Tetracycline-Resistant Escherichia coli in a Small Stream Receiving Fish Hatchery Effluent

We examined the impact of the effluent discharged from a freshwater (trout and related species) fish hatchery on the presence of antibiotic-resistant microorganisms in a small stream. There had been no documented use of antibiotics in the hatchery for at least 6 months prior to our study, although a variety of biocides were employed routinely for cleaning. Heterotrophic bacteria and Escherichia coli were isolated from both water column and sediment samples at sites above and below the discharge of the hatchery effluent as well as from the hatchery effluent itself. Randomly chosen isolates (≥96 isolates per site) were tested for their resistance to ampicillin, cephalexin, erythromycin, and tetracycline. Resistance to at least one antibiotic was found in greater than 30% of both the heterotrophic isolates and the E. coli isolates from each of the sites. There were no significant differences among the sites in the proportion of the heterotrophic isolates resistant to any specific antibiotic. The proportion of E. coli isolates resistant to tetracycline in the hatchery effluent and in both the downstream water and sediment samples was significantly higher than in either the upstream water or sediment. These results support the possibility of the hatchery as a source of tetracycline-resistant microorganisms even in the absence of recent use of this antibiotic.     

Antibiotic Resistance, Gene Transfer, and Water Quality Patterns Observed in Waterways near CAFO Farms and Wastewater Treatment Facilities

We examined water quality indicators (pH, temperature, turbidity, total phosphorus, and fecal coliform density) and bacterial antibiotic resistance (prevalence, conjugative transfer, and genetic linkage of resistance elements) at locations impacted by confined animal feeding operations (CAFOs) and compared them to nearby reference sites. Sites located upstream and downstream of two wastewater treatment facilities were also compared. Sites near CAFO farms had poor water quality (elevated total phosphorus and turbidity), while water quality remained relatively good downstream of wastewater treatment plants. High proportions of antibiotic-resistant bacteria were observed at all study sites, and frequent conjugative transfer of resistance was observed in laboratory assays. Out of a total of 830 environmental bacterial isolates, 77.1% were resistant to only ampicillin, while 21.2% were resistant to combinations of antibiotics including ampicillin (A), kanamycin (K), chlorotetracycline (C), oxytetracycline (O), and streptomycin (S). Multi-drug-resistant bacteria were significantly more common at sites impacted by CAFO farms. In conjugation assays, 83.3% of the environmental isolates transferred one or more antibiotic resistance genes to a laboratory strain of Salmonella typhimurium. A subset of multi-drug-resistant (A, C, and O) isolates was screened for specific tetracycline resistance genes and class I and II integrons. None of the screened isolates (n?=?22) were positive for integrons, while 13 isolates contained resistance genes for tet (B) and tet (C). Our results indicate that CAFO farms not only impair traditional measures of water quality but may also increase the prevalence of multi-drug-resistant bacteria in natural waters.     

Detection of R-plasmids in Salmonella isolated from clams and marine waters of Kuwait

A total of 24 strains of Salmonella isolated from clams and marine waters of Kuwait were speciated and investigated for drug resistance. Six of these were resistant to antibiotics. Their resistance patterns were ACT (4 isolates) and ACKST (2 isolates). These were examined for the transferability of their resistance. They had autotransferable R-plasmids and 25% of the transconjugants had an enbloc transfer at high frequency. The co-relation between Salmonella isolates from marine environment and clinical cases was established.     

Heavy metal-antibiotic resistant bacteria in a lake recreational area

We undertook this study to determine the impact of urbanization on the microbial content of waters of a major recreational area of Lake Champlain. We followed changes in the numbers of total coliforms, fecal coliforms, fecal streptococci, and ascertained resistance of coliforms to heavy metals and antibiotics. We determined the distribution of these bacterial indicators of pollution by examining water and sediment samples. We also surveyed changes in physical parameters such as pH, temperature, turbidity, and wind direction to evaluate their effect on the examined aquatic microbial population. The level of all standard microbial indices of pollution frequently exceeded acceptable levels for recreational waters. Sediment samples indicated prolonged deposition of microbial pollutants, but efforts to demonstrate their release from sediment by dislocation and resuspension were inconclusive. We confirmed the presence of a population of heavy-metal and antibiotic-resistant coliforms isolated from harbor waters. Eighty-four percent of the total coliforms isolated from harbor water samples were resistant to one or more antibiotics and 96% were resistant to two or more heavy metals. The presence of these antibiotic resistant properties in coliforms, as well as elevated fecal coliform and fecal streptococci counts in both water and sediment samples, suggests to us that the bay is polluted by human fecal matter. This pollution probably emanates from south harbor sources and affects the shores of the major northern bathing area. It constitutes a potential hazard to public health because of the geophysical nature of the harbor and the location of the pollution source.     

