Antibiotic resistant bacteria/genes dissemination in lacustrine sediments highly increased following cultural eutrophication of Lake Geneva (Switzerland)

This study investigates faecal indicator bacteria (FIB), multiple antibiotic resistant (MAR), and antibiotic resistance genes (ARGs), of sediment profiles from different parts of Lake Geneva (Switzerland) over the last decades. MARs consist to expose culturable Escherichia coli (EC) and Enterococcus (ENT) to mixed five antibiotics including Ampicillin, Tetracycline, Amoxicillin, Chloramphenicol and Erythromycin. Culture-independent is performed to assess the distribution of ARGs responsible for, β-lactams (blaTEM; Amoxicillin/Ampicillin), Streptomycin/Spectinomycin (aadA), Tetracycline (tet) Chloramphenicol (cmlA) and Vancomycin (van). Bacterial cultures reveal that in the sediments deposited following eutrophication of Lake Geneva in the 1970s, the percentage of MARs to five antibiotics varied from 0.12% to 4.6% and 0.016% to 11.6% of total culturable EC and ENT, respectively. In these organic-rich bacteria-contaminated sediments, the blaTEM resistant of FIB varied from 22% to 48% and 16% to 37% for EC and ENT respectively, whereas the positive PCR assays responsible for tested ARGs were observed for EC, ENT, and total DNA from all samples. The aadA resistance gene was amplified for all the sediment samples, including those not influenced by WWTP effluent water. Our results demonstrate that bacteria MARs and ARGs highly increased in the sediments contaminated with WWTP effluent following the cultural eutrophication of Lake Geneva. Hence, the human-induced changing limnological conditions highly enhanced the sediment microbial activity, and therein the spreading of antibiotic resistant bacteria and genes in this aquatic environment used to supply drinking water in a highly populated area. Furthermore, the presence of the antibiotic resistance gene aadA in all the studied samples points out a regional dissemination of this emerging contaminant in freshwater sediments since at least the late nineteenth century.     

Lethal and sublethal effects of the pyrethroid,bifenthrin, on grass shrimp (Palaemonetes pugio) and sheepshead minnow (Cyprinodon variegatus)

This study investigated the lethal and sublethal effects of the pyrethroid insecticide bifenthrin on adult and larval grass shrimp, Palaemonetes pugio, and adult sheepshead minnows, Cyprinodon variegatus. The effects were determined by conducting 96-h aqueous static renewal tests and 24-h static tests with sediment. Oxidative stress biomarkers, lipid peroxidation, glutathione, and catalase were also assessed. The 96-h aqueous LC₅₀ value for adult shrimp was 0.020 μ g/L (95% CI: 0.015–0.025 μ g/L) and for larval shrimp was 0.013 μ g/L (95% CI: 0.011–0.016 μ g/L). The 96-h aqueous LC₅₀ for adult sheepshead minnow was 19.806 μ g/L (95% CI: 11.886–47.250 μ g/L). The 24-h sediment LC₅₀ for adult shrimp was 0.339 μ g/L (95% CI: 0.291–0.381 μ g/L) and for larval shrimp was 0.210 μ g/L (95% CI: 0.096–0.393 μ g/L). The oxidative stress assays showed some increasing trends toward physiological stress with increased bifenthrin concentrations but they were largely inconclusive. Given the sensitivity of grass shrimp to this compound in laboratory bioassays, additional work will be needed to determine if these exposure levels are environmentally relevant.     

Toxicity of carbaryl,diquat dibromide,and fluoranthene,individually and in mixture,to larval grass shrimp,Palaemonetes pugio

This study examined the toxicity of two pesticides (carbaryl and diquat dibromide) and one polycyclic aromatic hydrocarbon (fluoranthene), both singly and in mixture, to grass shrimp larvae (Palaemonetes pugio). These three chemicals are all present in coastal environments and can easily enter estuarine ecosystems. Fluoranthene was the most toxic chemical with a 96-h LC₅₀ value of 32.45 μ g/L, followed by carbaryl (43.02 μ g/L) and diquat dibromide (1624 μ g/L). In the chemical mixture tests, the binary carbaryl/diquat dibromide mixture and the ternary carbaryl/diquat dibromide/fluoranthene mixture had additive results.     

