Avian Haemosporidian Infection in Wildlife Rehabilitation Centres of Portugal: Causes, Consequences, and Genetic Diversity.

In the last decade, over 40% of bird species in Europe have experienced poor and bad conservation status, with more than 30% of bird species in mainland Portugal threatened with extinction. Along with anthropogenic factors, parasites and pathogens such as avian haemosporidians have been suggested to be responsible for these avian population declines. Wildlife rehabilitation centres play an essential role in species conservation and preservation. Moreover, animals admitted for rehabilitation can provide valuable information regarding transmission and pathogenicity of many diseases that affect wild birds that are rarely sampled in nature. However, reports of haemosporidians in captive birds are still limited. Here, we explored the prevalence and genetic diversity of avian haemosporidians in 89 birds from 29 species admitted to rehabilitation centres in Portugal, showing an overall infection prevalence of 30.3%. The prevalence of infection was higher in Strigiformes and in birds admitted to rehabilitation centres due to debilitating diseases. Remarkably, 30% of the infected bird species have not been found to harbour malaria parasites in preceding studies. We detected 15 different haemosporidian lineages infecting a third of bird species sampled. Notably, 2 out of these 15 detected haemosporidian lineages have not been obtained previously in other studies. Furthermore, we also identified nine new host-parasite interactions representing new host records for these haemosporidian parasites. Finally, our results revealed that birds infected with haemosporidians require longer rehabilitation treatments, which increase the economic costs for rehabilitation and may impair their survival prospects. These findings emphasise the importance of integrating haemosporidian infection considerations into rehabilitation protocols, highlighting the challenges posed by these infections in avian conservation and rehabilitation, including economic and logistical demands.

Effect of salinity on scytonemin yield in endolithic cyanobacteria from the Atacama Desert.

Cyanobacteria inhabiting extreme environments constitute a promising source for natural products with biotechnological applications. However, they have not been studied in-depth for this purpose due to the difficulties in their isolation and mass culturing. The Atacama Desert suffers one of the highest solar irradiances that limits the presence of life on its hyperarid core to endolithic microbial communities supported by cyanobacteria as primary producers. Some of these cyanobacteria are known to produce scytonemin, a UV-screening liposoluble pigment with varied biotechnological applications in cosmetics and other industries. In this work we carried out a strain selection based on growth performance among 8 endolithic cyanobacteria of the genera Chroococcidiopsis, Gloeocapsa and Gloeocapsopsis isolated from non-saline rocks of the Atacama Desert. Then we investigated the influence of NaCl exposure on scytonemin production yield. Results in the selected strain (Chroococcidiopsis sp. UAM571) showed that rising concentrations of NaCl lead to a growth decrease while triggering a remarkable increase in the scytonemin content, reaching maximum values at 20 g L-1 of NaCl over 50-fold higher scytonemin contents than those obtained without NaCl. Altogether, these findings point out to cyanobacteria from the Atacama Desert as potentially suitable candidates for pilot-scale cultivation with biotechnological purposes, particularly to obtain scytonemin.

Dynamics and effects of plastic contaminants' assimilation in gulls.

Polybrominated diphenyl ethers are persistent disrupters assimilated by organisms, yet little is known about their link to plastic ingestion and health effects. In an experiment, two groups of yellow-legged/lesser black-backed gulls (Larus michahellis/Larus fuscus) were fed plastics with BDE99 to assess leaching into brain, preen oil, liver and fat tissues and evaluate effects on health and stress parameters. Although most plastic was regurgitated, we observed a clear relation between plastic ingestion and chemical leaching. BDE99 exhibited higher levels in brain tissue of gulls from the plastic groups. Also, only values of cholinesterases measured in plasma were significantly reduced in the 'plastic' groups. Cholinesterase activity in the brain also tended to decrease, suggesting a negative effect in gulls' neurofunction. Results indicate that chemical leaching occurs, even when plastics stay in the stomach for a short period of time and showed that this can affect gulls' health.

