Antibiotic resistance of Enterococcus spp. isolated from wastewater and sludge of poultry slaughterhouses

Antibiotimicrobial resistance was investigated in 537 Enterococcus spp. isolates recovered from 22 samples of crude inflow, treated effluent and sludge collected in wastewater treatment plants of eight poultry slaughterhouses of Portugal. No significant differences (P > 0.05) were found in the resistance to each antimicrobial agent with regards to the origin of the sample (inflow, sludge and effluent). Many of the isolates displayed resistance to tetracycline (85.7%), erythromycin (45.7%), nitrofurantoin (34.0%) and rifampicin (17.8%). Resistance was also observed, but to a lesser extent, to ciprofloxacin (10.2%), ampicillin (8.0%), chloramphenicol (4.6%), vancomycin (0.9%) and gentamicin (0.4%). Resistance to three or more antimicrobial classes was present in 37.1% of the isolates. Wastewater treatment resulted in viable enterococci decrease between less than 1 log and 4 log; nevertheless, more than 4.4 x 10(5) colony forming units (CFU)/100 mL were present in the outflow of the plants and thus resistant enterococci are not prevented from reaching the general environment.     

The effect of wastewater treatment on antibiotic resistance in Escherichia coli and Enterococcus sp.

The effects of wastewater treatment on the proportion of Escherichia coli and Enterococcus sp. resistant to specific antibiotics were investigated at two facilities in Davis County, Utah, one of which received hospital waste. Samples were taken from the influent, effluent before disinfection, and secondary anaerobic sludge digester effluent. There was very little difference in antibiotic resistance among E. coli in the inflow waters of the plants but the plant receiving hospital waste had a significantly higher proportion of antibiotic resistant Enterococcus. The effect of wastewater treatment on antibiotic resistance was more pronounced on enterococci than E. coli. Although some increases in antibiotic resistance were observed, the general trend seemed to be a decrease in resistance, especially in the proportion of multidrug resistant Enterococcus sp.     

Enterotoxin production and antimicrobial susceptibility in Staphylococci isolated from traditional raw milk cheeses in Serbia

This study was undertaken to determine the prevalence of coagulase positive staphylococci (CPS) by examining a total of 71 raw milk cheeses. Additionally, enterotoxigenicity, antimicrobial susceptibility and the presence of mecA and mecC genes in the staphylococcal isolates were investigated. The isolation and enumeration procedure of CPS followed the International Organization for Standardization (ISO) standard. The presumptive staphylococci were identified by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) using the VITEK MS system. VIDAS® Staph enterotoxin II assay was used for the detection of classical enterotoxins. Antimicrobial susceptibility testing (AST) was accomplished performing the disk diffusion method. All suspected methicillin resistant staphylococci were investigated for the presence of mecA and mecC genes by PCR assay. A high prevalence (87.32%) of CPS was detected in the cheeses at contamination levels up to 5.58 log CFU g^?1. Among 47 staphylococcal isolates screened for enterotoxin production, only one isolate, identified as S. hyicus, was confirmed as being enterotoxigenic. Resistance to penicillin (63.70%) was the most common resistance among the tested Staphylococcus aureus isolates. The dominant phenotypic resistance patterns in coagulase negative staphylococci (CNS) were resistance to ofloxacin and fusidic acid. All CNS isolates were susceptible to the clinically important antibiotics clindamycin, chloramphenicol, gentamicin, linezolid, rifampicin and trimethoprim-sulfamethoxazole. The mecA and mecC genes were not detected. To the best of our knowledge, this is the first study concerning evaluation of the presence of methicillin resistant staphylococci (MRS) in dairy products in Serbia.     

