Molecular characterization of antibiotic resistance in enterococci recovered from seagulls (Larus cachinnans) representing an environmental health problem

Antimicrobial resistance and the mechanisms implicated were studied in 54 enterococci recovered from 57 seagull fecal samples. Almost 78% of the recovered enterococci showed resistance against one or more antibiotics and these isolates were identified to the species level. E. faecium was the most prevalent species (52.4%). High percentages of erythromycin and tetracycline resistances were found among our isolates (95.2%), and lower percentages were identified to other antibiotics. Most of the tetracycline-resistant strains carried the tet(M) and/or tet(L) genes. Genes associated with Tn916/Tn1545 and/or Tn5397 transposons were detected in 45% of tetracycline-resistant isolates. The erm(B) gene was detected in 65% of erythromycin-resistant isolates. The vat(D) and vat(E) genes were present in 5.9% and 11.8% of quinupristin/dalfopristin-resistant isolates, respectively. The ant(6)-Ia gene was identified in 57.1% of streptomycin-resistant isolates. All nine kanamycin-resistant isolates carried the aph(3)'-IIIa gene. The cat(A) gene was found in two chloramphenicol-resistant isolates. Seagulls should be considered a risk species for spreading in the environment antimicrobial resistant enterococci and can serve as a sentinel for antibiotic pressure from the surrounding farm and urban setting.     

Speciation and antimicrobial resistance of Enterococci isolated from recreational beaches in Malaysia

We report the first study on the occurrence of antibiotic-resistant enterococci in coastal bathing waters in Malaysia. One hundred and sixty-five enterococci isolates recovered from two popular recreational beaches in Malaysia were speciated and screened for antibiotic resistance to a total of eight antibiotics. Prevalence of Enterococcus faecalis and Enterococcus faecium was highest in both beaches. E. faecalis/E. faecium ratio was 0.384:1 and 0.375:1, respectively, for isolates from Port Dickson (PD) and Bagan Lalang (BL). Analysis of Fisher’s exact test showed that association of prevalence of E. faecalis and E. faecium with considered locations was not statistically significant (p?<?0.05). Chi-square test revealed significant differences (χ ²?=?82.630, df?=?20, p?<?0.001) in the frequency of occurrence of enterococci isolates from the considered sites. Resistance was highest to nalidixic acid (94.84 %) and least for chloramphenicol (8.38 %). One-way ANOVA using Tukey–Kramer multiple comparison test showed that resistance to ampicillin was higher in PD beach isolates than BL isolates and the difference was extremely statistically significant (p?<?0.0001). Frequency of occurrence of multiple antibiotic resistance (MAR) isolates were higher for PD beach water (64.29 %) as compared to BL beach water (13.51 %), while MAR indices ranged between 0.198 and 0.48. The results suggest that samples from Port Dickson may contain MAR bacteria and that this could be due to high-risk faecal contamination from sewage discharge pipes that drain into the sea water.     

Antibiotic resistance profile of bacterial isolates from animal farming aquatic environments and meats in a peri-urban community in Daejeon, Korea

The increasing usage of antibiotics in the animal farming industry is an emerging worldwide problem contributing to the development of antibiotic resistance. The purpose of this work was to investigate the prevalence and antibiotic resistance profile of bacterial isolates collected from animal farming aquatic environments and meats in a peri-urban community in Daejeon, Korea. In an antibacterial susceptibility test, the bacterial isolates showed a high incidence of resistance (～26.04%) to cefazolin, tetracycline, gentamycin, norfloxacin, erythromycin and vancomycin. The results from a test for multiple antibiotic resistance indicated that the isolates were displaying an approximately 5-fold increase in the incidence of multiple antibiotic resistance to combinations of two different antibiotics compared to combinations of three or more antibiotics. Most of the isolates showed multi-antibiotic resistance, and the resistance patterns were similar among the sampling groups. Sequencing data analysis of 16S rRNA showed that most of the resistant isolates appeared to be dominated by the classes Betaproteobacteria and Gammaproteobacteria, including the genera Delftia, Burkholderia, Escherichia, Enterobacter, Acinetobacter, Shigella and Pseudomonas.     

