La conservazione dell’ambiente e della biodiversità rappresenta una delle maggiori sfide della scienza odierna. Nella strategia della conservazione, che comporta l’implementazione delle basi conoscitive, l’identificazione del microbiota intestinale di organismi wild (come le iguane terrestri delle Isole Galapagos e le tartarughe marine delle Isole Eolie) può essere utilizzato sia come dato conoscitivo di base sia come strumento applicativo: in toto lo studio e la definizione quantitativa e qualitativa dei micro-organismi che abitano l’apparato digerente di questi organismi, può fornire un quadro generale di grande interesse e per la gran parte ancora inesplorato, mentre il confronto del microbiota intestinale di organismi provenienti da aree antropizzate e non, può essere strumento di base per avviare una valutazione di impatto antropico. Marcatore della possibilità di uno scambio reciproco di microrganismi tra il comparto antropizzato e quello naturale, è il trasferimento delle antibiotico-resistenze. L’analisi dell’incidenza e della natura dei determinanti genici di antibiotico resistenza naturalmente presenti e/o acquisiti dai batteri enterici del microbiota intestinale di entrambi questi organismi, rappresenta un ulteriore strumento di valutazione dell’impatto antropico.

 

Il lavoro è stato avviato sulle iguane terrestri delle Galapagos che rappresenta un sistema particolarmente interessante in quanto le isole non soggette a impatto antropico possono fornire indicazioni su variabili della wildlife in assenza di perturbazioni, mentre le isole soggette a impatto antropico (indigeni e/o turisti) possono dare indicazioni sulle modifiche già avvenute o in fieri: una sorta di struttura “esperimento-controllo”. Il progetto ha previsto sia l'identificazione delle principali componenti del microbiota associato alla cloaca delle iguane terrestri e marine, sia l'incidenza e la natura dei determinanti di resistenza. Lo stesso lavoro è stato effettuato anche sulle iguane terrestri delle Antille Francesi nel Mar dei Caraibi.

 

Con lo stesso obiettivo è stato anche studiato il microbiota intestinale e l'incidenza e la natura dei determinanti di resistenza in tartarughe marine (Caretta caretta L.) campionate nelle Isole Eolie, nell’area dell’Isola di Filicudi. In questo caso sono state confrontate tartarughe in buone condizioni di salute con altre con disagi nell’alimentazione a causa di ami o buste di plastica ingeriti. Ancora, sulle stesse tartarughe marine è stato avviato uno studio pilota per valutare la struttura e la composizione dei colonizzatori del carapace: sono stati analizzati sia i macro-colonizzatori (macroalghe epifite, crostacei, ecc.) sia i micro-colonizzatori (cioè la comunità batterica) utilizzando il NGS, per cercare di definire un protocollo unitario per questo tipo di studi.

 

Questo lavoro è stato possibile grazie alle collaborazioni con: Gabriele Gentile del Dipartimento di Biologia dell'Università Tor Vergata, per le iguane; con Monica Blasi dell’associazione Filicudi Wildlife Conservation, di Filicudi (Messina), Daniela Mattei dell’Istituto Superiore di Sanità di Roma, e Valeria Alduina del Dipartimento di Scienze e Tecnologie Biologiche Chimiche e Farmaceutiche dell’Università di Palermo, per le tartarughe marine.

 

Blasi M.F., Rotini A., Bacci T., Targusi M., Bonanno Ferraro G., Vecchioni L., Alduina R. and Migliore L. (2021). ON CARETTA CARETTA’S SHELL: FIRST SPATIAL ANALYSIS OF MICRO- AND MACRO-EPIBIONTS ON THE MEDITERRANEAN LOGGERHEAD SEA TURTLE CARAPACE. Marine Biology Research, 17(7-8): 726-774. DOI: 10.1080/17451000.2021.2016840

 

The loggerhead sea turtle, Caretta caretta (Linnaeus, 1758), is the most common sea turtle species in the Mediterranean Sea, where it can experience severe anthropogenic impacts. Although C. caretta is known to host more than 200 epibiotic taxa (crustaceans, algae and cyanobacteria), no reports have included a detailed evaluation of the microbial community of its carapace scutes. Thus, this study aimed to determine the diversity and composition of the visible and invisible communities on the carapace scutes of wild loggerhead turtles from the Aeolian Archipelago (Southern Tyrrhenian Sea, Italy) by using a combined approach of morphological/spatial examination and molecular analyses. Altogether, our results displayed a higher abundance of crustaceans, macroalgae and Proteobacteria on the posterior carapace scutes, while Firmicutes were more abundant on the anterior scutes. For the first time, this study showed the complexity of the microbial (invisible) and visible epibionts of the loggerhead sea turtles from the Mediterranean Sea and suggests the importance of including evaluation of the microbial components when studying epibiont communities.

