Detection of Class I and II integrons for the assessment of antibiotic and multidrug resistance among Escherichia coli isolates from agricultural irrigation waters in Bulacan,Philippines

Contaminated irrigation water may greatly affect not only the quality of produce but also the people exposed to it. In this study, agricultural irrigation waters in Bulacan, Philippines were assessed and found to be contaminated with Escherichia coli (E. coli) ranging from 0.58 to 4.51 log₁₀ CFU/mL. A total of 79 isolates of E. coli were confirmed through polymerase chain reaction (PCR) amplifying the uidA gene and were tested for phenotypic resistance using 10 antimicrobials through the Kirby–Bauer disc diffusion method. Forty-six isolates (58.22%) were noted to be multidrug resistant (MDR) with high resistance rate to cephalothin, tetracycline, streptomycin, ampicillin, trimethoprim, nalidixic acid, and chloramphenicol. Moreover, this study also examined the prevalence of Class I and II integrons accounting to 67.39% and 17.39%, respectively, of the MDR E. coli strains using multiplex PCR. The results imply that the agricultural water used in Bulacan is contaminated with the fecal material of man or other animals present in the area, and the presence of MDR bacteria, which pose a potential threat to individuals in these areas, is alarming. In addition, detection of integrons could be a good marker for the identification of MDR isolates. Lastly, this study could develop strategies for the proper management of farming sites leading to the detection of food-borne pathogens and prevention of infectious diseases.     

Bioreduction of hexavalent chromium by Exiguobacterium indicum strain MW1 isolated from marine water of Paradip Port,Odisha, India

Hexavalent chromium-tolerant (1500?mg/L) bacterium MW1 was isolated from harbour water of Paradip Port and evaluated for Cr(VI) reduction potential. The isolate was identified as Exiguobacterium indicum by biochemical and 16S rRNA gene sequence methods. Salt tolerance of the bacterium was evaluated in a wide range of NaCl concentrations (0.5–13%, w/v). The Cr(VI) reduction of the strain was evaluated and optimised with varied Cr(VI) concentrations (100–1000?mg/L), pH (5.0–9.0), temperature (30–40°C) and shaking velocity (100–150?rpm) in two different minimal media (M9 and Acetate). Under optimised conditions, after 192?h of incubation nearly 92%, 50% and 46% reduction in the M9 minimal medium and 91%, 47% and 40% reduction in the acetate minimal medium were observed for 100, 500 and 1000?mg/L of Cr(VI), respectively. The exponential rate equation for Cr(VI) reduction yielded higher rate constant value, that is, 1.27?×?10^?2?h^?1 (M9) and 1.17?×?10^?2?h^?1 (Acetate) in case of 100?mg/L and became lower for 500 and 1000?mg/L Cr(VI) concentrations. Further, the association of bacterial cells with reduced product was ascertained by Fourier transform infrared spectrometer, UV–Vis–DRS and field-emission scanning electron microscope–energy-dispersive X-ray analyses. The above study suggests that the higher reducing ability of the marine bacterium E. indicum MW1 will be suitable for Cr(VI) reduction from saline effluents.     

活性污泥缺氧转化1-2-7-三氨基-8-羟基-3-6-萘二磺酸

1—2-7-三氨基-8-羟基-3—6-萘二磺酸（TAHNDS）作为偶氮染料的脱色产物很难被常规的厌氧-好氧染料废水处理工艺所去除。研究了未经驯化的活性污泥对TAHNDS的缺氧转化效果。结果表明,只有在特定的缺氧条件下（ORP在-50～-150mV之间）,TAHNDS才能被活性污泥所降解转化。当浓度在10—80mg／L范围内,TAHNDS可在72h内转化93％以上。加入100mg／L的硝酸盐和0．64mmol／L的氧化还原介体蒽醌-2-磺酸钠（AQS）可将40mg／L的TAHNDS的转化时间从84h缩短到36h。光谱及HPLC—MS分析表明,TAHNDS在缺氧条件下主要是通过脱氨基和脱磺酸作用生成已知可好氧生物降解的3,5-二氨基4-羟基萘-2-磺酸。因此,缺氧处理有望作为预处理工艺促进废水中TAHNDS的完全降解。     

