An annotated checklist of ladybeetle species (Coleoptera, Coccinellidae) of Portugal, including the Azores and Madeira Archipelagos

Abstract A comprehensive annotated checklist of the ladybeetle species of Portugal, including the Azores and Madeira archipelagos, is presented. The Coccinellidae fauna comprises a total of 101 species: 83 from the Mainland, 39 from Madeira, and 32 from the Azores. The listed species are distributed among 2 sub-families and 13 tribes: within the subfamily Microweiseinae, Madeirodulini (1 species), Serangiini (2 species), and within the subfamily Coccinellinae, Azyini (1 species), Chilocorini (4 species), Coccidulini (7 species), Coccinellini (30 species), Epilachnini (4 species), Hyperaspidini (7 species), Noviini (2 species), Platynaspini (1 species), Scymnini (37 species), Stethorini (3 species), and Sticholotidini (2 species). The Portuguese fauna comprises 10 exotic species: 5 present in the Mainland, 7 in Madeira, and 6 in the Azores. Harmoniaaxyridis (Pallas, 1773) from Madeira, Propyleaquatuordecimpunctata (Linnaeus, 1758) from the Azores, Delphastuscatalinae (Horn, 1895) from the Azores and Madeira, Nephus (Geminosipho) reunioni (Fürsch, 1974) and Nephus (Nephus) voeltzkowi Weise, 1910 from Madeira and Microserangium sp. from the Mainland, are reported for the first time. Some species are considered doubtful records, as explained in the text. These results were obtained by compiling information on the available literature regarding ladybeetle species on the Portuguese mainland and insular territories, and original data.

Ladybeetles comprise about 375 genera (Nedvěd 2020) and nearly 6000 species (Vandenberg 2002), distributed worldwide. They are characterised by a high diversity as regards to their life history, development, distribution, habitat, and food relationships (see Hodek et al. 2012 for review). This family of insects is very charismatic, in particular because most species are predators recognised as useful natural enemies of pests, including aphids (Aphidoidea), scale insects (Coccoidea), whiteflies (Aleyrodoidea) or mites (Acari). Largely used in biological control programmes, ladybeetles render important ecosystem services to agriculture and silviculture (Ameixa et al. 2018). In fact, the first successful case of classical biological control dates back to the late 1800s, when Novius (=Rodolia) cardinalis (Mulsant) was introduced in California from Australia to control the cottony cushion scale, Icerya purchasi Maskell, also of Australian origin (Caltagirone et al. 1989). The introduction of this ladybeetle in Portugal in 1897 was the first case of classical biological control in Europe (Amaro 1994). However, ladybeetles also have other food habits and a few species are herbivores causing heavy crop damages in Asia, America and Africa (Barrigossi et al. 2003;Beyene et al. 2007;Das et al. 2012).
Scientific evidence shows that human activities have huge negative impacts on biodiversity (e.g., Vitousek et al. 1997;Newbold et al. 2015;Jung et al. 2019). The structure and composition of insect communities have been particularly affected by these activities, with ecological domino effects along trophic chains (Dyer et al. 2003). Consequently, action in favour of insect conservation and recovery has been claimed (e.g., Harvey et al. 2020). Ladybeetle communities are no exception. For instance, several studies report long-term variations in the composition of ladybeetle communities fol-lowing the introduction of Harmonia axyridis Pallas around the world . Honӗk et al. (2014Honӗk et al. ( , 2017 included agricultural intensification, habitat (particularly urbanization) and climate changes as additional causes for ladybeetle community changes. In this context, and because it is important that each country should have an updated list of its fauna, from which we can detect changes in biodiversity and implement conservation and recovery programmes, we herein address the issue of the coccinellid fauna of Portugal, within the research project AZORESBIOPORTAL-PORBIOTA (ACORES-01-0145-FEDER-000072).
Based on the literature and unpublished data, we present an updated, comprehensive checklist of ladybeetles of Portugal, including the Azores and Madeira Archipelagos.

