Detection of antibiotic resistant bacteria inhabiting the sand of non-recreational marine beach

The present study examined the antibiotic resistance of heterotrophic bacteria, which were isolated from the sand of the beach located in the National Park of the southern Baltic Sea coast. The bacteria demonstrated low levels of antibiotic resistance. These microorganisms were the most resistant to cefaclor and clindamycin and the most sensitive to clarithromycin, doxycycline, gentamycin and oxytetracycline. The majority of bacteria inhabiting the sand of the studied beach were resistant to only one antibiotic out of 18 tested antibiotics in this study. The bacteria inhabiting the middle part of the beach and the dune were more antibiotic resistant than bacteria isolated from the seawater and the shoreline-seawater contact zone. Generally, there was no significant difference in antibiotic resistance between bacteria isolated from the surface and the subsurface sand layers. The bacterial antibiotic resistance level depends on the chemical structure of antibiotics.     

Ecosystem response to antibiotics entering the aquatic environment

Awareness of antibiotics in wastewaters and aquatic ecosystems is growing as investigations into alternate pollutants increase and analytical techniques for detecting these chemicals improve. The presence of three antibiotics (ciprofloxacin, norfloxacin and cephalexin) was evaluated in both sewage effluent and environmental waters downstream from a sewage discharge. Bacteria cultured from the sewage bioreactor and receiving waters were tested for resistance against six antibiotics (ciprofloxacin, tetracycline, ampicillin, trimethoprim, erythromycin and trimethoprim/sulphamethoxazole) and effects of short term exposure (24 h) to antibiotics on bacterial denitrification rates were examined. Antibiotics were detected entering the sewage treatment plant with varying levels of removal during the treatment process. Antibiotics were also detected in effluent entering receiving waters and detectable 500 m from the source. Among the bacteria cultured from the sewage bioreactor, resistance was displayed against all six antibiotics tested and bacteria cultured from receiving waters were resistant against two of the antibiotics tested. Rates of denitrification were observed to decrease in response to some antibiotics and not to others, though this was only observed at concentrations exceeding those likely to be found in the environment. Findings from this preliminary research have indicated that antibiotics are entering our aquatic systems and pose a potential threat to ecosystem function and potentially human health.     

Antibiotic resistant bacteria in fish from the Concepción Bay, Chile.

Antibiotic resistant bacteria from commercial demersal and pelagic fish captured in the Concepción Bay, Chile were investigated. Viable counts of antibiotic resistant bacteria isolated from gill and intestinal content samples showed high frequencies of resistance to ampicillin, streptomycin and tetracycline, while the proportion of chloramphenicol resistance was rather low. A high incidence of resistance to ampicillin, streptomycin, tetracycline and nitrofurantoin, as well as almost an absence of resistance to gentamicin, amikacin and cotrimoxazole was found among selected isolates which represented the resistant bacterial population. These strains mainly belonged to Vibrionaceae and Enterobacteriaceae and were predominantly resistant to 3 and 4 antibacterials. Isolates from demersal fish exhibited resistance to as many as 8-10 compounds, whereas those from pelagic fish were resistant to seven or fewer antibiotics. These results suggest that Chilean commercial fishes residing in waters near the disposals of urban sewage might play a role as carriers of antibiotic resistant bacteria prompting a health risk to public health for fish consumers.     

Bacterial flora and antibiotic resistance from eggs of green turtles Chelonia mydas: An indication of polluted effluents

Sea turtles migrate to various habitats where they can be exposed to different pollutants. Bacteria were collected from turtle eggs and their resistance to antibiotics was used as pollutant bio-indicators of contaminated effluents. Eggs were collected randomly from turtles when they were laying their eggs. A total of 90 eggs were collected and placed into sterile plastic bags (3 eggs/turtle) during June-December of 2003. The bacteria located in the eggshell, albumen and yolk were examined, and 42% of the eggs were contaminated with 10 genera of bacteria. Pseudomonas spp. were the most frequent isolates. The albumen was found to be the part of the egg to be the least contaminated by bacterial infection. Bacterial isolates tested with 14 antibiotics showed variations in resistance. Resistance to ampicillin was the highest. The presence of antibiotic resistant bacteria in eggs indicates that the green turtle populations were subjected to polluted effluents during some of their migratory routes and feeding habitats. Scanning electron microscopy revealed that Salmonella typhimurium penetrated all eggshell layers.     

