Hao-Chang Su

GROWTH-INHIBITING EFFECTS OF 12 ANTIBACTERIAL AGENTS AND THEIR MIXTURES ON THE FRESHWATER MICROALGA PSEUDOKIRCHNERIELLA SUBCAPITATA

The growth-inhibiting and binary joint effects of 12 antibacterial agents on the freshwater green alga Pseudokirchneriella subcapitata (Korschikov) Hindák were investigated over 72-h exposures. The toxicity values (the median inhibitory concentration value, in micromoles) in decreasing order of sensitivity were triclosan (0.0018)>triclocarban (0.054)>roxithromycin (0.056)>clarithromycin (0.062)>tylosin (0.20)>tetracycline (2.25)>chlortetracycline (3.49)>norfloxacin (5.64)>sulfamethoxazole (7.50)>ciprofloxacin (20.22)>sulfamethazine (31.26)>trimethoprim (137.78). Several of these antibacterial compounds would be toxic at the micrograms per liter concentrations reported in surface waters and sewage effluents. Simple additive effects were observed in binary mixtures of sulfonamides, and most tylosin, triclosan, or triclocarban combinations. Potentially synergistic effects were observed in binary mixtures of the same class, such as macrolides, tetracyclines, and fluoroquinolones, as well as in some combined drugs, such as trimethoprim and sulfonamides or tylosin and tetracyclines. Potentially antagonistic effects were only observed between tylosin and triclocarban, triclosan and norfloxacin, and triclocarban and norfloxacin. Although present at low concentrations in the aquatic environment, mixtures of these antibacterial agents can potentially affect algal growth in freshwater systems due to their combined action.

Simultaneous determination of four classes of antibiotics in sediments of the Pearl Rivers using RRLC-MS/MS

A simultaneous extraction and analysis method was developed for four classes of antibiotics: sulfonamides (SAs), macrolides (MLs), fluoroquinolones (FQs) and tetracyclines (TCs) in sediment. The antibiotics in the sediment were extracted by using an optimized mixed solution (citric buffer: acetonitrile, 50:50 v/v), and analyzed by rapid resolution liquid chromatography-tandem mass spectrometry (RRLC-MS/MS) equipped with electrospray ionization source in the positive mode. The recoveries for the antibiotics in the spiked sediment were: 66.3-123.5%, 48.2-72.0%, 61.8-160.4% and 60.6-151.9% for the SAs, TCs, FQs and MLs, respectively. The LOD ranged from 0.08 microg/kg to 4.2 microg/kg and LOQ from 0.26 microg/kg (erythromycin-H(2)O) to 14.01 microg/kg (norfloxacin). The developed analytical method was applied in the determination of the antibiotics in sediment samples collected from the Pearl Rivers, China. The antibiotics were found at the levels of microg/kg in the sediments of the three rivers. Ofloxacin was found to have the highest concentration of 1560 microg/kg in sediment. However, no significant seasonal variation could be found for the antibiotics in the sediments between wet season and dry season.

Simultaneous determination and assessment of 4-nonylphenol, bisphenol a and triclosan in tap water, bottled water and baby bottles

This study investigated the levels of 4-nonylphenol (4-NP), bisphenol A (BPA) and triclosan (TCS) in bottled water and tap water in Guangzhou and release of these chemicals from baby bottles using gas chromatography-mass spectrometry with negative chemical ionization. Results show that 4-NP was present in all the bottled water while 17 out of 21 contained BPA and 18 out of 21 contained TCS. Their concentrations in bottled water ranged from 108 to 298 ng/L, 17.6 to 324 ng/L and 0.6 to 9.7 ng/L, respectively. Five of the tap water samples from six drinking water plants were found to contain 4-NP and BPA both in June and December, while TCS was detected in the same five plants only in June. The highest concentrations in tap water for 4-NP, BPA and TCS were 1987, 317 and 14.5ng/L, respectively. Daily intakes of 4-NP, BPA and TCS of adults by drinking 2L of tap water were estimated to be 1410, 148 and 10 ng/day, respectively. BPA was found to be released within 24h from four brands of baby bottles at room temperature (24 degrees C), 40 degrees C and 100 degrees C. Increased temperature led to higher release of BPA from the baby bottles. Estimated daily intakes of 4-NP, BPA and TCS for infants were 705, 1340 and 5 ng/day, respectively, by drinking 1L of tap water from a baby bottle at 40 degrees C. This study showed that the exposure to the three compounds from drinking water is unlikely to pose a health risk.