人工湿地中抗生素抗性大肠杆菌和抗性基因的去除与分布

抗生素的滥用导致抗生素抗性菌和抗性基因随生活污水和养殖废水的排放在环境中肆意散播,其去除及环境行为越来越受到关注。采用K-B纸片法测定了9套不同工艺构型模拟人工湿地中大肠杆菌对7种抗生素的抗性率,并应用多重PCR检测磺胺类sul1、2、3与四环素tetA、B、C、D抗性基因,探究人工湿地对抗性菌的去除效率及抗性菌、抗性基因的分布规律。结果显示,人工湿地能有效去除污水中70%左右的抗性大肠杆菌,有效降低了细菌抗性的传播风险;共计分离出535株大肠肝菌中有378株对一种以上抗生素有抗性性,以四环素、磺胺类和氨苄西林抗性率最高,达到25%以上,其他4种抗性率较低,不足20%;2种抗性基因的检出率都在70%以上;对不同采样点大肠杆菌的抗性性及抗性基因的比较发现,各部分大肠杆菌的抗性水平、多重抗性指数(MRI)以及抗性基因(sul、tet)检出率和组合数表现出:基质≥出水>进水,推测抗性菌被湿地基质截留,在基质生物膜上发生抗性基因的重组,并释放抗性菌,提高了出水中抗性水平和抗性基因检出率。     

Toxicity of the Mosquito Control Pesticide Scourge® to Adult and Larval Grass Shrimp (Palaemonetes pugio)

Abstract

This study investigated the toxicity of various concentrations of technical resmethrin and Scourge® on adult and larval Palaemonetes pugio, a common grass shrimp species. Two types of tests were conducted for each of the resmethrin formulations using adult and larval grass shrimp life stages, a 96-h static renewal aqueous test without sediment, and a 24-h static nonrenewal aqueous test with sediment. For resmethrin, the 96-h aqueous LC₅₀ value for adult shrimp was 0.53 μg/L (95% confidence interval (CI): 0.46–0.60 μg/L), and for larval shrimp was 0.35 μg/L (95% CI: 0.28–0.42 μg/L). In the presence of sediment, technical resmethrin produced a 24-h LC₅₀ value for adult shrimp of 5.44 μg/L (95% CI: 4.52–6.55 μg/L), and for larval shrimp of 2.15 μg/L (95% CI: 1.35–3.43 μg/L). For Scourge®, the 96-h aqueous LC₅₀ for adult shrimp was 2.08 μg/L (95% CI: 1.70–2.54 μg/L), and for larval shrimp was 0.36 μg/L (95% CI: 0.24–0.55 μg/L). The 24-h sediment test yielded an LC₅₀ value of 16.12 μg/L (95% CI: 14.79–17.57 μg/L) for adult shrimp, and 14.16 μg/L (95% CI: 12.21–16.43 μg/L) for larvae. Adjusted LC₅₀ values to reflect the 18% resmethrin concentration in Scourge® are 0.37 μg/L (adult), 0.07 μg/L (larvae) for the 96-h aqueous test, and 2.90 μg/L (adult), 2.6 μg/L (larvae) for the 24-h sediment test. Larval grass shrimp were more sensitive to technical resmethrin and Scourge® than the adult life stage. The results also demonstrate that synergized resmethrin is more toxic to P. pugio than the nonsynergized form, and that the presence of sediment decreases the toxicity of both resmethrin and Scourge®     

Toxicity of the pyrethroid pesticide fenvalerate to freshwater catfish clarias gariepinus: Lethality,biochemical effects and role of dietary ascorbic acid

