Reduction of the edited domain of the mitochondrial A6 gene for ATPase subunit 6 in Trypanosomatidae

The sequence of mitochondrial A6 (MURF4) was compared for several trypanosomatids in order to assess the reduction of the edited domain (ED). The association between the ED reduction and the phylogenetic position of a species proved to be less tight than believed earlier. Compared with digenetic species, monogenetic ones more often displayed ED reduction and had smaller ED.     

Effects of species interactions on community organization along a trampling gradient

Abstract. Effects of interspecific interactions on the organization of two trampled communities, Eragrostio ferrugineae - Plantaginetum asiaticae (EP) and Eleusino indicae - Digitarietum violascentis (ED), were examined by 4-yr field experiments. We compared changes in the relative abundance of the main component species of the communities in monoculture and mixed culture along a trampling gradient. At no trampling in mixed culture, Ambrosia artemisiifolia var. elatior and Erigeron annuus (roadside herbs, RH) predominated and excluded the trampled community species. Severe trampling markedly reduced the predominance of these roadside herbs, promoted Eragrostis ferruginea (a perennial grass of EP), but suppressed Eleusine indica (an annual grass of ED). These results suggest that the differentiation of trampled and roadside herb communities under heavy trampling are caused by asymmetric competition between their main species. Several species pairs showed a constant rank order of relative cover at high trampling levels. Pennisetum alopecuroides (a perennial grass of both RH and EP) and Eragrostis coexisted, indicating symmetric competition. Eragrostis and Plantago asiatica (a perennial herb of EP) or Poa annua (a winter-annual grass of both EP and ED) coexisted through separation in phenology. Eragrostis promoted the survival of Plantago by moderating unpredictable drought (commensalistic positive interaction). This suggests that community composition is maintained by several interspecific interactions.     

The ED strategy: how species-level surrogates indicate general biodiversity patterns through an 'environmental diversity' perspective

Biodiversity assessment requires that we use surrogate information in practice to indicate more general biodiversity patterns. ‘ED’ refers to a surrogates framework that can link species data and environmental information based on a robust relationship of compositional dissimilarities to ordinations that indicate underlying environmental variation. In an example analysis of species and environmental data from Panama, the environmental and spatial variables that correlate with an hybrid multi‐dimensional scaling ordination were able to explain 83% of the variation in the corresponding Bray Curtis dissimilarities. The assumptions of ED also provide the rationale for its use of p‐median optimization criteria to measure biodiversity patterns among sites in a region. M.B. Araújo, P.J. Densham & P.H. Williams (2004, Journal of Biogeography 31 , 1) have re‐named ED as ‘AD’ in their evaluation of the surrogacy value of ED based on European species data. Because lessons from previous work on ED options consequently may have been neglected, we use a corroboration framework to investigate the evidence and ‘background knowledge’ presented in their evaluations of ED. Investigations focus on the possibility that their weak corroboration of ED surrogacy (non‐significance of target species recovery relative to a null model) may be a consequence of Araújo et al.'s use of particular evidence and randomizations. We illustrate how their use of discrete ED, and not the recommended continuous ED, may have produced unnecessarily poor species recovery values. Further, possible poor optimization of their MDS ordinations, due to small numbers of simulations and/or low resolution of stress values appears to have provided a possible poor basis for ED application and, consequently, may have unnecessarily favoured non‐corroboration results. Consideration of Araújo et al.'s randomizations suggests that acknowledged sampling biases in the European data have not only artefactually promoted the non‐significance of ED recovery values, but also artefactually elevated the significance of competing species surrogates recovery values. We conclude that little credence should be given to the comparisons of ED and species‐based complementarity sets presented in M.B. Araújo, P.J. Densham & P.H. Williams (2004, Journal of Biogeography 31 , 1), unless the factors outlined here can be analysed for their effects on results. We discuss the lessons concerning surrogates evaluation emerging from our investigations, calling for better provision in such studies of the background information that can allow (i) critical examination of evidence (both at the initial corroboration and re‐evaluation stages), and (ii) greater synthesis of lessons about the pitfalls of different forms of evidence in different contexts.     

