Faecal indicator organism concentrations and catchment export coefficients in the UK

Characterisation of faecal indicator organism (FIO) concentrations and export coefficients for catchments with particular combinations of land use and under specific climatic regimes is critical in developing models to predict daily loads and apportion sources of the microbial parameters used to regulate water quality. Accordingly, this paper presents a synthesis of FIO concentration and export coefficient data for the summer bathing season, with some comparative winter data, for 205 river/stream sampling points widely distributed across mainland UK. In terms of both geometric mean (GM) FIO concentrations and export coefficients (expressed as cfu km(-2) h(-1)), the results reveal (1) statistically significant elevations at high flow compared with base flow, with concentrations typically increasing by more than an order of magnitude and export coefficients by about two orders; (2) significantly higher values in summer than in winter under high-flow conditions; and (3) extremely wide variability between the catchments (e.g. four orders of magnitude range for GM faecal coliform concentrations), which closely reflects land use-with urban areas and improved pastures identified as key FIO sources. Generally, these two most polluting land uses are concentrated in lowland areas where runoff (m3 km(-2) h(-1)) is low compared with upland areas, which in the UK are dominated by rough grazing and forestry. Consequently, contrasts in export coefficients between land use types are less than for FIO concentrations. The GMs reported for most land use categories are based on 13 sites and exhibit quite narrow confidence intervals. They may therefore be applied with some confidence to other catchments in the UK and similar geographical regions elsewhere. Examples are presented to illustrate how the results can be used to estimate daily summer base- and high-flow FIO loads for catchments with different land use types, and to assess the likely effectiveness of certain strategies for reducing FIO pollutant loadings in areas with extensive areas of lowland improved pasture.     

Antimicrobial resistance and virulence genes of Arcobacter isolates recovered from edible bivalve molluscs

The goals of this study were to assess the prevalence, antimicrobial susceptibility and the presence of virulence genes of arcobacters recovered from edible bivalve molluscs. A total of 106 samples (21 clams, 18 mussels, 20 oysters, 20 razor clams, 11 scallops and 16 surf clams) were analysed by culture between 2010 and 2013. The obtained colonies were identified by multiplex PCR and PCR-RFLP and genotyped by ERIC-PCR. Furthermore, nine putative virulence genes (cadF, ciaB, cjl349, irgA, hecA, hecB, mviN, pldA and tlyA) were assessed by PCR and the antimicrobial resistance was tested by the dilution agar method. The global prevalence was 40.5%, with the highest value in surf clams (87.5%) followed by razor clams (65.0%), mussels (33.3%), clams (23.8%), scallops (18.0%) and oysters (15.0%). The most commonly found species was Arcobacter butzleri (62%) followed by Arcobacter cryaerophilus (21%), Arcobacter skirrowii (16%) and Arcobacter defluvii (1%). A high resistance was found to nalidixic acid and ampicillin, while the predominant detected virulence genes were mviN (83.8%), ciaB (82.8%) and tlyA (72.7%). Our results indicate a high prevalence of arcobacters in shellfish and the pathogenic potential of the recovered isolates suggests that this type of food could be a plausible transmission route of virulent strains to humans.     

Bio-accumulation of metals on reef associated organisms of Lakshadweep Archipelago

Tissue samples of marine organisms from the coastal waters of Agatti Island were subjected to analysis of metals (cadmium, cobalt, copper, iron, magnesium, manganese, nickel, lead and zinc), using Inductively Coupled Plasma Optical Emission Spectrometer (ICP-OES) for the assessment of the present condition of the Island ecosystem and compiling the baseline data for future monitoring, with respect of metal accumulation of marine organisms. Tissue samples of fish, shellfish and seaweed revealed that the metals have different levels of accumulation viz. Cd = 0.08-0.14, Co = 0.01-0.02, Cu = 0.16-0.98, Fe = 3.19-5.3, Mg = 86.73-152.45, Mn = 0.17-0.55, Ni = 0.06-0.26, Pb = 0.11-0.46 and Zn = 3.26-14.2 μg g⁻¹ dry wt. Metal concentrations were more in shellfish and less in finfish. Concentrations of toxic metals such as Cd, Co, Ni and Pb were well below the permissible limits proposed by the World Health Organization.     