The first full study of heavy metal(loid)s in western-European hedgehogs (Erinaceus europaeus) from Portugal.

The western-European hedgehog (Erinaceus europaeus) is an insectivore with a wide distribution in Portugal and a potential tool for biomonitoring relevant One Health hazards, including heavy metal(loid)s' pollution. The aim of this study was to positively contribute to the current knowledge about the metal(loid) pollution in Portugal. Forty-six hedgehogs (from rescue centres; with known provenance) were necropsied. Sex, age category and weight were determined. Spines, liver and kidney were collected, and metalloid concentrations were determined by inductively coupled plasma mass spectrophotometry (ICP-MS). In general, results did not present alarming metal(loid) concentrations, with the exception of cadmium (Cd) (in the kidneys) and copper (Cu). Hedgehogs from Viana do Castelo and Viseu showed elevated concentrations of arsenic (As) and Castelo Branco presented concerning values of cadmium (Cd). Adult and heavier hedgehogs tended to present higher levels of metal(loid)s. Sex does not seem to significantly affect the metal(loid)s' concentrations. Further analysis would be needed to prioritize areas with detail and allow the application of the necessary mitigation strategies.

New molecular data help clarify the taxonomy of central European avian dicrocoeliidae Looss, 1899 (Trematoda: Plagiorchiida).

Avian Dicrocoeliidae are difficult to identify, even in their adult stages. Molecular analyses have been conducted for only a few species, complicating the accurate identification of juvenile stages. The taxonomy of the family is unresolved, and the status of many dicrocoeliid species is uncertain. Sequences of nuclear and mitochondrial DNA loci of Central European avian Dicrocoeliidae were generated and analyzed. These included representatives of the genera Lyperosomum, Platynosomum, Stromitrema, Brachylecithum, Brachydistomum, and Lutztrema. All the sequences were obtained from morphologically identified adult specimens of dicrocoeliids isolated from avian hosts. Molecular support was obtained to validate Lyperosomum turdia, confirm the rejection of Lyperosomum dujardini and Lyperosomum alagesi, and resurrect Lyperosomum longicauda and Lyperosomum collurionis. The identity of European Platynosomum illiciens from avian hosts with American vouchers of the same species from avian and mammalian hosts was confirmed. Brachylecithum fringillae is not considered valid; the individuals that matched its diagnosis were subadult Brachydistomum ventricosum. Descriptions and comparative data for five new species are provided. These are Lyperosomum hirundinis sp. n., Lyperosomum tenori sp. n., Lyperosomum atricapillae sp. n., Stromitrema acrocephali sp. n., and Lutztrema atricapillae sp. n.. Based on the molecular data, suggestions are provided regarding the validity of dicrocoeliid species that parasitize Central European birds. Further research should address the polyphyletic status of Brachylecithum.

Phenotypic Characterization of Oral Mucor Species from Eurasian Vultures: Pathogenic Potential and Antimicrobial Ability.

Due to poisoning and decline in the food resources of Eurasian vultures, there has been a rise in the number of Griffon (Gyps fulvus) and Cinereous vultures (Aegypius monachus) needing veterinary care. In captivity, vultures often develop oral and other infectious diseases which can affect their survival and the probability of reintroduction in the wild. Therefore, it is important to characterize relevant microbial species present in the oral cavity of vultures, such as Mucor spp. In this work, seven Mucor spp. isolates previously obtained from Gyps fulvus and Aegypius monachus oral swabs collected at two rehabilitation centers in Portugal were characterized regarding their pathogenic enzymatic profile and antimicrobial activity. Isolates were identified by macro and microscopic observation, and PCR and ITS sequencing. Their antimicrobial activity was determined using a collection of pathogenic bacteria and two yeast species. Results showed that 86% of the isolates produced α-hemolysis, 71% expressed DNase, 57% produce lecithinase and lipase, 29% expressed gelatinase, and 29% were biofilm producers. Four isolates showed inhibitory activity against relevant human and veterinary clinical isolates, including Escherichia coli, Enterococcus faecium, Neisseria zoodegmatis, and Staphylococcus aureus. In conclusion, accurate management programs should consider the benefits and disadvantages of Mucor spp. presence in the oral mucosa.