Small-spatial scale changes in productivity of suprabenthic and infaunal crustaceans at the continental shelf of Ebro Delta (western Mediterranean)

Production (P) and production/biomass ratio (P/B) were estimated from 21 species of peracarid and eucarid crustaceans belonging to two different compartments (infauna: 10 species and suprabenthos: 11 species) using the Hynes size–frequency method. Calculations were based on 7 cruises performed between December 2002 and November 2003 in the continental shelf of the Ebro river delta (ED, Catalan Sea: northwestern Mediterranean). Within ED, P and P/B were compared between two neighbouring sites, one located on 47 m (S1), the other at 61 m (S2). The possible effects of natural variability on production were considered. Sediments in ED were muddy (%clay between 40 and 65%). No significant trend in production comparing S1 and S2 was detected, with 50% of species showing higher P value in S1, and 50% in S2. A significant trend emerged from P/B estimates, with 18 of 21 species showing higher P/B at S1. Similarly significant trends for infauna and suprabenthos were found after analyzing both compartments separately. It is suggested that the more significant P/B found on S1 both among infauna and suprabenthos could be caused by the higher temperature and organic matter content (%TOC) at S1. In multi-linear regression (MLR) models, differences in P between S1 and S2 were greater with increasing differences in the density of species (RD) between sites. Differences were greater in infauna than in suprabenthos, and greater among rare species (those with low dominance, %D). P/B differences between S1 and S2 were mainly explained by %D and RD, and to a lesser extent by differences between sites in the mean size of species (RW). In other words changes in P and P/B were not explained by removal of large size specimens. Some trawling activity occurred at S2, and it may alter the habitat of small macrofauna. In addition to natural variability, we discussed the possible influence of trawling disturbance on P and P/B, bearing in mind the feeding type, mobility, and dispersal capability of species. We suggest that amphipods of the genus Ampelisca, showing higher P/B on S1, characterized by low mobility (probably tube-dwellers), low trophic level (e.g. filter feeding strategies) and low dispersal capability (not free larvae), could be especially vulnerable to trawling impact.     

The Entner–Doudoroff pathway empowers Pseudomonas putida KT2440 with a high tolerance to oxidative stress

Glucose catabolism of Pseudomonas putida is carried out exclusively through the Entner–Doudoroff (ED) pathway due to the absence of 6‐phosphofructokinase. In order to activate the Embden–Meyerhof–Parnas (EMP) route we transferred the pfkA gene from Escherichia coli to a P. putida wild‐type strain as well as to an eda mutant, i.e. lacking 2‐keto‐3‐deoxy‐6‐phosphogluconate aldolase. PfkA^{E. coli} failed to redirect the carbon flow from the ED route towards the EMP pathway, suggesting that ED was essential for sugar catabolism. The presence of PfkA^{E. coli} was detrimental for growth, which could be traced to the reduction of ATP and NAD(P)H pools along with alteration of the NAD(P)H/NADP⁺ ratio. Pseudomonas putida cells carrying PfkA^{E. coli} became highly sensitive to diamide and hydrogen peroxide, the response to which is very demanding of NADPH. The inhibitory effect of PfkA^{E. coli} could in part be relieved by methionine, the synthesis of which relies much on NADPH. These results expose the role of the ED pathway for generating the redox currency (NADPH) that is required for counteracting oxidative stress. It is thus likely that environmental bacteria that favour the ED pathway over the EMP pathway do so in order to gear their aerobic metabolism to endure oxidative‐related insults.     

Proximate composition and energy density of nototheniid and myctophid fish in McMurdo Sound and the Ross Sea,Antarctica