 

Di Lallo G., D’Andrea M.M., Sennati S., Thaller M.C., Migliore L., Gentile G. (2021). EVIDENCE OF ANOTHER ANTHROPIC IMPACT ON IGUANA DELICATISSIMA FROM THE LESSER ANTILLES: THE PRESENCE OF ANTIBIOTIC RESISTANT ENTEROBACTERIA. Antibiotics, 10(8): 885. DOI: 10.3390/antibiotics10080885

 The improper use of antibiotics by humans may promote the dissemination of resistance in wildlife. The persistence and spread of acquired antibiotic resistance and human-associated bacteria in the environment, while representing a threat to wildlife, can also be exploited as a tool to monitor the extent of human impact, particularly on endangered animal species. Hence, we investigated both the associated enterobacterial species and the presence of acquired resistance traits in the cloacal microbiota of the critically endangered lesser Antillean iguana (Iguana delicatissima), by comparing two separate populations living in similar climatic conditions but exposed to different anthropic pressures. A combination of techniques, including direct plating, DNA sequencing and antimicrobial susceptibility testing allowed us to characterize the dominant enterobacterial populations, the antibiotic resistant strains and their profiles. A higher frequency of Escherichia coli was found in the samples from the more anthropized site, where multi-drug resistant strains were also isolated. These results confirm how human-associated bacteria as well as their antibiotic-resistance determinants may be transferred to wildlife, which, in turn, may act as a reservoir of antibiotic resistance.

 

Blasi M.F., Migliore L., Mattei D., Rotini A., Thaller M.C., Alduina R. (2020) - ANTIBIOTIC RESISTANCE OF GRAM-NEGATIVE BACTERIA FROM WILD CAPTURED LOGGERHEAD SEA TURTLES. Antibiotics, 9: 162. DOI: 10.3390/antibiotics9040162

Sea turtles have been proposed as health indicators of marine habitats and carriers of antibiotic-resistant bacterial strains, for their longevity and migratory lifestyle. Up to now, a few studies evaluated the antibacterial resistant flora of Mediterranean loggerhead sea turtles (Caretta caretta) and most of them were carried out on stranded or recovered animals. In this study, the isolation and the antibiotic resistance profile of 90 Gram negative bacteria from cloacal swabs of 33 Mediterranean wild captured loggerhead sea turtles are described. Among sea turtles found in their foraging sites, 23 were in good health and 10 needed recovery for different health problems (hereafter named weak). Isolated cloacal bacteria belonged mainly to Enterobacteriaceae (59%), Shewanellaceae (31%) and Vibrionaceae families (5%). Although slight differences in the bacterial composition, healthy and weak sea turtles shared antibiotic-resistant strains. In total, 74 strains were endowed with one or multi resistance (up to five different drugs) phenotypes, mainly towards ampicillin (~70%) or sulfamethoxazole/trimethoprim (more than 30%). Hence, our results confirmed the presence of antibiotic-resistant strains also in healthy marine animals and the role of the loggerhead sea turtles in spreading antibiotic-resistant bacteria.

 

 

 

Background: Antibiotic resistance, evolving and spreading among bacterial pathogens, poses a serious threat to human health. Antibiotic use for clinical, veterinary and agricultural practices provides the major selective pressure for emergence and persistence of acquired resistance determinants. However, resistance has also been found in the absence of antibiotic exposure, such as in bacteria from wildlife, raising a question about the mechanisms of emergence and persistence of resistant strains under similar conditions, and the implications for resistance control strategies. Since previous studies yielded some contrasting results, possibly due to differences in the ecological landscapes of the studied wildlife, we further investigated this issue in wildlife from a remote setting of the Galapagos archipelago.

Methodology/Principal Findings: Screening for acquired antibiotic resistance was carried out in commensal enterobacteria from Conolophus pallidus, the terrestrial iguana of Isla Santa Fe, where: i) the abiotic conditions ensure to microbes good survival possibilities in the environment; ii) the animal density and their habits favour microbial circulation between individuals; and iii) there is no history of antibiotic exposure and the impact of humans and introduced animal species is minimal except for restricted areas. Results revealed that acquired antibiotic resistance traits were exceedingly rare among bacteria, occurring only as non-dominant strains from an area of minor human impact.

Conclusions/Significance: Where both the exposure to antibiotics and the anthropic pressure are minimal, acquired antibiotic resistance traits are not normally found in bacteria from wildlife, even if the ecological landscape is highly favourable to bacterial circulation among animals. Monitoring antibiotic resistance in wildlife from remote areas could also be a useful tool to evaluate the impact of anthropic pressure.

 

 

 

The presence of Vibrio isolates was investigated in cloacal swabs from the Galápagos marine iguana (Amblyrhyncus cristatus). Such unique iguana is endemic to the Galápagos Archipelago, it is listed as vulnerable in the IUCN Red List (2009), and is strictly protected by CITES and Ecuador laws. Our results revealed an uneven isolation frequency of vibrios from animals living in different settings: maximal among the Santa Fe population, scarce at Bahía Tortuga but practically absent in the samples from Puerto Ayora and Plaza Sur. A 16S sequencing confirmed that the isolates belonged to the genus Vibrio, placing them within the V. alginolyticus group; the biochemical identification was, indeed, consistent with V. alginolyticus features. The reason of the observed discrepancy is not clear, but could be either linked to an higher pollution in the inhabited or more touristic places or to differential influence of chemical and physical parameters at a local scale. As V. alginolyticus is an opportunistic pathogen for man and it is known to cause disease in sea-living animals, the ability of these vibrios to enter and persist to a certain extent in the marine iguana gut should be regarded as a risk for health of both the animals and the human personnel involved in monitoring activities.