Environmental and Potential Health Effects of Growing Leafy Vegetables on Soil Irrigated Using Sewage Sludge and Effluent: A Case of Zn and Cu

Abstract

The use of sewage sludge and effluent as a source of nutrients and water for crop production is increasing worldwide. A study was conducted in 2001 at Pension farm (near Harare) to determine the effect of long term (>30 yrs) application of sewage sludge and effluent on Zn and Cu accumulation in top soil, uptake of these metals by lettuce (Lactuca sativa L.) and mustard rape (Brassica juncea L.), and dry matter yield. Application of sewage sludge/effluent significantly (p < 0.001) increased total Zn (13.7–1563.9 mg kg^?1) and Cu (2.5–133.3 mg kg^?1) in the top soil (0–20 cm depth) compared to the control. Sewage sludge/effluent addition significantly (p < 0.001) increased Zn uptake by both test crops, while Cu uptake was significant in the first crop of lettuce and the second crop of mustard rape. Based on the dietary patterns of poor urban households in Zimbabwe, the maximum possible intake of Cu will only constitute 40% the Maximum Daily Intake (MDI). The toxicological implications for Zn will however be more severe, exceeding the MDI by 77% through exposure by lettuce consumption and by 251% consumption of mustard rape. It was concluded that long-term addition of sewage sludge/effluent to soil at Pension farm had increased the concentration of Zn and Cu in top soil to levels that pose environmental concern. The consumption of leafy vegetables produced on these soils pose a health risk to poor communities that reside around the study site, especially children, through possible Zn toxicity.     

Prevalence and antimicrobial resistance of Shiga toxin-producing Escherichia coli in milk and dairy products in Egypt

Abstract

Food contaminated with Shiga toxin-producing Escherichia coli (STEC) represents a hazardous public health problem worldwide. Therefore, the present study was performed to elucidate the virulent and antimicrobial resistance characteristics of STEC isolated from milk and dairy products marketed in Egypt. A total of 125 samples (raw market milk, bulk tank milk, Kareish cheese, white soft cheese, and small scale-produced ice cream, 25 each) were collected for determination the prevalence and antimicrobial resistance profiling of STEC. Thirty-six STEC isolates were recovered from milk and dairy products. Serological analysis illustrated that three isolates were E. coli O157:H7 and 33 isolates belonged to different serotypes. Molecular examination indicated that all isolates harboured stx1 and/or stx2 genes, 14 isolates expressed eaeA gene and 3 isolates possessed rfbE gene. Antimicrobial resistance profiling of the isolates was both phenotypically and genetically examined. Interestingly, 31 out of 36 (86.11%) isolates were multidrug-resistant and harboured the extended-spectrum β-lactamase encoding genes, namely, bla_CTX-M-15, bla_SHV-12 and bla_CTX-M-14. Moreover, 12 isolates (33.33%) harboured plasmid-mediated quinolone resistant gene, qnrS. The overall conclusion of the current investigation indicated insufficient hygienic measures adopted during milking, handling, and processing leading to development of pathogenic and multidrug-resistant STEC.     

Antimicrobial resistance in Escherichia coli and Salmonella spp. isolates from fresh produce and the impact to food safety

Foodborne diseases associated with fresh produce consumption have escalated worldwide, causing microbial safety of produce of critical importance. Bacteria that have increasingly been detected in fresh produce are Escherichia coli and Salmonella spp., both of which have been shown to progressively display antimicrobial resistance. The study focused on the assessment of antimicrobial resistance of these enteric bacteria from different kinds of fresh produce from various open air markets and supermarkets in the Philippines. Using the disk diffusion assay on a total of 50 bacterial isolates obtained from 410 fresh produce surveyed, monoresistance to tetracycline was observed to be the most prevalent (38%), followed by multidrug resistance to tetracycline, chloramphenicol, ciprofloxacin, and nalidixic acid (4%), and lastly by dual resistance to tetracycline and chloramphenicol (2%). Using multiplex and simplex polymerase chain reaction (PCR) assays, tetA (75%) and tetB (9%) were found in tetracycline resistant isolates, whereas catI (67%) and catIII (33%) were detected in chloramphenicol resistant isolates. Sequence analysis of gyr and par genes from the ciprofloxacin and nalidixic acid resistant isolates revealed different mutations. Based on the results, fresh produce act as a reservoir of these antibiotic resistant bacteria which may pose health threat to consumers.     