Static bioassays were made in the laboratory to determine lethal concentration of the pyrethroid pesticide fenvalerate [(RS)-alpha-cyano-3-phenoxybenzyl (RS)-2-(4-chlorophenyl)-3-methylbutyrate] for the freshwater catfish Clarias gariepinus and effects of sublethal concentrations of the pesticide on some biochemical parameters of the fish. For exposure periods of 24 to 96 h, LC₅₀ values of fenvalerate ranged from 5.83–4.76 μ g/L and 4.24–2.94 μ g/L, respectively for water and acetone soluble fenvalerate. Two sublethal concentrations of fenvalerate were used in the bioassays for biochemical parameters: 2.1 μ g/L for 24 h and 1.4 μ g/L for 96 h exposure, both concentrations representing 50% of LC₅₀ value of acetone soluble fenvalerate for the respective exposure period. Hepatosomatic index, liver glycogen, alkaline phosphatase of liver and ascorbic acid of blood, liver, and kidney decreased while haemoglobin (Hb) %, plasma glucose levels and acid phosphatase level of liver increased after 24 h exposure to 2.1 μ g/L fenvalerate. Longer exposure (96 h) to even a lower concentration (1.4 μ g/L) of fenvalerate resulted in reduction of all the parameters (except Hb %) tested as compared with control. Fish previously fed for 60 days with a diet supplemented by a high level of ascorbic acid (100 mg/100 g diet) could reverse most of the effects caused by 24 h exposure to 2.1 μ g/L fenvalerate. A lower level of ascorbic acid (50 mg/ 100 g diet) supplement could not influence these effects of fenvalerate. Even the higher dose of ascorbic acid supplementation (100 mg/100 g diet) could not relieve the stress parameters, except for Hb% and HSI, when the pesticide was applied at 1.4 μ g/L for a longer time period (96 h).     

Inhibitory Effects on Esterase Enzymes Buche and Ache in Rainbow Trout (Oncorhyncus mykiss) Produced by the Slow Release Insecticide Chitosan Diethyl Phosphate

The study on the toxicity of chitosan diethyl phosphate (ChDP), a controlled release insecticide, on the activities of butyrylcholinesterase (BuChE) and acetylcholinesterase (AChE) in rainbow trout exposed to this pesticide was carried out. It was found that ChDP reduced BuChE activity in O. mykiss by a factor of eight at 6 days, with high fluctuation to the end of the exposition time at 12 days. The in vitro analysis of brain AChE treated with ChDP and Phenamiphos showed that it was competitively inhibited by both organophosphates. The values obtained for K_m and V_max for the AChE-ChDP (K_m: 21.23 μ M; V_max: 43.10 μ mol/min/g) and AChE-Phenamiphos (K_m: 38.62 μ M; V_max: 38.91 μ mol/min/g) systems were relatively low compared to values of the AChE (control) system (K_m: 62.99 μ M; V_max: 63.29 μ mol/min/g). Results reported in this study confirmed that chitosan diethyl phosphate performs similarly to organophosphate pesticides, producing inhibition in cholinesterases in rainbow trout.     

Elemental uptake by edible herbs and lettuce (Latuca sativa)

The total concentration of toxic elements (aluminum, cadmium, chromium and lead) and selected macro and micro elements (iron, manganese, copper and zinc) are reported in six leafy edible vegetation species, namely lettuce, spinach, cabbage, chards and green and red types of Amaranth herbs. Although spinach and chards had greater than 125 mv of iron, both the amaranthus herbs recorded > than 320 μ g g^{? 1} dry weight. In both the spinach and chard species, the Mn and Zn levels were appreciable recording > 225 μ g g^{? 1} and 150 μ g g^{? 1} dry weight, respectively. Aluminum concentrations were (in μ g g^{? 1} dry weight) lettuce (10), cabbage (11), spinach (167), chards (65), amaranthus green (293) and amaranthus red (233). All the micro and macro elements and the toxic elements (Ni, Cr, Cd and Pb) elements analyzed, were below the recommended maximum permitted levels (RMI) in vegetables. Further the elemental uptake and distribution of the nine elements, at three growth stages of the lettuce plant grown on soil bed under controlled conditions are detailed. In the soil, except for iron (16%), greater than 33% of the other cations were in exchangeable form. Generally in the lettuce plant, roots retained much of the iron (> 224 μ g g^{? 1}) and aluminum (> 360 μ g g^{? 1}), while leaves had less than 200 μ g g^{? 1} of iron and 165 μ g g^{? 1} of Al. Although the concentrations of elements marginally decreased with growth, the lettuce leaves had significant amounts of Mn (30 μ g g^{? 1}), Zn (50 μ g g^{? 1}) and Cu (3.6 μ g g^{? 1}). Some presence of lead in leaves (2.0 μ g g^{? 1}) was noticed, but all the toxic and other elements analyzed were well below the RMI values for the vegetables.     

