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Marine animals as sentinels to evaluate spread of carbapenemases producers
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Abstract(s)
Estirpes portadoras de carbapenemases (CP) constituem uma grande preocupação a nível
mundial, em virtude de inativarem os carbapenemos, e conferirem resistência não só a estes, que são
o último recurso para infeções nosocomiais, mas também a outros -lactâmicos. As carbapenemases
podem disseminar-se entre diferentes Géneros e Famílias e têm sido reportadas não só em estirpes
responsáveis por infecções nosocomiais, mas também em outros ambientes. Contudo, em animais
marinhos (MA) os dados são escassos. Este estudo teve por objectivo monitorizar a prevalência de
estirpes CP ou resistência a outros antimicrobianos incluindo metais pesados em MA provenientes da
Costa portuguesa.
Obtiveram-se 28 amostras de MA, numa lota de peixe da Região Centro de Portugal. A partir do
conteúdo visceral, com auxílio da técnica de pré-enriquecimento, procedeu-se à seleção de bacilos de
gram-negativo (GNB) Não-Fermentadores (NFB) e CP com recurso ao meio de cultura ChromID Carba
Smart Agar® e prova da oxidase. A identificação dos isolados foi realizada através da amplificação do
gene 16S rRNA por PCR (Polimerase Chain Reaction) e sequenciação. O estudo do perfil de
suscetibilidade incluiu nove antimicrobianos e realizou-se de acordo com as normas de referência
europeias; a pesquisa de carbapenemases e genes de tolerâncias a metais foi realizada por PCR.
Procedeu-se à análise estatística aplicando o teste exato de Fisher e recorrendo ao software
GRAPHPAD Prisma® (versão 8.4.2).
Foram selecionados 63 NFB: Pseudomonas spp., Aeromonas spp. e outros NFB (n=47/9/7,
respetivamente). Quanto ao perfil de resistência observou-se que: 100% eram resistentes à ticarcilina,
piperacilina, piperacilina-tazobactam, ceftazidima, ciprofloxacina e imipenemo (n=63/63); 27% ao
meropenemo (n=17/63); 13% à tobramicina (n=8/63: Aeromonas spp., Pseudomonas spp e NFB;
n=3/2/3, respetivamente); todos apresentaram suscetibilidade à amicacina. Nenhum dos seis isolados
suspeitos de serem PC (Pseudomonas spp., Aeromonas spp. e NFB: n=3/2/1, respetivamente) eram
portadores de genes codificadores das carbapenemases blaKPC, blaGES, blaIMP, blaNDM ou blaVIM. Os
genes merA (n=10/63) e silA (n=3/63), representaram uma prevalência de 15,87% e 4,76%,
respetivamente. Verificou-se que todos os isolados apresentaram perfil de MDR.
Os MA são portadores de bactérias MDR, mas não de estirpes CP. Os resultados realçam a
necessidade de monitorizar a AMR em ambiente marinho, particularmente no que respeita à estreita
ligação com a cadeia alimentar.