Antibiotic-resistant mutants identified from nodules of uninoculated soybeans grown in a strongly acidic soil

Two hypotheses that antibiotic-resistant nodule isolates from uninoculated soybeans grown in a strongly acidic soil were naturally occurring rhizobia which had acquired resistance to spectinomycin and streptomycin or were contaminants from adjacent, inoculated treatments, were tested in laboratory experiments. Soybean nodule isolates (166) as well as 48 cowpea and 89 Rhizobium japonicum strains were used in tests of resistance to six concentrations (0–500 μg ml^?1) of kanamycin, spectinomycin and streptomycin, tolerance of stresses of pH 4.6, with or without 50 μm Al, and serological cross-reactivity.More strains from the strongly acidic soil were resistant to the antibiotics than from slightly acidic soils, but resistance to antibiotics did not necessarily entail resistance to pH 4.6 or to 50 μm Al. Twenty-three nodule isolates which were resistant to spectinomycin or streptomycin cross-reacted with antisera of the inoculum strains, indicating that they were contaminants. None of 59 antibiotic-sensitive nodule isolates from uninoculated plants and none of 31 from inoculated plants cross-reacted with the antisera. All 53 antibiotic-resistant isolates from nodules of plants in inoculated plots cross-reacted with test antisera, indicating stability of the antibiotic markers.Cowpea rhizobia were generally more resistant to the antibiotics and more tolerant of pH 4.6 and 50 μm Al than were R. japonicum. Among strains of R. japonicum, slow growers were more resistant to antibiotics than moderately fast growers.     

Multiple antibiotic resistant fecal coliforms in raw sewage

Fecal coliforms in raw sewage from three localities in Bahrain were tested for resistance to 9 commonly available antibiotics. Nineteen percent of the isolates resistant to at least two antibiotics, showed resistance to Nalidixic acid and 10% were resistant to all other 8 antibiotics tested. Transmissability of antibiotic resistance was temperature dependent and was demonstrated in 86.0% of resistant isolates. The antibiotic resistant organisms also survived longer than sensitive isolates in seawater under laboratory conditions. Over the counter availability of antibiotics and a high incidence of multiple antibiotic resistant coliforms have possible implications on public health.     

Phenotypic Diversity of Multiple Antibiotic Resistant Enterococci with Emphasis on Enterococcus gallinarum Carrying vanA and vanB Genes

The prevalence and diversity of antibiotic resistant enterococci populations in samples collected four times from urban sewage treatment plant in Tehran, Iran between June 2005 and July 2006 were studied. Filtered samples were grown on mEnterococci medium containing 4 μg/ml vancomycin after which the enterococci isolates were identified to the species level. All strains were then tested for their resistance against nine antibiotics. Of the 131 isolates, 98 (75%) isolates were identified as Enterococcus gallinarum, followed by 24 (18%) and 9 (7%) for E. faecium and E. casseliflavus, respectively. All E. gallinarum isolates carried vanC1 gene with 64 (65%) and 14 (14%) isolates concomitantly harboured either vanA or vanB gene, respectively. Some E. casseliflavus concomitantly harboured vanA and vanC2 or vanB and vanC2. Typing the total enterococci isolates with a high resolution biochemical fingerprinting method showed a high diversity (D _i?=?0.91). We have shown by biochemical fingerprinting the presence of highly diverse glycopeptide resistant E. gallinarum and E. casseliflavus that have captured vanA and vanB genetic determinants under natural conditions. To our knowledge this is the first report in this geographical region showing high frequency antibiotic resistant enterococcal populations in particular E. gallinarum carrying assorted vancomycin resistance genes.     