Genotypic characterization of quinolone resistant-Escherichia coli isolates from retail food in Morocco

This study was conducted to assess the retail food as a possible vehicle for antimicrobial resistant, particularly quinolones resistant and pathogenic Escherichia coli. We determined the prevalence and characteristics of nalidixic acid (Nal) resistant E. coli isolates from diverse retail food samples. In all, 70 (28%) of 250 E. coli isolates studied were Nal-resistant E. coli and 91% of these were multi-drug resistant. Plasmid mediated quinolone resistance genes were identified in 32 isolates, including aac(6′)-Ib-cr (n = 16), qnrS1 (n = 11) and qnrB19 (n = 7). Mutations in gyr A and par C genes were detected among 80% of the isolates, and the isolates showed substitution Ser83-Leu and Asp87-Asn in gyrA and Ser80-Ile in parC. In addition, three different gene cassettes were identified (aadA1, aadA7, aac(3)-Id) in 18%. Virulence-associated genes stx1, eae, sfa, hlyA and stx2 were found in six (8%), three (4%), two (3%), three (4%) and three (4%) isolates, respectively. E. coli isolates of phylogenetic group A were dominant (64%, 45/70). Pulsed field gel electrophoresis revealed none epidemiological relationship between these isolates. The results of this work report the higher frequency of Nal-resistant E. coli isolates from Moroccan retail food samples including MDR and pathogenic isolates.     

活性污泥与生物膜复合工艺处理石化废水的研究

实验研究了活性污泥与生物膜复合工艺(HP)对石化废水的处理效果,并与活性污泥工艺(ASP)进行对比研究.结果表明,HP对石化废水中COD的去除效果略好于ASP,而对NH3-N的去除效果明显优于ASP.另外,两种工艺对硫化物和油的去除效果类似.提高进水负荷时,HP表现出较强的耐冲击负荷能力.以2L/h连续运行时,石化废水...     

Effect of Solids Concentration on Bacterial Leaching of Heavy Metals from Sewage Sludge

Abstract

Microbial metal leaching from sewage sludge (2-9% w/v) was carried out with the iron-oxidizing bacterium Thiobacillus ferrooxidans. Measurements of pH, oxidation-reduction potential, and concentration of Fe²⁺ indicated that T. ferrooxidans was effective in removing metals from an incubation bath containing less than 5% sludge solids concentration. Specifically, Cu leaching was completely suppressed at a high solids concentration of 9% (w/v). Results indicated that the deactivation of T. ferrooxidans at a high sludge content was mainly due to the presence of inhibiting materials such as organic matter. A mixed culture of sulfur-oxidizing bacteria was obtained by enrichment from anaerobically digested sewage sludge to enhance the efficiency of the microbial leaching process. These bacteria were much more effective in metal leaching than was iron-oxidizing T. ferrooxidans. At 9% (w/v) solids concentration, the leaching efficiencies of Zn and Cu were 78% (2.66 g/kg dry sludge) and 59% (1.36 g/kg dry sludge), respectively. Therefore, when removing heavy metals from the anaerobically digested sewage sludge, the indigenous sulfur-oxidizing bacteria isolated in the current study were more efficient than T. ferrooxidans, especially at high sludge solids concentrations.     

Case Studies on Biological Treatment of Tannery Effluents in India

Abstract

This paper presents a comparative assessment of the cost and quality of treatment of tannery wastewater in India by two common effluent treatment plants (CETPs) constructed for two tannery clusters, at Jajmau (Kanpur) and at Unnao in the state of Uttar Pradesh, India. The Jajmau plant is upflow anaerobic sludge blanket (UASB) process-based, while the Unnao plant is activated sludge process (ASP)-based. Investigations indicated that the ASP-based plant was superior in all respects. Total annualized costs, including capital and operation and maintenance costs, for the UASB and ASP plants were Rs. 4.24 million/million liters per day (MLD) and Rs. 3.36 million/MLD, respectively. Land requirements for the two CETPs were 1.4 hectares/MLD and 0.95 hectares/MLD, respectively. Moreover, the treated UASB effluent had higher biochemical and chemical oxygen demand (BOD/COD) and considerable amounts of other undesirable constituents, like chromium (Cr) and sulfide, as compared with the ASP effluent, which had lower BOD/COD and negligible concentration of sulfide and Cr. Sludge production from the UASB-based plant was also higher at 1.4 t/day/MLD, in comparison to the sludge production of 0.8 t/day/MLD for the ASP-based plant. Also, the entire sludge produced in the UASB-based plant was Cr-contaminated and, hence, hazardous, while only a small fraction of the sludge produced in the ASP-based plant was similarly contaminated. The results of this study are at variance with the conventional wisdom of the superiority of anaerobic processes for tannery waste-water treatment in tropical developing countries like India.     

