Taxonomic identification/inventory of Algerian Ants assisted by a molecular barcoding and phylogenetic approach using COX1 mitochondrial sequences

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Oussalah N, Salem Attia S, Ainouche A, Marniche F, 2025. Taxonomic identification/inventory of Algerian ants assisted by a molecular barcoding and phylogenetic approach using COX1 mitochondrial sequences. Museu de ciències naturals de Barcelona, checklistdataset: https://doi.org/

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關鍵字

Occurrence; Formicidae; Algerian ants; biodiversity; mitochondrial COXI gene subunit 1; DNA barcode; phylogenetic relationship; Specimen

聯絡資訊

地理涵蓋範圍

The samples were collected from various agricultural and urban ecosystems across different regions of Algeria (Figure 1). The specific sites are as follows: In the north, sampling was conducted at the Institute of Technology on Specialized Agriculture (ITMAS Heuraoua) in a 60-year-old Citrus orchard located in the humid eastern Sahel-Mitidja region, 30 km from Algiers. The agricultural area spans 17.25 hectares, with four hectares dedicated to around 180 sweet orange trees (Citrus sinensis) of four varieties. Another northern site was the Bainem forest near Algiers, covering approximately 500 hectares of hilly terrain (80-320 meters altitude). This ecosystem is dominated by Mediterranean species like Aleppo pine (Pinus halepensis), cork oak (Quercus suber), and maquis shrubs, and is rich in biodiversity but faces significant human pressures from fires, urbanization, and pollution. Further sampling took place in the southeastern region of Lakhdaria (Bouira Province), which has a sub-humid climate, focusing on a garden with rose bushes and an urban area. In the central region, a site was selected in the cold, dry steppes of the Saharan Atlas (Ouled Nail Mountains), about 2 km west of Djelfa. This extensive area of approximately 200,000 hectares is 60% covered by Aleppo pine, with associated plant species including evergreen oak, cade juniper, red juniper, rosemary, and alfa steppe at its border. In eastern Algeria, sampling focused on a cereal-growing area in Sidi Moussa Medjana (Bordj-Bou-Arréridj Province), located in a semi-arid high plain. Another site was the Chrea Forest, part of a National Park in the Tell Atlas (Blida Province), at an altitude of 1,200 to 1,800 meters. This massif is dominated by Atlas cedar (Cedrus atlantica), accompanied by oaks (Quercus spp.), Austrian black pines, and an undergrowth rich in aromatic and herbaceous plants. Sampling was also carried out within the Yakouren forest massif (Tizi-Ouzou Province), a dense forest with poorly developed undergrowth composed of mixed oaks: Afar oak (Quercus afares Pomel), Zeen oak (Q. canariensis), and Portuguese oak (Q. faginea Lam.). The final sampling site was in the Saharan region of Ouargla at the station of the Technical Institute for the Development of Saharan Agronomy (ITDAS), where ecological conditions are homogeneous and the vegetation is diverse, including underlying crops like alfalfa and plasticulture for vegetable crops. A total of 61 Algerian ant samples used in this study are listed in Table 1, with supplementary information on their taxonomic identification and origin in Table S1. Ant sampling was conducted over three years (2017-2019) using two complementary methods to capture a comprehensive representation of species and castes, including workers and queens. The first method, employed monthly at the ITMAS Heuraoua site in Bordj-Bou-Arréridj, involved standard pitfall trapping. Following established protocols (e.g., Hernández-Ruiz and Castaño-Meneses, 2006), ten metal containers (10.4 cm diameter) were deployed in a single line under trees at 5 m intervals. These traps, filled with water and a drop of liquid detergent to break surface tension, were left in situ for 48 hours before collection. The second method, active hand collection, was implemented in additional regions mentioned above during 2018 and 2019 to intentionally target a wider variety of ant species. All captured specimens from both methods were preserved in 70% alcohol for subsequent sorting and identification to species level in the laboratory based on morphological characters. Specimens were studied under a Leica stereomicroscope and identified morphologically using available keys (Cagniant, 1996, 1997, 2009; Bernard, 1968; Bolton, B. 1995; Seifert, 2021, 2023, 2024; Schifani, 2022; Menschetti, 2021) as well as the following Ant sites: http://www.antcat.org/ and https://www.antweb.org/contact.jsp. Individuals representing all samples were conserved in 90% ethanol under freezing temperature to prevent degradation of DNA until used for molecular analyses.