Nototheniid and myctophid fish are primary prey for marine piscivores, yet little is known about their nutritional value. In this study, we characterized the proximate composition [PC: water, fat (neutral lipids), crude protein (CP) and ash] and energy density (ED; kJ g⁻¹) of fifteen fish species from McMurdo Sound and the Ross Sea, Antarctica. We assayed the entire fish for all species except for the large Antarctic toothfish, Dissostichus mawsoni (muscle tissue only). On a wet mass basis (WM), fish were variable in composition: moisture content ranged from 64.9 to 87.3% WM, fat from 0.5 to 17.4% WM, CP from 7.7 to 16.7% WM, ash from 11.2 to 21.0% FFDM (fat-free dry mass), and ED from 2.9 to 10.3 kJ g⁻¹. Myctophids and pelagic nototheniids such as Pleuragramma antarcticum and D. mawsoni were high in fat content (7–17% WM), while a bathylagid and benthic nototheniids including most Trematomus spp. and Lepidonotothen squamifrons were low in fat (0.5–4% WM). The epibenthic Trematomus species (T. eulepidotus and T. lepidorhinus) were intermediate. Energy density tracked fat content, with highest values in myctophids and pelagic nototheniids. The variation in nutrient and energy density confirms that prey composition must be taken into account when modelling energy and nutrient fluxes within the Antarctic ecosystem. Further analyses of prey collected over a number of different locations and seasons are needed in order to determine how the nutritional value of certain species might affect annual or decadal variation in reproductive success or population size of top predators.     

Description of two new species of the genus Gobio (Teleostei: Cyprinidae) from the Black Sea coast of Turkey

Thesystematics of the genus Gobio, especially of the populations on the southern Black Sea coast, seems to be still far from being resolved. Seven species were rec-orded from the northern Black Sea coast while three species were recognized from the southern Black Sea coast. We examined in total 43 specimens from K?z?l?rmak River (southern Black Sea basin), 80 specimens from Çoruh River (the Western Cau-casus of the Black Sea basin), 14 specimens from Rioni River (the Western Caucasus of the Black Sea basin), and 6 specimens from Kherota and Shakhe Rivers (the Western Caucasus of the Black Sea basin). Based on an analysis of 32 metric and 5 meristic data as well as the number of vertebrae, we concluded that the K?z?l?rmak and Coruh populations are two distinct, unnamed species, and we describe them here as Gobio kizilirmakensis from the K?z?l?rmak River and G. artvinicus from the Çoruh River.

http://www.zoobank.org/urn:urn:lsid:zoobank.org:pub:1ED54FB1-588F-4412-B376-4BE4E4516EB1     

A transferable plasticity region in Campylobacter coli allows isolates of an otherwise non‐glycolytic food‐borne pathogen to catabolize glucose

Thermophilic Campylobacter species colonize the intestine of agricultural and domestic animals commensally but cause severe gastroenteritis in humans. In contrast to other enteropathogenic bacteria, Campylobacter has been considered to be non‐glycolytic, a metabolic property originally used for their taxonomic classification. Contrary to this dogma, we demonstrate that several Campylobacter coli strains are able to utilize glucose as a growth substrate. Isotopologue profiling experiments with ¹³C‐labeled glucose suggested that these strains catabolize glucose via the pentose phosphate and Entner‐Doudoroff (ED) pathways and use glucose efficiently for de novo synthesis of amino acids and cell surface carbohydrates. Whole genome sequencing of glycolytic C. coli isolates identified a genomic island located within a ribosomal RNA gene cluster that encodes for all ED pathway enzymes and a glucose permease. We could show in vitro that a non‐glycolytic C. coli strain could acquire glycolytic activity through natural transformation with chromosomal DNA of C. coli and C. jejuni subsp. doylei strains possessing the ED pathway encoding plasticity region. These results reveal for the first time the ability of a Campylobacter species to catabolize glucose and provide new insights into how genetic macrodiversity through intra‐ and interspecies gene transfer expand the metabolic capacity of this food‐borne pathogen.     

