Seawater carbonate chemistry and benthic marine community during experiments, 2011

Ocean acidification is predicted to impact all areas of the oceans and affect a diversity of marine organisms. However, the diversity of responses among species prevents clear predictions about the impact of acidification at the ecosystem level. Here, we used shallow water CO2 vents in the Mediterranean Sea as a model system to examine emergent ecosystem responses to ocean acidification in rocky reef communities. We assessed in situ benthic invertebrate communities in three distinct pH zones (ambient, low, and extreme low), which differed in both the mean and variability of seawater pH along a continuous gradient. We found fewer taxa, reduced taxonomic evenness, and lower biomass in the extreme low pH zones. However, the number of individuals did not differ among pH zones, suggesting that there is density compensation through population blooms of small acidification-tolerant taxa. Furthermore, the trophic structure of the invertebrate community shifted to fewer trophic groups and dominance by generalists in extreme low pH, suggesting that there may be a simplification of food webs with ocean acidification. Despite high variation in individual species' responses, our findings indicate that ocean acidification decreases the diversity, biomass, and trophic complexity of benthic marine communities. These results suggest that a loss of biodiversity and ecosystem function is expected under extreme acidification scenarios.

Global macrozoobenthos production and energy budget data base V150731, with link to database

I developed a new model for estimating annual production-to-biomass ratio P/B and production P of macrobenthic populations in marine and freshwater habitats. Self-learning artificial neural networks (ANN) were used to model the relationships between P/B and twenty easy-to-measure abiotic and biotic parameters in 1252 data sets of population production. Based on log-transformed data, the final predictive model estimates log(P/B) with reasonable accuracy and precision (r2 = 0.801; residual mean square RMS = 0.083). Body mass and water temperature contributed most to the explanatory power of the model. However, as with all least squares models using nonlinearly transformed data, back-transformation to natural scale introduces a bias in the model predictions, i.e., an underestimation of P/B (and P). When estimating production of assemblages of populations by adding up population estimates, accuracy decreases but precision increases with the number of populations in the assemblage.

Giant clams and rising CO2: light may ameliorate effects of ocean acidification on a solar-powered animal

Global climate change and ocean acidification pose a serious threat to marine life. Marine invertebrates are particularly susceptible to ocean acidification, especially highly calcareous taxa such as molluscs, echinoderms and corals. The largest of all bivalve molluscs, giant clams, are already threatened by a variety of local pressures, including overharvesting, and are in decline worldwide. Several giant clam species are listed as 'Vulnerable' on the IUCN Red List of Threatened Species and now climate change and ocean acidification pose an additional threat to their conservation. Unlike most other molluscs, giant clams are 'solar-powered' animals containing photosynthetic algal symbionts suggesting that light could influence the effects of ocean acidification on these vulnerable animals. In this study, juvenile fluted giant clams Tridacna squamosa were exposed to three levels of carbon dioxide (CO2) (control ~400, mid ~650 and high ~950 µatm) and light (photosynthetically active radiation 35, 65 and 304 µmol photons/m**2/s). Elevated CO2 projected for the end of this century (~650 and ~950 µatm) reduced giant clam survival and growth at mid-light levels. However, effects of CO2 on survival were absent at high-light, with 100% survival across all CO2 levels. Effects of CO2 on growth of surviving clams were lessened, but not removed, at high-light levels. Shell growth and total animal mass gain were still reduced at high-CO2. This study demonstrates the potential for light to alleviate effects of ocean acidification on survival and growth in a threatened calcareous marine invertebrate. Managing water quality (e.g. turbidity and sedimentation) in coastal areas to maintain water clarity may help ameliorate some negative effects of ocean acidification on giant clams and potentially other solar-powered calcifiers, such as hard corals.

Larval development of the barnacle Amphibalanus improvisus responds variably but robustly to near-future ocean acidification

Increasing atmospheric CO2 decreases seawater pH in a phenomenon known as ocean acidification. In two separate experiments we found that larval development of the barnacle Amphibalanus (Balanus) improvisus was not significantly affected by the level of reduced pH that has been projected for the next 150 years. After 3 and 6 days of incubation, we found no consistent effects of reduced pH on developmental speed or larval size at pH 7.8 compared with the control pH of 8.1. After 10 days of incubation, there were no net changes in survival or overall development of larvae raised at pH 7.8 or 7.6 compared with the control pH of 8.0. In all cases, however, there was significant variation in responses between replicate batches (parental genotypes) of larvae, with some batches responding positively to reduced pH. Our results suggest that the non-calcifying larval stages of A. improvisus are generally tolerant to near-future levels of ocean acidification. This result is in line with findings for other barnacle species and suggests that barnacles do not show the greater sensitivity to ocean acidification in early life history reported for other invertebrate species. Substantial genetic variability in response to low pH may confer adaptive benefits under future ocean acidification.

