A culture vessel for amniotic fluid cells allowing faster preparation of chromosome slides

A new culture vessel for amniotic fluid culture is presented (flaskette). It consists of a microscope slide, on top of which a culture chamber is mounted. Amniotic fluid cell cultures using in situ technique in the flaskette were compared to subcultured samples in ordinary (Falcon) tissue culture bottles. Working time was reduced by using this new culture vessel because of a very simple harvest procedure allowing simultaneous harvest of 15 samples. The interval between amniocentesis and harvest was shorter for the in situ technique than for the subcultivation technique. The frequency of aneuploidy in individual metaphases was higher with the subcultivation technique. while there was no difference in the frequency of structural anomalies.     

Arsenic accumulation in the shore crab Carcinus maenas: the influence of nutritional state,sex and exposure concentration

The accumulation of arsenate from seawater by the shore crab Carcinus maenas L. (collected from Odense Fjord, Denmark in 1991 and from Restronguet Creek, UK in 1991) was investigated in a series of laboratory experiments. A field study was also carried out to determine the effects of raised environmental arsenic concentrations on intra-organismal distribution and tissue concentrations. Studies on the influence of nutritional state and sex on accumulation of As(5) from seawater indicated that most of the arsenic taken up from seawater in laboratory experiments was retained in the gills and the midgut gland. Arsenic accumulation exhibited sex-dependent differences which were also evident in correlation analyses carried out between total lipid contents and total arsenic contents of midgut glands of individual crabs. Arsenic concentrations in the gonads of both sexes were strongly influenced by the nutritional state of the crabs. Elevated arsenic concentrations in seawater and food at an arsenic polluted site (Restronguet Creek) significantly influenced arsenic concentrations and distribution among the tissues of C. maenas. Arsenic concentrations and distribution patterns differed markedly from those crabs from an unpolluted site in Odense Fjord. The gills of the crabs from Restronguet Creek contained extremely high arsenic concentrations ranging from 179 to 483 g As g^-1 dry wt. These values were even higher than those measured in the gills of Odense crabs that had been exposed to 3 mgl^-1 As(5) for 2 wk in the laboratory. Arsenic concentrations in the exoskeleton of Odense Fjord crabs were 15 times lower than those measured in exoskeletons of Restronguet Creek crabs. Approximately 69% of the total body burden of arsenic was located in muscle tissue of crabs from Odense Fjord, whereas the major pool of arsenic (46%) in Restronguet Creek crabs was located in the exoskeleton.     

Echolocation in two very small bats from Thailand Craseonycteris thonglongyai and Myotis siligorensis

Summary The echolocation and hunting behavior of two very small bats, Craseonycteris thonglongyai (Hill) and Myotis siligorensis (Horsfield), from Thailand, were investigated using multiflash photographs, video, and high-speed tape recordings with a microphone array that allowed determination of distance and direction to the bats. C. thonglongyai is the world's smallest mammal and M. siligorensis is only slightly larger. Both bats hunted insects in open areas. The search signals of C. thonglongyai were 3.5 ms long multiharmonic constant frequency (CF) signals with a prominent second harmonic at 73 kHz repeated at around 22 Hz. The band width (BW) of the short terminal frequency modulated (FM) sweep increased during the very short approach phase. In the final buzz the CF component disappeared, the duration decreased to 0.2 ms, and the repetition rate increased to 215 Hz (Figs. 2, 3, 4). There was no drop in frequency in the buzz. The video recordings of C. thonglongyai indicated that it seizes insects directly with the mouth (Fig. 1). M. siligorensis produced 5.4 ms long CF search signals at 66 kHz. The repetition rate was around 13 Hz. In the approach phase an initial broad band FM sweep was added. The buzz consisted of two phases, buzz I and buzz II. Buzz 11 was characterized by short cry durations (around 0.3 ms), a constant high repetition rate (185 Hz), a distinct drop in frequency, and a prominent second harmonic (Figs. 5, 6, 7). The drop in frequency, apparently typical of vespertilionid bats, has been explained by physiological limitations in sound production. However, C. thonglongyai produced very short signals at very high repetition rates without any frequency drop. The drop may be of adaptive value since it enables M. siligorensis to produce very short signals with high sweep rates. The drop moves the pronounced second harmonic into the frequency range of most interest to the bat (Fig. 7D). The sweep rate in this frequency range may now increase to twice the maximum rate that the vocal cords can produce directly. C. thonglongyai and M. siligorensis belong to different superfamilies, Emballonuroidea and Vespertilionoidea, respectively. In spite of their phylogenetic distance they produce strikingly similar search signals of narrow BW around 70 kHz with high source levels (100–115 dB peSPL peak equivalent sound pressure level). We argue that the signal resemblance is due to the similarity in size and hunting behavior of the two bats both hunting insects in open areas. High frequencies are heavily attenuated in air, but because of their small size the bats are restricted to hunting small insects which only reflect echoes at high frequencies. Thus, the emitted frequency is probably the lowest possible given the prey size. Hence, the two bats can only maximize the range of their sonar by decreasing the BW and emitting high intensities. Correspondence to: A. Surlykke     