Using Multiple Antibiotic Resistance and land use characteristics to determine sources of fecal coliform bacterial pollution

Multiple Antibiotic Resistance (MAR) analysis and regression modeling techniques were used to identify surface water areas impacted by fecal pollution from human sources, and to determine the effects of land use on fecal pollution in Murrells Inlet, a small, urbanized, high-salinity estuary located between Myrtle Beach and Georgetown, South Carolina. MAR analysis was performed to identify areas in the estuary that are impacted by human-source fecal pollution. Additionally, regression analysis was performed to determine if an association exists between land use and fecal coliform densities over the ten-year period from 1989 to 1998. Land-use variables were derived using Geographic Information System (GIS) techniques and were used in the regression analysis.MAR analyses were conducted by comparing the frequency and patterns of antibiotic resistance found in Escherichia coli isolates derived from surface water samples and from sewage sources in the Murrells Inlet sewage collection system. The MAR results suggest that the majority of the fecal pollution detected in the Murrells Inlet estuary may be from non-human sources, including fecal coliforms isolated from areas in close proximity to high densities of active septic tanks.A MAR Index, which measures the frequency of antibiotic resistance, was calculated for each of twenty-three water samples and nine sewage samples. The antibiotic resistance pattern comparisons were performed using cluster analysis. Although the MAR indices indicated that several surface water sites had potential human-source contamination, the cluster analysis suggests that only one sampling site had MAR patterns that were similar to those found in the sewage samples. This site was in close proximity to several large pleasure boats as well as a sewage collection system lift station, but was not near areas with active septic tanks. The results of the regression analysis also suggest that sewage sources and rainfall runoff from urbanized areas may contribute to fecal pollution in the estuary.     

Assessment of physicochemical parameters and prevalence of virulent and multiple-antibiotic-resistant <Emphasis Type="Italic">Escherichia coli</Emphasis> in treated effluent of two wastewater treatment plants and receiving aquatic <Emphasis Type="Italic">milieu</Emphasis> in Durban,South Africa

The poor operational status of some wastewater treatment plants often result in the discharge of inadequately treated effluent into receiving surface waters. This is of significant public health concern as there are many informal settlement dwellers (ISDs) that rely on these surface waters for their domestic use. This study investigated the treatment efficiency of two independent wastewater treatment plants (WWTPs) in Durban, South Africa and determined the impact of treated effluent discharge on the physicochemical and microbial quality of the receiving water bodies over a 6-month period. Presumptive Escherichia coli isolates were identified using biochemical tests and detection of the mdh gene via PCR. Six major virulence genes namely eae, hly, fliC, stx1, stx2, and rfbE were also detected via PCR while antibiotic resistance profiles of the isolates were determined using Kirby-Bauer disc diffusion assay. The physicochemical parameters of the wastewater samples ranged variously between 9 and 313.33 mg/L, 1.52 and 76.43 NTUs, and 6.30 and 7.87 for COD, turbidity, and pH respectively, while the E. coli counts ranged between 0 and 31.2?×?10³ CFU/ml. Of the 200 selected E. coli isolates, the hly gene was found in 28 %, fliC in 20 %, stx2 in 17 %, eae in 14 %, with stx1 and rfbE in only 4 % of the isolates. Notable resistance was observed toward trimethoprim (97 %), tetracycline (56 %), and ampicillin (52.5 %). These results further highlight the poor operational status of these WWTPs and outline the need for improved water quality monitoring and enforcement of stringent guidelines.     

Antibiotic resistance analysis of fecal coliforms to determine fecal pollution sources in a mixed-use watershed

Antibiotic resistance analysis was performed on fecal coliform(FC) bacteria from a mixed-use watershed to determine thesource, human or nonhuman, of fecal coliform contamination. The study consisted of discriminant analysis of antibioticresistance patterns generated by exposure to fourconcentrations of six antibiotics (ampicillin, gentamicinsulfate, kanamycin, spectinomycin dihydrochloride,streptomycin sulfate, and tetracycline hydrochloride). Areference database was constructed from 1125 fecal coliformisolates from the following sources: humans, domestic animals(cats and dogs), agricultural animals (chickens, cattle, andhorses), and wild animals. Based on similar antibioticresistance patterns, cat and dog isolates were grouped asdomestic animals and horse and cattle isolates were grouped aslivestock. The resulting average rate of correctclassification (ARCC) for human and nonhuman isolates was94%. A total of 800 FC isolates taken from the watershedduring either a dry event or a wet event were classifiedaccording to source. Human sources contribute a majority(>50%) of the baseflow FC isolates found in the watershed inurbanized areas. Chicken and livestock sources areresponsible for the majority of the baseflow FC isolates foundin the rural reaches of the watershed. Stormwater introducesFC isolates from domestic (16%) and wild (21%) sourcesthroughout the watershed and varying amounts (up to 60%) fromchicken and livestock sources. These results suggest thatantibiotic resistance patterns of FC may be used to determinesources of fecal contamination and aid in the direction ofwater quality improvement.     