Manure placement depth impacts on crop yields and N retained in soil

The objective of this study was to determine the impact of manure placement depth on crop yield and N retention in soil. Experimental treatments were deep manure injection (45 cm), shallow manure injection (15 cm), and conventional fertilizer-based management with at least three replications per site. Water infiltration, and changes in soil N and P amounts were measured for up to 30 months and crop yield monitored for three seasons following initial treatment. Deep and shallow manure injections differed in soil inorganic N distributions. For example, in the manure slot the spring following application, NO₃-N in the surface 60 cm was higher (p < .01) when injected 15 cm (21.4 μ g/g) into the soil than 45 cm (11.7 μ g/g), whereas NH₄-N had opposite results with shallow injection having less (p = 0.045) NH₄-N (102 μ g/g) than deep (133 μ g/g) injection. In the fall one year after the manure was applied, NO₃-N and NH₄-N were lower (p = 0.001) in the shallow injection than the deep injection. The net impact of manure placement on total N was that deep injection had 31, 59, and 44 more kg N ha^{? 1} than the shallow injection treatment 12, 18, and 30 months after application, respectively. Deep manure injection did not impact soybean (Glycine max L.) yield, however corn (Zea mays L.) yield increased if N was limiting. The higher corn yield in the deep injected treatment was attributed to increased N use efficiency. Higher inorganic N amounts in the deep injection treatment were attributed to reduced N losses through ammonia volatilization, leaching, or denitrification. Results suggest that deep manure placement in glacial till soil may be considered a technique to increase energy, N use efficiency, and maintain surface and ground water quality. However, this technique may not work in glacial outwash soils due to the inability to inject into a rocky subsurface.     

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.     

Mixture Toxicity of the Antifouling Compound Irgarol to the Marine Phytoplankton Species Dunaliella tertiolecta

This study examined the toxicity of irgarol, individually and in binary mixtures with three other pesticides (the fungicide chlorothalonil, and the herbicides atrazine and 2,4-D), to the marine phytoplankton species Dunaliella tertiolecta. Standard 96-h static algal bioassays were used to determine pesticide effects on population growth rate. Irgarol significantly inhibited D. tertiolecta growth rate at concentrations ≥ 0.27 μ g/L. Irgarol was significantly more toxic to D. tertiolecta than the other pesticides tested (irgarol 96 h EC₅₀ = 0.7 μ g/L; chlorothalonil 96 h EC₅₀ = 64 μ g/L; atrazine 96 h EC₅₀ = 69 μ g/L; 2,4-D 96 h EC₅₀ = 45,000 μ g/L). Irgarol in mixture with chlorothalonil exhibited synergistic toxicity to D. tertiolecta, with the mixture being approximately 1.5 times more toxic than the individual compounds. Irgarol and atrazine, both triazine herbicides, were additive in mixture. The toxicity threshold of 2,4-D was much greater than typical environmental levels and would not be expected to influence irgarol toxicity. Based on these interactions, overlap of certain pesticide applications in the coastal zone may increase the toxicological risk to resident phytoplankton populations.     

