Chlorophenol tolerant and degradative bacteria isolated from a river receiving pulp mill discharges

A screening was carried out in a river receiving pulp mill discharges (BioBio river, central Chile) to study the bacterial population able to tolerate and degrade chlorophenols. In four selected stations and at different seasons, water samples were taken from the river. Total bacterial population and the number of chlorophenol tolerant bacteria were counted. Bacterial tolerance to 25, 50, 100, 200 and 400 μg/ml of 2,4‐dichlorophenol (DCP), 2,4,6‐trichlorophenol (TCP) and pentachlorophenol (PCP) was determined. Strains able to tolerate at least 400 μg/ml chlorophenols were found. Eighteen bacterial strains able to use chlorophenols as the sole carbon source were isolated: five of them were Pseudomonas fluorescens, two Alcaligenes sp., one Citrobacter freundii, one Aeromonas sp. and nine unidentified Gram negative bacilli. A good correlation (r ² = 0.758) between the logarithm of DCP tolerant bacteria and the content of adsorbable organic halogen (AOX) at low dilution in the river water was established. Four groups of tolerant bacteria were found: growing on PCP and DCP, growing on PCP and TCP, growing only on PCP and growing only on TCP.     

Multiple antibiotic resistance patterns of rhizospheric bacteria isolated from Phragmites australis growing in constructed wetland for distillery effluent treatment

Susceptibility patterns of 12 different antibiotics were investigated against rhizospheric bacteria isolated from Phragmites australis from three different zones i.e. upper (0-5 cm), middle (5-10 cm), lower (10-15 cm) in constructed wetland system with and without distillery effluent. The major pollutants of distillery effluent were phenols, sulphide, heavy metals, and higher levels of biological oxygen demand (BOD), chemical oxygen demand (COD) etc. The antibiotic resistance properties of bacteria were correlated with the heavy metal tolerance (one of distillery pollutant). Twenty-two species from contaminated and seventeen species from non-contaminated site were tested by agar disc-diffusion method. The results revealed that more than 63% of total isolates were resistance towards one or more antibiotics tested from all the three different zones of contaminated sites. The multiple-drug resistance property was shown by total 8 isolates from effluent contaminated region out of which 3 isolates were from upper zone, 3 isolates from middle zone and 2 isolates were from lower zone. Results indicated that isolates from contaminated rhizosphere were found more resistant to antibiotics than isolates from non-contaminated rhizosphere. Further this study produces evidence suggesting that tolerance to antibiotics was acquired by isolates for the adaptation and detoxification of all the pollutants present in the effluent at contaminated site. This consequently facilitated the phytoremediation of effluent, which emerges the tolerance and increases resistance to antibiotics.     

Effect of neem extract against the bacteria isolated from marine fish

Marine ornamental fishes are exceedingly valuable due to their high demand in domestic and international markets. There is a growing global interest to rear the fishes in captivity. But problem due to bacteria and fungi are the major hitch in captive condition. Since, the use of antibiotics is banned, an attempt was made to ascertain in vitro assay of the neem leaves extract against the bacterial pathogens isolated from infected fishes. Bacterial strains isolated from infected regions of the clown fishes Amphiprion sebae and A. ocellaris were identified as Aeromonas hydrophila, Enterobacter sp., E. coli, Pseudomonas aeruginosa, Proteus sp., Streptococcus sp., Vibrio cholerae, V. alginolyticus, V. parahaemolyticus and Yersinia enterocolitica. Ethanol and methanol extracts were highly inhibitory to the bacterial isolates when compared to other solvents. Ethanol extracts exhibited low minimum inhibitory concentration (75-250 microg ml(-1)) as compared to other extracts. The present finding revealed that the neem leaf extract significantly reduces the bacterial pathogens and their infection in marine ornamental fishes.     