Assessing estrogenic activity in surface water and sediment of the Liao River system in northeast China using combined chemical and biological tools.

This paper investigated some selected estrogenic compounds (4-t-octylphenol: 4-t-OP; 4-nonylphenols: 4-NP; bisphenol-A: BPA; diethylstilbestrol: DES; estrone: E1; 17β-estradiol: E2; 17α-Ethinylestradiol: EE2; triclosan: TCS) and estrogenicity in the Liao River system using the combined chemical and in vitro yeast screen bioassay and assessed their ecological risks to aquatic organisms. The estrogenic compounds 4-t-OP, 4-NP, BPA, E1, E2 and TCS were detected in most of the samples, with their concentrations up to 52.1 2065.7, 755.6, 55.8, 7.4 and 81.3 ng/L in water, and up to 8.6, 558.4, 33.8, 7.9, <LOQ and 33.9 ng/g in sediment, respectively. However, DES and EE2 were not detected in the Liao River. The estrogen equivalents (EEQ) of the water and sediment samples were also measured by the bioassay. High estrogenic risks to aquatic organisms were found in the river sections of metropolitan areas and the lower reach of the river system.

4-Nonylphenol, bisphenol-A and triclosan levels in human urine of children and students in China, and the effects of drinking these bottled materials on the levels.

4-Nonylphenol (4-NP), bisphenol-A (BPA) and triclosan (TCS) are three industrial chemicals used widely in daily products. This study investigated 4-NP, BPA and TCS levels in urine samples of 287 children and students aged from 3 to 24 years old in Guangzhou, China. Total (free and conjugated) amounts of 4-NP, BPA and TCS in the urine samples were detected using gas chromatography-mass spectrometry with negative chemical ionization. The detection rates of 4-NP, BPA and TCS were 100%, 100% and 93% respectively, given the detection limits of 3.8, 0.5 and 0.9 ng/L respectively. Data for 4-NP, BPA and TCS were presented in both creatinine-adjusted (microgram per gram creatinine) and unadjusted (microgram per liter) urinary concentrations. The geometric mean (GM) concentrations of urinary 4-NP, BPA and TCS were 15.92 μg/g creatinine (17.40 μg/L), 2.75 μg/g creatinine (3.00 μg/L) and 3.55 μg/g creatinine (3.77 μg/L) respectively. Multiple regression models considering age, gender, preferred drinking bottle and log-transformed creatinine were used to calculate the adjusted least square geometric mean (LSGM). Among these subjects, the females had higher LSGM concentrations of 4-NP, BPA and TCS than the males; and the only statistically significant difference was found for the LSGM concentrations of triclosan (p=0.031). Participants who reported to use ceramic cups more frequently had significantly lower LSGM concentrations of BPA than those who used plastic cups (p=0.037). Meanwhile, a three-week test of using polycarbonate bottles and ceramic cups to drink bottled water and boiled tap-water was carried out among 12 graduate students of 25 years old. The GM concentrations of urinary BPA at the end of the first week after using ceramic cups to drink bottled water were 7.16 μg/g creatinine, then decreased significantly to 3.49 μg/g creatinine after the second week of using ceramic cups to drink boiled tap-water (p<0.05), and finally increased to 4.15 μg/g creatinine after the third week of using polycarbonate bottles in drinking boiled tap-water. The results indicate that in daily life the use of polycarbonate bottles or drinking of bottled water is likely to increase the ingestion of BPA, resulting in an increase in urinary BPA levels.