The carbapenemase-producing (CP) strains are a great concern worldwide, due to their capacity to inactivate the carbapenems, and conferring resistance not only to this group which is one of the lastresort antibiotics for nosocomial infections, but also to other groups of -lactams. The carbapenemases can be spread between various Genus and Families and have not only been reported, in nosocomial infections, but also in other environments. However, the available data in marine animals (MA) are scarce. This study aimed to monitor the prevalence of CP strains or resistance to other antimicrobials. Twenty-eight samples were recovered from MA, from a fish auction in the Centre region of Portugal. From the visceral content, with the aid of the pre-enrichment technique, CP gram-negative bacilli (GNB) Non-Fermenting (NFB) were selected using the ChromID Carba Smart Agar® culture medium and oxidase test. The isolates identification was performed by the amplification of the 16S rRNA gene by PCR (Chain Reaction Polymerase) and sequencing. The study of the susceptibility profile included nine antimicrobials and was carried out according to the European reference standards; research of carbapenemases and metal tolerance genes were performed by PCR. The statistical analysis was performed using the Fisher’s exact test and GRAPHPAD Prisma® software (version 8.4.2). Sixty-Three NFB were recovered: Pseudomonas spp., Aeromonas spp. and other NFB (n=47/9/7, respectively). As for the resistance profile, it was observed: 100% for ticarcillin, piperacillin, piperacillintazobactam, ceftazidime, ciprofloxacin and imipenem (n=63/63); 27% to meropenem (n=17/63); 13% to tobramycin (n=8/63: Aeromonas spp., Pseudomonas spp. and NFB; n=3/2/3, respectively); all were susceptible to amikacin. In none of the six representative isolates suspected of being CP (Pseudomonas spp., Aeromonas spp. and NFB, n=3/2/1, respectively) were observed blaKPC, blaGES, blaIMP, blaNDM or/and blaVIM encoding carbapenemases genes. The merA (n=10/63) and silA genes (n=3/63) represented a prevalence of 15,9% and 4,8%, respectively. It was noted that all the isolates carried a Multidrug-Resistant (MDR) profile. The MA harbour bacteria MDR, but not CP strains, thus allows to conclude that carbapems resistance was mediated by other mechanisms. The results highlight the need to monitor the antimicrobial resistance in the marine environment, particularly concerning the close connection with the food chain.
The carbapenemase-producing (CP) strains are a great concern worldwide, due to their capacity to inactivate the carbapenems, and conferring resistance not only to this group which is one of the lastresort antibiotics for nosocomial infections, but also to other groups of -lactams. The carbapenemases can be spread between various Genus and Families and have not only been reported, in nosocomial infections, but also in other environments. However, the available data in marine animals (MA) are scarce. This study aimed to monitor the prevalence of CP strains or resistance to other antimicrobials. Twenty-eight samples were recovered from MA, from a fish auction in the Centre region of Portugal. From the visceral content, with the aid of the pre-enrichment technique, CP gram-negative bacilli (GNB) Non-Fermenting (NFB) were selected using the ChromID Carba Smart Agar® culture medium and oxidase test. The isolates identification was performed by the amplification of the 16S rRNA gene by PCR (Chain Reaction Polymerase) and sequencing. The study of the susceptibility profile included nine antimicrobials and was carried out according to the European reference standards; research of carbapenemases and metal tolerance genes were performed by PCR. The statistical analysis was performed using the Fisher’s exact test and GRAPHPAD Prisma® software (version 8.4.2). Sixty-Three NFB were recovered: Pseudomonas spp., Aeromonas spp. and other NFB (n=47/9/7, respectively). As for the resistance profile, it was observed: 100% for ticarcillin, piperacillin, piperacillintazobactam, ceftazidime, ciprofloxacin and imipenem (n=63/63); 27% to meropenem (n=17/63); 13% to tobramycin (n=8/63: Aeromonas spp., Pseudomonas spp. and NFB; n=3/2/3, respectively); all were susceptible to amikacin. In none of the six representative isolates suspected of being CP (Pseudomonas spp., Aeromonas spp. and NFB, n=3/2/1, respectively) were observed blaKPC, blaGES, blaIMP, blaNDM or/and blaVIM encoding carbapenemases genes. The merA (n=10/63) and silA genes (n=3/63) represented a prevalence of 15,9% and 4,8%, respectively. It was noted that all the isolates carried a Multidrug-Resistant (MDR) profile. The MA harbour bacteria MDR, but not CP strains, thus allows to conclude that carbapems resistance was mediated by other mechanisms. The results highlight the need to monitor the antimicrobial resistance in the marine environment, particularly concerning the close connection with the food chain.
Description
Keywords
Animais marinhos Antimicrobianos Multiresistência Metais pesados Cabapenemases Marine animals Antimicrobials Multi drug resistant Heavy metals Carbapenemases