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  • The potential of Aeromonas spp. from wildlife as antimicrobial resistance indicators in aquatic environments
    Publication . Grilo, Miguel; Santos, Carla Sousa; Robalo, Joana; Oliveira, Manuela
    Declines in freshwater fish populations are occurring at a fast rate, increasing the importance of ex-situ conservation programs supported by sound knowledge of population dynamics and life-history traits of the target species. We analysed the growth and age structure of wild and captive stocks of the western ruivaco Achondrostoma occidentale, a Portuguese endangered cyprinid, targeted for captive breeding and restocking since 2007. Specifically, we compared maximum size, longevity, and length-at-age among captive and wild populations, restocked and non-restocked. We found considerable variation in length-at-age and longevity between captive-bred and wild fish, with the former generally growing faster and living longer. Analysis of length-age distributions among wild populations suggested a positive effect of restocking actions in fish abundance. Results highlight the value of captive breeding for the conservation of endangered cyprinids, and reinforce the need for detailed data on life-history trait variation between captive and wild stocks to assess the efficiency of ex-situ conservation programs.
  • Molecular epidemiology, virulence traits and antimicrobial resistance signatures of aeromonas spp. in the critically endangered iberochondrostoma lusitanicum follow geographical and seasonal patterns
    Publication . Grilo, Miguel; Isidoro, Sara; Chambel, Lélia; Marques, Carolina S.; Marques, Tiago A.; Sousa-Santos, C.; Robalo, Joana; Oliveira, Manuela
    Despite the fact that freshwater fish populations are experiencing severe declines worldwide, our knowledge on the interaction between endangered populations and pathogenic agents remains scarce. In this study, we investigated the prevalence and structure of Aeromonas communities isolated from the critically endangered Iberochondrostoma lusitanicum, a model species for threatened Iberian leuciscids, as well as health parameters in this species. Additionally, we evaluated the virulence profiles, antimicrobial resistance signatures and genomic relationships of the Aeromonas isolates. Lesion prevalence, extension and body condition were deeply affected by location and seasonality, with poorer performances in the dry season. Aeromonas composition shifted among seasons and was also different across river streams. The pathogenic potential of the isolates significantly increased during the dry season. Additionally, isolates displaying clinically relevant antimicrobial resistance phenotypes (carbapenem and fluroquinolone resistance) were detected. As it inhabits intermittent rivers, often reduced to disconnected pools during the summer, the dry season is a critical period for I. lusitanicum, with lower general health status and a higher potential of infection by Aeromonas spp. Habitat quality seems a determining factor on the sustainable development of this fish species. Also, these individuals act as reservoirs of important antimicrobial resistant bacteria with potential implications for public health.
  • Aeromonas spp. Prevalence, virulence, and antimicrobial resistance in an ex situ program for tThreatened freshwater fish—A pilot study with protective measures
    Publication . Grilo, Miguel; Amaro, Guadalupe; Chambel, Lélia; Marques, Carolina S.; Marques, Tiago A.; Gil, Fátima; Sousa-Santos, Carla; Robalo, Joana; Oliveira, Manuela
    Ex situ breeding programs are important conservation tools for endangered freshwater fish. However, developing husbandry techniques that decrease the likelihood of disease, antimicrobial resistance, and virulence determinants acquisition during this process is challenging. In this pilot study, we conducted a captivity experiment with Portuguese nase (Iberochondrostoma lusitanicum), a critically endangered leuciscid species, to investigate the influence of simple protective measures (i.e., material disinfection protocols and animal handling with gloves) on the dynamics of a potential pathogenic genus, Aeromonas, as well as its virulence profiles and antimicrobial resistance signatures. Our findings show that antimicrobial resistance in Aeromonas spp. collected from I. lusitanicum significantly increased during the extent of the assay (5 weeks), with all isolates collected at the end of the study classified as multidrug-resistant. Additionally, humans handling fishes without protective measures were colonized by Aeromonas spp. The use of protective measures suggested a decreasing trend in Aeromonas spp. prevalence in I. lusitanicum, while bacterial isolates displayed significantly lower virulence index values when virulence phenotypical expression was tested at 22 °C. Despite this study representing an initial trial, which needs support from further research, protective measures tested are considered a simple tool to be applied in ex situ breeding programs for aquatic animals worldwide. Furthermore, current results raise concern regarding antimicrobial resistance amplification and zoonotic transmission of Aeromonas spp. in aquatic ex situ programs.