The effects of marine heatwaves on acute heat tolerance in corals

Scleractinian coral populations are increasingly exposed to conditions above their upper thermal limits due to marine heatwaves, contributing to global declines of coral reef ecosystem health. However, historic mass bleaching events indicate there is considerable inter- and intra-specific variation in thermal tolerance whereby species, individual coral colonies and populations show differential susceptibility to exposure to elevated temperatures. Despite this, we lack a clear understanding of how heat tolerance varies across large contemporary and historical environmental gradients, or the selective pressures that underpin this variation. Here we conducted standardised acute heat stress experiments to identify variation in heat tolerance among species and isolated reefs spanning a large environmental gradient across the Coral Sea Marine Park. We quantified the photochemical yield (F_v/F_m) of coral samples in three coral species, Acropora cf humilis, Pocillopora meandrina, and Pocillopora verrucosa, following exposure to four temperature treatments (local ambient temperatures, and + 3°C, +6°C and + 9°C above local maximum monthly mean). We quantified the temperature at which F_v/F_m decreased by 50% (termed ED50) and used derived values to directly compare acute heat tolerance across reefs and species. The ED50 for Acropora was 0.4–0.7°C lower than either Pocillopora species, with a 0.3°C difference between the two Pocillopora species. We also recorded 0.9°C to 1.9°C phenotypic variation in heat tolerance among reefs within species, indicating spatial heterogeneity in heat tolerance across broad environmental gradients. Acute heat tolerance had a strong positive relationship to mild heatwave exposure over the past 35 years (since 1986) but was negatively related to recent severe heatwaves (2016–2020). Phenotypic variation associated with mild thermal history in local environments provides supportive evidence that marine heatwaves are selecting for tolerant individuals and populations; however, this adaptive potential may be compromised by the exposure to recent severe heatwaves.     

Testing the effectiveness of discrete and continuous environmental diversity as a surrogate for species diversity

Biodiversity surrogates are needed because detailed data on the distributions of species and communities is very limited. Among alternative surrogate strategies there is an environmental diversity (ED) framework, which uses the p-median model to sample environmental space as evenly as possible. The underlying idea is that such a sample would represent underlying species diversity well. However, tests of the effectiveness of ED as a surrogate for species diversity have been inconclusive, and there is a debate concerning the usefulness of different implementations of ED. In particular, it has been argued that tests of the ED framework are flawed because they used discrete algorithms (calculating the p-median from an observed environmental space), while continuous versions of ED should be preferred (i.e. calculating the p-median from a theoretical, continuously spaced, environmental space). Unfortunately, progress has been hampered by lack of independent testing of the two ED approaches. Here, we provide the first empirical test of the effectiveness of both continuous and discrete ED using European distributions of amphibians and reptiles. Analyses were implemented considering two different extents: (1) western Europe and (2) the Iberian Peninsula. In both cases, implementations of ED represented species at a lower rate than expected by chance (P < 0.05). Unlike suggested by some authors, the continuous ED was not consistently superior to the discrete form: continuous implementations of ED performed slightly better when fewer areas were selected and discrete ED performed better when the whole of the western European region was considered. Our results support findings that ED has only limited value as a surrogate for biodiversity and invite the interpretation that failure of ED is more likely to be related with oversimplification of assumptions underlying the model than to the particular p-median algorithm used.     

The origin and diversity of Drilus Olivier, 1790 (Elateridae: Agrypninae: Drilini) in Crete based on mitochondrial phylogeny

We investigated the phylogeography and speciation of Drilus in the Mediterranean, with focus on the Aegean and especially Crete. Altogether 12 species were sequenced for two fragments of the cytochrome c oxidase subunit I gene (cox1–3′ and cox1–5′ mtDNA) and provided 1381 nucleotides. Both fragments were analysed under the maximum likelihood criterion and Bayesian inference separately and concatenated as a single dataset. The Drilus species from the Peloponnese, the Ionian islands and Crete did not form a monophylum. Drilus sp. E from the Peloponnese and Zakynthos was sister to D. mauritanicus Lucas from Spain in most cases. The remaining Greek Drilus species formed a robustly supported clade in all analyses; however, the species from Crete do not seem to be monophyletic. Estimating species divergences using BEAST, we found out that the key dates in the west Aegean Drilus phylogeography appeared to be the Tortonian Crete–Peloponnese separation (12–9 Mya), the desiccation of the Mediterranean basin during the Messinian Salinity Crisis (5.96–5.33 Mya), and the repeated fragmentation of Crete during the Pliocene and Pleistocene. Within the Drilus lineages, we obtained a substitution rate estimate of 2.75% divergence per million years, which is in excellent agreement with previous studies. A Generalized Mixed Yule Coalescent (GMYC) analysis suggested the presence of six Drilus species in Crete (seven species in total, since the DNA sequences were not available for D. creticus Pic); however, we formally identified and (re)described only four which can be morphologically defined: D. creticus, D. longulus Kiesenwetter, D. horasfakionus sp. nov. and D. baenai sp. nov. These species are endemic to Crete and surrounding islets. Their diagnostic characters are illustrated and an identification key to males of these species is provided. The intraspecific variability, distribution and ecology of all species are discussed and suggestions for further research are given.