PIXE analysis of biological bodies in Uranouchi bay

We have investigated shellfish called Japanese littleneck clam (Ruditapes philippinarum) which were collected from Uranouchi bay (Kochi, Japan) for analysis of elemental concentration by Particle Induced X-ray Emission (PIXE). A 4 MeV He⁺⁺ ion beam was used to analyze shellfish in the ion beam laboratory of Kochi University of Technology (Japan). In the present study, shellfish have been chosen as a representative of the biological bodies in the Uranouchi bay. The concentration of heavy metals in shellfish has been quantified and the results are compared with those obtained by analyzing Market shellfish. Analyzing shellfish collected from three different places in the bay Cu, Zn, Br, Sr and Zr are detected as heavy metals. Our results suggest that in comparison with Market shellfish the Uranouchi bay shellfish contain more heavy metals.     

Removal of Escherichia coli, Enterococcus fecalis, coliphage MS2, poliovirus, and hepatitis A virus from oysters (Crassostrea virginica) and hard shell clams (Mercinaria mercinaria) by depuration

Filter-feeding bivalve mollusks (shellfish) can bioaccumulate pathogenic microorganisms in up to 1000-fold higher levels than overlying waters, and therefore disease risks are associated with consuming raw or partially cooked shellfish. Many of these shellfish-borne diseases are due to enteric bacteria and viruses associated with fecal contamination. To control shellfish-borne diseases, guidelines for shellfish harvest waters and shellfish meat have been devised, which include cleansing of contaminated shellfish by depuration in controlled systems, heat pasteurization, or relay to clean waters. This study examines the depuration of oysters (Crassostrea virginica) and hard shell clams (Mercinaria mercinaria) in a flow-through depuration system under variable temperature (12 °C, 18 °C, and 25 °C), salinity (8 ppt, 18 ppt, and 28 ppt), turbidity (< 1 NTU, 10 NTU, and 20 NTU), pH (pH 7 and pH 8), and algae conditions (0 cells/mL and 50,000 cells/mL), with constant dissolved oxygen (5-7 mg/L). Oysters and hard shell clams were artificially contaminated with enteric microorganisms: Escherichia coli, Enterococcus faecalis, coliphage MS2, Poliovirus type-1 and Hepatitis A virus HM-175 (HAV), then depurated in 5-day trials with daily sampling. In oysters, optimizing environmental parameters of water temperature improved E. coli, MS2, poliovirus and HAV depuration, and optimized salinity improved E. coli, E. faecalis, and MS2 depuration rates. In hard shell clams, salinity improved E. coli and E. faecalis depuration rates. Adjusting turbidity, pH or algae did not improve microorganism depuration in either oysters or hard shell clams, with the exception of turbidity on E. faecalis in hard shell clams. Microorganism depuration rates in oysters from greatest to least were: MS2 > E. coli > E. faecalis > poliovirus > HAV, and in clams depuration rates from greatest to least were: E. coli > E. faecalis > HAV > MS2 > poliovirus. Because E. coli and E. faecalis were removed at faster rates than HAV and poliovirus, these fecal bacteria appear to be poor process indicators of the virological quality of depurated oysters and hard shell clams.     

免疫亲和柱净化-高效液相色谱-三重四极杆质谱法测定双壳类水产中软骨藻酸

目的 采用免疫亲和柱净化双壳类水产中的软骨藻酸,建立高效液相色谱-三重四级杆质谱串联（LC-MS/MS）方法检测双壳类水产中的软骨藻酸,为水产品中的软骨毒素检测提供方法依据。方法 选用InfinityLab Poroshell 120 EC-C18色谱柱,以10 mmol/L甲酸铵-0.1%甲酸-甲醇为流动相,采用梯度洗脱进行分离。样品用80%甲醇水提取,上清液加入PBS缓冲液后经免疫亲和柱净化,将洗脱液氮吹至干定容后上机测定。多重态反应监测（MRM）方式检测。结果 软骨藻酸的线性范围为20.0~200 ng/ml,检出限为0.002 μg/g,回收率在75.2% ~ 85.7%之间。结论 本方法特异性强、提取效果好、无基质抑制效应,适用于双壳类水产中软骨藻酸的痕量检测。     

Occurrence and distribution of Vibrio parahaemolyticus in retail oysters in Sao Paulo State, Brazil

