigation, meth-
odology, validation, writing—review & editing. L.d.l.V., S.A., E.M.: data curation, investigation and methodology.
L.S.J.: soware, review & editing. D.L.: review & editing. M.U.: conceptualization, data curation, formal analysis,
investigation. A.G.: conceptualization, investigation, methodology, supervision, validation, writing—review &
editing.
Competing interests
e authors declare no competing interests.
Additional information
Supplementary Information e online version contains supplementary material available at https:// doi. org/
10. 1038/ s41598- 021- 03581-5.
Correspondence and requests for materials should be addressed to F.R., M.L.-V.orA.G.
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... It also causes respiratory allergies in humans. A study in Cantabria, Spain, showed that C. selloana produces pollen later than the local grasses (which cause allergies in 30% of the population), thereby substantially increasing the period of grass allergies in the region for about three months and increasing the overall impact of respiratory allergies on the human population (Gobierno de Cantabria 2018; Rodríguez et al. 2021). ...
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Cortaderia selloana (Schult. & Schult. f.) Asch. & Graebn. (Pampas grass) is a perennial grass native to temperate and subtropical regions of South America. The species was introduced to western Europe for ornamental purposes during the nineteenth century, where it has become naturalized in anthropogenic and natural habitats, especially in sandy, open, and disturbed areas. Female plants of C. selloana produce thousands of seeds that are dispersed over long distances by wind and germinate readily. Its invasive success is also attributed to its ability to adapt and tolerate a wide range of environmental conditions, such as high salinity levels, long droughts, and soil chemical pollution. Cortaderia selloana usually invades human-disturbed habitats where it encounters little competition with other plants and high resource availability. However, the species can invade natural habitats, especially those with high light availability, causing biodiversity loss and changes in ecosystem functioning (e.g. alteration of succession and nutrient dynamics). The species may cause negative socio-economic impacts by reducing productivity of tree plantations, causing respiratory allergies, and decreasing the recreational value of invaded areas. Control costs are high due to the extensive root system that C. selloana develops and the high resprouting ability following physical damage. Although herbicides are effective control measures, their use is not allowed or is undesirable in all situations where the plant occurs (e.g. near riverbanks, natural protected sites). No biological control agents have been released on C. selloana to date, but the planthopper Sacchasydne subandina and the gall midge Spanolepis selloanae are promising targets.
... & Graebn. is a tall grass widely used in gardening for the ornamental value of its large tussocks and tall inflorescences, that has become a global invader (DiTomaso et al., 2010;Domènech & Vilà, 2006;Houliston & Goeke, 2017;Pausas et al., 2006). The impacts of C. selloana includes disturbance of native ecosystems, increasing fire risk and causing allergies to people, among others (Cires et al., 2022;Rodríguez et al., 2021). Cortaderia selloana is sexually dioecious: female plants produce ovaries and lack stamens, while hermaphroditic plants generate and release large amounts of pollen and rarely produce viable seeds (Astegiano et al., 1995;Testoni & Linder, 2017). ...
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The ability to balance the allocation of resources between growth and reproduction as a response to stress factors, can be an advantage for plants in disturbed environments. Invasive alien plants (IAPs) often show high levels of phenotypic variability in resource allocation, a key trait that plays a crucial role in their success to invade new areas. Control management for IAPs must consider this capacity in the development of effective strategies. In this study, we performed continuous measures of leaf growth and reproductive traits of Cortaderia selloana , an IAP of global concern, and applied generalised linear models (GLMs) to evaluate trade‐offs between vegetative growth, leaf composition and reproductive success at different cutting moments. Cutting moment, but not flowering, affected the length of the vegetative growth period (VGP) and average growth rate (AGR), and the interaction with flowering affected AGR and final leaf length (vegetative growth total, VGT). Specific leaf area (SLA), leaf nitrogen (N) content and the isotopic value of δ ¹³ C were affected by cutting, and N was also affected by flowering and the interaction with cutting time. Silica also showed a negative correlation with leaf carbon (C) depicting a trade‐off between both structural components. Cortaderia selloana successfully adapted its leaf growth and composition to cutting moment, but this was also modulated by flowering. Moreover, the species is dioecious, and its response may differ between female and hermaphroditic plants. This suggests flexible trade‐offs in resource allocation, therefore the time for cutting must be precisely scheduled to suppress flowering.