http://zoobank.org/urn:lsid:zoobank.org:pub:F8DDBCC1-C2D1-48F7-BE99-55D9ED4C2234     

New species of Epitoniidae (Gastropoda: Epitonioidea) from the Red Sea

Based on material stored in the Zoological Museum of the University of Bologna (MZB), four new species of epitoniids from the shallow waters of the Red Sea are described: Epitonium (Laeviscala) yemenita n. sp., Epitonium (Laeviscala) morassii n. sp., Epitonium (Labeoscala) zabargadense n. sp. and Epitonium (Labeoscala) aranea n. sp. Scanning electron microscope illustrations of type specimens of Scalaria gracilis Sowerby, 1844, Scalaria fucata Pease, 1861, Graciliscala histricosa Jousseaume, 1912, Graciliscala rostrata Jousseaume, 1912, Scala rissoinaeformis Melvill, 1903 and Scala thelcteria Melvill & Standen, 1903 are given. Systematic remarks on some species belonging to the subgenus Laeviscala are treated in additional notes. A lectotype of Graciliscala histricosa is designated.

http://zoobank.org/urn:lsid:zoobank.org:pub:8304A24D-5985-43D5-AEBF-1A7ED39A1F1C     

Applying Landscape Metrics to Characterize Potential Habitat of Bonobos (Pan paniscus) in the Maringa-Lopori-Wamba Landscape, Democratic Republic of Congo

To conserve areas and species threatened by immediate landscape change requires that we make planning decisions for large areas in the absence of adequate data. Here we study the utility of broad-scale landscape metrics as predictors of species occurrence, especially for remote areas where there is a need to make the most of limited spatial and biological data. Bonobos (Pan paniscus) are endangered great apes endemic to lowland forests of the Democratic Republic of Congo. They are threatened by bushmeat hunting that is exacerbated by habitat fragmentation through slash-and-burn agriculture and timber harvest. We developed four landscape metrics —edge density (ED), COHESION, CONTAGION, and class area (CA)— that may serve as surrogates for measuring accessibility of areas to hunting in order to predict relative bonobo-habitat suitability. We calculated the metrics for the Maringa-Lopori-Wamba (MLW) landscape and evaluated them for utility in predicting bonobo-nest occupancy based on 2009 field data. Cross-validations showed that all four metrics performed similarly. However, forest ED was arguably the best predictor, with an overall classification accuracy of 72.1% in which 85% of known nest blocks (N = 124) were classified correctly. We demonstrated that for a relatively intact landscape and a mobile forest-dwelling species that is fairly tolerant of forest openings, forest fragmentation can still be an important predictor of species occurrence. We suggest that ED can be helpful when mapping bonobo habitat in MLW and can aid landscape-planning and conservation efforts. Our approach may be applied to other edge-sensitive species, especially where high-resolution data are deficient.     

Enrichment of Geobacter Species in Response to Stimulation of Fe(III) Reduction in Sandy Aquifer Sediments

