Browsing by Author "Belas, Adriana"
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- Biocide and antimicrobial susceptibility of methicillin-resistant staphylococcal isolates from horsesPublication . Couto, Natacha; Belas, Adriana; Tilley, Paula; Couto, Isabel; Gama, Luís T.; Kadlec, Kristina; Schwarz, Stefan; Pomba, ConstançaThe aim of this study was to evaluate the biocide and antimicrobial susceptibility of methicillin-resistant staphylococcal isolates from horses. Fourteen methicillin-resistant staphylococci (MRS) were subjected to an extensive genotype characterization, including SCCmec, dru, spa, PFGE and MLST typing. Antimicrobial susceptibility testing was performed and resistance genes were detected by PCR. Minimum bactericidal concentrations (MBCs) of four biocides [chlorhexidine acetate (CHA), benzalkonium chloride (BAC), triclosan (TCL) and glutaraldehyde (GLA)] were determined following the recommendations of document NF EN 1040. The presence of qac and sh-fabI genes was investigated by PCR. Several antimicrobial resistance patterns and genes were detected. When MRS strains were exposed for a longer period of time, a lower concentration of the biocide was needed to achieve lethality. TCL had the lowest MBC values. All MBC values were lower than the recommended in-use concentrations for veterinary medicine. S. haemolyticus and S. cohnii subsp. cohnii carried plasmid-borne qacA and sh-fabI or qacB and a qacH-like genes, respectively. Biocides appear to be a reliable antiseptic option against MRS, since even in the presence of bacterial efflux mechanisms, the recommended concentration is much higher than the in vitro MBC.
- Clonal relatedness of Proteus mirabilis strains causing urinary tract infections in companion animals and humansPublication . Marques, Cátia Filipa Saraiva; Belas, Adriana; Aboim, Catarina; Trigueiro, Graça; Cavaco-Silva, Patrícia; Gama, Luis; Pomba, C.ABSTRACT - Proteus mirabilis is a major cause of urinary tract infection (UTI) in humans and companion animals. This study aimed to evaluate the antimicrobial resistance, virulence and clonal relatedness of P. mirabilis isolated from dogs, cats and humans with UTI. P. mirabilis isolated from companion animals (N = 107) and humans (N = 76) with UTI were compared by PFGE analysis after overnight Nod macro-restriction using Dice/UPGMA with a 1.5% tolerance. Strains were characterized for antimicrobial resistance by disk diffusion. Twenty-four resistance genes and four virulence genes were screened by PCR. Thirty-nine clusters (similarity > 80%) and 73 single pulse-types were detected. Nine clusters included P. mirabilis isolated from community and hospital patients, including strains with 100% similarity. A high number of clusters (43.6%, n = 17/39) included strains from companion animals and humans. Similarity between some companion animal and human strains varied between 80-100%. One strain from a dog was 100% similar to one human community-acquired P. mirabilis. One P. mirabilis from a cat was found to be 94.7% and 92.4% similar to community and hospital patient strains, respectively. P. mirabilis CMY-2-producers did not cluster all together. Nevertheless, cluster C36 included five P. mirabilis from companion animals (similarity 85.8%-95.7%), of which, four (80%) were multidrug-resistant CMY-2-producers. This study shows that companion animals and humans become infected with closely related P. mirabilis strains. The high number of clusters containing companion animals and human strains points to the zoonotic nature of P. mirabilis. These results underline the potential role of companion animals as reservoirs and in the dissemination of uropathogenic P. mirabilis to humans and vice versa.
- Epidemiological study of pesticide poisoning in domestic animals and wildlife in Portugal : 2014–2020Publication . Grilo, Andreia; Moreira, Anabela S.; Carrapiço, Belmira; Belas, Adriana; Ferreira São Braz, BertaABSTRACT - Nowadays the intentional poisoning of domestic and wild animals is a crime in the European Union (EU), but as in the past the poison is still used in rural areas of a number of European countries to kill animals that were considered harmful for human activities. From January 2014 up until October 2020, the Laboratory of Pharmacology and Toxicology of the Faculty of Veterinary Medicine (LFT-FMV) has done the analytical detection of poisoning substances in 503 samples of wildlife and domestic animals and pesticides residues were found in 239 of the samples analyzed. In this retrospective study, toxicology results from domestic species (dog, cat, sheep, cows, and horses), wildlife species (red foxes, birds of prey, lynx, and wild boar), and food baits, are presented. During this period the samples analyzed at the LFT-FMV, were received from all over the country. Analytical detections were performed via solvent extraction followed by thin layer chromatography. Molluscicides (47%, n = 109) and Carbamates (24%, n = 57) were found to be the first category of pesticides involved in intoxications, in both domestic and wild animals, followed by rodenticides (13%, n = 30)—in this group second and third generation, were the most represented; Strychnine is the third (11%, n = 26) even though this pesticide has been banned in Portugal since 1988 and in the European Union since 2006 and finally Organophosphates (5%, n = 11) in the small number. This study allowed to realize that a great number of positive samples involved banned pesticides (i.e., Aldicarb and Strychnine) but, at the same time, many positives cases were due to the exposure to commercially available products (i.e., Methiocarb and Anticoagulant rodenticides). Also, it’s possible to identify the areas where domestic species are the most affected (i.e., Setubal and Lisboa) and the areas where the wild animals are the mainly affected species (i.e., Faro, Castelo Branco, and Bragança).