Materials and methods
In this work we follow the suprageneric classification of Coccinellidae proposed by Che et al. (2021). This recent revised classification recognises three subfamilies: Microweiseinae, Monocoryninae stat. nov., and Coccinellinae. It should be noted that the tribe Coccidulini sensu Seago et al. (2011), which comprises several species for Portugal, was redefined by Che et al. (2021) in which Scymnini and Stethorini were split independently.
Current taxonomic affiliation follows Kovář (2007). Species cited for Portugal after Kovář (2007) or those for which the taxonomic position has changed as a result of more recent studies, are indicated here following more recent publications.
The species are listed in alphabetic order according to their valid tribe and genera. Species of the genera Nephus and Scymnus are listed under the respective subgenera because they are particularly important for their identification. Synonymy is mentioned but is restricted to the original name.
For each species, we specify the regions where the species were recorded (Mainland, Azores and/or Madeira), under "Distribution" and we provide brief notes about the status of the species in each region, possible taxonomic incongruences, and the current biogeographical (limits as in Löbl and Smetana 2007) distribution under "Comments". New records are marked with a black spot ( • ), doubtful records with an asterisk (*) and exotic species with a dollar ($). The bibliographic references associated with each species recorded for Portugal are presented in Table 1.
Comments. Currently established in Palearctic, Afrotropical, Nearctic and Neotropical regions. This exotic species of Australian origin, used around the world for biological control since the 19 th century, was introduced from France in the early 20 th century for the biological control of the citrus mealybug, Planococcus citri (Risso) ), but the first record in Europe was in Italy, 1908 (Roy and Migeon 2010).
Comments. This is an endemic species of the Iberian Peninsula, associated with maritime pine forests, and is a specialist predator of the maritime pine bast scale, Matsucoccus feytaudi Ducasse Tavares et al. 2015a;Tavares et al. 2015b). Adults and mostly larvae were shown to be attracted by the sex pheromone of their prey .
Comments. Palaearctic, Nearctic and Australian distribution. This exotic species of Australian origin was introduced in Europe in the 1980´s for the biological control of scale insects (Coccoidea), and became established in different countries, including Italy, France, Greece and Albania (Roy and Migeon 2010;Soares et al. 2018). The first record in Europe was in Italy in 1982 (Roy and Migeon 2010). It was recently recorded in the Azores .

Coccinella septempunctata Linnaeus, 1758
Distribution. Mainland, Madeira and Azores*. Comments. Palearctic, Afrotropical, Nearctic and Oriental distribution. Relatively important species in cereal crops in the Azores, especially in the first half of the 20 th century, having eventually disappeared when these crops became scarce Soares et al. 2017). The taxonomic status of C. algerica has been under discussion because of its morphological similarities to the geographically widespread Coccinella septempunctata L. Although Lecompte et al. (2016) revealed a high genetic structuring pattern, with an Algerian rear-edge population highly differentiated, consistent with their morphological distinctiveness, a recent study by Romanowski et al. (2019) demonstrated that individuals from Canarian populations, usually classified as C. algerica, can hybridise with individuals from European populations of C. septempunctata giving rise to fertile F1 descendants. These authors therefore propose to synonymise C. algerica with C. septempunctata but, taking into account the morphological peculiarities of the North African and the Canarian populations, they consider that this species is a subspecies: Coccinella septempunctata algerica Kovář, 1977.
Comments. Palaeartic, Australian and Nearctic distribution. In the Azores, it is a threatened species due to anthropogenic pressures on the coastal areas .
Comments. Palearctic and Afrotropical distribution. Although previously recorded in the Madeira archipelago, there are doubts regarding its present occurrence. It might have been introduced but did not establish (Franquinho Aguiar, personal communication).
Comments. Palearctic distribution. Although previously recorded for the Madeira archipelago, there are doubts as to its present occurrence. It might have been introduced but did not establish (Franquinho Aguiar, personal communication).
Comments. Of Neotropical origin where it is very common. Although previously reported for the Azores, it did not become established (fide A. O. Soares, after intensive surveys).
Comments. Palearctic distribution.  Comments. Established in Palearctic/Madeira, originally from Afrotropical region. The first record for Europe was in 2002, in Madeira archipelago (Roy and Migeon 2010). In Madeira, it was introduced as a biological control agent against Insignorthezia insignis (Browne 1887).
Comments. Palearctic distribution. According to Eizaguirre (2015), this species does not exist in the Iberian Peninsula and has been mistaken for the endemic species, H. illecebrosa.