Abstract Engineered stimulation of Fe(III) has been proposed as a strategy to enhance the immobilization of radioactive and toxic metals in metal-contaminated subsurface environments. Therefore, laboratory and field studies were conducted to determine which microbial populations would respond to stimulation of Fe(III) reduction in the sediments of sandy aquifers. In laboratory studies, the addition of either various organic electron donors or electron shuttle compounds stimulated Fe(III) reduction and resulted in Geobacter sequences becoming important constituents of the Bacterial 16S rDNA sequences that could be detected with PCR amplification and denaturing gradient gel electrophoresis (DGGE). Quantification of Geobacteraceae sequences with a PCR most-probable-number technique indicated that the extent to which numbers of Geobacter increased was related to the degree of stimulation of Fe(III) reduction. Geothrix species were also enriched in some instances, but were orders of magnitude less numerous than Geobacter species. Shewanella species were not detected, even when organic compounds known to be electron donors for Shewanella species were used to stimulate Fe(III) reduction in the sediments. Geobacter species were also enriched in two field experiments in which Fe(III) reduction was stimulated with the addition of benzoate or aromatic hydrocarbons. The apparent growth of Geobacter species concurrent with increased Fe(III) reduction suggests that Geobacter species were responsible for much of the Fe(III) reduction in all of the stimulation approaches evaluated in three geographically distinct aquifers. Therefore, strategies for subsurface remediation that involve enhancing the activity of indigenous Fe(III)-reducing populations in aquifers should consider the physiological properties of Geobacter species in their treatment design. Received: 19 October 1999; Accepted: 28 December 1999; Online Publication: 25 April 2000     

Variation in the sensitivity to metalaxyl of Pythium spp. isolated from carrot and other sources

Over a 3-yr period 261 isolates of 17 species of Pythium were tested for sensitivity to metalaxyl at concentrations of 5, 50 or 100 μ/ml. A wide range of responses was observed, from isolates where growth ceased at 5 μg/ml to those where growth at 100 μg/ml was similar to that of the untreated controls. In further tests isolates of 11 different species had ED50's < 1 μg/ml. A lower sensitivity was detected in isolates of six Pythium spp. where values in the range 1–10 μg/ml were obtained. This lower sensitivity was not related to previous known use of metalaxyl. Three isolates of Pythium dissotocum from sites where the fungicide had been used repeatedly had ED50's > 100 μg/ml and were considered resistant. The resistance was stable over a 2-yr period and isolates were cross-resistant to furalaxyl, benalaxyl, ofurace, cyprofuram and oxadixyl. Increasing concentrations of metalaxyl reduced or prevented the production of zoospores by four species of Pythium, although when zoospores were produced, this was followed by the normal processes of encystment and germination. Culturing P. dissotocum on different sub-lethal concentrations of metalaxyl for 18 wk did not induce a high level of resistance to the fungicide.     

A study on nitrate reductase activity (NRA) of geophytes from Mediterranean environment

Nitrate reductase activity (NRA) in different compartments of 14 Mediterranean geophytes (bulbous, tuberous and rhizomatous) and actual mineral nitrogen (NO₃⁻ and NH₄⁺) in their soils were investigated. The nitrate reduction capacities of each species were determined as NRA per total plant material. Differences among compartments for NRA were significant in all species. The highest NRA was found in leaves of tuberous species (Anemone coronaria, Cyclamen coum) and of most bulbous species (Allium flavum, Allium guttatum, Bellevelia sarmatica, Galanthus plicatus, Leucojum aestivum, Ornithogalum nutans, Tulipa sylvestris). Therefore, in this group of species the contribution of the leaves to total plant NRA was the highest. The other bulbous species (Allium scorodoprasum, Crocus chrysanthus, Fritillaria bithynica, Muscari neglectum) and one rhizomatous taxon (Iris suaveolens) have a different NRA distribution within the plants. In these species the highest values of NRA were found in different organs. For example, in Allium scorodoprasum the highest NRA was in tunics, and in flowers in M. neglectum. Although leaves are the main compartments reducing nitrate in most of the studied geophytes, other compartments also contribute to total plant nitrate reduction.Our results show that the nitrate reduction capacity is different among geophyte species. Even if it roughly reflects the nitrogen supply in a habitat, differences in nitrate reduction capacities of different species collected from same sites indicate that the nitrate reducing capacity is species-specific.     

Effects of climate change on the distribution of Iberian tree species

Question: Will the predicted climate changes affect species distribution in the Iberian Peninsula? Location: Iberian Peninsula (Spain and Portugal). Methods: We modelled current and future tree distributions as a function of climate, using a computational framework that made use of one machine learning technique, the random forest (RF) algorithm. This algorithm provided good predictions of the current distribution of each species, as shown by the area under the corresponding receiver operating characteristics (ROC) curves. Species turnover, richness and the change in distributions over time to 2080 under four Intergovernmental panel on climate change (IPCC) scenarios were calculated using the species map outputs. Results and Conclusions: The results show a notable reduction in the potential distribution of the studied species under all the IPCC scenarios, particularly so for mountain conifer species such as Pinus sylvestris, P. uncinata and Abies alba. Temperate species, especially Fagus sylvatica and Quercus petraea, were also predicted to suffer a reduction in their range; also sub‐mediterranean species, especially Q. pyrenaica, were predicted to undergo notable decline. In contrast, typically Mediterranean species appeared to be generally more capable of migration, and are therefore likely to be less affected.     