降解SO2功能菌的16S rRAN基因测序及降解特性

用低浓度SO2诱导驯化方法获得高效脱硫菌群,并用分离培养与16S rRNA基因测序技术相结合的方法鉴定菌群种属,分析驯化过程中种群结构的动态变化,同时研究分离纯菌种的脱硫性能。结果表明,从诱导驯化7 d和14 d菌液中分别分离出23株菌和22株菌,16S rRNA序列分析发现这些菌归属于13个种,其中有6个种(Rhodococcus erythropolis、Pseudomonas putida、Microbacterium oxydans、Sphingomonas koreensis、Acinetobacter junii、Acinetobacter johnsonii)对SO2-3有较强的降解能力,并在持续驯化过程中稳定的生长传代,降解产物以硫酸根为主,还有极少量的单质硫。与含混合菌的驯化菌液降解SO2-3的能力相比,单一脱硫菌的脱硫性能较弱。脱硫功能菌株及其基本特性的研究为微生物处理SO2烟气提供了丰富的菌源信息和理论基础。     

假单胞菌好氧降解四溴双酚A的特性

为了解四溴双酚A(TBBPA)的好氧降解特性,采用选择富集法从活性污泥中分离出一株能够高效降解四溴双酚A的菌株。根据其形态、生理生化特性及16S rDNA核苷酸序列分析,该菌株被鉴定为假单胞菌属(Pseudomonas sp.)。研究结果表明,该菌株可通过好氧共代谢方式实现四溴双酚A的降解,葡萄糖是四溴双酚A降解的最佳碳源,其最优降解条件为葡萄糖8 g/L,牛肉膏0.5 g/L,pH值为7.0,培养温度为35℃,摇床转速为150 r/min。在该条件下其生物降解过程符合一级动力学模型,6 d后的降解率高达95.6%。LC-MS结果表明,四溴双酚A在好氧降解过程中会生成异丙苯酚类物质。     

Reduced Downward Mobility of Metolachlor and Metribuzin from Surfactant-Modified Clays

The study aims to prepare the organoclay complexes of metolachlor and metribuzin so as to reduce their downward mobility in soil profile. The organoclays were preadsorbed with phenyltrimethylammonium (PTMA) (50% of cation exchange capacity [CEC]) and hexadecyltrimethylammonium (HDTMA) (100% of CEC). Four metolachlor formulations, two for each organoclay, were prepared having 1% and 2% load of herbicide and were called PTMA-Metol-1%, PTMA-Metol-2%, HDTMA-Metol-1% and HDTMA-Metol-2%. Two metribuzin formulations were prepared and were called PTMA-Metri-1% and HDTMA-Metri-1%. Soil column leaching experiment showed that compared to their commercially available formulations, organoclay complexes were effective in reducing the downward mobility of both herbicides. Organoclay complexes of metolachlor and metribuzin prepared using PTMA-Montmorillonite performed better than the HDTMA-Montmorillonite complexes.     

Free Fatty Acids and Sterols in Swine Manure

Free fatty acids and sterols were assessed in fresh manure and anaerobic lagoon sludge from swine production facilities in North Carolina. Eight free fatty acids and five sterols were identified and quantified in both manure and sludge samples. Compound identification was performed by gas chromatography/mass spectroscopy (GC-MS), and compound quantities were determined by gas chromatography after solid phase extraction with a 50:50 mixture of diethyl ether and hexane. The free fatty acids occurring in greatest abundance in both fresh manure and lagoon sludge were palmitic, oleic, and stearic. Free fatty acid content in fresh manure ranged from approximately 3 μ g g^?1 dry weight (dw) to over 45 μ g g^?1 dw. In lagoon sludge, free fatty acid content ranged from about 0.8 μ g g^?1 dw to nearly 4 μ g g^?1 dw. Coprostanol and epicoprostanol were the sterols in largest concentrations in fresh manure and lagoon sludge samples. Total sterol content ranged from approximately 0.5 μ g g^?1 dw to around 11 μ g g^?1 dw in fresh manure and from 3.5 μ g g^?1 dw to almost 9 μ g g^?1 dw in lagoon sludge. Fresh manure and lagoon sludge both had high levels of inorganic cations (e.g., Ca, Mg, Fe) capable of binding free fatty acids and forming insoluble complexes, thereby potentially reducing fatty acid biodegradation. In anaerobic lagoons, sterols are an organic fraction of sludge that are resistant to bacterial degradation. In the case of fresh manure, fatty acids could represent a potential source of energy via the manufacture of biodiesel fuel, if efficient means for their extraction and transesterification can be devised.     