Valoración de Indicadores de calidad para el diseño e implementación de tecnologías de manejo en Luvisoles de la zona norte de la provincia de Las Tunas, Cuba

La calidad del suelo es una herramienta de evaluación que puede facilitar la adaptación de prácticas de manejo que promuevan sistemas agropecuarios sostenibles. La investigación de este trabajo se inició con un diagnóstico participativo en 12 comunidades rurales de la provincia de Las Tunas en el año 2009 en el cual los productores identificaron los puntos críticos de calidad de los suelos de la región y sirvieron de punta de partida para seleccionar las variables físicas, químicas y biológicas a determinar en cinco sistemas de uso agropecuario (arboleda, pasto natural, pasto cultivado y dos sistemas silvopastoriles) en la zona La Veguita, municipio Las Tunas. El sistema arboleda se utilizó como referencia de las propiedades naturales del suelo. El pasto natural se distingue por el desarrollo de especies de baja productividad, sin embargo el pasto cultivado está representado por Pennisetum purpureum vc CUBA CT-115, y constituye una contribución a la tecnología de bancos de biomasa, para utilizarse en el pastoreo durante la seca. Los sistemas silvopastoriles están representados por Leucaena leucocephala Lam. en franjas y Panicum maximun vc. Likoni, los que se diferencian en su diseño, manejo y propiedades mineralógicas. El objetivo fundamental fue valorar indicadores de calidad de los suelos Luvisoles háplicos sobre granitoides, para diseñar e implementar tecnologías de manejo que permitan incrementar la capacidad agroproductiva de los suelos. Mediante el análisis de componentes principales se obtuvo un conjunto mínimo de indicadores físicos, químicos y biológicos que proporcionaron información útil referente a los procesos edáficos y se integraron para determinar un índice de calidad. En el sistema de uso, caracterizado por el pasto cultivado (Pennisetum purpureum) se estableció, en parcelas experimentales, un ensayo de corta duración, en el que se comparó el laboreo tradicional y el laboreo sin inversión del prisma, con y sin aplicación de compost. En ambos sistemas de labranza se evaluó el desarrollo del cultivo e indicadores de calidad del suelo. Los resultados mostraron que del conjunto de indicadores edáficos estudiados se seleccionaron 6 en los que la capacidad de intercambio catiónico, materia orgánica, potasio intercambiable, contenido de arena, densidad aparente y biomasa de lombrices explicaron la mayor variabilidad y sirvieron de base para evaluar la calidad de estos suelos. Se establecieron valores umbrales de referencia de indicadores de calidad, que permitirán evaluar y monitorear los sistemas de uso y manejo de la región. El sistema Silvopastoril 2 resultó el de mayor índice de calidad de los suelos tomando como referencia la arboleda por su condición natural. El manejo silvopastoril influyó predominantemente en mejores resultados productivos pero las características edáficas principalmente físicas, deben definir su diseño y manejo. El sistema de pastos cultivados con Pennisetum purpureum vc CUBA CT 115, alcanzó la mayor acumulación de carbono orgánico, sin embargo, el manejo limitó su calidad física y el funcionamiento productivo del sistema. De manera general los sistemas de uso no garantizan un índice de la calidad del suelo, puesto que se ve afectado por las propiedades edáficas y las prácticas de manejo. En el ámbito biológico, las lombrices constituyeron los organismos más numerosos con predominio en los sistemas silvopastoriles y arboleda. Los valores superiores de densidad y biomasa de oligoquetos y mayor diversidad de otros individuos de la macrofauna, indican que la presencia de árboles en los pastizales de gramínea potencia y diversifican las comunidades de macroinvertebrados del suelo. El sistema de labranza sin inversión del prisma propicia una mejor calidad física del suelo, manteniendo el carbono e incrementando los rendimientos del Penisetum purpureum cv CUBA CT 115. La labranza tradicional, a base de aradura y grada, afecta a los contenidos de materia orgánica en el corto plazo y mantiene capas compactas en el horizonte subyacente, además influye desfavorablemente al flujo del aire, agua y al desarrollo radical de los pastos. La aplicación de compost favoreció mejores resultados productivos en ambas tecnologías de manejo. Los resultados alcanzados recomiendan la implantación de tecnologías de manejo conservacionistas y la aplicación