Association between heavy metals and antibiotic-resistant human pathogens in environmental reservoirs: A review

Heavy metals can act as co-selecting agents and promote antibiotic resistance. Most frequent resistances to heavy metals are observed for zinc and cadmium. P. aeruginosa and E. coli are commonly resistant to heavy metals and antibiotics. Heavy metals proliferate antibiotic resistance through co- and cross-resistance. Heavy metal and antibiotic resistances are common near anthropogenic activities. Antibiotic resistance in human pathogens can proliferate under selective pressures. Heavy metals in environmental reservoirs may contribute to selecting antibiotic-resistant strains. To determine the associations between heavy metals and antibiotic resistance, a literature review was conducted to systematically collect and categorize evidence for co-occurrence of resistance to heavy metals and antibiotics within human pathogenic bacteria in water, wastewater, and soil. In total, 42 publications adhered to inclusion criteria. Across the reservoirs, zinc and cadmium were the most commonly observed heavy metals associated with resistance to antibiotics. Pseudomonas aeruginosa and Escherichia coli were the most commonly studied bacteria with reported co-occurrence of resistance to several heavy metals and antibiotic classes. As co-selecting agents, prevalence of heavy metals in the environment can proliferate resistance to heavy metals and antibiotics through co-resistance and cross-resistance mechanisms. In comparing different reservoirs, soils and sediments harbor higher heavy metal and antibiotic resistances compared to water environments. Additionally, abiotic factors such as pH can affect the solubility and hence, the availability of heavy metals to bacterial pathogens. Overall, our review demonstrates heavy metals act as co-selecting agents in the proliferation of antibiotic resistance in human pathogens in multiple environmental reservoirs. More studies that include statistical data are needed to further describe the exposure-response relationships between heavy metals and antibiotic resistance in different environmental media. Moreover, integration of culture-based and molecular-based methods in future studies are recommended to better inform our understanding of bacterial co- and cross-resistance mechanisms to heavy metals and antibiotics.     

天津水产业磺胺类耐药细菌及其分布

水产养殖中抗生素的滥用可能会诱导水产品和周围环境中的耐药菌.针对天津市北辰区、西青区、东丽区和津南区的6个水产养殖场的底泥和水样,采用抗性平板筛选耐药细菌,通过对磺胺甲恶唑的耐药率分析,发现底泥中的耐药率要大大高于水中的耐药率,并且与国外的某些研究相比,底泥的细菌耐药率呈现较高的趋势.另外,利用16S-rDNA鉴定出8...     

Bioactive potential of seagrass bacteria against human bacterial pathogens

Study of marine organisms for their bioactive potential, being an important part of marine ecosystem, has picked up the rhythm in recent years with the growing recognition of their importance in human life. Investigation was carried out to isolate 32 strains of endo and epiphytic bacteria in 2 seagrass species viz., Syringodium isoetifolium and Cymodocea serrulata. Morphologically different bacterial strains were tested against 5 antibiotic resistant human bacterial pathogens, of which 10 associated bacteria shown inhibitory activity against one or more bacterial pathogens. Minimum inhibitory concentration (MIC) and Minimum bacterial concentration (MBC) determination with extracellular bioactive compounds from the associated bacteria reveals that, the strain ENC 5 showed inhibitory activity against all the bacterial pathogens with the maximum sensitivity against Pseudomonas aeruginosa at the MIC value of 500 microg ml(-1).     

水产养殖环境中抗生素抗性基因（ARGs）的研究及进展(Research Advancement of Antibiotics Resistance Genes（ARGs）in Aquaculture Environment)

抗生素在现阶段的水产养殖中具有不可替代的作用,但长期滥用抗生素会诱导水生动物体内产生携带抗性基因(ARGs)的细菌株,尤其是多重耐药性菌株的产生将使得各种疾病的治疗更加棘手.抗性基因一旦被排泄到水产环境中不仅会对养殖区域和周围的环境造成潜在的基因污染,而且还会通过各种可移动遗传元件如质粒、转座子、整合子等的水平迁移作用进入其他致病菌和环境细菌中,对人类的健康安全构成潜在的威胁.论文对抗生素抗性基因在水产养殖业中的来源、污染现状、潜在的传播途径和相关的检测方法进行了综述,指出了开展水产养殖业中抗生素抗性基因污染研究的必要性,建议政府和有关部门尽快进行水产养殖业抗生素抗性基因的污染机理与控制对策研究.     

