Multiple Antimicrobial Resistance of Gram-Negative Bacteria from Natural Oligotrophic Lakes Under Distinct Anthropogenic Influence in a Tropical Region

Pontes, Daniela Santos; Pinheiro, F. A.; Lima-Bittencourt, Cláudia I.; Guedes, Rafael Lucas Muniz; Cursino, Luciana; Barbosa, Francisco A. R.; Santos, Fabrício R.; Chartone-Souza, Edmar; Nascimento, Andréa M. A.

doi:10.1007/s00248-009-9539-3

Cited by 53 publications

(48 citation statements)

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“…This result is in agreement with previous studies of bacterial diversity in freshwater, which have reported the occurrence of these phyla (Hiorns et al 1997, Berg et al 2009). Moreover, the dominance of Proteobacteria and the relatively small numbers of Bacteroidetes and Deinococcus-Thermus have also been reported in previous studies (Lemke et al 2009, Pontes et al 2009). Enterobacter, Moraxella, Staphylococcus and Acinetobacter have also been detected in lentic environments (Berg et al 2009, Lemke et al 2009).…”

Section: Discussionsupporting

confidence: 77%

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Phylogeny of cultivable heterotrophic bacteria derived from mixed colonies

Costa¹,

Lima-Bittencourt²,

Barbosa³

et al. 2012

Aquat. Microb. Ecol.

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In nature, bacteria interact with each other in ways that do not occur in pure laboratory cultures. However, when bacteria are purified from environmental samples, the resulting colonies occasionally harbor diverse bacterial isolates, which we have termed 'associated isolates'. Bacteria were obtained from a freshwater lake along a euphotic gradient (100, 10 and 1% light penetration). Surprisingly, 76 of the 1196 resulting colonies harbored between 2 and 5 associated isolates (for a total of 168 associated isolates), and 121 of these associated isolates were recovered as pure cultures. A portion of the isolates (47/168) was non-viable after re-streaking, suggesting an inability to survive and reproduce in the absence of their associates. Partial sequencing of 16S rDNA revealed that these isolates were affiliated with Proteobacteria, Firmicutes, Bacteroidetes, Actinobacteria and Deinococcus-Thermus, and represented 26 genera. Colonies harboring associated isolates and those that harbored the largest number of isolates were primarily sampled from the 1% euphotic gradient. Significant differences in the distribution of associated isolates along the euphotic gradient suggest that environmental factors are driving these associations.KEY WORDS: Associated bacteria · Intracolony · Diversity · 16S rDNA · Freshwater Resale or republication not permitted without written consent of the publisherAquat Microb Ecol 66: [95][96][97][98][99][100][101][102][103][104][105][106] 2012 through several stages of re-streaking to obtain pure cultures. Moreover, in many cases, when the physical separation of associated isolates is accomplished, one or all of them does not survive, suggesting their physiological inability to survive and reproduce in the absence of certain associations. The objective of this study was to provide insight into the phylogeny of isolates derived from an original colony. We looked for the presence of 2 or more different taxa from a single colony, performing a molecular and phylogenetic characterization of these isolates. These bacteria were collected from a natural freshwater lake in a tropical region. MATERIALS AND METHODS Study area and sampling siteCarioca Lake is a natural body of water situated in the middle of the Rio Doce Basin of Brazil. It is located in a Conservation Unit (Parque Estadual do Rio Doce, PERD, 19°29' 24'' to 19°48' 18'' S and 42°28' 18'' to 42°38' 30'' W) that is the largest remnant of Atlantic Forest in the state of Minas Gerais. Carioca Lake is meso trophic, round, shallow (11.8 m of maximum depth) and relatively small, with an area of 14.1 ha (Bortoluzzi et al. 2004, Bezerra-Neto et al. 2010). Sampling and bacterial isolationWater samples (500 ml) across a euphotic gradient in the limnetic (Lim) zone (Carioca Lake) were taken with Van Dorn bottles. The same water samples were used for physical and chemical measurements and bacterial culture. Collections took place on 23 June and 27 August 2007, which is the dry season. Sampling was conducted in the water column at diffe...

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Section: Discussionsupporting

confidence: 77%

“…The DNA was purified as previously described by Dramsi et al (1995). The complete 16S rRNA gene was amplified by touchdown PCR according to Pontes et al (2009), using the conserved primer set PA (5'-TCC TGG CTC AGA TTG AAC GC-3'), modified from Kuske et al (1997), and U2 (5'-ATC GGY TAC CTT GTT ACG ACT TC-3'), described by Lu et al (2000). …”

mentioning

confidence: 99%

Phylogeny of cultivable heterotrophic bacteria derived from mixed colonies

Costa¹,

Lima-Bittencourt²,

Barbosa³

et al. 2012

Aquat. Microb. Ecol.

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“…Due to the extensive human and veterinary utilization and the potentials to cause the selective proliferation of antibiotic resistant bacteria, antibiotics have raised increasing concerns recently (Pontes et al, 2009;Wilkinson et al, 2016). Antibiotics, including their parent compounds and transformation products, are being continuously discharged into the environment during manufacturing, consumption and disposal (Daughton and Ternes, 1999;Lin et al, 2010;Sharma et al, 2016).…”

Section: Introductionmentioning

confidence: 99%

Occurrence and removal of antibiotics in ecological and conventional wastewater treatment processes: A field study