Vibrio parahaemolyticus is a potentially pathogenic bacterium that occurs naturally in estuarine environments worldwide, and is often associated with gastroenteritis in humans following consumption of raw bivalve mollusks, especially raw oysters. The occurrence of total and pathogenic V. parahaemolyticus in 74 samples of raw oysters collected in restaurants, supermarkets, groceries and beach huts in Sao Paulo State, was monitored between February 2006 and January 2007. Enumeration of V. parahaemolyticus was performed according to the most probable number (MPN) procedure. Five to ten typical colonies were selected from thiosulfate-citrate-bile salts-sucrose (TCBS) agar plates for confirmation by the presence of the species-specific gene tlh and the virulence genes tdh and trh by multiplex PCR. V. parahaemolyticus was detected in 100% of samples. The densities of total V. parahaemolyticus varied from 1.78 to 6.04 log₁₀ (MPN/g), with higher densities being detected in fall and summer, and lower densities in winter (P < 0.05). There was no statistical difference among densities of V parahaemolyticus regarding the site of collection. None of the 1943 V. parahaemolyticus isolates contained tdh and/or trh. These data provide information for the assessment of exposure to V. parahaemolyticus in oysters consumed in Sao Paulo, State, Brazil.     

多环芳烃蒽在两种贝类中的检测及生物降解研究

目的比较月亮贝(Neolamprologuscalliurus)和竹蛏(Solenstrictu)对多环芳烃蒽的富集作用,筛选获得高效蒽降解菌株。方法采用气相色谱-质谱法(gas chromatohraphy-mass spectromety,GC-MS)检测两种贝类中蒽的含量,采用高效液相色谱法(high performance liquid chromatography,HPLC)比较6种菌株和上述菌株等体积混合得到的菌群对蒽的降解率,并对高效菌株或菌群的降解条件进行优化。结果两种贝类均能够对蒽进行富集,在0.5和5μg/L蒽处理条件下,月亮贝对蒽的富集比竹蛏高2.75和3.34倍。6种菌株对蒽的降解率可达92.11%~95.90%,而混合菌群对蒽的降解率最高可达97.17%。菌群生物降解蒽的最佳条件为pH 6、温度27℃、降解时间15 d。另外,添加30 mg/L Mg;促进了混合菌群对蒽的降解。结论月亮贝对蒽的富集高于竹蛏,混合菌群对蒽的降解率最高,添加Mg;可以促进蒽的生物降解。     

海产贝类食品安全预警研究

为防控海贝类食源性污染、食物中毒的发生,通过开展贝类生物海洋污染本底调查、进行贝类食用安全性分析评价、在海区布设预警站点、实施贝毒素预警监测、建设海贝类食品安全示范基地等方式,探索出一条适合当地沿海实际的贝类食品安全预警保障模式,并对贝类食品依法行政、贝类的清洁生产与健康养殖等提出了见解。     

Changes in the ultrastructure and texture of prawn muscle (Macrobrachuim rosenbergii) during cold storage

Changes in the ultrastructure and proteolytic activity of prawn muscle were determined during storage at 5 °C, in order to better understand changes in physical and sensory texture measurements. Progressive deterioration of myofibril structure was observed during refrigeration of prawn for 14 d. The loss of density and order in Z-line alignment was first detected after 3 d of storage. Progressive disruption of Z-line, I-bands and M-lines was observed after 4-6 d of storage. Muscle degradation included pronounced disruption of the mitochondria as revealed by swollen cristae, loss of cristae material, and membrane breakage. Along with ultrastructural changes, decreased shear force values and mean textures scores were measured. An initial shear force value of 18.21 N/g decreased to 14.50, 12.46, and 10.79 N/g on days 3, 6, and 14, respectively. Mean texture scores indicated that prawn muscle maintained firm texture during 0-3 d of storage, and became soft during 4-6 d of storage. After 6 d of storage, the prawn texture was very soft. Increased deterioration of the muscle ultrastructure coincided with the increase of proteolytic microorganisms and salt soluble muscle proteins found by Sodium dodecyl sulfate-polyacrylamide gel electrophoresis. It is concluded that the weakening of the ultrastructure is related to proteolytic activity and results in a more soft texture in prawns.