Response of zooxanthellae in symbiosis with the Mediterranean corals Cladocora caespitosa and Oculina patagonica to elevated temperatures

Scleractinian symbiotic corals living in the Ligurian Sea (NW Mediterranean Sea) have experienced warm summers during the last decade, with temperatures rapidly increasing, within a few days, to 3–4°C above the mean value of 24°C. The effect of elevated temperatures on the photosynthetic efficiency of zooxanthellae in symbiosis with temperate corals has not been well investigated. In this study, the corals, Cladocora caespitosa and Oculina patagonica were collected in the Ligurian Sea (44°N, 9°E), maintained during 2 weeks at the mean summer temperature of 24°C and then exposed during 48 h to temperatures of 24 (control), 27, 29 and 32°C. Chlorophyll (chl) fluorescence parameters [F _v/F _m, electron transport rate (ETR), non-photochemical quenching (NPQ)] were measured using pulse amplitude modulated (PAM) fluorimetry before, during the thermal increase, and after 1 and 7 days of recovery (corals maintained at 24°C). Zooxanthellae showed a broad tolerance to temperature increase, since their density remained unchanged and there was no significant reduction in their maximum quantum yield (F _v/F _m) or ETR up to 29°C. This temperature corresponded to a 5°C increase compared to the mean summer temperature (24°C) in the Ligurian Sea. At 32°C, there was a significant decrease in chl contents for both corals. This decrease was due to a reduction in the chl/zooxanthellae content. For C. caespitosa, there was also a decrease in ETR_max, not associated with a change in F _v/F _m or in the non-photochemical quenching (NPQ); for O. patagonica, both ETR_max and F _v/F _m significantly decreased, and NPQ_max showed a significant increase. Damages to the photosystem II appeared to be reversible in both corals, since F _v/F _m values returned to normal after 1 day at 24°C. Zooxanthellae in symbiosis with the Mediterranean corals investigated can therefore be considered as resistant to short-term increases in temperature, even well above the maximum temperatures experienced by these corals in summer.     

Co-evolution-driven cuticular hydrocarbon variation between the slave-making ant Rossomyrmex minuchae and its host Proformica longiseta (Hymenoptera: Formicidae)

Summary. Proformica longiseta exists as two populations in the Sierra Nevada Mountains in Spain, only one of which is parasitized by the slave-maker ant Rossomyrmex minuchae. To investigate the possible effect of co-evolutionary pressures on cuticular hydrocarbon (CHC) profiles (the presumed nestmate recognition cues), we performed a comparative analysis of the CHC of R. minuchae and P. longiseta colonies from both the allopatric and sympatric populations; the latter includes samples of enslaved as well as free-living workers. Discriminant analyses based on these chemical profiles showed two clear profile groups: the first comprised R. minuchae and both enslaved and free-living P. longiseta from the sympatric population; and the second the allopatric P. longiseta workers. As expected, the profiles of the two sympatric P. longiseta groups (enslaved and free-living) were distinct; but, interestingly, those of the enslaved P. longiseta and its parasite R. minuchae were also distinguishable. This indicates that despite their cohabitation each species maintains its own chemical identity. Profile similarity between the sympatric free-living P. longiseta and its parasite may explain the lower than expected aggression observed during raids. We further speculate that in view of the differences between the sympatric and allopatric population of P. longiseta, co-evolutionary pressures have driven changes in the profile of the former to better match that of its parasite R. minuchae. Such an adjustment may have enabled nests of the sympatric P. longiseta to endure multiple raids by the parasite (due to the reduced aggression) and thus to continue to reproduce despite the damage inflicted by the raids.     

Food influence on colonial recognition and chemical signature between nestmates in the fungus-growing ant <Emphasis Type="Italic">Acromyrmex subterraneus subterraneus</Emphasis>

Summary. Neotropical Fungus-growing leaf-cutting ants (tribe Attini) live in obligatory symbiosis with a fungus, which they grow on fresh leaves harvested by workers. Colonial recognition is likely based on chemical cues provided by cuticular hydrocarbons that have been found to be partly influenced by environmental odor sources. The diet breadth of Acromyrmex subterraneus subterraneus enabled us to test the impact of different plant diets on colonial recognition. The intermediary of the fungus in the ants feeding habit adds a special angle to the question. From a queenright (QR) mother colony of A. s. subterraneus we formed several groups of queenless (QL) workers with fungus (approx. 700 ants). The QR colony and two of the QL-groups were fed with the same diet of fresh bramble leaves. Two other QL-groups were fed with privet leaves and two with rose flowers. After 4 months, QR workers were significantly more aggressive towards the QL-group fed with rose flowers or privet leaves than towards workers of the QL-groups fed with fresh bramble leaves. Rose-fed QL workers were aggressive towards privet-fed QL workers and vice versa, but never towards workers of their counterpart group that fed on the same diet. These results suggest that the absence of the queen or the separation time between groups played a minor role in shaping nestmate recognition cues as compared to the diet. The behavioral studies were supplemented by chemical analyses of cuticles, postpharyngeal glands (PPG) and plant-food extracts revealing profiles variations that were correlated with the dietary changes. However, although the plant extract contained several hydrocarbons there was no congruency between the plant profile and the respective diet-group ants. These results support the hypothesis that the diet influences indirectly the chemical profiles and consequently the recognition cues in A. s. subterraneus.     