Manure as a Source of Antibiotic-Resistant Escherichia coli and Enterococci: a Case Study of a Wisconsin, USA Family Dairy Farm

The spread of antibiotic-resistant bacteria in the environment is raising serious public health concerns, and manure is being increasingly recognized as a major source of antibiotic-resistant bacteria. In this research, we isolated Escherichia coli and enterococci from manure produced in a Wisconsin, USA family dairy farm to determine their resistance to six representative antibiotics. The average densities for E. coli and enterococci were 6.37(±4.38)?×?10⁷ colony formation units (CFU)?g^?1 and 1.60(±1.57)?×?10⁴ CFU g^?1, respectively. The E. coli isolates were found to be resistant to cephalothin, ampicillin, tetracycline, and erythromycin. In addition to these four antibiotics, the Enterococcus isolates were also resistant to gentamicin and ciprofloxacin. Additionally, we examined the survival and growth of E. coli and enterococci in dairy manure over a period of ~3 days. While the densities of enterococci remained stable over the study period, the concentrations of E. coli on average increased by 1.5 log10 units. Further tests of the bacterial antibiotic resistance over time showed no significant changes in the prevalence of antibiotic resistance. This result indicated that slightly aged manure could represent a larger source of antibiotic-resistant E. coli than fresh manure and the accumulation of antibiotic-resistant E. coli and enterococci in the agricultural fields must be accounted for in the modeling of the spread of antibiotic-resistant bacteria in the environment.     

牛粪沼渣中多重耐药细菌的环境风险

为研究牛粪厌氧发酵沼渣中多重耐药细菌的环境污染风险，选用畜禽养殖中常用的5种抗生素(红霉素、氨苄青霉素、金霉素、链霉素、环丙沙星)对牛粪沼渣中可培养抗生素抗性细菌进行筛选。结果表明：多重耐药细菌的比例高达76.5%,其中，抗5种和4种抗生素的细菌分别有11,21株。所有多重耐药细菌均对氨苄青霉素具有抗性，抗红霉素、金霉素和链霉素的细菌分别占总多重耐药细菌数的92.0%,89.3%,61.3%。通过细菌16S rRNA测序鉴定，32株具有4种以上抗性的细菌分别属于福氏志贺氏菌、摩根氏菌和假中间苍白杆菌，均为重要的临床致病菌。使用全基因组测序对7株典型多重耐药细菌携带的抗性基因进行分析，共检测出28种抗生素抗性基因，对应9种抗生素抗性类型。通过分析抗性基因及插入序列所在位点信息发现，多重耐药细菌普遍携带含有抗性基因和插入序列共存的质粒，表明抗性基因具有高度的可移动性和较强的传播风险。综上所述，牛粪沼渣中含有大量的多重耐药细菌，可视作抗生素抗性基因的储存库，应重视沼渣农业资源化利用过程中的环境风险监测与评估。     

Prevalence and Antimicrobial Resistance of Enterococcal Species in Sewage Treatment Plants in Iran

We investigated for the first time the occurrence, stability and antibiotic resistance of 593 enterococci in six samples collected from three urban sewage treatment plants (STPs) located in the north, south and west part of Tehran, Iran between October 2004 and September of 2005. Isolates were typed with a biochemical fingerprinting method (the PhPlate system) and tested for their resistance to six antibiotics. The most prevalent species in all three STPs were E. faecium followed by E. hirae and E. faecalis accounting for 93% of the total isolates examined. In all, 317 (55%) isolates were susceptible to all six antibiotics tested and the remaining isolates were resistant to between 1 and 6 antibiotics. Biochemical fingerprinting with PhPlate system showed a high diversity for E. faecalis (D _i?=?0.95), E. hirae (D _i?=?0.93) and E. faecium (D _i?=?0.95) populations with an overall diversity of D _i?=?0.97 for the whole enterococcal populations found in all three STPs. Our data indicate a high degree of polyclonality among the enterococci populations of human origin. This study suggest that the municipal wastewaters might be an important source of dissemination of antibiotic-resistant enterococci in Iran.     