Drug-resistance transfer in Escherichia coli in New York Bight sediment

Sediment from the sewage sludge dump site area in the New York Bight contains bacteria resistant to antibiotics and heavy metals. The purpose of this study was to determine if antibiotic resistance could be transferred from donor to recipient Escherichia coli strains inoculated into glass vessels containing sediment and seawater obtained near the dump site. Temperature was maintained at 10°C, the mean winter temperature of benthic water at the dump site. Transconjugants (recipients which inherit donor genes for tetracycline resistance) were isolated from the seawater and sediment within one hour after inoculation and were found in all subsequent sediment samples for one month. Donor and recipient E. coli remained viable in the sediment for at least one month. Our results indicate that sewage sludge polluted sediment may serve as an environment conducive to conjugal transfer of antibiotic resistance genes.     

Coincidence of cadmium and antibiotic resistance in New York Bight apex benthic microorganisms

Microorganisms resistant to 1–500 ppm cadmium were isolated over an 11 month period from sediments at the sewage sludge, dredge spoils and industrial acid waste disposal sites, as well as at an estuarine outflow to the New York Bight apex. Tests for antibiotic resistance in these isolates revealed that 94% of the isolates were resistant to one or more antibiotics and that 91% of the original isolates could be characterized by multiple drug resistance. Different selective pressures may account for the various genera and antimicrobial resistance patterns observed. The possibility of extrachromosomal linkage of cadmium and streptomycin resistance is discussed.     

Untersuchungen zum Vorkommen von R+-Bakterien im Flußwasser

With colony numbers of 2 · 10⁴… 3 · 10⁵ and coliform contents of 2 · 10³… 1 · 1 · 10⁴ per ml, in the investigated river section 2.1% of colonies were resistant to tetracycline, 0.3% to chloramphenicol, 7.5% to kanamycin, 5.4% to trimethoprim, 12.7% to ampicillin and 0.14% to gentamycin. For the coliforms the resistances were 5.1% to tetracycline, 2.1% to chloramphenicol, 2.7% to kanamycin and 0.15% to gentamycin. There were isolated 63 different resistance types. Of the resistant Klebsiella strains 75% were resistant to three or more antibiotics. Of the coliform bacteria (Escherichia coli, Klebsiella, Enterobacter, Citrobacter) 31.6… 51.9% of the strains transferred their antibiotics-resistance to Escherichia coli K-12, for the antibiotics-resistant strains of Pseudomonas and Aeromonas a transfer of resistance was not observed at all.     

Microbial degradation of petroleum at low temperature

Bacteria were isolated from littoral sediments collected in Chedabucto Bay, Nova Scotia, and from oil-contaminated soil adjacent to a natural oil seep at Cape Simpson, Alaska. Data are presented on the range of hydrocarbon utilization and growth temperature of two bacteria, which suggest that bacteria existing in these environments play a significant role in the biodegradation of pollutant hydrocarbons.     

Gilthead seabream (Sparus aurata) carrying antibiotic resistant enterococci. A potential bioindicator of marine contamination?

Antibiotic resistance in bacteria is a growing problem that is not only restricted to the clinical setting but also to other environments such as marine species that harbor antibiotic resistant bacteria and therefore may serve as reservoirs for antibiotic-resistance genetic determinants. The aim of this study was to evaluate antibiotic resistance phenotypes in enterococci isolated from fecal samples of gilthead seabream and the associated mechanisms of resistance. A collection of 118 samples were analyzed and 73 enterococci were recovered. The strains showed high percentages of resistance to erythromycin and tetracycline (58.9% and 17.8%, respectively). Lower level of resistance (<13%) was detected for quinupristin-dalfopristin, ampicillin, high-level-gentamicin, high-level-streptomycin, high-level-kanamycin, ciprofloxacin and chloramphenicol. The erm(B), tet(L) or tet(M), aac(6′)-aph(2″) and aph(3′)-IIIa genes were shown in isolates resistant to erythromycin, tetracycline, high-level gentamicin and high-level kanamycin, respectively. Antibiotic resistance in natural microbiota is becoming a concern of human and environmental health.     