Detection of antibiotic resistance and tetracycline resistance genes in Enterobacteriaceae isolated from the Pearl rivers in South China.

This study investigated antibiotic resistance profiles and tetracycline resistance genes in Enterobacteriaceae family isolates from the Pearl rivers. The Enterobacteriaceae isolates were tested for susceptibility to seven antibiotics ampicillin, chloramphenicol, ciprofloxacin, levofloxacin, sulphamethoxazole/trimethoprim, tetracycline and trimethoprim. In Liuxi reservoir, with an exception to ampicillin resistant strains (11%) no other antibiotic resistance bacterial strains were detected. However, multiple drug resistance in bacterial isolates from the other sites of Pearl rivers was observed which is possibly due to sewage discharge and input from other anthropogenic sources along the rivers. Four tetracycline resistance genes tet A, tet B, tet C and tet D were detected in the isolates from the rivers. The genes tet A and tet B were widely detected with the detection frequencies of 43% and 40% respectively. Ciprofloxacin and levofloxacin resistant enteric bacteria were also isolated from the pig and duck manures which suggest a wider distribution of human specific drugs in the environment. This investigation provided a baseline data on antibiotic resistance profiles and tetracycline resistance genes in the Pearl rivers delta.

Spatial and seasonal distribution of selected antibiotics in surface waters of the Pearl Rivers, China

The distribution and occurrence of 15 antibiotics in surface water of the Pearl River System (Liuxi River, Shijing River and Zhujiang River) and effluents of four wastewater treatment plants (WWTPs) were investigated in two sampling events representing wet season and dry season by using rapid resolution liquid chromatography–electrospray tandem mass spectrometry (RRLC-MS/MS) in positive ionization mode. Only eight antibiotics (sulfadiazine, sulfapyridine, sulfamethazine, sulfamethoxazole, trimethoprim, roxithromycin, erythromycin-H2O and norfloxacin) were detected in the water samples of the three rivers and the effluents. The detection frequencies and levels of antibiotics in the dry season were higher than those in the wet season. This could be attributed to the dilution effects in the wet season and relatively lower temperature in the dry season under which antibiotics could persist for a longer period. The levels of the detected antibiotics in different sites are generally in a decreasing order as follows: Shijing River ⩾WWTP effluent ⩾Zhujiang River ⩾ Liuxi River. Risk assessment based on the calculated risk quotients showed that only erythromycin-H2O and roxithromycin detected in the Pearl Rivers might have adverse effects on aquatic organisms.

Evaluation of triclosan and triclocarban at river basin scale using monitoring and modeling tools: implications for controlling of urban domestic sewage discharge.

Triclosan (TCS) and triclocarban (TCC) are two commonly used personal care products. They may enter into aquatic environments after consumption and pose potential risks to aquatic organisms. We investigated the occurrence and fate of TCS and TCC in five large rivers (the Liao River, Hai River, Yellow River, Zhujiang River and Dongjiang River) in China, and compared the monitoring data with the predicted results from Level III fugacity modeling. TCS and TCC were detected in the five large rivers with the detection frequencies of 100% or close to 100% in surface water and sediments of almost every river. TCS and TCC were found at concentrations of up to 478 ng/L and 338 ng/L in surface water, and up to 1329 ng/g and 2723 ng/g in sediments. Cluster analysis indicated that the sites with higher concentrations were usually located in or near urban area. Meanwhile, principal component analysis also suggested that the mass inventories of TCS and TCC in water and sediment were significantly influenced by the factors such as the total or untreated urban domestic sewage discharge at river basin scale. The concentrations and mass inventories from the fugacity modeling were found at the same order of magnitude with the measured values, suggesting that the fugacity modeling can provide a useful tool for evaluating the fate of TCS and TCC in riverine environments. Both monitoring and modeling results indicated that the majority of mass inventories of TCS and TCC were stored into sediment, which could be a potential pollution source for river water. The wide presence of TCS and TCC in these large rivers of China implies that better controlling of urban domestic sewage discharge is needed.