High Levels of Heavy Metal(loid)s Related to Biliary Hyperplasia in Hedgehogs (Erinaceus europaeus).

Heavy metal(loid) pollution of ecosystems is a current One Health problem. The liver is one of the most affected organs in cases of acute or chronic exposure to abnormal amounts of these substances, inducing histopathologic lesions. In order to assess the influence of heavy metal(loids), forty-five European hedgehogs (Erinaceus europaeus) were submitted to necropsy, and liver samples were collected for a routine histopathology exam and metal(loid)s determination (As, Cd, Co, Cr, Cu and Pb) by ICP-MS. Age was estimated during the necropsy exam. Biliary hyperplasia was the most frequent lesion observed (16/45; 35.56%). No statistically significant associations were found between biliary hyperplasia and age or sex. Metal(loid)s' concentrations were higher in animals with biliary hyperplasia (except for As). There was a statistically significant difference for both Cd and Co. For As, Cd and Co, cubs and juveniles animals showed significantly lower concentrations than elder individuals. Only for Pb were significant differences found between females and males. As described in the literature, exposure to metal(loid)s may be a cause of biliary hyperplasia, although further research (including the use of biochemical methods) is needed to support these results. To the authors' knowledge, this is the first report of this association in hedgehogs.

Pacientes críticos COVID-19. ¿Han variado el manejo y los resultados en la UCI tras un año de pandemia? Estudio multicéntrico, prospectivo, observacional / Critical patients COVID-19 has changed the management and outcomes in the ICU after 1 year of the pandemic? A multicenter, prospective, observational study

Objetivo: Comparar las características clínicas, los tratamientos y la evolución de los pacientes críticos con neumonía por COVID-19 atendidos en unidades de cuidados intensivos (UCI) tras un año de pandemia. Metodología: Estudio multicéntrico, prospectivo, en el que se incluyó pacientes críticos COVID-19 en 9 UCI del noroeste de España. Se compararon las características clínicas, los tratamientos y la evolución de pacientes ingresados en UCI durante los meses de marzo-abril de 2020 (periodo1) con pacientes ingresados en enero-febrero de 2021 (periodo2). Resultados: Se incluyeron 337 pacientes (98 en el periodo1 y 239 en el periodo2). En el periodo2 menos pacientes requirieron ventilación mecánica invasiva (VMI) (65% vs 84%, p<0,001), utilizándose con mayor frecuencia cánulas nasales de alto flujo (CNAF) (70% vs 7%, p<0,001), ventilación mecánica no invasiva (VMNI) (40% vs 14%, p<0,001), corticoides (100% vs 96%, p=0,007) y posición de decúbito prono tanto en pacientes despiertos (42% vs 28%, p=0,012) como en pacientes intubados (67% vs 54%, p=0,034). Los días de VMI, de estancia en UCI y hospitalaria fueron inferiores en el periodo2. La mortalidad fue similar en los dos periodos estudiados (16% vs 17%). Conclusiones: Tras un año de pandemia, observamos que en los pacientes ingresados en UCI se ha utilizado con mayor frecuencia CNAF, VMNI, uso del decúbito prono y corticoides, disminuyendo los pacientes en VMI, así como los tiempos de estancia en UCI y de estancia hospitalaria. La mortalidad ha sido similar en los dos periodos a estudio.(AU)