Antibiotic resistance and plasmid profiling of <Emphasis Type="Italic">Vibrio</Emphasis> spp. in tropical waters of Peninsular Malaysia

Vibrio species isolated from four different sampling stations in the west coast of Peninsular Malaysia were screened for their antimicrobial resistance and plasmid profiles. A total of 138 isolates belonging to 15 different species were identified. Vibrio campbellii, V. parahaemolyticus, V. harveyi, and V. tubiashii were found to predominance species at all stations. High incidence of erythromycin, ampicillin, and mecillinam resistance was observed among the Vibrio isolates. In contrast, resistance against aztreonam, cefepime, streptomycin, sulfamethoxazole, and sulfonamides was low. All the Vibrio isolates in this study were found to be susceptible to imipenem, norfloxacin, ofloxacin, chloramphenicol, trimethoprim/sulfamethoxazole, and oxytetracycline. Ninety-five percent of the Vibrio isolates were resistant to one or more different classes of antibiotic, and 20 different resistance antibiograms were identified. Thirty-two distinct plasmid profiles with molecular weight ranging from 2.2 to 24.8 kb were detected among the resistance isolates. This study showed that multidrug-resistant Vibrio spp. were common in the aquatic environments of west coast of Peninsular Malaysia.     

Fecal source tracking by antibiotic resistance analysis on a watershed exhibiting low resistance

The ongoing development of microbial source tracking has made it possible to identify contamination sources with varying accuracy, depending on the method used. The purpose of this study was to test the efficiency of the antibiotic resistance analysis (ARA) method under low resistance by tracking the fecal sources at Turkey Creek, Oklahoma exhibiting this condition. The resistance patterns of 772 water-isolates, tested with nine antibiotics, were analyzed by discriminant analysis (DA) utilizing a five-source library containing 2250 isolates. The library passed various representativeness tests; however, two of the pulled-sample tests suggested insufficient sampling. The resubstitution test of the library individual sources showed significant isolate misclassification with an average rate of correct classification (ARCC) of 58%. These misclassifications were explained by low antibiotic resistance (Wilcoxon test P < 0.0001). Seasonal DA of stream E. coli isolates for the pooled sources human/livestock/deer indicated that in fall, the human source dominated (P < 0.0001) at a rate of 56%, and that human and livestock respective contributions in winter (35 and 39%), spring (43 and 40%), and summer (37 and 35%) were similar. Deer scored lower (17–28%) than human and livestock at every season. The DA was revised using results from a misclassification analysis to provide a perspective of the effect caused by low antibiotic resistance and a more realistic determination of the fecal source rates at Turkey Creek. The revision increased livestock rates by 13–14% (0.04 ≤ P ≤ 0.06), and decreased human and deer by 6–7%. Negative misclassification into livestock was significant (0.04 ≤ P ≤ 0.06). Low antibiotic resistance showed the greatest effect in this category.     

Aeromonas and Pseudomonas: antibiotic and heavy metal resistance species from Iskenderun Bay, Turkey (northeast Mediterranean Sea)

We studied the susceptibility patterns to 15 different antibiotics and six heavy metals in Aeromonas spp. and Pseudomonas spp. isolated from Iskenderun Bay, Turkey (northeast Mediterranean Sea). A high percentage of Aeromonas isolates showed resistance to cefazolin (66.6%) and trimethoprim–sulphamethoxazole (66.6%). Amongst the Pseudomonas isolates, there was a high incidence of resistance to nitrofurantoin (86.2%), cefazolin (84.8%) and cefuroxime (71.7%). Most isolates showed tolerance to different concentrations of heavy metals, and minimal inhibition concentrations ranged from 25 to >3,200 μg/ml. The Aeromonas spp. and Pseudomonas spp. showed high resistance to copper of 98.3% and 75.4%, respectively, and low resistance to lead of 1.7% and 7.2%, respectively. Our results show that antibiotic and heavy metal resistant Aeromonas spp. and Pseudomonas spp. were widespread in Iskenderun Bay in 2007 and 2008. The increasing presence of antibiotic and heavy metal resistant Aeromonas spp. and Pseudomonas spp. may become a potential human health hazard.     