The effect of pesticides and metals on acetylcholinesterase (AChE) in various tissues of blue mussel (Mytilus trossulus L.) in short-term in vivo exposures at different temperatures

The purpose of the study was to assess the impact of short-term exposure to selected toxicants as well as metal accumulation upon acetylcholinesterase (AChE) in the blue mussel, Mytilus trossulus L., in laboratory in vivo experiments. Mussels were exposed for up to 48 hours to a mixture of copper (Cu^{2 +}, 400 μ g L^?1) and cadmium (Cd^{2 +}, 200 μ g L^?1), to dichlorvos (DDVP, 100 μ g L^?1), and to carbaryl (100 μ g L^?1) at two temperatures: 5°C and 20°C. Samples were collected after 0, 6, 12, 24, and 48 hours of exposure, and AChE activity and metal concentration (where applicable) were analysed in gills, digestive gland, mantle+muscles, and the whole soft tissue. Very strong AChE inhibition was observed in response to the dichlorvos treatment, mainly in gills. Carbaryl and the metals caused a short-term inhibition effect. Considerable differences in AChE activity between the two temperatures were noticed. In particular, the metals were accumulated much faster at 20°C than at 5°C, especially in gills. No correlation between AChE activity and metal concentration was found. Gills turned out to be the optimal tissue for AChE activity analysis in short-term studies.     

Differences in response of two model estuarine crustaceans after lethal and sublethal exposures to chlorpyrifos

This study assessed the in vitro and in vivo effects of an acetylcholinesterase enzyme inhibitor (chlorpyrifos) in two estuarine crustaceans: grass shrimp (Palaemonetes pugio) and mysid (Americamysis bahia). The differences in response were quantified after lethal and sublethal exposures to chlorpyrifos and in vitro assays with chlorpyrifos-oxon. Results from the in vitro experiments indicated that the target enzyme, acetylcholinesterase (AChE), in the two species was similar in sensitivity to chlorpyrifos inhibition with IC50s of 0.98 nM and 0.89 nM for grass shrimp and mysids, respectively. In vivo experiments showed that mysids were significantly more sensitive to chlorpyrifos-induced AChE inhibition after 24 h of exposure. The in vivo EC50s for AChE inhibition were 1.23 μg L^?1 for grass shrimp and 0.027 μg L^?1 for mysids.

Median lethal concentrations (24h LC50 values) were 1.06 μg L^?1 for grass shrimp and 0.068 μg L^?1 for mysids. The results suggest that differences in the response of these two crustaceans are likely related to differences in uptake and metabolism rather than target site sensitivity.     

Fungicidal Activity of Artemisia herba alba Asso (Asteraceae)

The antifungal activity of Artemisia herba alba was found to be associated with two major volatile compounds isolated from the fresh leaves of the plant. Carvone and piperitone were isolated and identified by GC/MS, GC/IR, and NMR spectroscopy. Antifungal activity was measured against Penicillium citrinum (ATCC 10499) and Mucora rouxii (ATCC 24905). The antifungal activity (IC₅₀) of the purified compounds was estimated to be 5 μ g/ml, 2 μ g/ml against Penicillium citrinum and 7 μ g/ml, 1.5 μ g/ml against Mucora rouxii carvone and piperitone, respectively.     

Analysis and residue levels of forchlorfenuron (CPPU) in watermelons

This paper describes the application of liquid chromatography-tandem mass spectrometry (LC/MS-MS) for analysis of residues of forchlorfenuron (CPPU), a new plant growth regulator, in watermelons, after a sample preparation step based on the buffered Quick, Easy, Cheap, Effective, Rugged and Safe extraction method. Analytical determinations were carried out in a triple quadrupole system fitted with an electrospray interphase operating in the positive ionisation mode (ESI+). Three simultaneous MS-MS transitions of the quasi-molecular ion m/z 248 (precursor ion) were monitored for data adquisition (248 > 129, 248 > 155, and 248 > 248), using the transition 248 > 129 for quantitation. Recovery studies on watermelons at levels of 1–200 μ g/kg, performing five replicates at each level and using bracketing single-level calibration with matrix-matched standards for quantitation, gave forchlorfenuron mean recoveries ranging from 82 to 106% with relative standard deviations (RSD) lower than 18%. The limit of determination was established at 1 μ g/kg. The method was applied to evaluate the persistence of forchlorfenuron residues in watermelons grown in plastic greenhouses, after applying an individual spray treatment to the flower ovary at the anthesis stage (45 μ g/flower and 60 μ g/flower for cv “Extazy” and cv “reina de corazones” watermelons, respectively). One month after treatment, 20 “Extazy” watermelon units (1.5–2 kg/unit) and 20 “Reina de corazones” watermelon units (4–5 kg/unit) were collected and analyzed individually. None of the samples contained forchlorfenuron residues higher than 1 μ g/kg.     