Objective: To compare the clinical characteristics, treatments, and evolution of critical patients with COVID-19 pneumonia treated in intensive care units (ICU) after one year of pandemic. Methodology: Multicenter, prospective study, which included critical COVID-19 patients in 9 ICUs in northwestern Spain. The clinical characteristics, treatments, and evolution of patients admitted to the ICU during the months of March-April 2020 (period1) were compared with patients admitted in January-February 2021 (period2). Results: 337 patients were included (98 in period1 and 239 in period2). In period2, fewer patients required invasive mechanical ventilation (IMV) (65% vs. 84%, P<.001), using high-flow nasal cannulas (CNAF) more frequently (70% vs. 7%, P<.001), ventilation non-invasive mechanical (NIMV) (40% vs. 14%, P<.001), corticosteroids (100% vs. 96%, P=.007) and prone position in both awake (42% vs. 28%, P=.012), and intubated patients (67% vs. 54%, P=.034). The days of IMV, ICU stay and hospital stay were lower in period2. Mortality was similar in the two periods studied (16% vs. 17%). Conclusions: After one year of pandemic, we observed that in patients admitted to the ICU, CNAF, NIMV, use of the prone position, and corticosteroids have been used more frequently, reducing the number of patients in IMV, and the length of stay in the ICU and hospital stay. Mortality was similar in the two study periods.(AU)

Critical patients COVID-19 has changed the management and outcomes in the ICU after 1 year of the pandemic? A multicenter, prospective, observational study.

OBJECTIVE: To compare the clinical characteristics, treatments, and evolution of critical patients with COVID-19 pneumonia treated in Intensive Care Units (ICU) after one year of pandemic. METHODOLOGY: Multicenter, prospective study, which included critical COVID-19 patients in 9 ICUs in northwestern Spain. The clinical characteristics, treatments, and evolution of patients admitted to the ICU during the months of March-April 2020 (period 1) were compared with patients admitted in January-February 2021 (period 2). RESULTS: 337 patients were included (98 in period 1 and 239 in period 2). In period 2, fewer patients required invasive mechanical ventilation (IMV) (65% vs 84%, p < 0.001), using high-flow nasal cannulas (CNAF) more frequently (70% vs 7%, p < 0.001), ventilation non-invasive mechanical (NIMV) (40% vs 14%, p < 0.001), corticosteroids (100% vs 96%, p = 0.007) and prone position in both awake (42% vs 28%, p = 0.012), and intubated patients (67% vs 54%, p = 0.034). The days of IMV, ICU stay and hospital stay were lower in period 2. Mortality was similar in the two periods studied (16% vs 17%). CONCLUSIONS: After 1 year of pandemic, we observed that in patients admitted to the ICU, CNAF, NIMV, use of the prone position, and corticosteroids have been used more frequently, reducing the number of patients in IMV, and the length of stay in the ICU and hospital stay. Mortality was similar in the two study periods.

[Critical patients COVID-19 has changed the management and outcomes in the ICU after 1 year of the pandemic? A multicenter, prospective, observational study]. / Pacientes críticos COVID-19. ¿Han variado el manejo y los resultados en la UCI tras un año de pandemia? Estudio multicéntrico, prospectivo, observacional.

Objective: To compare the clinical characteristics, treatments, and evolution of critical patients with COVID-19 pneumonia treated in intensive care units (ICU) after one year of pandemic. Methodology: Multicenter, prospective study, which included critical COVID-19 patients in 9 ICUs in northwestern Spain. The clinical characteristics, treatments, and evolution of patients admitted to the ICU during the months of March-April 2020 (period 1) were compared with patients admitted in January-February 2021 (period 2). Results: 337 patients were included (98 in period 1 and 239 in period 2). In period 2, fewer patients required invasive mechanical ventilation (IMV) (65% vs. 84%, P < .001), using high-flow nasal cannulas (CNAF) more frequently (70% vs. 7%, P < .001), ventilation non-invasive mechanical (NIMV) (40% vs. 14%, P < .001), corticosteroids (100% vs. 96%, P = .007) and prone position in both awake (42% vs. 28%, P = .012), and intubated patients (67% vs. 54%, P = .034). The days of IMV, ICU stay and hospital stay were lower in period 2. Mortality was similar in the two periods studied (16% vs. 17%). Conclusions: After one year of pandemic, we observed that in patients admitted to the ICU, CNAF, NIMV, use of the prone position, and corticosteroids have been used more frequently, reducing the number of patients in IMV, and the length of stay in the ICU and hospital stay. Mortality was similar in the two study periods.