Incidence of metal and antibiotic resistance in Pseudomonas spp. from the river water, agricultural soil irrigated with wastewater and groundwater

A total of 144 isolates of Pseudomonas spp. (48 each from the Yamuna River water, wastewater irrigated soil and groundwater irrigated soil) were tested for their resistance against certain heavy metals and antibiotics. Minimum inhibitory concentrations (MICs) of Hg^2?+?, Cd^2?+?, Cu^2?+?, Zn^2?+?, Ni^2?+?, Pb^2?+?, Cr^3?+? and Cr^6?+? for each isolate were also determined. A maximum MIC of 200 ??g/ml for mercury and 3,200 ??g/ml for other metals were observed. The incidences of metal resistance and MICs of metals for Pseudomonas isolates from the Yamuna water and wastewater irrigated soil were significantly different to those of groundwater irrigated soil. A high level of resistance against tetracycline and polymyxin B (81.2%) was observed in river water isolates. However, 87.5% of Pseudomonas isolates from soil irrigated with wastewater showed resistance to sulphadiazine, whereas 79.1% were resistant to both ampicillin and erythromycin. Isolates from soil irrigated with groundwater exhibited less resistance towards heavy metals and antibiotics as compared to those of river water and wastewater irrigated soil. Majority of the Pseudomonas isolates from water and soil exhibited resistance to multiple metals and antibiotics. Resistance was transferable to recipient Escherichia coli AB2200 strains by conjugation. Plasmids were cured with the curing agent ethidium bromide and acridine orange at sub-MIC concentration.     

Prevalence of antibiotic-resistant bacteria in three different aquatic environments over three seasons

The objective of this study was to evaluate the impact of urbanization and seasonal changes on the prevalence of antibiotic-resistant bacteria in different aqueous environments. To this end, bacteria were isolated from three different water sources: the River Hooghly in Kolkata, River Kangsabati and groundwater from Kharagpur, West Bengal over three seasons: post-monsoon, winter and summer in 2012–2013. A total of 163 Gram-negative bacteria were isolated from the River Hooghly (n?=?138), River Kangsabati (n?=?13) and groundwater (n?=?12). Antibiotic susceptibility testing was done using 12 antibiotic discs. The percentages of multiple antibiotic-resistant (MAR) bacteria at the three sampling locations were found to be 71.01 % (98/138) for River Hooghly, 15.38 % (2/13) for River Kangsabati and 8.33 % (1/12) for groundwater. Prevalence of MAR bacteria with respect to the three seasons were the following: 73.58 % in post-monsoon, 59.26 % in winter and 53.57 % in summer. Antibiotic resistance index (ARI) was calculated for each location and each season. In general, ARI values for all the River Hooghly samples were >0.2 while those for the River Kangsabati and groundwater in Kharagpur were always <0.2 indicating greater exposure to antibiotics and subsequent resistance in bacteria from the River Hooghly compared to the other two locations. In addition, percentage of MAR and ARI values followed the trend: post-monsoon?>?winter?>?summer. This may be due to the additional terrestrial resistants that get swept along with surface runoff during the monsoons.     

Prevalence of heavy metal resistance in bacteria isolated from tannery effluents and affected soil

