Ingestion of microplastics and microfibers by the invasive blue crab Callinectes sapidus (Rathbun 1896) in the Balearic Islands, Spain.

The blue crab Callinectes sapidus Rathbun, 1896 is native to the western coasts of the Atlantic Ocean and is currently considered an invasive species in the Mediterranean Sea. In this study, we examined the stomach contents of C. sapidus to determine the frequency of occurrence of microplastics (MPs) and microfibers (MFs) in the Balearic Islands archipelago in the western Mediterranean Sea. A total of 120 individuals were collected from six locations between 2017 and 2020. Overall, 65.8% of the individuals had MPs and/or MFs particles with an average of 1.4 ± 1.6 particles ind.-1 of which an average of 1.0 ± 1.3 items ind.-1 were MFs and an average of 0.4 ± 0.8 items ind.-1 were MPs. In terms of type, fragments were the dominant type of MPs and the most common size of items ranged from between 0.5 and 1 mm (40%) followed by 1-5 mm (31%). The most prevalent polymers were low-density polyethylene (39%) and high-density polyethylene (26%). In terms of links to human activities, MP ingestion was positively correlated with an increase in drain pipes, whereas MF ingestion was positively correlated with an increase in sewage pipelines, providing evidence of potential sources and the bioavailability of these particles in various environments. This study confirms the widespread presence of MP and MF particles, even in areas that are currently managed under different protection statuses, in the stomach contents of invasive blue crab species throughout coastal communities.

Microplastic Presence in the Digestive Tract of Pearly Razorfish Xyrichtys novacula Causes Oxidative Stress in Liver Tissue.

Plastic pollution in the oceans is a growing problem, with negative effects on exposed species and ecosystems. Xyrichtys novacula L. is a very important fish species both culturally and economically in the Balearic Islands. The aim of the present study was to detect and categorise the presence of microplastics (MPs) in the digestive tract of X. novacula, as well as the existence of oxidative stress in the liver. For this purpose, the fish were categorised into two groups based on the number of MPs observed in the digestive tracts: a group with no or low presence of MPs (0-3 items) and a group with a higher presence of MPs (4-28 items). MPs were found in 89% of the specimens analysed, with a dominance of fibre type and blue colour. Regarding the type of polymer, polycarbonate was the most abundant, followed by polypropylene and polyethylene. For the group with a greater presence of MPs, the activities of the antioxidant enzymes glutathione peroxidase and glutathione reductase, as well as the phase II detoxification enzyme glutathione s-transferase, were higher than the activities observed in fish with little to no presence of MPs. The activities of catalase and superoxide dismutase and the levels of malondialdehyde did not show significant differences between both groups. In conclusion, these results demonstrate the presence of MPs in the digestive tract of X. novacula and the existence of an antioxidant and detoxification response, mainly based on the glutathione-based enzymes.

Oxidative stress and behavioral responses of moorish geckos (Tarentola mauritanica) submitted to the presence of an introduced potential predator (Hemorrhois hippocrepis).

Stressful situations induce an increase in the production of reactive oxygen species (ROS) which can lead to molecular damage and alteration of cell function. The introduction of new potential predators induces physiological stress in native fauna. However, behavioral responses have been reported in preys, demonstrating an induction of the defenses against alien species. Behavioral and antioxidant enzyme responses in the moorish gecko, Tarentola mauritanica, against the invasive predator horseshoe whip snake (Hemorrhois hippocrepis) were assessed. Behavior was recorded and a tissue sample from the tail was collected after placing the gecko in a terrarium with previous absence or presence of the snake in 'Control' and 'H. hippocrepis' groups, respectively. Fifteen behavioral variables were examined, including tongue flick (TF) and locomotion patterns. Antioxidant enzyme activities -catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione reductase (GR)-, and the levels of reduced (GSH) and oxidized glutathione (GSSG), glutathione/glutathione disulfide ratio (GSH/GSSG) and malondialdehyde (MDA) concentrations were measured in the tissue sampled. Geckos exposed to the snake's odor showed a higher number of TF, longer amounts of time remaining motionless or moving in slow motion and they spent less time on the ground in comparison to the 'Control' group. The presence of the snake produced a significant increase in the activities of CAT, SOD and GR and a decrease in the GSH/GSSG ratio in T. mauritanica individuals exposed to the snake's scent. Thus, both behavioral responses and oxidative stress biomarkers clearly showed that T. mauritanica is able to recognize H. hippocrepis as a potential predator, despite being a recently introduced snake at the Balearic Islands.