Airborne observations of vegetation and implications for biogenic emission characterization

Measuring hydrocarbons from aircraft represents one way to infer biogenic emissions at the surface. The focus of this paper is to show that complementary remote sensing information can be provided by optical measurements of a vegetation index, which is readily measured with high temporal coverage using reflectance data. We examine the similarities between the vegetation index and in situ measurements of the chemicals isoprene, methacrolein, and alpha-pinene to estimate whether the temporal behavior of the in situ measurements of these chemicals could be better understood by the addition of the vegetation index. Data were compared for flights conducted around Houston in August and September 2000. The three independent sets of chemical measurements examined correspond reasonably well with the vegetation index curves for the majority of flight days. While low values of the vegetation index always correspond to low values of the in situ chemical measurements, high values of the index correspond to both high and low values of the chemical measurements. In this sense it represents an upper limit when compared with in situ data (assuming the calibration constant is adequately chosen). This result suggests that while the vegetation index cannot represent a purely predictive quantity for the in situ measurements, it represents a complementary measurement that can be useful in understanding comparisons of various in situ observations, particularly when these observations occur with relatively low temporal frequency. In situ isoprene measurements and the vegetation index were also compared to an isoprene emission inventory to provide additional insight on broad issues relating to the use of vegetation indices in emission database development.     

Compartment modeling of MTBE in the generic environment and estimations of the aquatic MTBE input in Germany using the EQC model

The use of the gasoline additive methyl tert-butyl ether (MTBE) has caused serious concern about groundwater and surface water contamination. The behavior of MTBE in the two most relevant compartments, surface water and air in a generic environment and in a simulated German environment is investigated using the equilibrium criterion (EQC) model. Due to lack of literature data, the half-life time of MTBE in river water is estimated to about 80-120 d (105 d) at 18 degrees C and roughly 1.5 a (year)(533 d) at 4 degrees C from a batch experiment. The EQC model considers four compartments, air, surface water, soil and sediment in an environment of typically 100,000 km2 with about 10% of the area covered with water. The user can progress through the tiered sequence of Level I to III with increasing complexity which reveals more information about the the fate of the considered chemical. The equilibrium mass distribution of MTBE calculated with the Level I model shows that 87% partitions into air and 13% into surface water at 10 degrees C. The results of the Level II calculations indicate that 50% of MTBE in the air is transported from the system and 38% in the air is degraded at 10 degrees C. The resulting total persistence time of 3 d for MTBE in the generic environment of the Level II model can be compared to the calculated value for chlorobenzene. The MTBE input into water is significantly more sensitive to the 'mode of entry' than input into air. The MTBE concentration in surface water is almost exclusively the result of direct emission into water, whereas the atmosphere can additionally be loaded by volatilization from water. The total aquatic MTBE emission in Germany and the average MTBE concentration in German surface waters were roughly estimated to 20-80 t a(-1) (tons per year)(50 t a(-1)) and 50 ng L(-1), respectively. Surface water concentrations calculated with the underlying assumptions of the model can neither be explained by exposure through waste water and industrial effluents nor with an estimated loss of industrially used MTBE in Germany. For the year-round scenario at 10 degrees C, MTBE concentrations of 19 ng L(-1) (surface water) and 167 ng m(-3) (air) result. However, it remains unclear whether the assumptions of the model, the lack of analyses from industrial effluents or both are responsible for the difference. Additional aquatic emission sources could result from gasoline transport on and storage near rivers. The comparison of winter and summer scenarios shows that in summer, atmospheric (25%) and aqueous (50%) concentrations are lower than in winter due to higher degradation rates.     

Trace elements and organochlorines in the shoalgrass community of the lower Laguna Madre,Texas

Our objectives were to measure concentrations of seven trace elements and 14 organochlorine compounds in sediment and biota of the shoalgrass (Halodule wrightii) community of the lower Laguna Madre of south Texas and to determine whether chemicals associated with agriculture (e.g. mercury, arsenic, selenium, organochlorine pesticides) were highest near agricultural drainages. Arsenic, mercury, selenium, lead, cadmium, and organochlorines were generally at background concentrations throughout the lower Laguna Madre. Nickel and chromium concentrations were exceptionally high in shrimp and pinfish (Lagodon rhomboides), which is difficult to explain because of no known anthropogenic sources for these trace elements. For sediment and blue crabs (Callinectes sapidus), mercury was highest near agricultural drainages. Also, DDE was more frequently detected in blue crabs near agricultural drainages than farther away. In contrast, selenium concentrations did not differ among collecting sites and arsenic concentrations were lowest in shoalgrass, blue crabs, and brown shrimp (Penaeus aztecus) near agricultural drainages.     