Impact of heavy metals of industrial plant wastewater on benthic communities of Bizerte Lagoon (Tunisia)

ABSTRACT

The aim of this study was to assess the consequences of human impact on the characteristics of sediments heavy metal concentration, grain size and its influence on the structure of the microbial and meiofaunal community assemblages. A survey was carried out in July 2013 within six sites located in the Bizerte Lagoon (Tunisia), both downstream and upstream of industrial effluents. The highest total sediment metal concentrations were detected in stations located close to the industrial sewage discharge points. In these stations, the lowest densities of the total meiofauna (33?±?13?ind/10?cm^?2) and conversely the highest densities of cultivable bacteria that are heavy metal resistant have been reported (16?±?80.34?CFU?g^?1). Univariate (ANOVA) and multivariate (MDS/CCA) analyses demonstrate high dissimilarity (0.06) in meiofaunal and bacterial community structures between downstream and upstream industrial sewages. Furthermore, canonical correspondence analysis CCA results indicated that heavy metal sediment contamination promoted bacteria that are resistant to heavy metals, while heterotrophic bacteria supported the development of meiofauna taxa. The results highlight the importance of bacteria/meiofauna interactions, as both meiofaunal and microbial communities give indications of the ecological impact of heavy metal contamination in sediment.     

Characterization of boron tolerant bacteria isolated from a fly ash dumping site for bacterial boron remediation

Boron is an essential micronutrient for plants, but can above certain concentrations be toxic to living organisms. A major environmental concern is the removal of boron from contaminated water and fly ash. For this purpose, the samples were collected from a fly ash dumping site, Nagasaki prefecture, Japan. The chemical characteristics and heavy metal concentration of the samples were performed by X-ray fluorescent analysis and leaching test. For bacterial analysis, samples were collected in sterile plastic sheets and isolation was carried out by serial dilution method. The boron tolerant isolates that showed values of maximum inhibitory concentration toward boron ranging from 100 to 260 mM level were screened. Based on 16S rRNA sequencing and phylogenetic analysis, the isolates were most closely related to the genera Bacillus, Lysinibacillus, Microbacterium and Ralstonia. The boron tolerance of these strains was also associated with resistant to several heavy metals, such as As (III), Cr (VI), Cd, Cu, Pb, Ni, Se (III) and Zn. Indeed, these strains were arsenic oxidizing bacteria confirmed by silver nitrate test. These strains exhibited their salt resistances ranging from 4 to 15 % were determined in Trypticase soy agar medium. The boron tolerant strains were capable of removing 0.1–2.0 and 2.7–3.7 mg l^?1 boron from the medium and fly ash at 168 h. Thus, we have successfully identified the boron tolerant and removal bacteria from a fly ash dumping site for boron remediation.     

In-vitro antimicrobial activity and synergistic/antagonistic effect of interactions between antibiotics and some spice essential oils

Spices and herbs have been used for many years by different cultures. The aim of the present study is (1) to investigate in-vitro antimicrobial effects of different spices and herbs (5 species: Rosmarinus officinalis (Rosemary), Coriandrum sativum (coriander), Micromeria fruticosa (L.) Druce subsp. Brachycalyx P.H. Davis (White micromeria), Cumium cyminum (cumin), Mentha piperita (Peppermint) against different bacteria and fungi species, and (2) to discuss the in-vitro possible effects between the plants and antibiotics. The microorganisms used were Micrococcus luteus LA 2971, Bacillus megaterium NRS, Bacillus brevis FMC 3, Enterococcus faecalis ATCC 15753, Pseudomonas pyocyaneus DC 127, Mycobacterium smegmatis CCM 2067, Escherichia coil DM, Aeromonas hydrophila ATCC 7966, Yersinia enterocolitica AU 19, Staphylococcus aureus Cowan 1, Streptococcus faecalis DC 74 bacteria, and Saccharomyces cerevisiae WET 136, Kluvyeromyces fragilis DC 98 fungi in this study. The results indicated that essential oils of Rosmarinus officinalis, Coriandrum sativum L., Micromeria fruticosa (L.) Druce subsp. brachycalyx P.H. Davis, Cumium cyminum L., Mentha piperita L. were shown antimicrobial activity in the range of 7-60 mm 2 microl(-1) inhibition zone to the microorganisms tested, using disc diffusion method. Standard antibiotic such as Gentamicin (10 microg), Cephalothin (30 microg), Ceftriaxone (10 microg), Nystatin (10 U) discs were used for comparison with the antimicrobial activities of essential oils of these plants. In addition, antibacterial activity of essential oils of these plants was researched by effects when it was used together with these standard antibiotics in vitro. However, antibacterial activity changed also by in vitro interactions between these standard antibiotics and essential oils of these plants. Synergic, additive or antagonist effects were observed in antibacterial activity.     