de materiales orgánicos que restituyan los elementos nutricionales requeridos por los pastos, por lo que no se justifica la continuidad del uso de prácticas tradicionales de laboreo con inversión del prisma que se realizan actualmente. ABSTRACT The soil quality is an assessment tool, which could facilitate the adaptation of management practices that promote sustainable agricultural systems. The present investigation was carried out with a participatory diagnostic in twelve rural communities from Las Tunas province in 2009, in which producers identified the critical soil quality points of region and served as a starting point to select the physical, chemical and biological variables, in order to determine on five agricultural used systems (grove, natural grass, cultivated grass and two silvopastoral systems) in La Veguita zone from municipality Las Tunas. The system grove was used as reference of natural soil properties. The natural grass is distinguished by the development of low-productivity species, however the cultivated grass is represented by Pennisetum purpureum vc CUBA CT-115, and is a contribution to the biomass banks technology, in order to use in grazing during the dry season. The silvopastoral systems are represented by Leucaena leucocephala Lam. in stripes and Panicum maximum cv. Likoni, which differ in their design, handling and mineralogical properties. The main aim of this study was to assess the quality indicators for haplic Luvisols on granitoids for designing and implementing management technologies in order to increase the agroproductive capacity of soils. A minimal set of physical, chemical and biological indicators by Principal Component Analysis was obtained, which provided some useful information regarding soil processes and their integration for determining an index of quality. In the use system, characterized for the cultivated grass (Pennisetum purpureum) a short term assay in experimental plots was established, where the traditional and prism without inversion tillage were compared with and without compost application. In both tillage systems were evaluated the crop development and soil quality indicators. The results showed that the studied soil indicators set, six were selected, specifically the ones with exchangeable cationic capacity, organic matter, interchangeable potassium, sand content, bulk density and earthworm biomass, which explained the higher variability and served as the basis for evaluating the soil quality. The Reference threshold values of quality indicators for evaluating and monitoring the use and management systems from the region were established. The silvopastoral system 2 had the highest quality soil index, taking of reference the grove system for its natural condition. The silvopastoral management influenced on better productive results, but the soil characteristics, particularly the physical properties to be defined its design and management. However, the cultivated grass system with Pennisetum purpureum vc CUBA CT 115, reached the greatest accumulation of organic carbon. However, the management limited its physical quality and productive performance of the system. In addition, the use systems do not guarantee an index of soil quality, since it is affected by soil properties and management practices. From the biological aspect, the earthworms are the most numerous organisms on the silvopastoral systems and grove. The higher values of oligochaetes biomass and density and the greater diversity of other organisms from macrofauna indicate that the tree presence on the pasture grasses allows enhancing and diversifying soil macro invertebrate communities. The non-inversion prism tillage system provides a better physical quality of soil, maintaining the carbon content and increasing the yields of Penisetum purpureum vc CUBA CT 115. The traditional tillage, using the plowing and harrowing affects the organic matter content in a short term and keeps on compact layers of underlying horizon, and adversely affects the air and water flow, and pasture radical development. The compost application favored the best production results in both management technologies. The results obtained recommend the implementation of conservation management technologies and the application of organic materials that restore the nutritional elements required by the pasture, so it does not justify the continued use of traditional tillage practices with prism investment that are currently being made.