Role of shellfish hatchery as a reservoir of antimicrobial resistant bacteria

The main aim of this study was to determine the occurrence of resistant bacteria in florfenicol-treated and untreated scallop larval cultures from a commercial hatchery and to characterize some selected florfenicol-resistant strains. Larval cultures from untreated and treated rearing tanks exhibited percentages of copiotrophic bacteria resistant to florfenicol ranging from 0.03% to 10.67% and 0.49–18.34%, respectively, whereas florfenicol resistance among oligotrophic bacteria varied from 1.44% to 35.50% and 3.62–95.71%, from untreated and treated larvae, respectively. Florfenicol resistant microbiota from reared scallop larvae mainly belonged to the Pseudomonas and Pseudoalteromonas genus and were mainly resistant to florfenicol, chloramphenicol, streptomycin and co-trimoxazole. This is the first study reporting antimicrobial resistant bacteria associated to a shellfish hatchery and the results suggest that a continuous surveillance of antimicrobial resistance even in absence of antibacterial therapy is urgently required to evaluate potential undesirable consequences on the surrounding environments.     

Antibiotic Resistance of Gram Negatives isolates from loggerhead sea turtles (Caretta caretta) in the central Mediterranean Sea

Previous studies on fish and marine mammals support the hypothesis that marine species harbor antibiotic resistance and therefore may serve as reservoirs for antibiotic-resistance genetic determinants. The aim of this study was to assess the resistance to antimicrobial agents of Gram negative strains isolated from loggerhead sea turtles (Caretta caretta). Oral and cloacal swabs from 19 live-stranded loggerhead sea turtles, with hooks fixed into the gut, were analyzed. The antimicrobial resistance of the isolates to 31 antibiotics was assessed using the disk-diffusion method. Conventional biochemical tests identified Citrobacter spp., Proteus spp., Enterobacter spp., Escherichia spp., Providencia spp., Morganella spp., Pantoea spp., Pseudomonas spp. and Shewanella spp. Highest prevalences of resistance was detected to carbenicillin (100%), cephalothin (92.6%), oxytetracycline (81.3%) and amoxicillin (77.8%). The isolates showing resistance to the widest range of antibiotics were identified as Citrobacter freundii, Proteus vulgaris, Providencia rettgeri and Pseudomonas aeruginosa. In this study, antibiotic resistant bacteria reflect marine contamination by polluted effluents and C. caretta is considered a bioindicator which can be used as a monitor for pollution.     

Antibiotic Resistance and Transferable R Plasmids in Coliforms from a Polluted Estuary

The aim of this work was to find out how far the Nervion Estuary serves as a reservoir of R plasmids and/or for their dissemination. The fecal pollution bacterial indicator population was estimated along the estuary and 167 coliform isolates were assayed for resistance to 16 antibiotics. About sixty-nine per cent of the coliform isolates were antibiotic-resistant and forty-two per cent of these were simultaneously resistant to three or more of 16 antibiotics tested. Furthermore, 31.5% of enterobacterial strains resistant to any of the antibiotics tested could transfer their resistance determinants in conjugal matings. Most of the transferred plasmids confer resistance to ampicillin or to other β-lactamic antibiotics and several codify resistance to tetracyclin, chloramphenicol of fosfomycin. Taking account of the coliforms density found and the ratio of these isolates able to transfer R plasmids, an average of 5 × 10³ coliform strains able to transfer R plasmids in 100 ml of water was obtained. This amount must indicate a significant potential public health risk because these polluted waters get into an area with intensive fishing and recreational use.     