Dong

Yuan

Wang

et al. 2016

Journal of Environmental Management

150

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a b s t r a c tThe occurrence and removal of 19 antibiotics (including four macrolides, eight sulfonamides, three fluoroquinolones, three tetracyclines, and trimethoprim) were investigated in two ecological (constructed wetland (CW) and stabilization pond (SP)) and two conventional wastewater treatment processes (activated sludge (AS) and micro-power biofilm (MP)) in a county of eastern China. All target antibiotics were detected in the influent and effluent samples with detection frequencies of >90%. Clarithromycin, ofloxacin, roxithromycin and erythromycin-H 2 O were the dominant antibiotics with maximum concentrations reaching up to 6524, 5411, 964 and 957 ng/L, respectively; while the concentrations of tiamulin, sulfamerazine, sulfathiazole, sulfamethazine, sulfamethizole and sulfisoxazole were below 10 ng/L. Although the mean effluent concentrations of target antibiotics were obviously lower than the influent ones (except ciprofloxacin), their removals were usually incomplete. Principal component analysis showed that the AS and CW outperformed the MP and SP processes and the AS performed better than the CW process in terms of antibiotics removal. Both the AS and CW processes exhibited higher removal efficiencies in summer than in winter, indicating biological degradation could play an important role in antibiotics removal. Because of the incomplete removal, the total concentration of detected antibiotics increased in the mixing and downstream sections of a local river receiving the effluent from a typical wastewater treatment facility practicing AS process. Nowadays, ecological wastewater treatment processes are being rapidly planned and constructed in rural areas of China; however, the discharge of residual antibiotics to the aquatic environment may highlight a necessity for optimizing or upgrading their design and operation.

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“…Recent studies reported that the bla TEM-1 gene was detected in most of the Gram-negative isolates resistant to ampicillin recovered from lakes in Brazil or from wastewater treatment plants in China (24,40). Furthermore, several studies identified extended-spectrum ␤-lactamases (ESBLs) from water samples.…”

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confidence: 99%

Diversity of Clavulanic Acid-Inhibited Extended-Spectrum β-Lactamases in Aeromonas spp. from the Seine River, Paris, France

Girlich

Poirel

Nordmann

2011

Antimicrob Agents Chemother

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Environmental Aeromonas sp. isolates resistant to ceftazidime were recovered during an environmental survey performed with water samples from the Seine River, in Paris, France, in November 2009. Selected isolates were identified by sequencing of the 16S rRNA and rpoB genes. PCR and cloning experiments were used to identify broad-spectrum-␤-lactamase-encoding genes and their genetic context. Clavulanic acid-inhibited extended-spectrum-␤-lactamase (ESBL) genes were identified in 71% of the Aeromonas sp. isolates. A variety of ESBL genes were detected, including bla VEB-1a , bla SHV-12 , bla PER-1 , bla PER-6 , bla TLA-2 , and bla GES-7 , suggesting an aquatic reservoir of those ESBL genes. Moreover, the repeated elements and different insertion sequences were identified in association with the bla PER-6 and the bla VEB-1a genes, respectively, indicating a wide diversity of mobilization events, making Aeromonas spp. a vehicle for ESBL dissemination.Bacteria with intrinsic or acquired resistance to antibiotics are commonly found in aquatic environments, where Pseudomonas, Serratia, and Aeromonas are commonly identified (2,17). Recent studies reported that the bla TEM-1 gene was detected in most of the Gram-negative isolates resistant to ampicillin recovered from lakes in Brazil or from wastewater treatment plants in China (24,40). Furthermore, several studies identified extended-spectrum ␤-lactamases (ESBLs) from water samples. For instance, the clinically relevant ESBL PER-1 was identified from Aeromonas media in a Swiss lake (32), and CTX-M-14 and TEM-52 were detected in Escherichia coli recovered from a South Korean river (21). Besides, nonclinically relevant Ambler class A ␤-lactamases TLA-2 and PER-6 were identified from uncultured bacteria (43,12) and Aeromonas allosaccharophila isolates, respectively (15).In addition, other studies reported on the identification of carbapenem-hydrolyzing ␤-lactamases in the environment, such as the Ambler class A ␤-lactamases IMI-2 and BIC-1 from Enterobacter asburiae (1, 5) and Pseudomonas fluorescens (14), respectively; the Ambler class B ␤-lactamase VIM-2 from Pseudomonas pseudoalcaligenes (41); and the class D ␤-lactamase OXA-23 from Acinetobacter baumannii (13).The present study was designed to evaluate the diversity of broad-spectrum ␤-lactam resistance determinants in Aeromonas species in water samples from the Seine River, Paris, France. The working hypothesis was that Aeromonas spp. could be an important reservoir for ESBLs. MATERIALS AND METHODSBacterial strains and plasmids. Three water samples (100 ml) were collected from approximately 0.2 m below the water surface in sterile bottles in November 2009 and filtered through nitrocellulose membranes (pore size, 0.45 m; Millipore), and the filters were resuspended in 3 ml of sterile water. Aliquots (0.1 ml) were plated on MacConkey agar plates supplemented with ceftazidime (2 g/ ml), and 10-fold dilutions were plated (0.1 ml) in parallel on antibiotic-free MacConkey agar plates for bacterial counts. Samples were proces...

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Multiple Antimicrobial Resistance of Gram-Negative Bacteria from Natural Oligotrophic Lakes Under Distinct Anthropogenic Influence in a Tropical Region

Cited by 53 publications

References 53 publications

Phylogeny of cultivable heterotrophic bacteria derived from mixed colonies

Phylogeny of cultivable heterotrophic bacteria derived from mixed colonies

Occurrence and removal of antibiotics in ecological and conventional wastewater treatment processes: A field study

Diversity of Clavulanic Acid-Inhibited Extended-Spectrum β-Lactamases in Aeromonas spp. from the Seine River, Paris, France

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