Evaluating the Presence of Marine Litter in Cetaceans Stranded in the Balearic Islands (Western Mediterranean Sea).

The global distribution and presence of plastic, at all levels of the water column, has made plastic debris one of today's greatest environmental challenges. The ingestion and entanglement of plastic-containing marine debris has been documented in more than 60% of all cetacean species. In light of the increasing pressure on cetaceans, and the diversity of factors that they face, the aim of this work is to provide evidence of the impact of plastic debris on stranded cetaceans, in terms of ingestion and entanglement, in the Balearic Islands for the first-time. Detailed examinations, necropsies, and plastic debris analysis were performed on 30 of the 108 cetaceans stranded between 2019 and 2022. Specimens belonging to five different species, Stenella coeruleoalba, Tursiops truncatus, Grampus griseus, Balaenoptera physalus, and Physeter macrocephalus, were evaluated. Ten percent of the cetaceans (N = 3) presented plastic debris in their stomach, with one case of obstruction and perforation. Fishery gear fragments (ropes and nets) were found in two adults of T. truncatus, whereas packaging debris (plastic bag, packing straps, and plastic sheets) were found in a juvenile P. macrocephalus. Plastic items analysed by Fourier transform infrared spectroscopy (FT-IR) reported three polymer types: polypropylene, polyamide, and high-density polypropylene. A total of seven cases of entanglement were recorded during the study, affecting four different species (S. coeruleoalba, T. truncatus, P. macrocephalus, and Megaptera novaeangliae). Only two individuals were freed from the nets, although one died after a week, whereas the rest were already found dead. In conclusion, data collected in the present study provided evidence of plastic ingestion and entanglement in cetaceans of the Balearic Islands for the first-time, thus highlighting the need for the regular examination of stranded cetaceans (as they are top predators) in future research to better understand the effects of these pollutants.

Effects of Human Activity on Markers of Oxidative Stress in the Intestine of Holothuria tubulosa, with Special Reference to the Presence of Microplastics.

Pollution in the seas and oceans is a global problem, which highlights emerging pollutants and plastics, specifically microplastics (MPs), which are tiny (1 µm to 5 mm) ubiquitous plastic particles present in marine environments that can be ingested by a wide range of organisms. Holothurians are benthic organisms that feed on sediment; therefore, they can be exposed to contaminants present in the particles they ingest. The objective was to evaluate the effects of human activity on Holothuria tubulosa through the study of biomarkers. Specimens were collected in three different areas throughout the island of Eivissa, Spain: (1) a highly urbanized area, with tourist uses and a marina; (2) an urbanized area close to the mouth of a torrent; (3) an area devoid of human activity and considered clean. The results showed a higher presence of microplastics (MPs) in the sediments from the highly urbanized area in relation to the other two areas studied. Similarly, a higher number of MPs were observed in the digestive tract of H. tubulosa from the most affected area, decreasing with the degree of anthropic influence. Both in the sediment and in the holothurians, fibers predominated with more than 75% of the items. In the three areas, mesoplastics were analyzed by means of FTIR, showing that the main polymer was polypropylene (27%) followed by low-density polyethylene (17%) and polystyrene (16%). Regarding the biomarkers of oxidative stress, the intestine of H. tubulosa from the most impacted areas showed higher catalase, superoxide dismutase (SOD), glutathione reductase (GRd), and glutathione S-transferase (GST) activities and reduced glutathione (GSH) levels compared to the control area. The intermediate area only presented significant differences in GRd and GST with respect to the clean area. The activities of acetylcholinesterase and the levels and malondialdehyde presented similar values in all areas. In conclusion, human activity evaluated with the presence of MPs induced an antioxidant response in H. tubulosa, although without evidence of oxidative damage or neurotoxicity. H. tubulosa, due to its benthic animal characteristics and easy handling, can be a useful species for monitoring purposes.

Microplastic presence in the pelagic fish, Seriola dumerili, from Balearic Islands (Western Mediterranean), and assessment of oxidative stress and detoxification biomarkers in liver.