抗生素、重金属和杀生剂抗性共选择机制

细菌抗生素抗性已被世界卫生组织认定为对公众健康的重要威胁。细菌抗性除了受到抗生素的直接选择压力,也受到其他物质如重金属和杀生剂的共同影响。研究发现,细菌可以通过外排泵系统、细胞膜通透性改变、作用靶标点的改变、酶修饰或降解作用、应激反应改变生理特性、金属螯合和生物转化等抗性机制对这些选择压力产生抗性,或者通过协同抗性、交叉抗性和共调控抗性的机制产生多重抗性。本文综述了细菌对抗生素、重金属和杀生剂的单一抗性机制、多重抗性机制,以及重金属和杀生剂对细菌抗生素抗性的影响机制,并分析了目前研究的不足之处。     

Combined effects of heavy metal (Hg) concentration and algal (Chlorella vulgaris) food density on the population growth of Brachionus calyciflorus (Rotifera: Brachionidae)

The combined effects of two food levels (0.5 x 10(6) and 1.5 x 10(6) cells ml(-1) of Chlorella vulgaris) and five concentrations (0, 0.000625, 0.00125, 0.0025, 0.005 mg l(-1) of HgCl2) of mercury on the population growth of the rotifer Brachionus calyciflorus was evaluated. The growth experiments were conducted for 18 days at 23+/-1 degrees C under continuous fluorescent illumination. For each food level - heavy metal combination, we maintained 3 replicates. Our data showed that regardless of food level, increase in the heavy metal concentration in the medium resulted in decreased population growth of B. calyciflorus. At any given heavy metal concentration, B. calyciflorus grown under higher food levels had higher population abundance. The rate of population increase was significantly influenced by both the heavy metal concentration and the algal level. The highest population growth rate (0.435+/-0.003 per day) was observed in controls at 1.5 x 10(6) cells ml(-1). The results of this study were discussed in relation to the protective role of algal density against heavy metal toxicity.     

太湖流域3种氯酚类化合物水质基准的探讨

按照美国地面水水质基准制定的程序和规范,筛选了太湖流域广泛存在的水生生物物种并收集了相应的基础毒性数据,探讨了五氯酚(PCP)、2,4-二氯酚(2,4-DCP)和2,4,6-三氯酚(2,4,6-TCP)在我国太湖地区的水生态基准的定值.同时采用蒙特卡罗构建物种敏感度分布(SSD)曲线和生态毒理模型方法预测了3种氯酚类化合物对太湖水生生物的急性基准浓度(CMC)和慢性基准浓度(CCC).结果表明,基于EPA规范方法和急慢性毒性比率得到的PCP、2,4-DCP和2,4,6-TCP3种氯酚类化合物的CMC值分别为25、908和594μg·L-1,CCC值分别为12、176和162μg·L-1;基于SSD曲线得到的CMC值分别为25、818和648μg·L-1,CCC值分别为6、75和198μg·L-1;基于生态毒理模型得到的CCC值分别为4、15和67μg·L-1,显示出3种方法得到的氯酚类化合物的CMC或CCC在同一个数量级上,但在数值上由生态毒理模型得出的CCC要小于其它两种方法,并且除PCP的急慢性基准值与美国EPA推出的水生态基准值相近外,其它两种氯酚类化合物的急慢性基准值均低于美国EPA推出的急慢性基准值.研究结果希望能为我国水质基准的制定提供一些有用的线索.     