... C. selloana has many negative impacts on native biodiversity (e.g., changes soil nutrient properties and community structure; creates barriers to the movement of fauna, uses the resources available to other species.; Domènech et al., 2006), economies (e.g., forestry production systems; Gadgil et al., 1992) and human health and well-being (e.g., allergies and respiratory diseases, Rodríguez et al., 2021). Indeed, it achieves the highest impact score across scales (using Generic Impact Scoring System -GISS) when compared with other alien grasses (Nkuna et al., 2018). ...
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Deep learning has advanced the content analysis of digital data, unlocking opportunities for detecting, mapping, and monitoring invasive species. Here, we tested the ability of open source classification and object detection models (i.e., convolutional neural networks: CNNs) to identify and map the invasive plant Cortaderia selloana (pampas grass) in mainland Portugal. CNNs were trained over citizen science images and then applied to social media content (from Flickr, Twitter, Instagram, and Facebook), allowing to classify or detect the species in over 77% of situations. Images where the species was identified were mapped, using their georeferenced coordinates and time stamp, showing previously unreported occurrences of C. selloana, and a tendency for the species expansion from 2019 to 2021. Our study shows great potential from deep learning, citizen science and social media data for the detection, mapping, and monitoring of invasive plants, and, by extension, for supporting follow-up management options.
... The following considerations about the plants that attained PAV values considered high and very high are related to the herbaceous species Cortaderia selloana and Parietaria judaica. Cortaderia selloana is on the National List of Invasive Species (Decree-Law n. • 92/ 2019, July 10, 2019) and in the last few decades, this Poaceae originated from South American, commonly known by Pampas grass, has expanded worldwide in several countries, including those in Western Europe (Rodríguez et al., 2021). According to these authors, C. selleona might cause respiratory allergies to a similar extent to local grasses and it pollinates later than the local grasses, which would extend the period of grass allergies in the region for about three months every year. ...
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Background and Aims: Cortaderia selloana, commonly known as Pampas Grass, manifests as an invasive plant across numerous countries with predominant studies focused on its control measures (physical, biological, and chemical approaches). Native to South America, this perennial tussock grass negatively impacts economic, environmental, and human health. This study aims to explore the diverse uses of Cortaderia selloana in geographical, cultural and ecological contexts, to provide insights into its applications, and to contribute to socio-economic and ecological understanding. Methods: This study comprises a systematic literature review based on the PRISMA 2020 guidelines. The search was conducted in EBSCOhost, ScienceDirect and Web of Science using the search question “("Cortaderia selloana") AND ("use" OR "purpose") NOT (“control”)”. Two researchers independently reviewed the titles and abstracts, applying predefined inclusion and exclusion criteria and extracting data on various aspects covered in the selected studies. Key results: A total of 88 articles were retrieved of which 16 were included in this systematic review. This study described diverse applications attributed to Cortaderia selloana, including wastewater treatment, composite synthesis, traditional medicine, bioremediation, biorefining, product development in the automobile industry and decoration. The utilization of this invasive species demonstrated socio-economic and environmental benefits, providing a novel perspective on transforming something harmful into a resource with various applications. Furthermore, the necessity for more research is emphasized, to enhance understanding of known applications and explore new potential uses. This study's main limitation is that it only includes peer-reviewed articles from selected databases. Conclusions: This research provides valuable insights into the diverse uses of Cortaderia selloana across geographical, cultural, and ecological contexts. These findings underscore the importance of considering both the beneficial applications and the challenges posed by this invasive species in order to inform balanced and sustainable management practices.
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Exotic allergenic species constitute an important element of global change and are an emergent health issue in Europe due to their potential allergenicity. The grass pollen season is of great importance from the allergic point of view because it includes pollen from ubiquitous species which are responsible for high sensitization rates. In this study, we used flowering phenology data for dominant grass species in the city of Madrid (Spain) and airborne pollen data to explore differences between native and exotic species and their potential contribution to the observed peaks of pollen exposure. We found that exotic grasses flowered later than Mediterranean native grasses, and that ornamental grass species (such as Cortaderia selloana and Pennisetum villosum) cause an unusual second pollen season in autumn with implications for public health. These results support the need to coordinate the efforts of plant ecologists and aerobiologists to protect the population by identifying sources of allergenic pollen and sustain the appropriate urban plans.