Examination of hydroxamate-siderophore production by neritic eukaryotic marine phytoplankton

Species of neritic eukaryotic marine phytoplankton were investigated during 1982 for hydroxamate-type siderophore production under iron-sufficient and iron-deficient culture conditions. Three of the 5 Prorocentrum species examined produced siderophores. Prorocentrin, the extracellular hydroxamate-type siderophore isolated from P. minimum, was also produced by P. mariae-lebouriae and P. gracile. P. maximum and P. micans grew poorly in iron-deficient medium and did not produce intracellular or extracellular hydroxamate-type siderophores. Thalassiosira pseudonana and Dunaliella tertiolecta produced extracellular siderophores under iron-deficient conditions, but siderophore production was not detected in the other two species, Skeletonema costatum and Olisthodiscus luteus. Each species which produced extracellular Csaky-positive hydroxamate showed a similar pattern of production. Under iron-sufficient conditions there was no measurable siderophore found either intracellularly or extracellularly. Under iron-deficient culture conditions hydroxamate-type siderophore was produced 1 to 2 d after the cessation of growth in the stationary phase. Production was over a short period of time (1 to 2 d) and the siderophore did not remain in the medium. The rate of siderophore disappearance from the medium was similar to the rate of production. Each species which produced siderophores showed an increase in in vivo fluorescence coincidental with the disappearance of the extracellular siderophore from the culture medium. There was no corresponding increase in in vivo fluorescence in iron-sufficient cultures. It is suggested that in vivo fluorescence may be used as a screening procedure for determining hydroxamate-type siderophore production in eukaryotic phytoplankton. An hypothesis on the iron uptake mechanism is proposed.     

Alpha-fetoprotein and acetylcholinesterase are not predictive of fetal junctional epidermolysis bullosa,Herlitz variant

Junctional epidermolysis bullosa, Herlitz variant (junctional EB-Herlitz) is a lethal autosomal recessive skin disorder currently amenable to prenatal diagnosis only by direct analysis of fetal skin. However, elevated levels of alpha-fetoprotein, as well as the presence of acetylcholinesterase in amniotic fluid, have been associated with other severe fetal genodermatoses. Fetal skin samplings were performed in ten pregnancies at risk for fetal junctional EB-Herlitz, with three fetuses affected on the basis of electron microscopic detection of blisters within the lamina lucida and abnormal hemidesmosomes. In neither affected nor unaffected pregnancies were maternal serum or amniotic fluid alpha-fetoprotein levels elevated. Moreover, alphafetoprotein levels in both maternal serum and amniotic fluid were not statistically different comparing affected and unaffected fetuses. Acetylcholinesterase was not present in the amniotic fluid samples of the three affected pregnancies. Unlike other severe fetal genodermatoses, neither alpha-fetoprotein nor acetylcholinesterase was predictive of junctional EB-Herlitz.     

Substance Flow Analysis and Source Mapping of Chemical UV-filters

Chemical ultraviolet (UV)-filters are used in sunscreens to protect the skin from harmful UV radiation which may otherwise cause sunburns and skin cancer. Commonly used chemical UV-filters are known to cause endocrine disrupting effects in both aquatic and terrestrial animals as well as in human skin cells. Here, source mapping and substance flow analysis were applied to find the sources of six UV-filters (oxybenzone, avobenzone, 4-methylbenzylidene camphor, octyl methoxycinnamate, octyl dimethyl PABA and homosalate) and to identify the most dominant flows of these substances in Denmark. Urban water, composed of wastewater and surface waters, was found to be the primary recipient of UV-filters, whereby wastewater received an estimated 8.5–65 tonnes and surface waters received 7.1–51 tonnes in 2005. In wastewater treatment plants, their sorption onto sludge is perceived to be an important process and presence in effluents can be expected due to a lack of biodegradability. In addition, the use of UV-filters is expected to continue to increase significantly. Not all filters (e.g., octyl dimethyl PABA and homosalate) are used in Denmark. For example, 4-MBC is mainly associated with self-tanning liquids and private import of sunscreens.     