The fate of lindane in the conventional activated sludge treatment process

A pilot-scale treatability study was performed to evaluate the fate of lindane (gamma-hexachlorocyclohexane) in wastewater treatment plants operating in the conventional activated sludge mode. Different types of wastewater (industrial and municipal) spiked with variable lindane concentrations were used at different dosing rates in order to determine distribution and removal under various operational conditions. The major amount (67-91%) of lindane inputs to the treatment process was found to concentrate in primary sludge. A significant linear correlation between the compound's partition coefficient (logKp) and the organic fraction of primary sludge (foc) was found. Sorption on primary sludge solids was concluded to be the major removal mechanism. Only 0.1-2.8% of lindane inputs was concentrated in activated sludge. Lindane losses in primary treatment were low (4-26%). Higher losses (up to 61%) were observed during the biological treatment probably due to biodegradation. These losses were negatively correlated with the inflow rate of lindane into the aeration tank. Activated sludge aged about 23 d presented the maximum loss of lindane. Increased sludge age was associated with increased percentages of lindane in the final effluent.     

Antibiotic resistance,virulence factors and biofilm formation ability in Escherichia coli strains isolated from chicken meat and wildlife in the Czech Republic

Attachment of pathogenic bacteria to food contact surfaces and the subsequent biofilm formation represent a serious threat for the food industry, since these bacteria are more resistant to antimicrobials or possess more virulence factors. The main aim of this study was to investigate the correlation between antibiotic resistance against 13 antibiotics, distribution of 10 virulence factors and biofilm formation in 105 Escherichia coli strains according to their origin. The high prevalence of antibiotic resistance that we have found in wildlife isolates could be acquired by horizontal transfer of resistance genes from human or domestic or farm animals. Consequently, these commensal bacteria might serve as indicator of antimicrobial usage for human and veterinary purposes in the Czech Republic. Further, 46 out of 66 resistant isolates (70%) were able to form biofilm and we found out statistically significant correlation between prevalence of antibiotic resistance and biofilm formation ability. The highest prevalence of antibiotic resistance was observed in weak biofilm producers. Biofilm formation was not statistically associated with any virulence determinant. However, we confirmed the correlation between prevalence of virulence factors and host origin. Chicken isolates possessed more virulence factors (66%), than isolates from wildlife (37%). We can conclude that the potential spread of antibiotic resistance pattern via the food chain is of high concern for public health. Even more, alarming is that E. coli isolates remain pathogenic potential with ability to form biofilm and these bacteria may persist during food processing and consequently lead to greater risks of food contamination.     

Removal and formation of chlorinated triclosan derivatives in wastewater treatment plants using chlorine and UV disinfection

Triclosan, a common antimicrobial agent, may react during the disinfection of wastewater with free chlorine to form three chlorinated triclosan derivatives (CTDs). This is of concern because the CTDs may be photochemically transformed to tri- and tetra-chlorinated dibenzo-p-dioxins when discharged into natural waters. In this study, wastewater influent, secondary (pre-disinfection) effluent, and final (post-disinfection) effluent samples were collected on two occasions each from two activated sludge wastewater treatment plants, one using chlorine disinfection and one using UV disinfection. Concentrations of triclosan and three CTDs were determined using ultra performance liquid chromatography-triple quadrupole mass spectrometry with isotope dilution methodology. Triclosan and the CTDs were detected in every influent sample at levels ranging from 453 to 4530 and 2 to 98 ng L⁻¹, respectively, though both were efficiently removed from the liquid phase during activated sludge treatment. Triclosan concentrations in the pre-disinfection effluent ranged from 36 to 212 ng L⁻¹, while CTD concentrations were below the limit of quantification (1 ng L⁻¹) for most samples. In the treatment plant that used chlorine disinfection, triclosan concentrations decreased while CTDs were formed during chlorination, as evidenced by CTD levels as high as 22 ng L⁻¹ in the final effluent. No CTDs were detected in the final effluent of the treatment plant that used UV disinfection. The total CTD concentration in the final effluent of the chlorinating treatment plant reached nearly one third of the triclosan concentration, demonstrating that the chlorine disinfection step played a substantial role in the fate of triclosan in this system.     