Multiple Metal Resistant Transferable Phenotypes in Bacteria as Indicators of Soil Contamination with Heavy Metals (6 pp)

Background, Aim and Scope   Environmental contamination by heavy metals affects microbial communities. The number of single and multiple heavy metal resistant bacteria may be an indictor of the level of contamination. This paper details the isolation and characterisation of metal resistant microorganisms isolated from rhizosphere/soil samples obtained from an abandoned zinc, lead and copper mine and a local unaffected site. This data was compared to the level of heavy metal in the soils to establish the effect of metals on the microbial community and to determine the relationship between pollutant levels and resistant strains. This paper outlines the diversity of transferable resistance determinants between both sites and details the levels of heavy metal resistant bacteria and those expressing transferable multiple heavy metal tolerance.Methods   The sample sites were located in Co. Galway, Ireland. The first sample site (site A) was a former lead, zinc and copper mine, which was closed in 1961 due to exhaustion of ore. The second site (site B) was located two and a half kilometres from the mining site and was not affected by the mining operations. Composite soil samples were characterised for general soil matrix composition, organic content, pH and general chemical parameters. The soil was also enumerated for the total viable heterotrophic counts and tested on Pseudomonas selective agar (PSA) for total Pseudomonas counts and Sucrose Asparagine (SA), which is semi-selective for fluorescent Pseudomonas.Results and Discussion   Samples from both site A and site B were analysed by atomic absorption spectrophotometry for the presence of heavy metals. In the case of copper, which has a Dutch list recommended minimum permissible level of 190 µg/Kg dry weight, the levels detected at site A were 1270 µg/Kg dry weight while site B was detected at 36 µg/Kg. The arsenic levels detected at site A were eight times the permissible level (416 µg/Kg) while only half the permissible level was found at site B (13 µg/Kg). Zinc concentrations were also high at site A (4460 µg/Kg) while at site B (553 µg/Kg) they were well below the Dutchlist guidelines (720 µg/Kg). A large number of heavy metal tolerant strains were isolated from both sites. 270 isolates (site A (170) and site B (100)) were screened against 8 metals to examine the extent of multiple resistance. 82% of the strains from site A were found to be resistant to 5 metals. A total of 18% showed resistance to all 8 metals and of those examined only 4% were resistant to only one metal. In contrast isolates from site B showed no multiple resistance to more than 5 metals, while 62% showed resistance to individual metals only. Site A had a higher level of multiple heavy metal resistance strains. Stains isolated from site A had 23 (14%) isolates resistant to zinc, copper, nickel, arsenic and cobalt and site B had no bacteria resistant to all five of these selected metals. The transferability of heavy metal resistance was investigated in the case of 60 multiple heavy metal resistant isolates taken from site A and 50 multiple resistance isolates from site B. Transfer was only detected in isolates from site A, 13% showed transfer and expression of copper, zinc and arsenic resistance determinants. In most cases the transconjugants only expressed resistance to copper, zinc and arsenic, which were the metals, used for selection. This co-transfer of all three determinants suggests a genetic link between these resistance determinants.Conclusion   Heavy metal resistant bacteria are present in both sites, however, the number and presence of multiple transferable resistance phenotypes are confined to the isolates from the heavy metal contaminated site. The presence of high levels of heavy metals selects these multiple resistance phenotypes. Within these communities there seems to be little diversity between the microorganisms, which provides a hugely preferable environment for gene transfer of such metal resistant determinants.Recommendation and Perspective   The experiments have shown a microbes ability to mobilize heavy metal determinants and a relationship between heavy metal resistance and metal contamination has been identified These multiple heavy metal resistant bacteria could eventually be used for detection and qualification of the level of heavy metal-polluted soil/water environments.     

Diversity and antibiotic-producing potential of cultivable marine-derived actinomycetes from coastal sediments of Turkey

Purpose

Marine environments, especially sediments, are rich sources of actinomycetes that provide many bioactive compounds, primarily antibiotics. The goal of this study was to investigate the diversity of cultivable actinomycetes and their potential to produce antibiotics from sediments collected from the coastal zones of Turkey.

Materials and methods

Thirty sediment samples were collected from nine different coastal sites in three seas surrounding the Anatolian Peninsula of Turkey. Of the samples, 6 were collected from one site in the Black Sea, 18 from seven sites in the Aegean Sea, and 6 from one site in the Mediterranean Sea. Strains of pure actinomycetes were isolated by modified actinomycetes isolation agar (MAIA), M1 agar, M6 agar, and modified R2A agar. Ethyl acetate extracts and fermentation broths were used for the evaluation of antimicrobial activity against antibiotic resistant test microorganisms. The identification of the isolates was undertaken by 16S rRNA gene sequencing.