Individual and combined effect of mercury and manganese on phenol and proline content in leaf and stem of mungbean seedlings

Mungbean (Vigna radiata L. Wilczelk cv. Pusa Baisakhi seedlings were raised in individual (0, 1, 10, 100 and 1000 ppm) and combined solutions (1 : 1, 10: 1, 1: 0 ppm Hg : Mn) of mercury and manganese for 6 days. Phenol and proline were found to accumulate in leaves in response to treatment with heavy metals. The magnitude of accumulation correlated with concentration of metals. However, a reverse trend was noticed in stem for phenol. Accumulation of phenol in response to heavy metal treatment was organ specific and occurred at higher rate in plant parts, which faced the stress mostly. However, accumulation of proline helped the plant to survive stress situation. In combined solutions, amelioration of mercurial toxicity by manganese was recorded.     

Determination of atmospheric heavy metals using two lichen species in Katni and Rewa cities, India

A biomonitoring study was conducted to assess the levels of atmospheric heavy metal pollution in Katni and Rewa cities of Madhya Pradesh, state in central India. The Pyxine cocoes and Phaeophyscia hispidula, two epiphytic foliose lichen were used as bioindicators in the present study and seven metals (As, Al, Cd, Cr, Fe, Zn, Pb) were analyzed in naturally growing thallus. The concentrations of these metals was observed to be in higher range as maximum values of Al, Cd, Cr and Zn were reported from the lichen samples from Rewa city which was 561.8 +/- 2.4, 6.8 +/- 0.8, 35.2 +/- 1.4, 214.6 +/- 2.0 microg g(-1) dry weight respectively. Whereas As, Fe and Pb were reported maximum in the lichen samples collected from Katni city areas with 33.4 +/- 0.05, 689.4 +/- 2.6, 13.3 +/- 0.5 microg g(-1) dry weight respectively. However the accumulation of Cd and Pb from both the cities are more or less similar in concentration. The selectivity sequence of metals were Fe>Al>Zn>As>Cr>Pb>Cd in Katni city, and Al>Fe>Zn>Cr>As>Pb>Cd in Rewa city. The findings of this study indicates that extent of heavy metal pollution in the atmosphere of the two cities which may lead to adverse health affects.     

Modified dehydrogenase enzyme assay for evaluation of the influence of Hg,Cd, and Zn on the bacterial community structure of a wastewater treatment plant

In this study, a traditional assay was modified to evaluate the effect of Hg, Cd, and Zn on the bacterial community of a sequencing batch reactor and activated sludge plants and heavy metal-resistant bacterial species were determined. After the isolation of metal-resistant bacteria, their 16S rRNA gene fragments were sequenced. The BLAST program was used to compare the resulting 16S rRNA sequences with those in GenBank database to identify the isolated bacterial species. Hg was found to be the most toxic metal for both bacterial communities investigated. Sequence batch reactor bacteria were comparatively more resistant to Hg, Cd, and Zn than those from activated sludge. The resistant strains were close to the members of genus Pseudomonas, Kocuria, Stenotrophomonas, Enterococcus, and Staphylococcus. The modified dehydrogenase enzyme assay seems to be simple, robust, and competent for evaluation of the impact of metals on bacterial activity. Sequencing batch reactor systems should be preferred over activated sludge when wastewaters containing hazardous metals are to be treated.     

Assessment of deposition of MEMC (seed dressing fungicide) in exposed rice grain through study of seedling growth parameters

Rice seed grains exposed to 240 ppm of seed dressing fungicide, MEMC showed increase in deposition of mercury with increase in period of observation (0h to 48h) in 765+/-5 mg of intact endosperm. The gradual increase in mercury deposition in intact endosperm caused simultaneous decrease in mercury content of the seed coat. Exposure periods of 5 to 20 min with 240 ppm of MEMC showed 5.5 to 8.72 microg of mercury deposition in 765+/-5 mg of intact endosperm. Increase in period of exposure (3 to 20 min) to 240 ppm of MEMC and also increase in chronic treatment to lower concentrations (1.5 ppm to 24 ppm) to seed grains have shown similar results in seedling growth parameters. Lower period of exposure or exposure to lower concentrations, both showed a significant inhibition of root growth, however the shoot growth showed stimulation. But higher exposure period or exposure to higher concentrations brought inhibition of both root and shoot growth. The exposure period showing 50% of root growth inhibition of 72h old seedling was calculated to be 10 min So also the chronic treatment showing the 50% of root growth inhibition was calculated to be 9 ppm. Ten min exposure of rice grain to 240 ppm of MEMC showed deposition of 7.11 microg of mercury in 765+/-5 mg of intact endosperm at 48h of observation (optimal phase of germinating process). Regression analysis indicated that by the time of 72h the intact endosperm and developing seedling might have acquired with a minimum amount of 10.23 microg of mercury from the seed coat. Finally it can be said that 10.23 microg of MEMC might be bringing 50% root growth inhibition in 72h old seedlings and 16.71 microg of mercury causing 50% inhibition of both root and shoot growth of 120h old seedlings.     