Recent large-scale philophthalmosis outbreak in Portugal: inefficacy of common antihelminthic agents.

BACKGROUND: Parasitic conjunctivitis caused by Philophthalmus spp. is a common ophthalmic disease in birds, with localized outbreaks occurring worldwide. There is no consensus on treating this disease; mechanical removal is considered a standard recommendation, but is associated with disease relapses within days or weeks. METHODS: From 2015 to 2020, we examined 4295 Larus michahellis and Larus fuscus gulls in southern Portugal for the presence of Philophthalmus spp. Due to the need to treat dozens of infected gulls in the rescue station, we tested three treatment regimens aimed at targeting Philophthalmus lucipetus in the infected gulls: (I) the ophthalmic application of levamisole; (II) the oral application of milbemycin in combination with praziquantel; and (III) the subcutaneous application of ivermectin. RESULTS: The outbreak of philophthalmosis in gulls in southern Portugal has been ongoing since the first cases were reported in 2015-2016. The prevalence of philophthalmosis has fluctuated annually, peaking a maximum of 10.3% in L. fuscus in 2017 and at 2.1% in L. michahellis in 2016. The infection intensity peaked at a median of 11.5 eye-flukes per host bird in L. fuscus in 2016 and a median of six eye-flukes per host bird in L. michahellis in 2017. Nine gulls were infected with >50 eye-flukes. None of the treatment options were effective at treating P. lucipetus infections: the numbers of eye-flukes in the infected birds did not decrease, and the clinical signs of the disease did not change. CONCLUSIONS: An outbreak of philophthalmosis in southern Portugal has massively affected two species of gulls in the region. Two previously suggested philophthalmosis treatments (ocular levamisole and praziquantel given orally), as well as a third mode of treatment with a previously failed compound (ivermectin administered subcutaneously) were used. However, the treatments did not affect the numbers of P. lucipetus in the eyes of the treated gulls. Further research should address ophthalmic gel formulations or sub-conjunctival delivery mode for antihelminthic drugs that are effective against Philophthalmus spp. in vitro.

Co-infection by classic MYXV and ha-MYXV in Iberian hare (Lepus granatensis) and European wild rabbit (Oryctolagus cuniculus algirus).

Myxomatosis is an emergent disease in the Iberian hare (Lepus granatensis). In this species, the disease is caused by a natural recombinant virus (ha-myxoma virus [MYXV]) identified for the first time in 2018 and has since been responsible for a large number of outbreaks in Spain and Portugal. The ha-MYXV, which harbours a 2.8 Kb insert-disrupting gene M009L, can also infect and cause disease in wild and domestic rabbits, despite being less frequently identified in rabbits. During the laboratory investigations of wild leporids found dead in Portugal carried out within the scope of a Nacional Surveillance Plan (Dispatch 4757/17, MAFDR), co-infection events by classic (MYXV) and naturally recombinant (ha-MYXV) strains were detected in both one Iberian hare and one European wild rabbit (Oryctolagus cuniculus algirus). These two cases were initially detected by a multiplex qPCR detection of MYXV and ha-MYXV and subsequently confirmed by conventional PCR and sequencing of the M009L gene, which contains an ha-MYXV-specific insertion. To our knowledge, this is the first documented report of co-infection by classic MYXV and ha-MYXV strains either in Iberian hare or in European wild rabbit. It is also the first report of infection of an Iberian hare by a classic MYXV strain. These findings highlight the continuous evolution of the MYXV and the frequent host range changes that justify the nonstop monitoring of the sanitary condition of wild Leporidae populations in the Iberian Peninsula.