Microplastics (MPs) are characterized by their high persistence in marine ecosystems, and due to their small size, they can be easily ingested by very diverse organisms. Although the presence of MPs in wild fish is well documented, there is still limited information on their potential to induce adverse effects. Pelagic fish species, because of their wide distribution, are considered good bioindicators for monitoring environmental pollution of marine ecosystems. This study investigated the presence of MPs in the gastrointestinal tract of the predatory pelagic fish (Seriola dumerili) in the Balearic Islands (Mediterranean Sea), and the possible relationship with oxidative stress through the analysis of biomarkers in liver tissue. The results showed the presence of MPs in 98% of total samples examined (n = 52) with an average of 12.2 ± 1.3 MPs/individual. A greater amount of fibre-like particles was isolated compared to fragments. No correlation between the presence of MPs in the gastrointestinal contents and the size of the fishes was noted. Antioxidant enzymes (superoxide dismutase and catalase) and the phase II detoxification enzyme glutathione-S-transferase showed increased activities in fish with higher MPs load. The activity ethoxyresorufin-O-deethylase and the levels of malondialdehyde were similar in both groups. In conclusion, the present results provide an important database on the assessment of the presence of MP debris in S. dumerili gastrointestinal tract and, the potential capability to cause oxidative stress.

First detection of microplastics in Xyrichtys novacula (Linnaeus 1758) digestive tract from Eivissa Island (Western Mediterranean).

Plastic waste and its ubiquity in the oceans represent a growing problem for marine life worldwide. Microplastics (MPs) are ubiquitous in the sea and easily enter food webs. Xyrichtys novacula L. is one of the main target species of recreational fishing in the Balearic Islands, Spain. In the present study, the quantity of MPs in gastrointestinal tracts of X. novacula from two different areas (a marine protected area (MPA) and a non-protected area) of Eivissa Island (in the Balearic archipelago) has been assessed, as well as MPs evaluation within the sediment of both areas. The results showed that over 80% of sampled individuals had MPs in their gut with an average of 3.9 ± 4.3 plastic items/individual. Eighty percent of these plastics were fibres, while the rest were fragments. Although the sediment of the non-protected area had a significant higher presence of MPs, no significant differences in the number of MPs were observed in X. novacula from both areas. The µ-FT-IR analysis showed that the main polymers in the sediments were polycarbonate (PC) and polypropylene (PP), whereas in the digestive tract of fish PC, PP, polyethylene, polystyrene and polyester. In conclusion, practically all X. novacula specimens presented MPs in their digestive tract regardless if the capture zone was in a MPAs or not. These results highlight the ubiquity of MPs in coastal marine areas, and further studies might be necessary to evaluate further implications of MP presence in this species.

Ingestion and characterization of plastic debris by loggerhead sea turtle, Caretta caretta, in the Balearic Islands.

Plastic waste has become ubiquitous pollutants in seas and oceans and can affect a wide range of species. For some marine species, plastic debris could pose a considerable threat through entanglement, ingestion, and habitat degradation and loss. Sea turtles are one of the most sensitive species, as their migratory behaviour and multifaceted life cycles make these reptiles especially vulnerable to the negative effects of plastic debris. The present study aimed to assess the amount and composition of plastic debris ingested by loggerhead turtles (Caretta caretta, Linnaeus, 1758) in the Balearic Islands Sea, thusly providing new information to complete the knowledge for this topic. In this work, 45 stranded dead C. caretta specimens were necropsied, and their digestive tract content analysed for the presence of plastic debris. Plastic objects were observed in 27 individuals (60.0%), with an average of 12.7 ± 4.7 plastic items per turtle. Litter in the faecal pellet was also monitored in 67 living individuals, observing plastic elements in 46 (68.7%) of the specimens, reporting an average of 9.7 ± 3.3 plastic elements per individual. Overall, 785 plastic items were found, measured, weighed and categorized according to size, colour, shape, and type of polymer. The main elements ingested were plastic sheets that were found in 65.3% of the turtles analysed, being white (42.7%) and transparent (29.2%) the most predominant colours. Most elements were macroplastics (59.3%), while microplastics were not found. Fourier Transform Infrared Spectrometry (FT-IR) analysis showed that high-density polyethylene and polypropylene were the main polymer plastics, representing 42.3% and 33.8% of the total, respectively. In conclusion, the high occurrence of plastic debris determined in the present study evidenced for the first time plastic ingestion in loggerhead turtles in the Balearic Islands, and highlights C. caretta as a bioindicator organism for marine pollution.