The OECD Validation Program of the H295R Steroidogenesis Assay for the Identification of In Vitro Inhibitors and Inducers of Testosterone and Estradiol Production. Phase 2: Inter-Laboratory Pre-Validation Studies (8 pp)

Background, Goals and Scope In response to concerns that have been raised about chemical substances that may alter the function of endocrine systems and result in adverse effects on human health, an OECD initiative was undertaken to develop and validate in vitro and in vivo assays to identify chemicals that may interfere with endocrine systems of vertebrates. Here we report on studies that were conducted to develop and standardize a cell-based screening assay using the H295R cell line to prioritize chemicals that may act on steroidogenic processes in humans and wildlife. These studies are currently ongoing as part of the ‘Special Activity on the Testing and Assessment of Endocrine Disruptors’ within the OECD Test Guidelines Program to review, develop, standardize, and validate a number of in vitro and in vivo toxicological assays for testing and assessment of chemicals concerning their potential to interact with the endocrine system of vertebrates. Study Design Six laboratories from five countries participated in the pre-validation studies. Each laboratory tested the effects of three model chemicals on the production of testosterone (T) and estradiol (E2) using the H295R Steroidogenesis Assay. Chemicals tested were well described inducers or inhibitors of steroidogenic pathways (forskolin, prochloraz and fadrozole). All experiments were conducted in 24 well plates following standard protocols. Six different doses per compound were analyzed in triplicate per plate. A quality control (QC) plate was run in conjunction with the chemical exposure plate to account for inter-assay variation. Each chemical exposure was conducted two or three times. Results All laboratories successfully detected increases and/or decreases in hormone production by H295R cells after exposure to the different model compounds and there was good agreement in the pattern of response for all groups. Forskolin increased both T and E2 while fadrozole and prochloraz decreased production of both hormones. All chemicals affected hormone production in a dose-dependent manner with the exception of fadrozole which caused maximum inhibition of E2 at the two least concentrations tested. Some inter-laboratory differences were noted in the alteration of hormone production measured in chemically exposed cells. However, with the exception of the production of T measured at one laboratory in cells exposed to forskolin, the EC₅₀s calculated were comparable (coefficients of variation 34–49%) for all hormones. Discussion and Perspectives The results indicated that the H295R Steroidogenesis Assay protocol was robust, transferable and reproducible among all laboratories. However, in several instances that were primarily related to one laboratory there were unexplained minor uncertainties related to the inter-laboratory hormone production variation. Based on the findings from this Phase 2 prevalidation study, the H295R Steroidogenesis Assay protocol is currently being refined. The next phase of the OECD validation program will test the refined protocol among the same group of laboratories using an extended set of chemicals (∼30) that will include positive and negative chemical controls as well as a broad spectrum of different potential inducers and inhibitors of steroidogenic pathways. Submission Editor: Dr. Carsten Brühl (bruehl@uni-landau.de)     

Amniotic fluid microvillar enzyme activities in the early detection of fetal abnormalities

Measurement of the microvillar enzymes, γ-glutamyltranspeptidase (GGTP), aminopeptidase M (APM) and alkaline phosphatase (ALP), in amniotic fluid supernatant has been proposed as a method for the early prenatal diagnosis of cystic fibrosis. The activities of these enzymes in a series of other fetal abnormalities have now been examined. GGTP activities were below the 5th percentile in 28 out of 54 cases of trisomy 21 and 9 of 14 cases of trisomy 18, while APM values were below this cut-off in 26 of 54 cases of trisomy 21 and 8 of 14 cases of trisomy 18. Abnormal ALP isoenzyme ratios were found in 6 of 54 cases of trisomy 21 and 4 of 14 cases of trisomy 18. If prenatal cytogenetic studies are routinely carried out on amniotic fluid cells, the occasional confounding effect of abnormal microvillar enzymes associated with fetal trisomies rather than with cystic fibrosis should be avoided.     

Prenatal diagnosis of fragile X syndrome: Management of the male fetus with a premutation

Direct detection of the fragile X mutation by DNA analysis has greatly simplified prenatal diagnosis of this disease. However, women carrying a fragile X premutation may pass their expanded trinucleotide repeat to sons without expansion to a full mutation. Such sons are predicted to be intellectually normal. In this situation, the accuracy with which the fetal status can be inferred from analysis of chorionic villus sample (CVS) DNA is unclear. We describe such a case, in which it was felt necessary to proceed to fetal blood sampling despite technically unambiguous DNA results from the CVS. The lack of prospective data means that this dilemma may be expected to recur over the next few years when performing prenatal diagnosis on fragile X premutation carriers.     

Umbilical cord pseudocyst in trisomy 18

Prenatal diagnosis of cord defects by means of ultrasound examination is possible and highly accurate. Although this is a rare pathological finding, we report two cases in which umbilical cord pseudocysts were associated with trisomy 18. These observations underscore the need of umbilical blood sampling for establishing the karyotype in fetuses with such umbilical cord anomalies and the importance of careful examination of placentas and infants born with such defects.