Histochemical demonstration of mercury in the intestine,nephridia and epidermis of the marine polychaete Nereis virens exposed to inorganic mercury

Marine polychaetes Nereis virens (Sars) were exposed to 9 ppb Hg as mercuric chloride in the aquarium water. Concentrations of mercury in the water and in the worms were monitored using radiolabelled mercury. After 11 d, the average mercury concentration in the nereids was 8.41 ppm Hg and the bioconcentration factor was calculated to be 930. Accumulations of mercury in the tissues were made visible for light and electron microscopy by autometallographic silver enhancement. Mercury was demonstrated in the intestine, nephridia, epidermis and cuticula. In the intestine, mercury was found to be located predominantly in the apical part of the epithelial cells. In the nephridia, mercury deposits were apparent in the peritoneal cells of the nephridial tubules. Silver-enhanced mercury was also observed at the epicuticula, in the collagen fibres of the endocuticula and in the epidermal basal cells. Intracellularly, the observed mercury accumulations were localised predominantly in lysosomes. Extracellularly, mercury was observed in the basal laminae of the intestine and the epidermis, and also in the intestinal peritrophic membrane. How N. virens copes with mercury toxicity is discussed.     

High recovery of culturable bacteria from the surfaces of marine algae

The culturability of heterotrophic marine bacteria obtained from the surfaces of two species of marine algae (Lobophora variegata andHalimeda copiosa) was assessed by comparing total DAPI-stained cell counts to colony-forming bacterial counts on two agar media. The colony-forming bacterial counts on a low-nutrient medium (LN) consisting of seawater and agar were significantly greater for both algal species than counts obtained on a high-nutrient medium (HN) similar in composition to that typically used for the isolation of heterotrophic marine bacteria. On average, 14 and 58%, respectively, of the total bacteria fromL. variegata andH. copiosa were culturable on LN. These recovery rates far exceed those typically reported for marine bacteria. Of 119 LN strains obtained in pure culture, 55% failed to grow on HN. The yeast extract component of HN (1.5 gl^-1) was responsible for the majority of the observed inhibition, suggesting that this nutrient can be highly toxic to marine bacteria. Eighty-nine percent of the strains inhibited by HN were capable of growth when the nutrients in this medium were diluted by a factor of 100 with seawater. Of 66 epiphytic strains, 30 (45%) initially inhibited by HN showed the ability to adapt to this medium after a period of laboratory handling. The initial inability of low-nutrient-adapted bacteria to grow on high-nutrient media may be due to nutrient shock. The results presented here indicate that the culturability of specific populations of marine bacteria can be dramatically improved by the use of low-nutrient media. Further, the importance of developing new medium formulations that eliminate traditional nutrients, some of which are clearly toxic to bacteria, is demonstrated.     

A comparison of amniotic fluid alpha-fetoprotein and acetylcholinesterase in the prenatal diagnosis of open neural tube defects and anterior abdominal wall defects

Amniotic fluid samples received for routine prenatal diagnosis of open neural tube defects were used for a study to compare amniotic fluid acetylcholinesterase (AChE) determination using a monoclonal antibody (4F19) enzyme antigen immunoassay and amniotic fluid alpha-fetoprotein (AFP) measurement as diagnostic tests for open neural tube defects. The study was based on 9964 women with singleton pregnancies and known outcome (including 6 with anencephaly and 18 with open spina bifida) having an amniocentesis at 14–23 weeks of gestation. The AChE immunoassay yielded detection rates for anencephaly of 100 per cent (95 per cent confidence interval (CI) 54·07–100 per cent), for open spina bifida of 100 per cent (95 per cent CI 81·47–100 per cent), for anterior abdominal wall defects of 20 per cent (95 per cent CI 0-51-71-64 per cent), and a false-positive rate of 0·22 percent (95 per cent CI 0·14–0·34 per cent) excluding anencephaly, open spina bifida, and anterior abdominal wall defects. For similar detection rates the false-positive rate of the AFP test was significantly higher, 0·74 per cent (95 per cent CI 0·58–0·94 per cent). On the basis of these findings, it is recommended that the technically simple AChE immunoassay should be used on all samples; the AFP test should only be used on the 0·5 per cent of the samples with concentrations of AChE activity ⩾ 8·5 nkat/1 for clear samples and blood-stained samples becoming clear after centrifugation, and ⩾ 25·0 nkat/1 for blood-stained samples that are discoloured after centrifugation; an AFP cut-off level of 2·0 MOM is recommended for this policy. Thereby, the detection rates for anencephaly, open spina bifida, and anterior abdominal wall defects would be 100, 100, and 20 per cent, respectively (95 per cent CIs 54·07–100, 81·47–100, and 0·51–71·64 per cent, respectively), and the false-positive rate would be 0·08 per cent (95 per cent CI 0·03–0·16 per cent) (excluding anencephaly, open spina bifida, and anterior abdominal wall defects).     

Fetal tissue involvement in the late infantile type of neuronal ceroid lipofuscinosis

Prenatal diagnosis in a pregnancy at risk for late infantile neuronal ceroid lipofuscinosis (Batten's disease) was undertaken at 17 weeks' gestation by ultrastructural examination of amniotic fluid cells. The presence of curvilinear profiles indicated an affected fetus and the diagnosis was confirmed, after the pregnancy was terminated, by the finding of many typical curvilinear profiles in multiple tissues which included skin, amnion, umbilical vessels, blood, liver, and brain. Comparison between the involved cells in the amniotic fluid and fetal tissues suggests that these cells are probably derived from the periderm, and possibly also from the amnion. The prominent presence of cytosomes in the periderm and intermediate cells of the fetal epidermis and occasionally also in the endothelial cells of the dermis suggests that fetal skin may be a useful alternative site for assessing fetal involvement. Control specimens of the amniotic fluid, fetal skin, amnion, and liver showed no similar cytosomes. However, some control amniotic fluid samples did contain cells with large collections of irregular trilaminar membranes, and these could be open to misinterpretation. It is important that only typical curvilinear profiles are considered as an indication of an affected pregnancy.     

An Optimised Direct Lysis Method for Viral RNA Extraction and Detection of Foodborne Viruses on Fruits and Vegetables

Detection of norovirus (NoV) and hepatitis A virus (HAV) on fruits and vegetables using current standard methodologies can be inefficient. Method optimisat