Morphological, molecular, and biochemical study of cyanobacteria from a eutrophic Algerian reservoir (Cheffia).

The cyanobacteria management in water bodies requires a deep knowledge of the community composition. Considering the reliable and thorough information provided by the polyphasic approach in cyanobacteria taxonomy, here we assess the cyanobacterial community structure of the Cheffia reservoir from Algeria. Cyanobacteria were identified on the basis of morphological traits and next-generation sequencing (NGS); toxins-related genes were localized in addition to the identification of toxins; temperature and nutrient level of water samples were also determined. The polyphasic approach was essential for cyanobacteria investigation; 28 genera were identified through 16S rRNA metabarcoding with the dominance of taxa from Microcystis (34.2%), Aphanizomenon (20.1%), and Planktothrix (20.0%), and morphological analysis revealed the association in this water body of five species within the genus Microcystis: M. aeruginosa, M. novacekii, M. panniformis, M. ichthyoblabe, and M. flos-aquae. The presence of mcyE genotypes was detected; moreover, HPLC-PDA and LC-ESI-MS/MS revealed the production of microcystin-LR. Results obtained in our study are very important since this ecosystem is used for water supply and irrigation; as a consequence, a good water management plan is essential.

Prevalence of Antibodies to Toxoplasma gondii in Different Wild Bird Species Admitted to Rehabilitation Centres in Portugal.

Toxoplasma gondii is a worldwide zoonotic parasite. According to the "One Health" approach, studies on toxoplasmosis are essential since it affects humans and domestic and wild animals. In the present study, antibodies to T. gondii were determined in serum samples from 263 wild birds located in five wildlife rehabilitation centres in mainland Portugal by using the modified agglutination test (MAT) with a cut-off titre of 20. An overall seroprevalence of 36.5% (95% confidence interval [CI]: 30.7-42.6) was observed. For the first time, antibodies to T. gondii were detected in some avian species, including pallid swift (Apus pallidus) (33.3%), black-backed gull (Larus fuscus) (39.3%), European turtle-dove (Streptopelia turtur) (100%), bee-eater (Merops apiaster) (50.0%), carrion crow (Corvus corone) (33.3%), and Egyptian vulture (Neophron percnopterus) (100%), which expands the list of intermediate hosts of T. gondii. A lower seroprevalence was found in juvenile birds (31.9%) compared to adults (48.7%) (p = 0.016). The central region of Portugal was considered a risk factor for T. gondii infection in wild birds (odds ratio: 3.61; 95% CI: 1.09-11.91). This pioneer study calls attention to the need for further studies, to provide a clearer understanding of T. gondii epidemiology in Portugal, because it reflects wide dispersion of T. gondii oocysts in the environment.

Paralytic and Amnesic Shellfish Toxins Impacts on Seabirds, Analyses and Management.

Marine biotoxins have been frequently implicated in morbidity and mortality events in numerous species of birds worldwide. Nevertheless, their effects on seabirds have often been overlooked and the associated ecological impact has not been extensively studied. On top of that, the number of published studies confirming by analyses the presence of marine biotoxins from harmful algal blooms (HABs) in seabirds, although having increased in recent years, is still quite low. This review compiles information on studies evidencing the impact of HAB toxins on marine birds, with a special focus on the effects of paralytic and amnesic shellfish toxins (PSTs and ASTs). It is mainly centered on studies in which the presence of PSTs and/or ASTs in seabird samples was demonstrated through analyses. The analytical techniques commonly employed, the tissues selected and the adjustments done in protocols for processing seabird matrixes are summarized. Other topics covered include the role of different vectors in the seabird intoxications, information on clinical signs in birds affected by PSTs and ASTs, and multifactorial causes which could aggravate the syndromes. Close collaboration between seabird experts and marine biotoxins researchers is needed to identify and report the potential involvement of HABs and their toxins in the mortality events. Future studies on the PSTs and ASTs pharmacodynamics, together with the establishment of lethal doses in various seabird species, are also necessary. These studies would aid in the selection of the target organs for toxins analyses and in the postmortem intoxication diagnoses.