Results and discussion

A total of 261 strains of actinomycetes were isolated, of which 66 (25 %) were active against at least one antibiotic-resistant microorganism. Sixty-five of the actinomycetes isolates with antimicrobial activity were Streptomyces spp. and one was Nocardia sp., which implied that genus Streptomyces was predominant. Whereas MAIA agar was the best medium to recover actinomycetes, M6 agar was superior to others for the isolation of antibiotic-producing strains.

Conclusions

Extensive screening of the extracts from the 261 isolates for antimicrobial activities revealed considerable potential to produce antibiotics. These findings imply that actinomycetes from marine sediments of the Anatolian Peninsula coasts have potential for the discovery of novel bioactive compounds.     

Chromium-resistant microorganisms isolated from evaporation ponds of a metal processing plant

Chromium occurs naturally at trace levels in most soils and water, but disposal of industrial waste and sewage sludge containing chromium compounds has created a number of contaminated sites, which could pose a major environmental threat. This study was conducted to enumerate and isolate chromium-resistant microorganisms from sediments of evaporation ponds of a metal processing plant and determine their tolerance to other metals, metalloids and antibiotics. Enumeration of the microbiota of Cr-contaminated sediments and a clean background sample was conducted by means of the dilution-plate count method using media spiked with Cr(VI) at concentrations ranging from 10 to 1000 mg L^?1. Twenty Cr(VI) tolerant bacterial isolates were selected and their resistance to other metals and metalloids, and to antibiotics was assessed using a plate diffusion technique. The number of colony-forming units (cfu) of the contaminated sediments declined with increasing concentrations from 10 to 100 mg L^?1 Cr(VI), and more severely from 100 to 1000 mg L^?1 Cr(VI). The background sample behaved similarly to 100 mg L^?1 Cr(VI), but the cfu declined more rapidly thereafter, and no cfu were observed at 1000 mg L^?1 Cr(VI). Metals and metalloids that inhibited growth (from the most to least inhibitory) were: Hg > Cd > Ag > Mo = As(III) at 50 μg mL^?1. All 20 isolates were resistant to Co, Cu, Fe, Ni, Se(IV), Se(VI), Zn, Sn, As(V), Te and Sb at 50 μg mL^?1 and Pb at 100 μg mL^?1. Eighty-five percent of the isolates had multiple antibiotic resistance. In general, the more metal-tolerant bacteria were among the more resistant to antibiotics. It appears that the Cr-contaminated sediments may have enriched for bacterial strains with increased Cr(VI) tolerance.     

Investigation of outgrowths of Klebsiella spp in distributed potable water

The boundary of the study in this organisation involved laboratory experiments on the facets of the nature of coliform (Klebsiella) bacteria in parts of the Sydney water system which supplies 3.3 × 10⁶ people. After September 1976 they appeared increasingly throughout the 18 000 km of reticulation system. Non-thermotolerant Klebsiella spp were persistent despite swabbing and flushing, their survival being enhanced by deposits in mains and turbidity greater than one unit in 75 % of 813 water samples. A multiple approach (biochemistry, genetics, typing, antibiotic and metallic sensitivity, and plasmid transfer) was adopted using 848 isolates in 1980 from 38 sampling runs. While the species persisted from 1980 to 1982, their designation of oxytoca remained unclear despite usage of 57 biochemical tests and G + C of 56.3. Cultures from both dates were bacteriocin typed, half being untypable as were the majority of 73 environmental cultures collected between Traralgon and Townsville in adjoining states. Antibiotic sensitivity testing by diffusion and agar dilution procedures demonstrated multi-resistance to twenty antibiotics that exceeded or equalled results reported from other habitats. Heavy metal resistance was considerably elevated. These two sets of markers are plasmid home and were more actively transferred at 37 than 20° C by environmental Klebsiella to E. coli K12. Because of the inability of the typing method, the pattern or origin of the contamination could not be found. The clinical significance of this activity at 37° C would require another study elsewhere because these bacteria are ingested.     