Molecular evolution of integrins: Genes encoding integrin β subunits from a coral and a sponge

The integrin family of cell surface receptors is strongly conserved in higher animals, but the evolutionary history of integrins is obscure. We have identified and sequenced cDNAs encoding integrin β subunits from a coral (phylum Cnidaria) and a sponge (Porifera), indicating that these proteins existed in the earliest stages of metazoan evolution. The coral βCn1 and, especially, the sponge βPo1 sequences are the most divergent of the “β1-class” integrins and share a number of features not found in any other vertebrate or invertebrate integrins. Perhaps the greatest difference from other β subunits is found in the third and fourth repeats of the cysteine-rich stalk, where the generally conserved spacings between cysteines are highly variable, but not similar, in βCn1 and βPo1. Alternatively spliced cDNAs, containing a stop codon about midway through the full-length translated sequence, were isolated from the sponge library. These cDNAs appear to define a boundary between functional domains, as they would encode a protein that includes the globular ligand-binding head but would be missing the stalk, transmembrane, and cytoplasmic domains. These and other sequence comparisons with vertebrate integrins are discussed with respect to models of integrin structure and function.

Midgut-specific immune molecules are produced by the blood-sucking insect Stomoxys calcitrans

We have cloned and sequenced two defensins, Smd1 and Smd2, from anterior midgut tissue of the blood-sucking fly Stomoxys calcitrans. The DNA and N-terminal protein sequences suggest both are produced as prepropeptides. Smd1 differs from the classic defensin pattern in having an unusual six-amino acid-long N-terminal sequence. Both Smd1 and Smd2 have lower pI points and charge than insect defensins derived from fat body/hemocytes. Northern analysis shows both of these defensin molecules are tissue specific; both are produced by the anterior midgut tissue and, unlike the other insect defensins reported to date, neither appears to be expressed in fat body or hemocytes. Northern analysis also shows that mRNAs for both defensins are constitutively produced in the anterior midgut tissues and that these transcripts are up-regulated in response to sterile as well as a lipopolysaccharide-containing blood meal. However, anti-Gram-negative biological activity in the midgut is substantially enhanced by lipopolysaccharide. These findings suggest that the insect midgut has its own tissue-specific immune mechanisms and that this invertebrate epithelium is, like several vertebrate epithelia, protected by specific antibacterial peptides.

Three novel families of miniature inverted-repeat transposable elements are associated with genes of the yellow fever mosquito, Aedes aegypti

Three novel families of transposable elements, Wukong, Wujin, and Wuneng, are described in the yellow fever mosquito, Aedes aegypti. Their copy numbers range from 2,100 to 3,000 per haploid genome. There are high degrees of sequence similarity within each family, and many structural but not sequence similarities between families. The common structural characteristics include small size, no coding potential, terminal inverted repeats, potential to form a stable secondary structure, A+T richness, and putative 2- to 4-bp A+T-biased specific target sites. Evidence of previous mobility is presented for the Wukong elements. Elements of these three families are associated with 7 of 16 fully or partially sequenced Ae. aegypti genes. Characteristics of these mosquito elements indicate strong similarities to the miniature inverted-repeat transposable elements (MITEs) recently found to be associated with plant genes. MITE-like elements have also been reported in two species of Xenopus and in Homo sapiens. This characterization of multiple families of highly repetitive MITE-like elements in an invertebrate extends the range of these elements in eukaryotic genomes. A hypothesis is presented relating genome size and organization to the presence of highly reiterated MITE families. The association of MITE-like elements with Ae. aegypti genes shows the same bias toward noncoding regions as in plants. This association has potentially important implications for the evolution of gene regulation.

Nerve growth factor acutely reduces chemical transmission by means of postsynaptic TrkA-like receptors in squid giant synapse

Tyrosine phosphorylation has been shown to be an important modulator of synaptic transmission in both vertebrates and invertebrates. Such findings hint toward the existence of extracellular ligands capable of activating this widely represented signaling mechanism at or close to the synapse. Examples of such ligands are the peptide growth factors which, on binding, activate receptor tyrosine kinases. To gain insight into the physiological consequences of receptor tyrosine kinase activation in squid giant synapse, a series of growth factors was tested in this preparation. Electrophysiological, pharmacological, and biochemical analysis demonstrated that nerve growth factor (NGF) triggers an acute and specific reduction of the postsynaptic potential amplitude, without affecting the presynaptic spike generation or presynaptic calcium current. The NGF target is localized at a postsynaptic site and involves a new TrkA-like receptor. The squid receptor crossreacts with antibodies generated against mammalian TrkA, is tyrosine phosphorylated in response to NGF stimulation, and is blocked by specific pharmacological inhibitors. The modulation described emphasizes the important role of growth factors on invertebrate synaptic transmission.