Dissemination of antibiotic resistance genes in representative broiler feedlots environments: identification of indicator ARGs and correlations with environmental variables.

Livestock operations are known to harbor elevated levels of antibiotic resistance genes (ARGs) that may pose a threat to public health. Broiler feedlots may represent an important source of ARGs in the environment. However, the prevalence and dissemination mechanisms of various types of ARGs in the environment of broiler feedlots have not previously been identified. We examined the occurrence, abundance and variation of ARGs conferring resistance to chloramphenicols, sulfonamides and tetracyclines in the environments of two representative types of broiler feedlots (free range and indoor) by quantitative PCR, and assessed their dissemination mechanisms. The results showed the prevalence of various types of ARGs in the environmental samples of the broiler feedlots including manure/litter, soil, sediment, and water samples, with the first report of five chloramphenicol resistance genes (cmlA, floR, fexA, cfr, and fexB) in broiler feedlots. Overall, chloramphenicol resistance genes and sulfonamides sul genes were more abundant than tetracyclines tet genes. The ARG abundances in the samples from indoor boiler feedlots were generally different to the free range feedlots, suggesting the importance of feeding operations in ARG dissemination. Pearson correlation analysis showed significant correlations between ARGs and mobile genetic element genes (int1 and int2), and between the different classes of ARGs themselves, revealing the roles of horizontal gene transfer and coselection for ARG dissemination in the environment. Further regression analysis revealed that fexA, sul1 and tetW could be reliable indicator genes to surrogate anthropogenic sources of ARGs in boiler feedlots (correlations of fexA, sul1 and tetW to all ARGs: R = 0.95, 0.96 and 0.86, p < 0.01). Meanwhile, significant correlations were also identified between indicator ARGs and their corresponding antibiotics. In addition, some ARGs were significantly correlated with typical metals (e.g., Cu, Zn, and As with fexA, fexB, cfr, sul1, tetW, tetO, tetS: R = 0.52-0.71) and some environmental parameters (e.g., TOC, TN, TP, NH3-N with fexA, fexB, cfr, sul1, tetW, tetO, tetQ, tetS: R = 0.53-0.87) (p < 0.01). Further redundancy analysis demonstrated that the distribution and transportation of ARGs from the boiler feedlots to the receiving environments were correlated with environmental variables. The findings highlight the contribution of some chemicals such as antibiotics and metals to the development of ARGs in broiler feedlots environments; and the observed ARG dissemination mechanism in the broiler feedlots facilitates the development of effective mitigation measures.

Tissue-specific bioaccumulation of human and veterinary antibiotics in bile, plasma, liver and muscle tissues of wild fish from a highly urbanized region

We investigated the bioaccumulation of antibiotics in bile, plasma, liver and muscle tissues of wild fish from four rivers in the Pearl River Delta region. In total, 12 antibiotics were present in at least one type of fish tissues from nine wild fish species in the four rivers. The mean values of log bioaccumulation factors (log BAFs) for the detected antibiotics in fish bile, plasma, liver, and muscle tissues were at the range of 2.06-4.08, 1.85-3.47, 1.41-3.51, and 0.48-2.70, respectively. As the digestion tissues, fish bile, plasma, and liver showed strong bioaccumulation ability for some antibiotics, indicating a different bioaccumulation pattern from hydrophobic organic contaminants. Human health risk assessment based on potential fish consumption indicates that these antibiotics do not appear to pose an appreciable risk to human health. To the best of our knowledge, this is first report of bioaccumulation patterns of antibiotics in wild fish bile and plasma.