Functional Constraints of 6-Phosphogluconate Dehydrogenase (6-PGD) Based on Sequence and Structural Information

The pentose phosphate cycle is considered as a major source of NADPH and pentose needed for nucleic acid biosynthesis. 6-Phosphogluconate dehydrogenase (6PGD), an enzyme participating in this cycle, catalyzes the oxidative decarboxylation of 6PGD to ribulose 5-phosphate with the subsequent release of CO₂ and the reduction of NADP. We have determined the amino acid sequence of 6PGD of Bactrocera oleae and constructed a three-dimensional model based on the homologous known sheep structure. In a comparative study of 6PGD sequences from numerous species, all the conserved and variable regions of the enzyme were analyzed and the regions of functional importance were localized, in an attempt promoted also by the direct involvement of the enzyme in various human diseases. Thus, analysis of amino acid variability of 37 6PGD sequences revealed that all regions important for the catalytic activity, such as those forming the substrate and coenzyme binding sites, are highly conserved in all species examined. Moreover, several amino acid residues responsible for substrate and coenzyme specificity were also found to be identical in all species examined. The higher percentage of protein divergence is observed at two regions that accumulate mutations, located at the distant parts of the two domains of the enzyme with respect to their interface. These peripheral regions of nonfunctional importance are highly variable and are predicted as antigenic, thus reflecting possible regions for antibody recognition. Furthermore, locating the differences between diptera 6PGD sequences on the three-dimensional model suggests probable positions of different amino acid residues appearing at B. oleae fast, intermediate, and slow allozymic variants. Abbreviations used: 6Pgd, 6-Phosphogluconate dehydrogenase gene; 6PGD, 6-Phosphogluconate dehydrogenase enzyme; NADPH, nicotinamide adenine dinucleotide phosphate; ADP, adenine dinucleotide phosphate; TNBS, 2,4,6 trinitrobenzensulfonic acid.     

A SURVEY OF THE STEROL COMPOSITION OF THE MARINE DINOFLAGELLATES KARENIA BREVIS,KARENIA MIKIMOTOI,AND KARLODINIUM MICRUM: DISTRIBUTION OF STEROLS WITHIN OTHER MEMBERS OF THE CLASS DINOPHYCEAE1

The sterol composition of different marine microalgae has been examined to determine the utility of sterols as biomarkers to distinguish members of various algal classes. For example, members of the class Dinophyceae possess certain 4‐methyl sterols, such as dinosterol, which are rarely found in other classes of algae. The ability to use sterol biomarkers to distinguish certain dinoflagellates such as the toxic species Karenia brevis Hansen and Moestrup, responsible for red tide events in the Gulf of Mexico, from other species within the same class would be of considerable scientific and economic value. Karenia brevis has been shown by others to possess two major sterols, (24S)‐4α‐methyl‐5α‐ergosta‐8(14),22‐dien‐3β‐ol (ED) and its 27‐nor derivative (NED), having novel structures not previously known to be present in other dinoflagellates. This prompted the present study of the sterol signatures of more than 40 dinoflagellates. In this survey, sterols with the properties of ED and NED were found in cultures of K. brevis and shown also to be the principal sterols of Karenia mikimotoi Hansen and Moestrup and Karlodinium micrum Larsen, two dinoflagellates closely related to K. brevis. They are also found as minor components of the more complex sterol profiles of other members of the Gymnodinium/Peridinium/Prorocentrum (GPP) taxonomic group. The distribution of these sterols is consistent with the known close relationship between K. brevis, K. mikimotoi, and K. micrum and serves to limit the use of these sterols as lipid biomarkers to a few related species of dinoflagellates.