- Evidence of sharing of Klebsiella pneumoniae strains between healthy companion animals and cohabiting humansPublication . Marques, Cátia Filipa Saraiva; Belas, Adriana; Aboim, Catarina; Cavaco-Silva, Patrícia; Trigueiro, Graça; Gama, Luis; Pomba, C.This study aimed to characterize the fecal colonization and sharing of Klebsiella pneumoniae strains between companion animals and humans living in close contact. Fecal samples were collected from 50 healthy participants (24 humans, 18 dogs, and 8 cats) belonging to 18 households. Samples were plated onto MacConkey agar (MCK) plates with and without cefotaxime or meropenem supplementation. Up to five K. pneumoniae colonies per participant were compared by pulsed-field gel electrophoresis (PFGE) after XbaI restriction. K. pneumoniae strains with unique pulse types from each participant were characterized for antimicrobial susceptibility, virulence genes, and multilocus sequence type (MLST). Fecal K. pneumoniae pulse types were compared to those of clinical K. pneumoniae strains from animal and human patients with urinary tract infections (n = 104). K. pneumoniae colonization was detected in nonsupplemented MCK in around 38% of dogs (n = 7) and humans (n = 9). K. pneumoniae strains isolated from dogs belonged to sequence type 17 (ST17), ST188, ST252, ST281, ST423, ST1093, ST1241, ST3398, and ST3399. None of the K. pneumoniae strains were multidrug resistant or hypervirulent. Two households included multiple colonized participants. Notably, two colonized dogs within household 15 (H15) shared a strain each (ST252 and ST1241) with one coliving human. One dog from H16 shared one PFGE-undistinguishable K. pneumoniae ST17 strain with two humans from different households; however, the antimicrobial susceptibility phenotypes of these three strains differed. Two main virulence genotypes were detected, namely fimH-1 mrkD ycfM entB kfu and fimH-1 mrkD ycfM entB kpn. These results highlight the potential role of dogs as a reservoir of K. pneumoniae to humans and vice versa. Furthermore, to our best knowledge, this is the first report of healthy humans and dogs sharing K. pneumoniae strains that were undistinguishable by PFGE/MLST.
- Extended–spectrum–beta-lactamases, cephalosporinases and carbapenemase-producing Escherichia coli in the human-dog interfacePublication . Belas, Adriana; Pomba, Maria Constança Matias Ferreira; Carriço, João André Nogueira CustódioExtended–spectrum–beta-lactamases (ESBLs), cephalosporinases (encoded by the ESBLs and Ampc genes, respectively) and carbapenemase–producing Escherichia coli have become a major public health concern to both human and animal health. Urinary tract infections (UTI) are one of the most frequent bacterial infections in both human and companion animals. Uropathogenic E. coli (UPEC), belonging to extraintestinal pathogenic E. coli (ExPEC), is the most common bacterium isolated from companion animals. Moreover, the close contact of companion animals with humans creates opportunities for interspecies transmission of resistant bacteria and genes. E. coli from companion animals with UTI were found to harbour important antimicrobial resistance mechanisms and to belong to high-risk human clonal lineages, namely third-generation cephalosporin (3GC)-resistant E. coli O25b:H4-B2-ST131-H30/H30Rx, CC23 and ST648. In this work E. coli O25b:H4-ST131-H30/H30Rx was described for the first time in Europe in companion animals. Furthermore, the blaCMY-2 producing E. coli ST648 is the most common high-risk clonal lineage causing UTI in companion animals from the Lisbon area. Companion animals also seem to be reservoirs of bacteria and clinically important resistance genes, such β-lactams genes (classe A and C) which supports their role as reservoirs. The detection of faecal high-risk clone OXA-181-producing- E. coli ST410 strains that were closely related to uropathogenic clinical human strains was also an important finding and to our best knowledge was the first description in Portugal and Europe. These studies highligth the importance of companion animals as reservoirs of pathogenic E. coli harbouring important antimicrobial resistant genes. The emergence and spread of multidrug-resistant (MDR) E. coli in the natural environment by companion animal faecal contamination is also a concern towards animal and human health. These results point to need for control measures to prevent the dissemination of MDR ESBLs/AmpC and carbapenemases – producing bacteria from companion animals.