Acute and chronic toxicity of organophosphate monocrotophos to Daphnia magna

The acute and chronic toxicity of monocrotophos (MCP), the binary joint toxicity of MCP and bifenthrin (BF), and sodium dodecyl benzene sulfonate (SDBS) to Daphnia magna (D. magna) was evaluated. The 24 h-median effective concentration (24 h-EC₅₀) and 48 h-median lethal concentration (48 h-LC₅₀) of MCP towards D. magna were 161 and 388 μ g/L, respectively. In addition, the lowest-observed effective concentration (LOEC) and non-observed effective concentration (NOEC) of MCP to D. magna were 10 and 5 μ g/L, respectively. Furthermore, the chronic value (ChV) of MCP against D. magna was 7 μ g/L and the acute chronic ratio (ACR) was 55. The number of offspring per female and the intrinsic rate of natural increase (r) were identified as the parameters that were most sensitive to MCP. In addition, toxic unit (TU) analysis was employed to evaluate the joint toxicities. The calculated TU_mix values of binary equitoxic mixtures of MCP + BF and MCP + SDBS were 1.47 and 1.63, respectively, which suggests that both equitoxic mixtures exert a limited antagonistic effect. The results of this study revealed that the toxic threshold of MCP towards D. magna is higher than its reported highest residue (4 μ g/L) in the ordinary aquatic environment, and that concurrent exposure to BF or SDBS may exert a slight antagonistic effect.     

Relationship between antibiotic residues and occurrence of resistant bacteria in Nile tilapia (Oreochromisniloticus) cultured in cage-farm

The aim of this study was to investigate the relationship between antibiotic residues found in the muscle of cage-farm-raised Nile tilapia (Oreochromisniloticus), the occurrence of resistant bacteria, and the sanitary practices adopted by farmers in Ilha Solteira reservoir, Brazil. Nine fish (three small fish, 40–200 g; three medium-sized fish, 200–500 g; and three large fish, 500–800 g) were collected from four cage farms every three months from April 2013 to January 2014. Ten antibiotic residues were determined using liquid chromatography-mass spectrometry, and bacteria were isolated and tested for antibiotic resistance to determine the multiple antibiotic resistance (MAR) index. Only three antibiotics (oxytetracycline, tetracycline, and florfenicol) were detected in the muscle of Nile tilapia, and their residues were the highest in small fish; however, the MAR index was higher in large fish. In addition, a direct positive relationship between the MAR index and the concentration of antibiotic residues in Nile tilapia was found. Overall, the adoption of prophylactic management practices improved the sanitary status of cage farms, reducing bacterial infections and hampering the development of antibiotic-resistant bacteria.     

Aquatic toxicity of cartap and cypermethrin to different life stages of Daphnia magna and Oryzias latipes