Natural Products: Repellency and Toxicity of Wild Tomato Leaf Extracts to the Two-Spotted Spider Mite,Tetranychus urticae Koch

The potential of using phytochemicals from leaves of wild tomato for controlling the two-spotted spider mite, Tetranychus urticae Koch, is explored in this study as a promising alternative to the use of synthetic pesticides. Wild tomato accessions of Lycopersicon hirsutum plants that are not consumed by humans were planted under greenhouse conditions for mass production of leaves. Crude extracts from leaves of three accessions of L. hirsutum, six accessions of L. hirsutum f. glabratum, and one accession each of L. pennellii and L. pimpinellifolium were prepared in chloroform, ethanol and hexane. Two spider mite bioassays, one a measure of antibiosis and the other a measure of repellency, were utilized to determine the acaricidal performance of the crude extracts. The bioassay for antibiosis was a 6-h no-choice test. The bioassay for repellency utilized a ring bioassay. Chloroform leaf extracts of L. hirsutum f. glabratum accessions (PI-251304, PI-134417, PI-134418, and PI-126449) exhibited greatest antibiotic activity on two-spotted spider mites; the hexane extracts exhibited greatest repellency. Extracts from PI-251304, PI-126449, PI-134417, and PI-134418 were especially lethal (chloroform) or repellent (hexane). We investigated differences in chemical composition of the crude leaf extracts that may explain the observed differences in mortality and repellency among the different accessions. Major chemical compounds (α -curcumene, α -zingiberene, trans-caryophyllene, 2-undecanone, and 2-tridecanone) known to have pesticidal efficacy were detected and quantified in the crude leaf extracts using a gas chromatograph (GC) equipped with a mass spectrometer (GC/MS). Lethality of extracts was mainly associated with the presence of high concentrations of 2-tridecanone; repellency of extracts was mainly associated with the presence of trans-caryophyllene. Leaf extracts of L. hirsutum f. glabratum accessions that contain significant quantities of 2-tridecanone and/or trans-caryophyllene could be useful for managing populations of spider mites, which could reduce reliance on synthetic pesticides.     

Enrichment and Isolation of a Mixed Bacterial Culture for Complete Mineralization of Endosulfan

In the present study, we isolated three novel bacterial species, namely, Staphylococcus sp., Bacillus circulans–I, and Bacillus circulans–II, from contaminated soil collected from the premises of a pesticide manufacturing industry. Batch experiments were conducted using both mixed and pure cultures to assess their potential for the degradation of aqueous endosulfan in aerobic and facultative anaerobic condition. The influence of supplementary carbon (dextrose) source on endosulfan degradation was also examined. After four weeks of incubation, mixed bacterial culture was able to degrade 71.82 ± 0.2% and 76.04 ± 0.2% of endosulfan in aerobic and facultative anaerobic conditions, respectively, with an initial endosulfan concentration of 50 mg l^?1. Addition of dextrose to the system amplified the endosulfan degradation efficiency by 13.36 ± 0.6% in aerobic system and 12.33 ± 0.6% in facultative anaerobic system. Pure culture studies were carried out to quantify the degradation potential of these individual species. Among the three species, Staphylococcus sp. utilized more beta endosulfan compared to alpha endosulfan in facultative anaerobic system, whereas Bacillus circulans–I and Bacillus circulans–II utilized more alpha endosulfan compared to beta endosulfan in aerobic system. In any of these degradation studies no known intermediate metabolites of endosulfan were observed.     

Growth Inhibition by Nitrocompounds of Selected Uric Acid-Utilizing Microorganisms Isolated from Poultry Manure

The overall objective of this study was to evaluate the potential ability of nitrocompounds to reduce ammonia volatilization by inhibiting uric acid–utilizing microorganisms. Experiment I was conducted to evaluate the effects of nitrocompounds on the growth of uric acid–utilizing microorganisms isolated from poultry manure during six-hour incubation. There were five treatments: (1) control, (2) 50 mM nitroethane, (3) 50 mM nitroethanol, (4) 50 mM nitropropanol, and (5) 50 mM nitropropionic acid. Optical density values of nitrocompounds were significantly lower than that of control at two, four, and six hours. Plate counts of uric acid–utilizing microorganisms after six-hour incubation exhibited that nitrocompounds greatly reduced the growth of these microorganisms except for the nitroethane (P < 0.05). The nitropropanol and nitropropionic acid treatments showed significantly higher inhibitory effects compared to the nitroethanol. Experiments II and III were conducted to evaluate inhibitory effects of nitrocompounds on growth of uric acid–utilizing microorganisms compared to non-nitrocompounds such as ethanol, propanol, and propionic acid. Experiments II and III consisted of seven treatments: (1) control, (2) nitroethanol, (3) nitropropanol, (4) nitropropionic acid, (5) ethanol, (6) propanol, and (7) propionic acid. The incubation times of Experiments II and III were 6 and 24 h, respectively. The nitrocompounds were significantly more successful in inhibiting growth of uric acid–utilizing microorganisms compared to those non-nitrocompounds. These results suggest that nitrocompounds exhibit potential to reduce ammonia volatilization in poultry manure by inhibiting growth of uric acid–utilizing microorganisms.