Microbial colonizers of microplastics in an Arctic freshwater lake.

Microplastics (MPs) have been found everywhere as they are easily transported between environmental compartments. Through their transport, MPs are quickly colonized by microorganisms; this microbial community is known as the plastisphere. Here, we characterized the plastisphere of three MPs, one biodegradable (PHB) and two non-biodegradables (HDPE and LDPE), deployed in an Arctic freshwater lake for eleven days. The plastisphere was found to be complex, confirming that about a third of microbial colonizers were viable. Plastisphere was compared to microbial communities on the surrounding water and microbial mats on rocks at the bottom of the lake. Microbial mats followed by MPs showed the highest diversity regarding both prokaryotes and eukaryotes as compared to water samples; however, for fungi, MPs showed the highest diversity of the tested substrates. Significant differences on microbial assemblages on the three tested substrates were found; regarding microbial assemblages on MPs, bacterial genera found in polar environments such as Mycoplana, Erythromicrobium and Rhodoferax with species able to metabolize recalcitrant chemicals were abundant. Eukaryotic communities on MPs were characterized by the presence of ciliates of the genera Stentor, Vorticella and Uroleptus and the algae Cryptomonas, Chlamydomonas, Tetraselmis and Epipyxis. These ciliates normally feed on algae so that the complexity of these assemblages may serve to unravel trophic relationships between co-existing taxa. Regarding fungal communities on MPs, the most abundant genera were Betamyces, Cryptococcus, Arrhenia and Paranamyces. MPs, particularly HDPE, were enriched in the sulI and ermB antibiotic resistance genes (ARGs) which may raise concerns about human health-related issues as ARGs may be transferred horizontally between bacteria. This study highlights the importance of proper waste management and clean-up protocols to protect the environmental health of pristine environments such as polar regions in a context of global dissemination of MPs which may co-transport microorganisms, some of them including ARGs.

Mitochondrial sequences of Rhipicephalus and Coxiella endosymbiont reveal evidence of lineages co-cladogenesis.

Rhipicephalus ticks are competent vectors of several pathogens, such as Spotted Fever Group Rickettsiae (SFGR) and many Babesia species. Within this genus, different R. sanguineus s.l. lineages show an unequal vector competence and resistance regarding some pathogenic strains. Current literature supports that tick endosymbionts may play an essential role in the transmission ability of a vector. Indeed, the microbial community of Rhipicephalus seems to be dominated by Coxiella-like endosymbionts (CLE). Still, their co-evolutionary associations with the complicated phylogeny of Rhipicephalus lineages and their transmissible pathogens remain unclear. We performed a phylogenetic congruence analysis to address whether divergent R. sanguineus s.l. lineages had a different symbiont composition. For that, we applied a PCR based approach to screen part of the microbial community present in 279 Rhipicephalus ticks from the Iberian Peninsula and Africa. Our analyses detected several qPCR-positive signals for both SFGR and Babesia species, of which we suggest R. sanguineus-tropical lineage as a natural vector of Babesia vogeli and R. sanguineus-temperate lineage of SFGR. The acquisition of 190 CLE sequences allowed to evaluate co-phylogenetic associations between the tick and the symbiont. With this data, we observed a strong but incomplete co-cladogenesis between CLE strains and their Rhipicephalus tick lineages hosts.

Response of Endolithic Chroococcidiopsis Strains From the Polyextreme Atacama Desert to Light Radiation.