In the present study, a total of 198 bacteria were isolated, 88 from the tannery effluents and 110 from agricultural soil irrigated with the tannery effluents. Tannery effluents and soils were analyzed for metal concentrations by atomic absorption spectrophotometer. The tannery effluents and soil samples were found to be contaminated with chromium, nickel, zinc, copper, and cadmium. All isolates were tested for their resistance against Cr^6?+?, Cr^3?+?, Ni^2?+?, Zn^2?+?, Cu^2?+?, Cd^2?+?, and Hg^2?+?. From the total of 198 isolates, maximum bacterial isolates were found to be resistant to Cr^6?+? 178 (89.9%) followed by Cr^3?+? 146 (73.7%), Cd^2?+? 86 (43.4%), Zn^2?+? 83 (41.9%), Ni^2?+? 61 (30.8%), and Cu^2?+? 51 (25.6%). However, most of the isolates were sensitive to Hg^2?+?. Among the isolates from tannery effluents, 97.8% were resistant to Cr^6?+? and 64.8% were resistant to Cr^3?+?. Most of the soil isolates were resistant against Cr^6?+? (83.6%) and Cr^3?+? (81.8%). All isolates were categorized into Gram-positive and Gram-negative bacteria. In a total of 114 Gram-positive isolates, 91.2% were resistant to Cr^6?+? followed by 73.7% to Cr^3?+?, 42.1% to Zn^2?+?, 40.4% to Cd^2?+?, and 32.5% to Ni^2?+?. Among Gram-negative isolates, 88.1% were found showing resistance to Cr^6?+?, 75.0% to Cr^3?+?, and 47.6% were resistant to Cd^2?+?. Majority of these metal-resistant isolates were surprisingly found sensitive to the ten commonly used antibiotics. Out of 198 isolates, 114 were found sensitive to all antibiotics whereas only two isolates were resistant to maximum eight antibiotics at a time. Forty-one and 40 isolates which constitute 20.7% and 20.2% were resistant to methicilin and amoxicillin, respectively.     

Antibiotic-resistant E. coli in surface water and groundwater in dairy operations in Northern California

Generic Escherichia coli was isolated from surface water and groundwater samples from two dairies in Northern California and tested for susceptibility to antibiotics. Surface samples were collected from flush water, lagoon water, and manure solids, and groundwater samples were collected from monitoring wells. Although E. coli was ubiquitous in surface samples with concentrations ranging from several hundred thousand to over a million colony-forming units per 100 mL of surface water or per gram of surface solids, groundwater under the influence of these high surface microbial loadings had substantially fewer bacteria (3- to 7-log₁₀ reduction). Among 80 isolates of E. coli tested, 34 (42.5 %) were resistant to one or more antibiotics and 22 (27.5 %) were multi-antibiotic resistant (resistant to ≥3 antibiotics), with resistance to tetracycline, cefoxitin, amoxicillin/clavulanic acid, and ampicillin being the most common. E. coli isolates from the calf hutch area exhibited the highest levels of multi-antibiotic resistance, much higher than isolates in surface soil solids from heifer and cow pens, flush alleys, manure storage lagoons, and irrigated fields. Among E. coli isolates from four groundwater samples, only one sample exhibited resistance to ceftriaxone, chloramphenicol, and tetracycline, indicating the potential of groundwater contamination with antibiotic-resistant bacteria from dairy operations.     

Determination of antimicrobial and heavy metal resistance profiles of some bacteria isolated from aquatic amphibian and reptile species

The aim of the present study was to determine the level of antibiotic resistance patterns and distribution of heavy metal resistance of bacterial isolates from aquatic animals (Lissotriton vulgaris, Pelophylax ridibundus, Emys orbicularis, Mauremys rivulata, and Natrix natrix) in Turkey (Kavak Delta). A total of 153 bacteria have been successfully isolated from cloaca and oral samples of the aquatic amphibians and reptilians which were found, namely, Aeromonas sp. (n?=?29), Plesiomonas sp. (n?=?7), Vibrio sp. (n?=?12), Citrobacter sp. (n?=?12), Enterobacter sp. (n?=?11), Escherichia sp. (n?=?22), Klebsiella sp. (n?=?22), Edwardsiella sp. (n?=?6), Hafnia sp. (n?=?1), Proteus sp. (n?=?19), Providencia sp. (n?=?8), and Pseudomonas sp. (n?=?4). In terms of antibiotic and heavy metal susceptibility testing, each isolate was tested against 12 antibiotics and 4 metals. There was a high incidence of resistance to cefoxitin (46.40 %), ampicillin (44.44 %), erythromycin (35.29 %), and a low incidence of resistance to gentamicin (6.53 %), kanamycin (8.49 %), chloramphenicol (9.15 %), and cefotaxime (10.45 %). The multiple antibiotic resistance index of each bacterial species indicated that bacteria from raised amphibians and reptiles have been exposed to tested antibiotics, with results ranging from 0 to 0.58. Most isolates showed tolerance to different concentrations of heavy metals, and minimal inhibition concentrations ranged from100 to >3,200 μg/mL. According to these results, a significant occurrence of bacteria in the internal organs of reptiles and amphibians, with a high incidence of resistance against antibiotics and heavy metals, may risk aquatic animals and the public health. These data appoint the importance of epidemiological surveillance and microbiological monitoring and reinforce the need to implement environment protection programs for amphibian and reptile species.     