Cartap and cypermethrin, which are among the most widely used pesticides in many countries, are considered safe because of their low mammalian toxicity and their low persistence in the environment. However, recent findings of endocrine-disrupting effects and developmental neurotoxicity have raised concerns about the potential ecological impacts of these pesticides. We evaluated the aquatic toxicity of cartap [S,S′-(2-dimethylaminotrimethylene) bis(thiocarbamate), unspecified hydrochloride] and cypermethrin [(RS)-alpha-cyano-3-phenoxybenzyl-(1RS,3RS,1RS,3SR)-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane carboxylate], both individually and combined, on different life stages of the freshwater cladoceran Daphnia magna and a freshwater teleost, Japanese medaka (Oryzias latipes). The 96-hr Daphnia median effective concentrations (EC50s) for cartap and cypermethrin were 91.0 μ g/L and 0.00061 μ g/L, respectively. Rapid recovery of Daphnia was observed after short-term pulsed exposure to cartap and cypermethrin; there were no adverse effects on reproduction or survival 20 d after a 24 hr exposure to cartap up to 1240 μ g/L and cypermethrin up to 1.9 μ g/L. Chronic continuous exposure (for 21 d) of 7-d-old Daphnia to cypermethrin significantly reduced the intrinsic population growth rate in a concentration-dependent manner. However, because the intrinsic population growth rates were all above zero, populations did not decrease even at the highest experimental concentration of 200 ng/L. Exposure of Daphnia neonates (< 24 hr old) to cypermethrin for 21 d caused significant, sub-lethal reproduction-related problems, such as increased time to first brood, reduced brood size, and reduced total brood number, at 0.0002, 0.002, and 0.2 ng/L cypermethrin, but the intrinsic population growth rate was not significantly affected. Oryzias latipes was relatively more resistant to both pesticides. In particular, embryos appeared to be more resistant than juveniles or adults, which may be partly due to the protective role of the chorion. The incidence of larval fish deformity was significantly higher after a 96 hr exposure to as low as 250 μ g/L of cartap or 40 μ g/L of cypermethrin. The mixture of both compounds showed no synergistic toxicity. The extremely high acute-to-chronic ratio suggests that the standard acute lethal toxicity assessment might not reflect the true environmental hazards of these frequently used pesticides. Ecological hazard assessments of long-term low dose or pulsed exposures to cartap and cypermethrin may reveal more realistic consequences of these compounds in surface water.     

Effects of T-2 toxin on histopathology,fatty acid and water distribution of shrimp (Litopenaeus vannamei) muscle

T-2 toxin (T-2), one of the naturally occurring mycotoxins, often accumulates in aquatic animals from contaminated feed. Shrimp (n?=?30 per group) were fed with different concentrations (0, 0.5, 1.5, 4.5 and 13.5?mg kg^?1) of T-2 for 20?days. Changes in histopathology, fatty acid and water distribution of shrimp muscle were analyzed. Histopathology of shrimp muscle showed dose-dependent marked degenerative and necrotic changes on exposure to dietary T-2. The T-2 significantly (P?<?0.05) affected the muscle fatty acid composition. ∑SFA, ∑MUFA and ∑PUFA initially decreased and then increased slowly in the high-dosed groups. C16:0, C18:1n-9 and C18:2n-6 were the main saturated fatty acid (SFA), monounsaturated fatty acid (MUFA) and polyunsaturated fatty acid (PUFA), respectively. Also, T-2 significantly affected water distribution in shrimp muscle. High doses of T-2 reduced free water content, resulting in a reduction in the water holding capacity and hence changes to the shrimp muscle quality. Collectively, these results illustrated that T-2 significantly affects the fatty acid and water distribution, and also muscle histopathology, all of which would result in a reduction in the quality and nutritional value of shrimp.     

Ciprofloxacin-sensitive and ciprofloxacin-resistant Campylobacter jejuni are equally susceptible to natural orange oil-based antimicrobials

A total of 10 ciprofloxacin-sensitive (ciprofloxacin minimum inhibitory concentration, MIC < 0.5 μ g/ml) and 10 ciprofloxacin-resistant (MIC 16 to 32 μ g/ml) presumptive C. jejuni were further characterized and evaluated for their inhibition by natural orange oil fractions. Partial species identification was performed by using a hippuricase gene-based polymerase chain reaction (PCR) assay. One of the isolates appeared to be atypical and failed to hydrolyze hippurate. Of the ciprofloxacin-resistant C. jejuni isolates tested, six were found to have their quinolone resistance determined by a C → T mutation in codon 86 of gyrA. Both groups of ciprofloxacin-sensitive and –resistant C. jejuni isolates were most susceptible to cold-pressed terpeneless Valencia orange oil (C4) which yielded inhibition zones from 44.0 ± 1.4 to 80 ± 0.0 mm. Less inhibitory responses were recorded for 5-fold concentrated Valencia orange oil (C3) and distilled d-limonene (C7) which exerted similar effects on both ciprofloxacin-sensitive and –resistant C. jejuni isolates. In general, ciprofloxacin-resistant and –sensitive C. jejuni isolates were equally susceptible to the respective orange oil fractions.