Hypotensive Effects of the Triterpene Oleanolic Acid for Cardiovascular Prevention.

BACKGROUND: Hypertension is a highly prevalent chronic disease worldwide and a major cardiovascular risk factor. Oleanolic acid (3ß-hydroxy-olea-12-en-28-oic acid) is a widely distributed bioactive pentacyclic triterpenoid with diverse biological activities such as anti-inflammatory, hepaprotective, anti-diabetic or anti-hypertensive. OBJECTIVE: The aim of this study was to review and highlight the available data regarding the antihypertensive activity of oleanolic acid and the described mechanisms of action. METHODS: Extensive searches were made in the available literature on oleanolic acid and the data investigating its antihypertensive effects were analysed. RESULTS: Most of the research has been performed on animal models of hypertension, ex vivo studies with aortic ring and some in vitro tests with cell cultures, whereas clinical trials are still lacking. Treatment of hypertensive animals with oleanolic acid significantly ameliorated the rise in the systolic blood pressure. In addition, the hypotensive effects of oleanolic acid are also related to a potent diuretic-natriuretic activity and nephroprotection. In vitro studies have characterized the participation of various signalling pathways that modulate the release of vasodilation mediators. CONCLUSION: In vitro and in vivo studies suggest that oleanolic acid effectively reduces blood pressure and could be an interesting co-adjuvant to conventional treatment of hypertension.

Physiological biomarkers in loggerhead turtles (Caretta caretta) as a tool for monitoring sanitary evolution in marine recovery centres.

The loggerhead turtle, Caretta caretta, is a very vulnerable species to human action which means that numerous specimens arrive at the recovery centres to be treated until they can be returned to the natural environment. The aim of the present study was to investigate the physiological evolution of C. caretta specimens that have entered a recovery centre by using oxidative stress biomarkers. Plasma and peripheral blood mononuclear cells of specimens were obtained at different periods: the day of arrival at the recovery centre (day 1), at 3, 9, and 30 days later, and a final sample collected before the animal was returned to the sea. The average residence time in the centre until the return to the sea was 58.5 ± 6.1 days. The activities of antioxidant enzymes - catalase (CAT), superoxide dismutase (SOD), glutathione reductase, and glutathione peroxidase activities in immune cells and CAT and SOD in plasma - progressively decreased throughout the recovery time. Similarly, H2O2 production by immune cells after lipopolysaccharide (LPS) and zymosan activation progressively decreased with the recovery process. Also, malondialdehyde (MDA), as a marker of lipid peroxidation, and the activity of the pro-oxidant myeloperoxidase were significantly decreased throughout the recovery process. In conclusion, the results evidenced that the turtles presented a high level of oxidative stress upon arrival at the recovery centre, which was normalized along with their rehabilitation. Oxidative stress biomarkers are a good tool to monitor the recovery process in C. caretta complementary to the veterinary control.

First report of heavy metal presence in muscular tissue of loggerhead turtles Caretta caretta (Linnaeus, 1758) from the Balearic Sea (Balearic Islands, Spain).

The concentrations of cadmium (Cd), mercury (Hg) and lead (Pb) were determined in muscular tissue of eleven loggerhead turtles (Caretta caretta) from the Balearic Islands (Spain, Western Mediterranean). The metal levels found in the present study were similar or lower than concentrations detected in Andalusia (mainland Spain), Italy, Canary Islands (Spain) or Japan. As the main source of metals in the loggerhead turtle is the diet, low metal burdens could be explained by its opportunistic feeding way. No significant differences were found in metal concentrations between juveniles and subadults in any of the heavy metals analysed. Furthermore, no significant correlation was detected between heavy metal concentrations and straight carapace length (SCL) of the studied individuals. These results could derive from the homogeneity in age and size of the turtles sampled, so further studies including adults are needed in order to assess the heavy metal accumulation with turtle growth.

Long-term exposure to microplastics induces oxidative stress and a pro-inflammatory response in the gut of Sparus aurata Linnaeus, 1758.