Mercury resistance among soil bacteria: Ecology and transferability of genes encoding resistance

A diverse population of bacteria, all with the common property of resistance to mercuric ions (Hg²⁺), was isolated from root-associated, and other soils. Each isolate was screened for its resistance to a variety of antibiotics and heavy metals as well as organomercurial compounds. Several isolates showed resistance values to individual antimicrobial agents that were significantly higher than the average resistance values with respect to those agents for the genus to which the isolate belonged. Of 76 distinct isolates tested, 4 were able to transfer the Hg^r phenotype to sensitive cells at environmental temperatures. “Mobilization” of putative Hg-resistant non-conjugative plasmids with the wide host range plasmid RPl resulted in the co-transfer of the RPl markers and resistance to Hg²⁺ in 9 cases. Pl transduetion showed that 7 of these transfers were due, at least in part. to the formation of RPl-Hg^r co-integrates. This suggests that translocatable DNA elements encoding resistance to Hg²⁺ may be partly responsible for the dissemination and maintenance of the Hg^r phenotype in microbial populations.     

Evaluation of a core collection of Brassica oleracea accessions for resistance to white rust of crucifers (Albugo candida) at the cotyledon stage

The identification of seedling resistance to white rust of crucifers was performed in a screening of a B. oleracea core collection with 400 accessions representing the genetic and geographic diversity of the species. Fifty seedlings per accession were tested against the Portuguese isolate Ac502 using the methodology and evaluation procedures developed by and . The percentage of resistant seedlings (%R) and the conventional rating criteria of the mean Disease Index (DI) based on the two different evaluation procedures of disease expression used, were compared and adopted as the criteria to rank the accessions for their interest as sources of resistance. A great variability of reactions was found between and within accessions of the core collection, ranging from complete resistance to full susceptibility. Sources of resistance were found namely among the cauliflowers, broccoli and tronchuda cabbages gene pools. Forty-seven accessions presented at least 20% of resistant seedlings. Nine accessions (the kales INRA18 and INRA62, the cauliflowers HRI4856, HRI4866 and HRI5424, the loose-head cabbage HRI11555, the savoy cabbage BRA848, the black broccoli HRI6318 and the Portuguese tronchuda cabbage ISA207) presented 50–78% of resistant seedlings and so they should be considered as potential and useful sources for direct use in breeding programs for white rust resistance. Fourteen inbred lines, representing the full range of disease expression, derived from resistant accessions of the core collection were also tested for resistance to other two Portuguese isolates (Ac503 and Ac504) and to a UK isolate. The results provided no evidence of differential reaction to the A. candida isolates tested.     

Antibiotic resistant coliform and faecal coliform bacteria in drinking water

Coliform and faccal coliform were isolated from drinking water samples obtained from tap water (TW) and storage tanks (ST) in Baghdad city. All isolates were identified and tested for their resistance to twelve antibiotics. Bacterial identification revealed that Ent. cloacae and Kl. pneumoniae were the predominant organisms from TW and ST water samples, respectively. Overall, 66% of the isolates from TW and 38% from ST were resistant to one or more of the drugs tested. Ampicillin, Carbenicillin, Cefalotin and Colistin resistance in particular were more frequent among bacterial isolates. The overall frequency of multiple antibiotic resistance among bacterial isolates in TW and ST samples were 47% and 33%, respectively. All bacterial strains isolated from drinking water were sensitive to Rifampicin. Results demonstrated the need for periodical bacteriological examination of drinking water and restriction in the use of antibiotics in this country.     

Genomic characterization of multidrug-resistant extraintestinal pathogenic Escherichia coli isolated from grain culture soils