Lytic bacteriophages specific to Flavobacterium columnare rescue catfish, Clarias batrachus (Linn.) from columnaris disease

This investigation was aimed to find out appropriate strategy against antibiotic resistant bacterial fish pathogen, F. columnare. This pathogen was found persistently associated with fishes causing columnaris disease and ensuing mass mortality in hatchery and culture system of Sub - Himalayan region. Nine lytic F. columnare phages (FCP1 - FCP9) specific to its fifteen isolates were isolated from the water and bottom sediments of various geo-climatic regions of North India. The F. columnare phage FCP1 (made of hexagonal head and non contractile long tail belonging to family Podovariedae, a member of DNA virus) exhibited broader host range to lyse 9 out of 15 isolates of F. columnare. Therapeutic ability of FCP1 phage was assessed in C. batrachus inoculated intramuscularly (im) with virulent bacterial isolate FC8 and post inoculated (PI) with FCP1 phage (@ 10(8) : 10(6):: cfu : pfu) through intramuscular (im), immersion (bath) and oral (phage impregnated feed) treatment. Significant (p < 0.001) reduction (less than 10(-3) cfu ml(-1)) in host bacterium in the sera, gill, liver and kidney of challenged fishes was noted after 6 hr of phage treatment. Quantum of phage played a significant role in bringing down bacterial population as in the sera of dose 1 (@ 4.55 x 10(6) pfu ml(-1)) and dose 2 (@ 9.15 x 10(6) pfu ml(-1)) treated fishes mean log10 cfu value reduced by 3 logs (58.39%) and 5 logs (73.77%) at 96 hr, respectively. Phage treatment led to disappearance of gross symptoms, negative bacteriological test, detectable phage and 100% survival in experimentally infected C. batrachus. Result of this study provides evidence of profound lytic impact of FCP1 phage and represents its interesting therapeutic importance against antibiotic resistant F. columnare.     

Prevalence of plasmid mediated pesticide resistant bacterial assemblages in crop fields

Three crop fields namely paddy sugarcane and tomato exposed to bavistin [Methyl (1H-benzimidazol-2-yl) carbomate], monocrotophos[Dimethyl(E)-1-methyl-2-(methyl-carbamoyl) vinyl phosphate] and kinado plus [(EZ)-2-chloro-3-dimethoxyphosphinoyloxy-X1, X1-diethylbut-2-enamide], respectively were chosen for the present investigation to know the bacterial population and degradation of pesticides. The chemical nature of the soil and water samples from the pesticide contaminated fields was analysed along with counting of the total heterotrophic bacteria (THB), Staphylococci and Enterococcci population. Mean calcium, phosphate and biological oxygen demand were maximum in tomato field water Field water recorded maximum phophate and silicate content, whereas, sugarcane field water elicited maximum dissolved oxygen content. On the other hand, available phosphate and exchangeable potassium were maximum is sugarcane field soil. Significant variations in the bacterial population were evident between the treatments in sugarcane field soil and tomato field water exposed to monocrotophos and kinado plus, respectively In addition, significant variations between THB, Staphlyococci and Enterococci population were also evinced in both the sugarcane andtomato fields. The dominant pesticide resistant bacteria, Staphylococcus aureus, Enterococcus faecalis and Pseudomonas aeuroginosa harboured plasmids and the resistant trait observed were found to be plasmid borne.