Bacterial transport experiments in fractured crystalline bedrock

The efficiency of contaminant biodegradation in ground water depends, in part, on the transport properties of the degrading bacteria. Few data exist concerning the transport of bacteria in saturated bedrock, particularly at the field scale. Bacteria and microsphere tracer experiments were conducted in a fractured crystalline bedrock under forced-gradient conditions over a distance of 36 m. Bacteria isolated from the local ground water were chosen on the basis of physicochemical and physiological differences (shape, cell-wall type, motility), and were differentially stained so that their transport behavior could be compared. No two bacterial strains transported in an identical manner, and microspheres produced distinctly different breakthrough curves than bacteria. Although there was insufficient control in this field experiment to completely separate the effects of bacteria shape, reaction to Gram staining, cell size, and motility on transport efficiency, it was observed that (1) the nonmotile, mutant strain exhibited better fractional recovery than the motile parent strain; (2) Gram-negative rod-shaped bacteria exhibited higher fractional recovery relative to the Gram-positive rod-shaped strain of similar size; and (3) coccoidal (spherical-shaped) bacteria transported better than all but one strain of the rod-shaped bacteria. The field experiment must be interpreted in the context of the specific bacterial strains and ground water environment in which they were conducted, but experimental results suggest that minor differences in the physical properties of bacteria can lead to major differences in transport behavior at the field scale.     

Molecular characterizations of oxytetracycline resistant bacteria and their resistance genes from mariculture waters of China

Oxytetracycline-resistant bacteria were isolated from a mariculture farm in China, and accounted for 32.23% and 5.63% of the total culturable microbes of the sea cucumber and the sea urchin rearing waters respectively. Marine vibrios, especially strains related to Vibrio splendidus or V. tasmaniensis, were the most abundant resistant isolates. For oxytetracycline resistance, tet(A), tet(B) and tet(D) genes were detected in both sea cucumber and sea urchin rearing ponds. The dominant resistance type for V. tasmaniensis-like strains was the combination of both tet(A) and tet(B) genes, while the major resistance type for V. splendidus-like strains was a single tet(D) gene. Most of the sea cucumber tet-positive isolates harbored a chloramphenicol-resistance gene, either cat IV or cat II, while only a few sea urchin tet-positive isolates harbored a cat gene, actually cat IV. The coexistence of tet and cat genes in the strains isolated from the mariculture farm studied was helpful in explaining some of the multi-resistance mechanisms.     

Prevalence and distribution of fecal indicator organisms in South Florida beach sand and preliminary assessment of health effects associated with beach sand exposure

Fecal indicator levels in nearshore waters of South Florida are routinely monitored to assess microbial contamination at recreational beaches. However, samples of sand from the surf zone and upper beach are not monitored which is surprising since sand may accumulate and harbor fecal-derived organisms. This study examined the prevalence of fecal indicator organisms in tidally-affected beach sand and in upper beach sand and compared these counts to levels in the water. Since indicator organisms were statistically elevated in sand relative to water, the study also considered the potential health risks associated with beach use and exposure to sand. Fecal coliforms, Escherichia coli, enterococci, somatic coliphages, and F(+)-specific coliphages were enumerated from sand and water at three South Florida beaches (Ft. Lauderdale Beach, Hollywood Beach, and Hobie Beach) over a 2-year period. Bacteria were consistently more concentrated in 100g samples of beach sand (2-23 fold in wet sand and 30-460 fold in dry sand) compared to 100ml samples of water. Somatic coliphages were commonly recovered from both sand and water while F(+)-specific coliphages were less commonly detected. Seeding experiments revealed that a single specimen of gull feces significantly influenced enterococci levels in some 3.1m(2) of beach sand. Examination of beach sand on a micro-spatial scale demonstrated that the variation in enterococci density over short distances was considerable. Results of multiple linear regression analysis showed that the physical and chemical parameters monitored in this study could only minimally account for the variation observed in indicator densities. A pilot epidemiological study was conducted to examine whether the length of exposure to beach water and sand could be correlated with health risk. Logistic regression analysis results provided preliminary evidence that time spent in the wet sand and time spent in the water were associated with a dose-dependent increase in gastrointestinal illness.     