Potential for CSO treatment with horizontal flow constructed wetlands: influence of hydraulic load,plant presence and loading frequency

This study aimed at analysing the performance of horizontal subsurface flow constructed wetlands (CWs) to treat combined sewer overflow (CSO). Four horizontal subsurface flow CWs, organized in two groups (A and B) each with a planted (Phragmites australis) and a non-planted bed, were loaded with simulated CSO, with group B receiving twice the hydraulic load of group A. Beds were monitored for pH, dissolved oxygen, conductivity, redox potential, chemical oxygen demand (COD), total suspended solids (TSS) and enterococci. Porosity variations were also estimated. Monitoring was conducted during spring and wintertime, with regular and irregular loading frequencies. Results showed an average treatment efficiency of 90–100 % for TSS, 60–90 % for COD and 2–6 log for enterococci. Removal rates were especially relevant in the first 24 h for COD and TSS. TSS and enterococci removal did not exhibit the influence of macrophytes or the applied hydraulic load while COD’s removal efficiency was lower in the higher load group and in planted beds.

     

造纸白泥和粉煤灰的添加对污泥消化液中氮磷回收的影响

污泥厌氧消化液中含有丰富的氮磷,若直接排放到环境中,将会对附近水体造成严重污染。由于消化液中Mg^2＋和Ca^2＋的含量很低,严重影响了氮磷的回收效果。把造纸白泥和粉煤灰引入到污泥厌氧消化液氮磷的回收当中,可以明显地提升消化液pH和提高PO4^3-P和NH3-N回收率。实验结果表明：当造纸白泥添加量为4g／（L·h）时,曝气12h后,pH可达10．19,此时PO4^3-P和NH3-N回收率分别达到64％和45％;而当粉煤灰添加量为4g／（L·h）时,曝气12h后,pH达到9．63,PO4^3-P和NH3-N回收率分别为46％和41％。但仅用曝气方式处理,12h后,pH值仅为8．52,PO4^3-P和NH3-N回收率分别只有20％和18％。实验结果还表明,水力停留时间（HRT）越大,pH上升速度越快,幅度越大,氮磷的回收效果就越好。     

Fate of Terpene Compounds in Activated Sludge Wastewater Treatment Systems

ABSTRACT

Terpene-based cleaners are being widely used in industrial cleaning formulations because of their ability to replace suspected ozone-depleting chemicals such as 1,1,1-trichloroethane and 1,1,2-trichloro-1,2,2-trifluoroethane (CFC-113). Substitution of chlorinated solvents with ter-pene-based cleaners, however, is expected to result in increased discharges to wastewater from industrial operations. A pilot-scale study was conducted at the U.S. Environmental Protection Agency's (EPA) Test & Evaluation Facility in Cincinnati, OH, to quantify the fate of specific terpene compounds in the activated sludge wastewater treatment process. Biodegradation rates of terpenes were estimated from the difference between the influent terpene mass flow rates and the amounts volatilized to air, partitioned to waste sludge, and passed through the treatment process unchanged. Any chemical transformation of the terpene compounds studied was attributed to biodegradation.

Analytical methods were developed to determine ter-pene concentrations in aqueous and gaseous media. The fate of two common terpene compounds (d-limonene and terpinolene) were evaluated in three identical pilot-scale systems: (1) a system with a high target spike range (2–10 mg/L), (2) a system with a low target spike range (0.5–2 mg/L), and (3) a control system (no spike).