- Human and companion animal proteus mirabilis sharingPublication . Marques, Cátia Filipa Saraiva; Belas, Adriana; Menezes, Juliana; da Silva, Joana Moreira; Cavaco-Silva, Patrícia; Trigueiro, Graça; Gama, Luis; Pomba, C.Proteus mirabilis is an important pathogen that is associated with urinary tract infections. This study aims to determine the colonization and sharing of P. mirabilis between healthy companion animals and humans that are living together and to evaluate the clonal relatedness of the fecal and clinical stains. Eighteen households (24 humans, 18 dogs, 8 cats) with at least one human–animal pair were studied. Fecal samples were plated onto MacConkey and Hektoen agar and P. mirabilis PFGE analysis (NotI; Dice/UPGMA; 1.5% tolerance) was conducted for the households with multiple positive participants. Antimicrobial-resistance was tested according to CLSI. The fecal P. mirabilis pulse-types were compared with uropathogenic clinical strains (n = 183). Forty-nine P. mirabilis were isolated from eight households. The percentage of colonization in the dogs (44.4%, n = 8/18) was significantly higher (p = 0.0329) than in the humans (12.5%, n = 3/24). Three households had multiple colonized participants. One human–dog pair shared related P. mirabilis strains, which clustered with a clinical strain of animal origin (82.5%). One fecal P. mirabilis strain, from a dog, clustered with two human community-acquired clinical strains (80.9%, 88.9%). To our knowledge, this is the first report of dogs and humans living in close contact and sharing related P. mirabilis strains. The high frequency of colonization in the dogs underlines their possible role as P. mirabilis reservoirs for humans and other dogs.
- Influence of Particle Size and Extraction Methods on Phenolic Content and Biological Activities of Pear PomacePublication . Ferreira, Joana; Tkacz, Karolina; Turkiewicz, Igor Piotr; Santos, Maria Isabel; Belas, Adriana; Lima, Ana; Wojdyło, Aneta; Sousa, IsabelThe main goal of this research was to investigate how particle size influences the characteristics of pear (Pyrus Communis L.) pomace flour and to examine the impact of different pre-treatment methods on the phenolic content and associated bioactivities. Pear pomace flour was fractionated into different particle sizes, namely 1 mm, 710 m, 180 m, 75 m and 53 m. Then two extraction methods, namely maceration with methanol and two-step extraction with hexane via Soxhlet followed by ultrasound extraction with methanol, were tested. Total phenolic and total flavonoid contents ranged from 375.0 to 512.9 mg gallic acid/100 g DW and from 24.7 to 34.6 mg quercetin/100 g DW, respectively. Two-step extraction provided antioxidant activity up to 418.8 (in FRAP assay) and 340.0 mg Trolox/100 g DW (in DPPH assay). In order to explore various bioactive properties, this study assessed the inhibitory effects of enzymes, specifically -amylase and -glucosidase (associated with antidiabetic effects), as well as angiotensin-converting enzyme (linked to potential antihypertensive benefits). Additionally, the research investigated antibacterial potential against both Gram-negative (E. coli) and Gram-positive (S. aureus) bacteria, revealing significant results (p < 0.05), particularly in the case of the two-step extraction method. This investigation underscores the substantial value of certain food industry wastes, highlighting their potential as bioactive ingredients within the framework of a circular economy.
- Oxa-181-producing extraintestinal pathogenic escherichia coli sequence type 410 isolated from a dog in PortugalPublication . Brilhante, Michael; Menzes, Juliana; Belas, Adriana; Feudi, Claudia; Schwarz, Stefan; Pomba, C.; Perretena, VincentTwo multidrug-resistant and carbapenemase-producing Escherichia coli clones of sequence type 410 were isolated from fecal samples of a dog with skin infection on admission to an animal hospital in Portugal and 1 month after discharge. Whole-genome sequencing revealed a 126,409-bp Col156/IncFIA/IncFII multidrug resistance plasmid and a 51,479-bp IncX3 blaOXA-181-containing plasmid. The chromosome and plasmids carried virulence genes characteristic for uropathogenic E. coli, indicating that dogs may carry multidrug-resistant E. coli isolates related to those causing urinary tract infections in humans.