Tribe NOVIINI $ Novius cardinalis (Mulsant, 1850), following Pang et al. (2020)
= Vedalia cardinalis Mulsant, 1850 Distribution. Mainland, Madeira and Azores. Comments. Established in Palaeartic, Afrotropical, Nearctic and Neotropical, Oriental. Native to Australian region. This exotic species was introduced in California and South Africa in the 1890´s for the control of the cottony cushion scale, Icerya purchasi Maskell (Roy and Migeon 2010). The first introduction in Europe was made in Portugal, 1897 (Amaro 1994;Roy and Migeon 2010). Rodolia cardinalis is the widely known name and it was included in the genus Novius by Pang et al. (2020).
Comments. Palearctic and Afrotropical distribution. This species was first reported as N. (bipunctatus) includens (Kirsch 1871) based on adults emerged from larvae collected in 1988 from citrus fruits infested with mealybugs, in the South of Portugal (Algarve) (Raimundo 1992). However, Eizaguirre (2015) indicates that N. (bipunctatus) includens is a junior synonym of N. conjuntus. Taking this into account and knowing that specimens previously collected in 1984 in the Algarve and identified as N. quadrimaculatus Herbst were in fact shown to be N. includens (Raimundo and Alves 1986;Magro et al. 1992;Raimundo 1992), we have to consider that the first report of this species dates from 1984. The distribution is apparently restricted to the Algarve . (Sicard, 1923) = Scymnus peyerimhoffi Sicard, 1923 Distribution. Mainland.
$ Nephus (Geminosipho) reunioni (Fürsch, 1974a) = Scymnus reunioni Fürsch, 1974a Distribution. Mainland, Azores and Madeira • . Comments. Palearctic and Afrotropical distribution. The first record in Europe was in France in 1983 (Roy and Migeon 2010). This species of Afrotropical origin was imported from France (laboratory rearing in Antibes) in the early 1980´s and released in Oeiras, in 1984. Its presence in Portugal was detected for the first time in 1990, in citrus orchards in the Setúbal region . In his Catalogue of the African species of the genus Nephus, Fürsch (2007) mentions that the distribution of N. reunioni is apparently restricted to Reunion Island and Mauritius, and that the references to its presence in South Africa and other Mediterranean countries, such as Portugal and Israel, are due to misidentifications by the author himself, which most likely correspond to N. derroni Fürsch 1974b, a species described from São Tomé Island and that is common in South Africa. However, Magro et al. (2020a) analysed specimens from Portugal and showed that they corresponded to the original description of N. reunioni made by Fürsch in Chazeau et al. (1974). Raimundo (1992), who first described N. reunioni for Portugal, also illustrated the external morphology and genitalia corresponding to the original description by Fürsch in Chazeau et al. (1974). In both cases, the observations showed that the specimens from the Portuguese population are distinct from N. derroni. In Madeira, collected by Aguiar and Jesus, in  Comments. Afrotropical origin. Very recently, two parthenogenetic populations of this species were found in the Azores and Mascarene archipelagos, becoming the first reported case of asexuality in the Coccinellidae . Observations by António Onofre Soares, in September of 1997, Madeira Island (Anjos; approximately at 32°41'15"N, 17°06'54"W; Faial approximately 32°47'24"N, 16°51'02"W; Caniçal approximately 32°44'49"N, 16°44'26"W), and Alexandra Magro and Miguel Sequeira in September 2018, from herbaceous plants (Anjos 32°69'11"N, 17°11'96"W; Ribeira de Natal, Caniçal 32°73'57"N, 16°74'62"W; Ribeira Brava 32°66'98"N, 17°06'09"W; Fajã dos Padres 32°65'45"N, 17°02'13"W). New record for Madeira.
Comments. Palearctic and Afrotropical distribution.
In Memoriam of Armando Américo Cardoso Raimundo (2.V.1942-9.Xi.2019): A farewell to colleague and friend. Thanks are due to Claudio Canepari for helping in the identification of Microserangium sp. and Jaroslav Větrovec of Harmonia axyridis Pallas from Madeira. This study was financed by FEDER in 85% and by Azorean Public funds by 15% through Operational Program Azores 2020, under the following projects AZORESBIOPORTAL -PORBIOTA (ACORES-01-0145-FEDER-000072), and under the project ECO2 -TUTA (ACORES-01-0145-FEDER-000081). The Forest Research Centre is funded by Fundação para a Ciência e a Tecnologia I.P. (FCT), Portugal (UIDB/00239/2020). Thanks are also due to FCT/MCES for financial support to CESAM (UIDB/50017/2020+UIDP/50017/2020) through national funds, and the co-funding by the FEDER, within the PT2020 Partnership Agreement and Compete 2020. OMCCA is funded by national funds (OE), through FCT, in the scope of the framework contract foreseen in the numbers 4, 5 and 6 of the article 23, of the Decree-Law 57/2016, of August 29, changed by Law 57/2017, of July 19. AM was supported by the "Laboratoires d'Excellence" LabEx TULIP (ANR-10-LABX-41). The Open Access of this manuscript was supported by the project FCT-UIDB/00329/2020-2024.