Multidrug-resistant (MDR) Escherichia coli, mainly extraintestinal pathogenic E. coli (ExPEC), has been widely reported in infections worldwide. In agricultural soils, manure is a hotspot for the dissemination of antimicrobial resistance genes (ARGs) and pathogenic bacteria; however, MDR bacteria have also been reported in soils with no history of manure use. In addition, cross-resistance and co-resistance have been described as responsible for the metal-driven selection of bacteria resistant to antimicrobials. Therefore, the aim of this study was to analyze three MDR E. coli isolates obtained from Brazilian grain culture soil samples with no history of manure use by whole-genome sequencing. The MDR E. coli isolates were recovered from soils from corn and coffee fields, and presented resistance to β-lactams, quinolones, aminoglycosides, tetracyclines, sulphonamides, and dihydrofolate reductase inhibitor. Resistome analysis showed ARGs to several antimicrobials (i.e., β-lactams, tetracyclines, aminoglycosides, sulphonamides, trimethoprim, phenicols, fosfomycin, and macrolides) as well as several metal resistance genes and antibacterial biocide resistance genes. In addition, known mutations in quinolone-resistance-determining regions of GyrA (Ser83Leu and Asp87Asn), ParE (Ser458Thr), and ParC (Ser80Ile) were also detected. Virulome analysis showed the presence of virulence genes (lpfA, mcmA, gad, mchF, iroN, cma, and iss) associated with ExPEC. Multidrug-resistant ExPEC isolates were assigned to phylogenetic group B1. The presence of MDR B1-ExPEC in soil samples shows the ability of these isolates to survive in soils. This study reports for the first time some sequence types (i.e., ST345, ST448, and ST1146) of MDR E. coli in Brazilian soils. Therefore, these findings contribute to the monitoring of antimicrobial resistance and surveillance studies based on whole-genome sequencing worldwide.     

Arsenic resistant microorganisms isolated from agricultural drainage water and evaporation pond sediments

Elevated levels of As in contaminated water and soil could pose a major threat to the environment. Relatively high levels of As have been reported in agricultural drainage water and in evaporation pond sediments in Kern County, California. The objective of this study was to enumerate and isolate As-resistant microorganisms from agricultural drainage water and evaporation pond sediments and to assess their tolerance to metals, metalloids and antibiotics. The culture medium was amended with arsenite (III), arsenate (V), methylarsonic acid (MAA), and dimethylarsinic acid (DMA). Among the water samples, As(V), MAA, and DMA added to the medium at concentrations from 0.1 to 1000 mg L^?1 showed no effect on the colony forming units (CFUs) compared with no As supplementation, while arsenite (III) (> 1.0 mg L^?1) inhibited the population. The sediments showed three trends: (i) no effect on CFUs in the presence of As(V) up to 1000 mg kg^?1, (ii) a decline in CFUs in the presence of > 100 mg kg^?1, As(III), and (iii) an increase in CFUs upon the addition of MAA or DMA at > 25 mg kg^?1, Arsenite (III) was much more toxic to the indigenous microflora than any other As species. Arsenite (III) inactivates many enzymes by having a high affinity for thiol groups of proteins. A plate diffusion method was used to assess the tolerance of the As-resistant bacteria to heavy metals, metalloids and antibiotics. Of 14 isolates tested, all were resistant to Co, Cu, Pb, Ni, Mo, Cr, Se(IV), Se(VI), As(III), As(V), Sb, Sn, and Ag (50 µg mL^?1). The most toxic trace elements were Cd followed by Hg>Te>Zn. Multiple antibiotic tolerance (resistance to 2 or more antibiotics) was found among 43% of the isolates. The As-resistant bacteria showed a high tolerance to metals and antibiotics.     

Intrinsic antibiotic resistance in relation to colony morphology in three populations of West African cowpea rhizobia

Rhizobia isolated from cowpeas (Vigna unguiculata (L.) Walp.) grown in three west African soils were examined for intrinsic resistance to five antibiotics and were scored for one of two colony morphologies. Half of the strains tested had a “wet”, slimy colony morphology and half had small discrete “dry”, non-slimy colonies. The populations as a whole were resistant to gentamicin (87%) but varied in their resistance to streptomycin, rifampicin, kanamycin and penicillin. Thirteen patterns of resistance were found for the 128 strains screened and strains within the same pattern usually had the same colony type. The most common pattern (32%) was resistance to all five antibiotics. Associations between resistances were random within populations and colony type. Each population was diverse, expressing from 6 to 8 patterns of resistance, but one population was relatively homogeneous, with 68% of its members exhibiting the same pattern. Correlations between intrinsic antibiotic resistance and colony type were demonstrated both for the west African cowpea rhizobia and for a broader group of cowpea miscellany rhizobia. The method was practical, rapid and reliable for identification of groups within populations.