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Alien plants in coastal habitats and their influence on natural vegetation are studied. After 5 years working on this subject in the Basque Country and surrounding areas, a number of results from the coastal ecosystems are presented. These ecosystems are one of the most threatened and affected by the invasion of alien plants, especially shore dunes, saltmarshes and cliffs. These kinds of habitats, especially the dunes, experience significant pressure from human activities which favours the expansion of some of these species: Arctotheca calendula, Sporobolus indicus and Oenothera spp. The presence and abundance of these invasive plants and others such as Baccharis halimifolia, Cortaderia selloana, Spartina patens and Carpobrotus edulis in the plant communities in an area between the French border and the western part of the region of Cantabria have been studied. The degree of invasion of each plant in each syntaxonomic unit has been analysed.
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Airborne Poaceae pollen counts are greatly influenced by weather-related parameters, but may also be governed by other factors. Poaceae pollen is responsible for most allergic reactions in the pollen-sensitive population of Galicia (Spain), and it is therefore essential to determine the risk posed by airborne pollen counts. The global climate change recorded over recent years may prompt changes in the atmospheric pollen season (APS). This survey used airborne Poaceae pollen data recorded for four Galician cities since 1993, in order to characterise the APS and note any trends in its onset, length and severity. Pollen sampling was performed using Hirst-type volumetric traps; data were subjected to Spearman's correlation test and regression models, in order to detect possible correlations between different parameters and trends. The APS was calculated using ten different methods, in order to assess the influence of each on survey results. Finally, trends detected for the major weather-related parameters influencing pollen counts over the study period were compared with those recorded over the last 30 years. All four cities displayed a trend towards lower annual total Poaceae pollen counts, lower peak values and a smaller number of days on which counts exceeded 30, 50 and 100 pollen grains/m(3). Moreover, the survey noted a trend towards delayed onset and shorter duration of the APS, although differences were observed depending on the criteria used to define the first and the last day of the APS.
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The Poaceae family comprises over 12,000 wind-pollinated species which release large amounts of pollen into the atmosphere. Poaceae pollen is currently regarded as the leading airborne biological pollutant and the chief cause of pollen-allergy worldwide. Sensitization rates vary by country, and those variations are reviewed here. Grass pollen allergens are grouped according to their protein structure and function. In Poaceae, although species belonging to different sub-families are characterized by distinct allergen subsets, there is a considerable degree of cross-reactivity between many species. Cross-reactivity between grass-pollen protein and fresh fruit pan-allergens is associated with the appearance of food allergies. The additional influence of urban pollution may prompt a more severe immunological response. The timing and the intensity of the pollen season is governed by species genetics, but plant phenology is also influenced by climate; as a result, climate changes may affect airborne pollen concentrations. This paper reviews the findings of worldwide research which has highlighted the major impact of climate change on plant phenology and also on the prevalence and severity of allergic disease. This article is protected by copyright. All rights reserved.
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The expansion histories of two South American species of Cortaderia , similar in morphology but differing profoundly in their breeding systems, were compared in California, USA. Both species were introduced to California in the mid‐1800s, but herbarium records indicate that the sexual C. selloana has expanded spatially at twice the rate of the asexual C. jubata . The invasiveness of C. selloana has increased over time, whereas that of C. jubata has remained relatively constant. Populations of C. selloana now occupy more vegetation types and more non‐ruderal habitats than C. jubata . Populations of C. selloana have experienced directional morphological change, whereas the morphology of C. jubata has been constant over the 90 years for which preserved specimens are available. The invasion of an alien species appears to be a malleable process, rather than a singular event. Species traits, such as inbreeding, can be advantageous at some stages but disadvantageous at others. Alien species also adjust over time to the novel and diverse selective regimes that they encounter as they expand spatially. Sexual species may have a greater ability to adjust to diverse selective landscapes relative to asexual species.
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