Assessment of animal impacts on bacterial water quality in a South Carolina, USA tidal creek system

Fecal pollution may adversely impact water quality in coastal ecosystems. The goal of this study was to determine whether cattle were a source of fecal pollution in a South Carolina watershed. Surface water samples were collected in June 2002 and February through March 2003 in closed shellfish harvesting waters of Toogoodoo Creek in Charleston County, SC. Fecal coliform concentrations in 70 % of the water samples taken for this study exceeded shellfish harvesting water standards. Ribotyping was performed in order to identify animal sources contributing to elevated fecal coliform levels. Escherichia coli isolates (n?=?253) from surface water samples were ribotyped and compared to a ribotype library developed from known sources of fecal material. Ribotypes from water samples that matched library ribotypes with 90 % maximum similarity or better were assigned to that source. Less than half of the unknown isolates (38 %) matched with library isolates. About half (53 %) of the matched ribotypes were assigned to cattle isolates and 43 % to raccoon. Ribotyping almost exclusively identified animal sources. While these results indicate that runoff from cattle farms was a likely source of fecal pollution in the watershed, wildlife also contributed. Given the small size of the library, ribotyping was moderately useful for determining the impact of adjacent cattle farms on Toogoodoo Creek. Increasing the number and diversity of the wildlife sources from the area would likely increase the usefulness of the method.     

Potential impacts of disinfection processes on elimination and deactivation of antibiotic resistance genes during water and wastewater treatment

Antibiotic resistance genes (ARGs), in association with antibiotic resistant bacteria (ARB), have been identified as widespread contaminants of treated drinking waters and wastewaters. As a consequence, concerns have been raised that ARB or ARG transport between aquatic compartments may enhance the spread of antibiotic resistance amongst non-resistant bacterial communities by means of horizontal gene transfer processes. Most often, discussion of horizontal gene transfer focuses on the probable role of conjugative plasmid or transposon exchange, which requires live ARB donor cells. Conventional water and wastewater disinfection processes generally provide highly effective means for mitigating the transport of live ARB; thereby minimizing risks of conjugative gene transfer. However, even if ARB present in a treated water are fully inactivated during a disinfection process, the possibility remains that intact remnants of DNA contained within the resulting cell debris could still confer resistance genotypes to downstream bacterial populations by means of natural transformation and/or transduction, which do not require live donor cells. Thus, a systematic evaluation of the capability of common disinfection technologies to ensure the destruction of bacterial DNA, in addition to pathogen inactivation, seems warranted. With that objective in mind, this review seeks to provide a concise introduction to the significance of ARB and ARG occurrence in environmental systems, coupled with a review of the role that commonly used water and wastewater disinfection processes may play in minimizing ARG transport and dissemination.     

Levels of some toxic elements and frequency of bacterial heavy metal resistance in sediment and sea water

The Golden Horn Estuary located in the Istanbul region of Turkey has been thought to be a heavily polluted area since the 1950s: the concentration of the elements, which include heavy metals such as Cu2+, Mn2+, Ni2+, Zn2+, Pb2+, Cd2+, and Fe2+, was therefore investigated in sediment and water samples of the area. The resistance of Enterobacteriaceae members to some heavy metal salts was investigated to determine levels of metal-resistant bacteria in the Golden Horn Estuary after the environmental restoration project in 1998. The sediment samples were collected with an Ekman-Grab in the period from November 2002 to February 2004 from depths of 4-15 m and analyzed by means of an atomic absorption spectrophotometer. Analyses of average heavy metals of sediment samples yielded the following results: 131.5 ppm Cu, 405.5 ppm Mn, 46.5 ppm Ni, 191.2 ppm Zn, 81.5 ppm Pb and 27668 ppm Fe. As with water, Ni concentrations in sediment were found lower than that in limit values. Frequency of heavy metals resistance to Cu, Zn, Fe, Ni, Mn, Pb and Cd was detected as an average of 65.0%, 64.4%, 62.5%, 38.4%, 37.3%, 36.2% and 28.4%, respectively in a total of 192 strains isolated from sediment samples. It was observed that there was no statistically significant difference among the results of analyses with respect to sampling dates. Despite the environmental restoration project in 1998, our study results showed that heavy metal levels were still high in the sediment and this situation induced the tolerance of bacteria to some heavy metals.     