Retinoic acid X receptor in the diploblast, Tripedalia cystophora

Nuclear hormone receptors comprise a characteristic family of transcription factors found in vertebrates, insects and nematodes. Here we show by cDNA and gene cloning that a Cnidarian, Tripedalia cystophora, possesses a retinoid receptor (jRXR) with remarkable homology to vertebrate retinoic acid X receptors (RXRs). Like vertebrate RXRs, jRXR binds 9-cis retinoic acid (Kd = 4 × 10−10 M) and binds to the DNA sequence, PuGGTCA as a monomer in vitro. jRXR also heterodimerizes with Xenopus TR beta on a thyroid responsive element of a direct repeat separated by 4 bp. A jRXR binding half-site capable of interacting with (His6)jRXR fusion protein was identified in the promoters of three T. cystophora crystallin genes that are expressed highly in the eye lens of this jellyfish. Because crystallin gene expression is regulated by retionoid signaling in vertebrates, the jellyfish crystallin genes are candidate in vivo targets for jRXR. Finally, an antibody prepared against (His6)jRXR showed that full-length jRXR is expressed at all developmental stages of T. cystophora except the ephydra, where a smaller form replaces is. These data show that Cnidaria, a diploblastic phylum ancestral to the triploblastic invertebrate and subsequent vertebrate lineages, already have an RXR suggesting that RXR is an early component of the regulatory mechanisms of metazoa.

Noninvasive measurement of gene expression in skeletal muscle

We have developed a noninvasive detection method for expression of viral-mediated gene transfer. A recombinant adenovirus was constructed by using the gene for arginine kinase (AK), which is the invertebrate correlate to the vertebrate ATP-buffering enzyme, creatine kinase. Gene expression was noninvasively monitored using 31P-magnetic resonance spectroscopy (31P-MRS). The product of the AK enzyme, phosphoarginine (PArg), served as an MRS-visible reporter of AK expression. The recombinant adenovirus coding for arginine kinase (rAdCMVAK) was injected into the right hindlimbs of neonatal mice. Two weeks after injection of rAdCMVAK, a unique 31P-MRS resonance was observed. It was observable in all rAdCMVAK injected hindlimbs and was not present in the contralateral control or the vehicle injected limb. PArg and phosphocreatine (PCr) concentrations were calculated to be 11.6 ± 0.90 and 13.6 ± 1.1 mM respectively in rAdCMVAK injected limbs. AK activity was demonstrated in vivo by monitoring the decreases in PArg and ATP resonances during prolonged ischemia. After 1 h of ischemia intracellular pH was 6.73 ± 0.06, PCr/ATP was decreased by 77 ± 8%, whereas PArg/ATP was decreased by 50 ± 15% of basal levels. PArg and PCr returned to basal levels within 5 min of the restoration of blood flow. AK activity persisted for at least 8 mo after injection, indicating that adenoviral-mediated gene transfer can produce stable expression for long periods of time. Therefore, the cDNA encoding AK provides a useful reporter gene that allows noninvasive and repeated monitoring of gene expression after viral mediated gene transfer to muscle.

The role of phosphoglycans in Leishmania–sand fly interactions

Leishmania promastigotes synthesize an abundance of phosphoglycans, either attached to the cell surface through phosphatidylinositol anchors (lipophosphoglycan, LPG) or secreted as protein-containing glycoconjugates. These phosphoglycans are thought to promote the survival of the parasite within both its vertebrate and invertebrate hosts. The relative contributions of different phosphoglycan-containing molecules in Leishmania–sand fly interactions were tested by using mutants specifically deficient in either total phosphoglycans or LPG alone. Leishmania donovani promastigotes deficient in both LPG and protein-linked phosphoglycans because of loss of LPG2 (encoding the Golgi GDP-Man transporter) failed to survive the hydrolytic environment within the early blood-fed midgut. In contrast, L. donovani and Leishmania major mutants deficient solely in LPG expression because of loss of LPG1 (involved in biosynthesis of the core oligosaccharide LPG domain) had only a slight reduction in the survival and growth of promastigotes within the early blood-fed midgut. The ability of the LPG1-deficient promastigotes to persist in the midgut after blood meal excretion was completely lost, and this defect was correlated with their inability to bind to midgut epithelial cells in vitro. For both mutants, when phosphoglycan expression was restored to wild-type levels by reintroduction of LPG1 or LPG2 (as appropriate), then the wild-type phenotype was also restored. We conclude, first, that LPG is not essential for survival in the early blood-fed midgut but, along with other secreted phosphoglycan-containing glycoconjugates, can protect promastigotes from the digestive enzymes in the gut and, second, that LPG is required to mediate midgut attachment and to maintain infection in the fly during excretion of the digested blood meal.