Environmental pollution from plastic debris is a major global concern, being a potential threat to marine organisms and ecosystems. The accumulation of microplastics (MPs) in the oceans has notable ecological implications due to their long persistence, their potential ecotoxicity, and their ability to adsorb other pollutants and act as vectors of pathogens. Nevertheless, whereas the number of investigations documenting the presence of MPs in wild fish has increased, less studies have addressed the toxicological effects associated with the ingestion of MPs in long-term laboratory conditions. The aim of the present study was to assess the physiological response of gilthead seabream (Sparus aurata) exposed to low-density polyethylene (LDPE) MPs during a 90-day exposure followed by an extra 30 days of depuration through the application of oxidative stress biomarkers in the gut. No changes were observed in the Fulton condition factor of fish associated with MP intake. The activities of antioxidant enzymes and glutathione s-transferase and the levels of reduced glutathione progressively increased throughout the study in the MPs-fed group compared to the control group, reaching the highest values at 90 days. Similarly, the activity of the pro-inflammatory enzyme, myeloperoxidase, and the levels of oxidative damage markers -malondialdehyde and protein carbonyls-also increased after 90 days of exposure to an enriched diet with MPs. During the 30-day depuration period, all the biomarkers analysed tended to normalize, with the majority recovering values similar to those of the control group. In conclusion, MPs exposure during 90 days to S. aurata induced oxidative stress and a pro-inflammatory response in gut, and were able to recover after the exposure to MPs was removed.

Antioxidant Defenses in Wild Growing Halophyte Crithmum maritimum from Inland and Coastline Populations.

Traditional Mediterranean diet includes the halophyte Crithmum maritimum L. (Apiaceae) which can be found in the coastline of the Balearic Islands but also inland. Both areas differed in the environmental conditions, mainly in salinity which can affect the oxidative status of this species. The aim was to evaluate the antioxidant enzyme activities, polyphenols and the lipid peroxidation in leaves of wild C. maritimum growing in a natural coastal area influenced by marine salinity and an inland area without marine influence. The activities of the antioxidant enzymes catalase, superoxide dismutase and glutathione peroxidase as well as polyphenol and reduced glutathione content were significantly higher in the samples from coastline population, whereas no significant differences were found in glutathione reductase activity and in malondialdehyde levels. The production of H2 O2 was also significantly higher in the population from coastline. In conclusion, C. maritimum adapt their antioxidant defense machinery to the different salinity conditions, avoiding the instauration of oxidative stress.

Potential Anti-inflammatory Effects of Hesperidin from the Genus Citrus.

BACKGROUND: The benefits of the Mediterranean diet for protecting against many diseases are usually attributed to high consumption of certain foods, characterized by the presence of bioactive substances such as polyphenols. Inflammation plays an important role in the pathogenesis of numerous diseases such as arthritis, allergies or neurodegenerative disorders. Dietary polyphenols constitute a large family of bioactive substances with potential beneficial effects against a broad group of diseases. Citrus fruits and juices are a rich source of vitamin C and flavonoids, with a potential effect on the inflammatory response. OBJECTIVE: The aim was to evidence the potential anti-inflammatory effects of the flavonoids hesperidin for its possible therapeutic application against diverse pathologies. METHOD: In the present review, available literature about the anti-inflammatory effects of hesperidin is reported and discussed. Moreover, we also discuss the chemistry, bioavailability and proposed mechanisms of action of hesperidin. RESULTS: Hesperidin is a flavonoid present in high concentration in citrus species and has numerous biological properties, principally antioxidant and anti-inflammatory. Several studies have been performed in order to evaluate the effects of hesperidin as anti-inflammatory agent using cellular and animal models and few clinical trials. Hesperidin treatment decreased inflammatory mediators and exerted significant antioxidant effects. The molecular basis for its anti-inflammatory effects seems to be mediated by signalling pathways especially the nuclear factor κß pathway. CONCLUSION: Although hesperidin evidenced anti-inflammatory effects, the specific mechanism of action is not completely known and additional studies are required for elucidation of the molecular targets.

Oxidative stress response in the seagrass Posidonia oceanica and the seaweed Dasycladus vermicularis associated to the invasive tropical green seaweed Halimeda incrassata.