Characterization of lead-resistant river isolate Enterococcus faecalis and assessment of its multiple metal and antibiotic resistance

Contamination of surface waters has a direct impact on the public health of entire communities. Microorganisms inhabiting contaminated surface waters have developed mechanisms of coping with a variety of toxic metals and drugs. Investigations were carried out to isolate and identify lead-resistant bacteria from the river K?z?l?rmak along the city of K?r?kkale, Turkey. Of the 33 lead-resistant isolates, one isolate with a minimal inhibitory concentration of 1,200 mg?L^?1 was isolated and identified as Enterococcus faecalis by using biochemical tests and 16S rRNA sequencing. Lead-resistant E. faecalis isolate was found out to be resistant to other heavy metals like aluminum, lithium, barium, chromium, iron, silver, tin, nickel, zinc, and strontium and to drugs like amikacin, aztreonam, and gentamicin. E. faecalis harbored four plasmids with the molecular sizes of 1.58, 3.06, 22.76, and 28.95 kb. Plasmid profile analyses of cured derivatives revealed that the lead resistance ability of E. faecalis was still existing despite the elimination of all the plasmids. Moreover, the antibiotic resistance pattern of the cured derivatives did not demonstrate any change from the parental strain. Our findings indicated that the lead resistance genes of E. faecalis were located on the chromosomal DNA rather than the plasmid.     

The residues and environmental risks of multiple veterinary antibiotics in animal faeces

To understand the residues and ecological risks of veterinary antibiotics (VAs) in animal faeces from concentrated animal feeding operations in northeastern China, 14 VAs were identified by high performance liquid chromatography, and the preliminary risks of six antibiotics were assessed using the hazard quotient (HQ). The investigated VAs occurred in 7.41 to 57.41 % of the 54 samples, and the levels ranged from 0.08 to 56.81 mg kg^?1. Tetracyclines were predominant with a maximum level of 56.81 mg kg^?1 mostly detected in pig faeces. Sulfonamides were common and detected with the highest concentration of 7.11 mg kg^?1. Fluoroquinolones were more widely detected in chicken faeces rather than in pig or cow faeces, which contained the dominant antibiotic enrofloxacin. In comparison, the residue of tylosin was less frequently found. The risk evaluations of the six antibiotics revealed that tetracyclines, especially oxytetracycline, displayed the greatest ecological risk because of its high HQ value of 15.75. The results of this study imply that multiple kinds of VAs were jointly used in animal feeding processes in the study area. These medicine residues in animal faeces may potentially bring ecological risks if the animal manure is not treated effectively.     

Isolation and Identification of Enterococci from Seawater Samples: Assessment of Their Resistance to Antibiotics and Heavy Metals

A hundred Enterococcus strains were isolated from seawater samples collected from coastal areas of Istanbul. Isolates were identified to the species level using standard biochemical tests specified by Facklam and Collins. The species distribution was as follows Enterococcus faecalis (96%), Enterococcus gallinarum (3%) and Enterococcus solitarius (1%). The resistance of bacteria to both heavy metals (zinc [Zn], iron [Fe], cadmium [Cd], chrome [Cr], cobalt [Co]) and antibiotics (ampicillin 10 μg [AP], penicillin G 10 Units [PG], gentamycin 10 μg [GM], streptomycin 10 μg [S], chloramphenicol 10 μg [C], erythromycin 15 μg [E], kanamycin 30 μg [K], amikacin 30 μg [AK], nalidixic acid 30 μg [NA], and vancomycin 30 μg [VA]) was evaluated. None of the strains was resistant to VA. It was found that among the 100 isolates, those that exhibit resistance to antibiotics, particularly NA, S and K, were also resistant all the heavy metals tested. To our knowledge this is the first report focusing on determination of resistance of environmental enterococci found in Istanbul against heavy metals and antibiotics. Thus, combined expressions of antibiotic and heavy metal resistance may help to reinforce ecological and epidemiological studies and to determine the role of these strains in antibiotic and heavy metal resistance dissemination.