Cyanobacteria exposed to high solar radiation make use of a series of defense mechanisms, including avoidance, antioxidant systems, and the production of photoprotective compounds such as scytonemin. Two cyanobacterial strains of the genus Chroococcidiopsis from the Atacama Desert - which has one of the highest solar radiation levels on Earth- were examined to determine their capacity to protect themselves from direct photosynthetically active (PAR) and ultraviolet radiation (UVR): the UAM813 strain, originally isolated from a cryptoendolithic microhabitat within halite (NaCl), and UAM816 strain originally isolated from a chasmoendolithic microhabitat within calcite (CaCO3). The oxidative stress induced by exposure to PAR or UVR + PAR was determined to observe their short-term response, as were the long-term scytonemin production, changes in metabolic activity and ultrastructural damage induced. Both strains showed oxidative stress to both types of light radiation. The UAM813 strain showed a lower acclimation capacity than the UAM816 strain, showing an ever-increasing accumulation of reactive oxygen species (ROS) and a smaller accumulation of scytonemin. This would appear to reflect differences in the adaptation strategies followed to meet the demands of their different microhabitats.

Unmasking the identity of toxigenic cyanobacteria driving a multi-toxin bloom by high-throughput sequencing of cyanotoxins genes and 16S rRNA metabarcoding.

Cyanobacterial harmful algal blooms (CyanoHABs) are complex communities that include coexisting toxic and non-toxic strains only distinguishable by genetic methods. This study shows a water-management oriented use of next generation sequencing (NGS) to specifically pinpoint toxigenic cyanobacteria within a bloom simultaneously containing three of the most widespread cyanotoxins (the hepatotoxins microcystins, MCs; and the neurotoxins anatoxin-a, ATX, and saxitoxins, STXs). The 2013 summer bloom in Rosarito reservoir (Spain) comprised 33 cyanobacterial OTUs based on 16S rRNA metabarcoding, 7 of which accounted for as much as 96.6% of the community. Cyanotoxins and their respective biosynthesis genes were concurrently present throughout the entire bloom event including: MCs and mcyE gene; ATX and anaF gene; and STXs and sxtI gene. NGS applied to amplicons of cyanotoxin-biosynthesis genes unveiled 6 toxigenic OTUs, comprising 3 involved in MCs production (Planktothrix agardhii and 2 Microcystis spp.), 2 in ATX production (Cuspidothrix issatschenkoi and Phormidium/Tychonema spp.) and 1 in STXs production (Aphanizomenon gracile). These toxigenic taxa were also present in 16S rRNA OTUs list and their relative abundance was positively correlated with the respective toxin concentrations. Our results point at MC-producing P. agardhii and ATX-producing C. issatschenkoi as the main contributors to the moderate toxin concentrations observed, and suggest that their distribution in Southern Europe is broader than previously thought. Our findings also stress the need for monitoring low-abundance cyanobacteria (<1% relative abundance) in cyanotoxicity studies, and provide novel data on the presence of picocyanobacteria and potentially ATX-producing benthic taxa (e.g., Phormidium) in deep thermally-stratified water bodies. This study showcases a straightforward use of amplicon metagenomics of cyanotoxin biosynthesis genes in a multi-toxin bloom thus illustrating the broad applicability of NGS for water management in risk-oriented monitoring of CyanoHABs.

Plastic ingestion in aquatic birds in Portugal.

In modern society, plastic items have become indispensable. The rapid growth of plastic production has led to an increase in the concentration of plastic waste in the environment and, consequently, wildlife has been severely affected. As wide-ranging foragers and predators, aquatic birds are ideal sentinels for monitoring changes in their environment. Plastic found in stomach contents of stranded aquatic birds collected throughout Portugal was examined. Out of the 288 birds processed, 12.9% ingested plastics. Six of the 16 species assessed showed evidence of plastic ingestion. The Lesser Black-backed Gull (18.7%) had the highest incidence while, among those that did ingest plastics, the Northern Gannet (4.8%) had the lowest. User plastics were the most common type of plastic ingested, while microplastics and off/white-clear were the most common size and colour respectively of plastics found. This study sets a first multispecies baseline for incidence of plastic ingestion by aquatic birds in Portugal.