分類群涵蓋範圍

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計畫資料

Algeria is well known for its eco-geographical diversity, which is regarded as a biodiversity hotspot that urgently needs particular attention for rational conservation of endangered organisms and ecosystems (Médail et al., 2001). Located in North Africa, in the southern and occidental part of the Mediterranean basin, Algeria experiences both continental and temperate climates. As a result of this combination, ecosystems are diverse and highly contrasted, including coastal and inland plains, mountains, steppes, deserts (Erg and Reg), and bioclimatic stages ranging from humid to arid (Dubost, 1989; ANAT 2004). This diversity of ecosystems provides habitats for a wide variety of species, particularly arthropods, including ants. Ants are the most species-rich of all social insects (Hölldobler and Wilson, 1990), with approximately 14,700 described species of ants (http://www.antcat.org/) and many more awaiting description. This group of insects is considered an indicator of biodiversity (Alonso, 2000). Indeed, they have the advantage of being abundant in most terrestrial ecosystems and have colonized all terrestrial regions except Greenland and Antarctica (Cagniant, 1973). The same author points out that ants can be found everywhere in Algeria, where it is not too cold: in forests, open areas, along permanent or temporary watercourses, in dry areas, on clay, and in rocky habitats. Ants are known to play a vital role in soil ecology, moving more soil than earthworms or termites (Pablo, 2004). The accumulation of organic matter within or near colonies also contributes to soil enrichment with nitrogen and phosphorus, which are essential for the growth of many plants (Beattie & Hughes, 2002). Bernard (1968) indicates that ants are essential for maintaining biological balances by acting as predators and parasites. Through these direct mechanisms—hunting and parasitizing other creatures—they effectively control the populations of other organisms, thereby promoting overall ecosystem stability. They can be considered ecosystem engineers (Folgarait, 1998), playing a key role in the animal and plant communities, they interact with (Dejean et al., 2007). Moreover, ants are easily transported by humans due to their small size and their nest type. Overall, more than 200 species have established populations outside their native zone, and over 600 have already been introduced away from their native range (Miravete et al., 2013). Many established non–native species (i.e., species that have completed the second step of the dispersal process or 'exotic established species according to Wittenborn and Jeschke (2011), possess invasive attributes and constitute a major part of the global environmental change. Such species have the potential to spread, often considerably, after establishment and can affect native ecosystems, causing devastating ecological consequences (e.g., Mack et al., 2000; Mooney and Hobbs, 2000; Pimentel et al., 2005). Therefore, continuously studying and updating our understanding of ant diversity, along with assessing how their distribution changes across space and time, is crucial for understanding their influence on the structure and dynamics of natural, agricultural, and urban ecosystems and communities. To date, relatively few studies have been conducted on Algerian myrmecofauna over the last hundred years. The first publications on entomology began with Seurat (1924) in his study "Forestry Zoology in Algeria," followed by the work of Cagniant (1966, 1968, 1969, 1970, 1973), Dartigues (1988), Doumandji and Mitiche (1988), Belkadi in the Kabylie region (1990). Barech (1999, 2005, 2014) studied ants in the Algiers Sahel, Bouzekri et al. (2011, 2014), Amara (2013) and Amara et al. (2020) in the steppe and southern Algeria, Chemala et al. (2017) and Chemala et al. (2019) in the northeastern Sahara, and Oussalah et al. (2019, 2021) investigated exotic ants. While interest in Algerian ants has grown in recent times, our knowledge and understanding of Algerian ants remain modest and limited, considering the vast geographical extent of the country and its remarkable diversity of ecosystems, which encompasses marine, coastal, forest, and mountain environments, as well as steppe, desert, wetland, and arid zone environments (Abdelguerfi et al., 2009). All these factors contribute to the great diversity of insect fauna, including ants (Cagniant, 1973), and the high degree of endemic and exotic species. The known Algerian myrmecofauna includes 218 species divided into eight subfamilies and 29 genera. The Myrmicinae subfamily is the richest and most diverse, containing 18 genera (61%) and 133 species (48%). Formicinae is the second leading ant group, with 10 genera (32%) and 70 species (24%). These two subfamilies alone account for over 93% of the total species. The other subfamilies appear to be less well represented or inhabit ecosystems in Algeria (Antweb, 2022). These data, while comprehensive, may not fully reflect the current diversity of ants in Algeria. Recent studies continue to reveal new records, leading to a steady increase in the documented number of species. For instance, Barech et al. (2016, 2020) not only contributed a list of the ant fauna from the Chott El Hodna salt lake area but also described a new species for science, Messor hodnii (Barech, Cagniant & Espadaler, 2020). Furthermore, Oussalah et al. (2018, 2021) reported two new exotic species for the country, Nylanderia jaegerskioeldi (Mayr) and Pheidole indica (Mayr). Similarly, Slimani et al. (2020) confirmed new reports of the highly invasive Argentine ant, Linepithema humile (Mayr, 1868). Compared to Algeria's neighboring countries with similar climatic conditions, it is of interest to notice that more species (237) belonging to 38 genera have been recognized in Morocco (Taheri et al., 2018), whereas a much higher number of species (295) was recorded in the Iberian Peninsula (Roig & Espadaler, 2010). Therefore, it can be expected that extensive fieldwork with a more systematic exploration of the diversity of habitats should allow us to supplement the ant inventories as much as possible and likely discover new populations and species in Algeria. This study is part of the efforts developed to improve knowledge and update inventories of the Algerian myrmecofauna, and more broadly of the North African region. In particular, the aim of this work consisted in the taxonomic identification of the diversity of ant specimens collected from various ecosystems, employing traditional morphological methods assisted here, for the first time in Algeria, by integrating a DNA barcoding and phylogenetic approach using sequences of the mitochondrial gene Cytochrome oxidase subunit 1 (COX1) (Hebert et al., 2003a, b). Such an approach not only has the advantage of combining different types of characters but of providing an integrative evolutionary framework for the circumscription, or even the discovery, of species or lineages in order to objectively evaluate their identification, their taxonomic assignment (or reassignment) and their evolutionary relationships and history (Hebert et al., 2003a, b; Johnson et al., 2003; Dayrat, 2005; Cardoso et al., 2009; Padial et al., 2010; Seifert et al., 2017).

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引用文獻

1. Oussalah N, Salem Attia S, Ainouche A, Marniche F, 2025. Taxonomic identification/inventory of Algerian ants assisted by a molecular barcoding and phylogenetic approach using COX1 mitochondrial sequences. Arxius de Miscel·lània Zooloògica 23, xxx-xxx. DOI: 10.32800/amz.2025.23.0xxx https://doi.org/10.32800/amz.2025.23.0xxx

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