Antibiotic resistance of bacteria isolated from shrimp hatcheries and cultural ponds on Donghai Island, China

The resistance of bacteria to 12 different antibiotics was investigated in shrimp farms on Donghai Island, China. Antibiotic-resistant bacteria were found to be widespread in shrimp farms, indicating a high environmental risk. Further, significant differences were found in bacterial strains among farms (ANOVA, p < 0.05), showing resistance to antibiotics such as ampicillin, trimethoprim, compound sinomi, tetracycline, chloramphenicol and cefazolin. No significant differences in antibiotic resistance were found among 6 hatcheries evaluated in this study (ANOVA, p > 0.05), between exalted and traditional shrimp ponds (ANOVA, p > 0.05), and between cultural ponds and corresponding control water source sites (T-test, p > 0.05). In cultural ponds, no significant difference in bacterial resistance to antibiotics was found between water and sediment (T-test, p > 0.05), and antibiotic resistance of bacteria from water showed a significant positive correlation with that from sediment (p < 0.05). Therefore, our study indicates that bacterial multiple antibiotic resistance (MAR) is more widespread in shrimp hatcheries than ponds.     

Isolates of Staphylococcus aureus and saprophyticus resistant to antimicrobials isolated from the Lebanese aquatic environment

The indiscriminate use of antimicrobials especially in developing countries has evoked serious bacterial resistance and led to the emergence of new and highly resistant strains of bacteria to commonly used antimicrobials. In Lebanon, pollution levels and bacterial infections are increasing at a high rate as a result of inadequate control measures to limit untreated effluent discharges into the sea or freshwater resources. The aim of this study was to isolate and molecularly characterize various Staphylococcus strains isolated from sea water, fresh water, sediments, and crab samples collected from representative communities along the coast of Lebanon. The results on the antimicrobial resistance indicated that the level of resistance of Staphylococcus aureus varied with various antimicrobials tested. The resistance patterns ranged between 45% in freshwater isolates and 54.8% in seawater ones. Fifty one percent of the tested isolates have shown resistance to at least one of the five tested antimicrobials; with seawater isolates exhibiting the highest rates of antimicrobial resistance.     

Impact of erosion and accretion on the distribution of enterococci in beach sands

Bacterial pathogens in coastal sediments may pose a health risk to users of beaches. Although recent work shows that beach sands harbor both indicator bacteria and potential pathogens, it is neither known how deep within beach sands the organisms may persist nor if they may be exposed during natural physical processes. In this study, sand cores of approximately 100 cm depth were collected at three sites across the beach face in Kitty Hawk, North Carolina, before, during, and after large waves from an offshore hurricane. The presence of DNA from the fecal indicator bacterium Enterococci was detected in subsamples at different depths within the cores by PCR amplification. Erosion and accretion of beach sand at the three sites were also determined for each sampling day. The results indicate that ocean beach sands with persisting enterococci signals could be exposed and redistributed when wind, waves, and currents cause beach erosion or accretion.     

Uptake of Dissolved and Oil Phase Organic Chemicals by Bacteria

Hydrophobic organic chemicals (HOCs) discharged into soil and ground water will partition into gaseous, aqueous, oil, and sorbed phases. Knowledge of how bacteria assimilate HOCs is important to individuals involved in evaluating intrinsic, or engineered, bioremediation. The majority of bacteria isolated from the subsurface are gram-negative. The outer membrane of gram-negative organisms acts as a selective barrier to many solutes, including hydrophobic chemicals. Thus, diffusional transport of a hydrophobic solute through the outer membrane may be the rate-limiting step in biodegradation. Bacteria may also produce biosurfactants that can facilitate cell-oil contact or assist solubilization of oil and sorbed phases.     

Pore water transport of enterococci out of beach sediments

Enterococci are used to evaluate the safety of beach waters and studies have identified beach sands as a source of these bacteria. In order to study and quantify the release of microbes from beach sediments, flow column systems were built to evaluate flow of pore water out of beach sediments. Results show a peak in enterococci (average of 10% of the total microbes in core) released from the sand core within one pore water volume followed by a marked decline to below detection. These results indicate that few enterococci are easily removed and that factors other than simple pore water flow control the release of the majority of enterococci within beach sediments. A significantly larger quantity and release of enterococci were observed in cores collected after a significant rain event suggesting the influx of fresh water can alter the release pattern as compared to cores with no antecedent rainfall.