The Mediterranean Sea is one of the most affected areas by the presence of invasive species. Halimeda incrassata (J Ellis) JV Lamoroux is newly arrived tropical seaweed in waters of the Mallorca Island (Balearic Islands, Western Mediterranean). The aim was to evaluate the effect of a potential competition between the invasive Halimeda incrassata, the native Posidonia oceanica and Dasycladus vermicularis, by means of antioxidant-related biomarkers in waters of Mallorca. The activities of the antioxidant enzymes-catalase, superoxide dismutase (SOD), glutathione peroxidase (GPx) and glutathione reductase (GRd)-, the levels of reduced glutathione (GSH) and malondialdehyde as indicator of lipid peroxidation were evaluated. The concentration of 3,6,7-trihydroxycoumarin (THC) was measured in D. vermicularis. P. oceanica biomarkers were not altered while D. vermicularis coexisting with the invader showed higher GSH levels (46%) and antioxidant enzyme activities (catalase 74%, SOD 65%, GPx 86% and GRd 98%), although without lipid damage. H. incrassata showed higher malondialdehyde and GSH levels (30% and 31%, respectively), and catalase, superoxide dismutase and glutathione reductase activities (51%, 35% and 84%, respectively) in presence of P. oceanica respect to being alone; and higher superoxide dismutase and glutathione reductase in the presence of D. vermicularis (22% and 42%, respectively). THC concentration in D. vermicularis was significantly higher (53%) in samples competing with H. incrassata. Altogether, native P. oceanica meadows seem unaffected by the alien H. incrassata-which suffered oxidative stress competing with the other species-; whereas increased antioxidant capacities were evidenced in D. vermicularis, possibly as an adaptation mechanism to the new stressful situation that reflect differences in the physiological activities of the three species. In conclusion, the presence of the invasive H. incrassata may be a competitor to be considered for D. vermicularis, while it does not seem to be a major problem for P. oceanica.

Cardioprotective Effects of the Polyphenol Hydroxytyrosol from Olive Oil.

BACKGROUND: The Mediterranean diet includes olive oil as its primary source of fat. This diet is frequently associated to longevity and a lower incidence of chronic diseases due to its biological activities and health effects. Apart from oleic acid, olive oil contains many bioactive components including polyphenols that have been reported to exert antioxidant and anti-inflammatory activities. Polyphenols may almost in part be responsible for the protective effects against cardiovascular diseases associated with olive oil. OBJECTIVE: To review and discuss the available literature on hydroxytyrosol effects as a cardioprotective agent. Moreover, we also discuss the chemistry, nutritional aspects and bioavailability of hydroxytyrosol. RESULTS: Hydroxytyrosol is one of the major phenolic compounds in olive oil and has demonstrated strong radical-scavenging properties. Several studies have been performed in order to look further into the effects of the polyphenol hydroxytyrosol in relation to cardiovascular events and illnesses in animal trials and in vitro. However, no clinical trials have focused on the specific action of hydroxytyrosol and cardiovascular diseases, although some are being undertaken to look at olive oil or olive leaf extract properties. CONCLUSION: Hydroxytyrosol from olive oil exerts antioxidant, anti-inflammatory, anti-platelet aggregation and ati-atherogenic activities in in vitro and animal models. However, its possible therapeutic use in humans requires additional clinical trials.

Invasive predator snake induces oxidative stress responses in insular amphibian species.

The presence of predators induces physiological stress responses in preys to avoid being captured. A stressful situation enhances reactive oxygen species production with potential damage to macromolecules and alterations in oxidant defences levels. The antioxidant enzyme response of the endemic Majorcan Midwife toad (Alytes muletensis) and the Balearic green toad (Bufotes balearicus) tadpoles against an invasive predator, the viperine snake (Natrix maura) was investigated. Tadpoles were introduced in aquaria containing N. maura exudates during 24h. Antioxidant enzyme activities - catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR), superoxide dismutase (SOD) - and reduced glutathione (GSH) and malondialdehyde (MDA) concentrations were measured in tadpoles. The presence of snake exudates induced a significant increase in CAT and GR activities and in GSH levels (p<0.05) in A. muletensis tadpoles, whereas no significant differences were reported in any of the parameters analysed in B. balearicus tadpoles. In conclusion, the presence of N. maura exudates is capable to induce an antipredatory response in the endemic A. muletensis tadpoles but not in B. balearicus.