The study showed that the primary removal mechanism for the terpene compounds in the activated sludge process is biodegradation. Typically, greater than 90% of the mass of terpenes entering the aeration basin of the activated sludge process biodegrades to other compounds; volatilization from the reaction basin accounts for less than 10%, while loss to waste activated sludge and the secondary clarifier effluent accounts for less than 1%.     

Occurrence and abundance of tetracycline,sulfonamide resistance genes,and class 1 integron in five wastewater treatment plants

To understand the transport and fate of antibiotic resistance genes in wastewater treatment plants, 12 resistance genes (ten tetracycline resistance genes, two sulfonamides genes) and class 1 integron gene (intI1) were studied in five wastewater treatment plants with different treatment processes and different sewage sources. Among these resistance genes, sulfonamides genes (sul1 and sul2) were of the most prevalent genes with detection frequency of 100 %. The effluent water contained fewer types of resistance genes than the influent in most selected plants. The abundance of five quantified resistance genes (tetG, tetW, tetX, sul1, and intI1) decreased in effluent of plants treating domestic or industrial wastewater with anaerobic/aerobic or membrane bioreactor (MBR) technologies, but tetG, tetX, sul1, and intI1 increased along the treatment units of plants treating vitamin C production wastewater by anaerobic/aerobic technology. In plant treating cephalosporins production wastewater by UASB/aerobic process, the quantities of tetG, tetX, and sul1 first decreased in anaerobic effluent water but then increased in aerobic effluent water.     

Antibiotic resistance and genetic diversity of Escherichia coli isolates from traditional and integrated aquaculture in South China

This study investigated antibiotic resistance profiles including antibiotic resistance frequencies, resistance genes and resistance patterns in Escherichia coli strains isolated from traditional and integrated aquaculture systems in South China by using antibiotic susceptibility testing and real time polymerase chain reaction (PCR) technique. The E. coli isolates were found to be resistant to at least one antibiotic among 12 antibiotics. Higher resistance frequencies to ampicillin, sulfamethoxazole, trimethoprime, streptomycin and tetracycline were found compared to the rest antibiotics. Among the 10 tetracycline resistance genes detected in the resistant isolates, the most prevalent tetracycline resistance genes were tetA, tetW and tetB with the frequency of 69.7%, 63.5% and 21.9%, respectively. Three sulfonamide resistance genes were detected in these resistant isolates, with their detection frequencies in the following order: sul2 (55.3%) > sul3 (28.2%) > sul1 (6.2%). Four resistance genes mainly encoding extended-spectrum β-lactamases (ESBLs) were detected in these resistant isolates, with the detection frequencies of blaTEM (28.4%) > blaOXA (9.7%) > blaCTX (9.3%) > blaCARB (5.2%) > blaSHV (0.0%). It was found that the integrated aquaculture system exhibited generally higher prevalence of antibiotic resistance than the traditional aquaculture system. An integrated aquaculture system could facilitate development of bacterial resistance and spread of the antibiotic resistance genes, and consequently become an important reservoir of resistance genes.     

Determination of three cresol isomers in sewage sludge by solid-liquid extraction with low temperature purification and gas chromatography-mass spectrometry

Abstract

Cresols are chemical contaminants derivative from phenol which can be found in sewage sludge. However, little attention has been given to monitoring these compounds in environmental matrices in the literature. Thus, the objective of this study was to develop a simple method based on solid-liquid extraction with low temperature purification for determining three cresol isomers in sludge. The quantification of these compounds was performed by gas chromatography coupled to mass spectrometry with a previous derivatization step. After a detailed study, the cresol recovery was higher than 91%, with relative standard deviation lower than 12% and a limit of quantification of 20?μg kg^?1. Linearity was achieved between 10 and 90?μg L^?1 (R² > 0.98) with the standard solutions prepared in matrix extracts due to the trouble caused by the matrix effect. The proposed method was applied with success for monitoring cresols in sewage sludge samples coming from six different wastewater treatment plants. All samples showed contamination by cresols, mainly p-cresol with values between 32.3 and 516.9?μg kg^?1. The majority of the analyzed samples showed a total sum of the isomers higher than the maximum residue limit established by Brazilian legislation (160?μg kg^?1).