Mineralisation studies of 14C-labelled metsulfuron-methyl, tribenuron-methyl, chlorsulfuron and thifensulfuron-methyl in one Danish soil and groundwater sediment profile.

Bacterial mineralisation of four sulfonylurea herbicides at 20 microg kg(-1) in a sandy soil from nine different depths in a sandy soil horizon (5-780 cm) was investigated in laboratory studies. Metsulfuron-methyl, chlorsulfuron, and tribenuron-methyl were 14C-labelled in the sulfonamide ring, while thifensulfuron-methyl was labelled in the thiophene ring. The highest mineralised amount in 126 days was observed for metsulfuron-methyl (40%) followed by tribenuron-methyl (25%), and thifensulfuron-methyl (11%). Chlorsulfuron showed low mineralisation in all the soils tested (<4%). Mineralisation of the herbicides metsulfuron-methyl and tribenuron-methyl varied according to soil depth (upper profile: 5-70 cm, and lower profile: 165-780 cm) and were proven faster in soil taken from depths 5-7 and 30-35 cm, and slower in depths 45-50 and 70-75 cm. Mineralisation was absent in the lower profile (165-780 cm). As an indicator of microbial activity bacterial counts were taken at the experimental start; these counts grouped in three levels: highest in the surface layer (5-7 cm), slightly lower in the depths 30-75 cm, and lowest in the lower profile (165-780 cm). Residual concentrations of metsulfuron-methyl correlated to the accumulated amount mineralised, with high residual concentrations in soil showing low mineralisation. Also chlorsulfuron showed high residual concentrations with increasing depth in the upper profile, but the relatively high dissipation at 30-35 cm and lower one at 45-50 cm could not be related with the lack of mineralisation. This shows that hydrolysis occurs, but mineralisation of the chloro-substituted sulfonamide is restricted. Tribenuron-methyl and thifensulfuron-methyl could not be detected due to interference with other compounds.     

A screening of liver,kidney, and thyroid gland morphology in organochlorine-contaminated glaucous gulls (Larus hyperboreus) from Svalbard

Concentrations of organochlorine (OC) contaminants and histomorphology of liver, kidney, and thyroid tissues were studied in nine adult and one subadult glaucous gulls (Larus hyperboreus) collected at Svalbard on 2 August 2011. Concentrations of liver polychlorinated biphenyls (PCB; range: 150–2820?ng?g^?1 ww), dichlorodiphenyltrichloroethane (DDT; range: 58–724?ng?g^?1 ww), and chlordanes (CHL; range: 11–126?ng?g^?1 ww) dominated the OC profile followed by hexachlorobenzene (HCB; range: 11–42?ng?g^?1 ww), mirex (range: 2–52?ng?g^?1 ww), and β-hexachlorocyclohexane (β-HCH; range: 1–7?ng?g^?1 ww). Histological examination of the liver showed mononuclear cell infiltrations and granulomas in 10 and 6 gulls, respectively, while intense intrahepatic lipid accumulation (steatosis) was found in two and focal necrosis in one gull. In kidney, glomerular sclerosis and adhesions was found in five and one gull, respectively. Thickening of the glomerular basement membranes and tubular necrosis was found in four and seven gulls, respectively, while mononuclear cell infiltrations were found in two individuals. In the thyroid gland, a high density of small follicles accompanied by follicular epithelial cell proliferation was observed in five glaucous gulls. Gulls with hepatic steatosis had significantly higher ΣDDT levels than the gulls without hepatic steatosis and a similar trend was found for ΣPCB. When normalizing OC concentrations for lipid content in liver, gulls with lipid granulomas had significantly lower β-HCH and significantly higher mirex levels, respectively, than gulls without lipid granulomas. Also; gulls with thickening of the glomerular basement membranes had non-significantly higher ΣPCB levels than gulls without. The histological findings were similar to those of controlled laboratory studies and OC-contaminated wildlife (e.g., polar bears; Ursus maritimus) and the data of this study therefore suggest that OC exposure may be a co-factor in the development of organ alterations in glaucous gulls. However, other environmental factors such as age, element exposure, and infectious micropathogens cannot be ruled out as co-factors, and it is uncertain if the tissue changes found exert adverse health effects on the population of Svalbard glaucous gulls.