Cryptic diversity and ecosystem functioning: a complex tale of differential effects on decomposition.

Marine ecosystems are experiencing accelerating population and species loss. Some ecosystem functions are decreasing and there is growing interest in the link between biodiversity and ecosystem functioning. The role of cryptic (morphologically identical but genetically distinct) species in this biodiversity-ecosystem functioning link is unclear and has not yet been formally tested. We tested if there is a differential effect of four cryptic species of the bacterivorous nematode Litoditis marina on the decomposition process of macroalgae. Bacterivorous nematodes can stimulate or slow down bacterial activity and modify the bacterial assemblage composition. Moreover, we tested if interspecific interactions among the four cryptic species influence the decomposition process. A laboratory experiment with both mono- and multispecific nematode cultures was conducted, and loss of organic matter and the activity of two key extracellular enzymes for the degradation of phytodetritus were assessed. L. marina mainly influenced qualitative aspects of the decomposition process rather than its overall rate: an effect of the nematodes on the enzymatic activities became manifest, although no clear nematode effect on bulk organic matter weight loss was found. We also demonstrated that species-specific effects on the decomposition process existed. Combining the four cryptic species resulted in high competition, with one dominant species, but without complete exclusion of other species. These interspecific interactions translated into different effects on the decomposition process. The species-specific differences indicated that each cryptic species may play an important and distinct role in ecosystem functioning. Functional differences may result in coexistence among very similar species.

Coexisting cryptic species of the Litoditis marina complex (Nematoda) show differential resource use and have distinct microbiomes with high intraspecific variability.

Differences in resource use or in tolerances to abiotic conditions are often invoked as potential mechanisms underlying the sympatric distribution of cryptic species. Additionally, the microbiome can provide physiological adaptations of the host to environmental conditions. We determined the intra- and interspecific variability of the microbiomes of three cryptic nematode species of the Litoditis marina species complex that co-occur, but show differences in abiotic tolerances. Roche 454 pyrosequencing of the microbial 16S rRNA gene revealed distinct bacterial communities characterized by a substantial diversity (85-513 OTUs) and many rare OTUs. The core microbiome of each species contained only very few OTUs (2-6), and four OTUs were identified as potentially generating tolerance to abiotic conditions. A controlled experiment in which nematodes from two cryptic species (Pm1 and Pm3) were fed with either an E. coli suspension or a bacterial mix was performed, and the 16S rRNA gene was sequenced using the MiSeq technology. OTU richness was 10-fold higher compared to the 454 data set and ranged between 1118 and 7864. This experiment confirmed the existence of species-specific microbiomes, a core microbiome with few OTUs, and high interindividual variability. The offered food source affected the bacterial community and illustrated different feeding behaviour between the cryptic species, with Pm3 exhibiting a higher degree of selective feeding than Pm1. Morphologically similar species belonging to the same feeding guild (bacterivores) can thus have substantial differences in their associated microbiomes and feeding strategy, which in turn may have important ramifications for biodiversity-ecosystem functioning relationships.

The effect of recombinant human bone morphogenetic protein-2 in single-level posterior lumbar interbody arthrodesis.

BACKGROUND: In this prospective, randomized controlled trial, our objective was to assess both the clinical and radiographic effect of recombinant human bone morphogenetic protein-2 (rhBMP-2) in patients treated with an instrumented single-level posterior lumbar interbody arthrodesis with polyetheretherketone cages. METHODS: Forty patients were randomized with a 1:1 ratio. Two patients who had a two-level arthrodesis (L4-L5 and L5-S1) were excluded. Patients completed the Oswestry Disability Index, the Short Form-36, and the visual analog scale preoperatively and postoperatively at three, six, twelve, and twenty-four months. Computed tomography scans with coronal and sagittal reconstructions were made at three, six, and twelve months postoperatively. Interbody arthrodesis was performed using polyetheretherketone cages, which were filled with 8 mg of rhBMP-2 in the study group and 2.5 mL of autologous bone in the control group. RESULTS: Baseline demographic data showed no significant difference between groups, except for the body mass index, which was higher in the study group (p = 0.032). There were no significant differences in the clinical results (visual analog scale, Oswestry Disability Index, and Short Form-36) between the groups at each postoperative visit. At three months, end-plate resorption was noted around the cages filled with rhBMP-2 in all patients in the study group. No cage migration or subsidence was observed. Bridging trabecular bone scale scores and bone density measures were significantly lower in the study group. Osteolysis and ectopic bone formation occurred in seven of nineteen patients in the study group and did not occur in the control group. This did not result in radicular symptoms within the time span of this study. At one year, computed tomography scans showed osseous healing in all patients. There were no revision procedures. CONCLUSIONS: This trial showed no clinical difference when rhBMP-2 was used in posterior lumbar interbody arthrodesis compared with autologous bone. On computed tomography scans, fusion was equally achieved, but trabecular bone formation occurred at a slower rate and interbody bone density was lower within the first year after surgery when rhBMP-2 was used. End-plate resorption, osteolysis, and ectopic bone formation were frequently noted in the rhBMP-2 group.

Investigating the ecology and evolution of cryptic marine nematode species through quantitative real-time PCR of the ribosomal ITS region.

The presence of morphologically similar but genetically distinct species has impacted biogeographical and ecological paradigms. In marine sediments, free-living nematodes form one of the most abundant and diverse faunal groups. Inferring the importance of nematode diversity for ecosystem functioning requires species-level identification, which is hampered by the lack of easily observable diagnostic characters and the presence of cryptic species. New techniques are urgently needed to adequately study the ecology and evolution of cryptic species. The aim of the present study was to evaluate the potential of a quantitative real-time PCR (qPCR) assay using the internal transcribed spacer (ITS) region of the ribosomal DNA to detect and quantify cryptic species of the R. (P.) marina complex. All primer pairs proved to be highly specific, and each primer pair was able to detect a single juvenile in a pool of 100 nematodes. C(t) values were significantly different between developmental stages for all species except for PmIII. Despite differences between developmental stages, a strong correlation was observed between the amount of extracted DNA and the number of nematodes present. Relative and absolute quantification estimates were comparable and resulted in strong positive correlations between the qPCR estimate and the actual number of nematodes present in the samples. The qPCR assay developed here provides the ability to quickly identify and quantify cryptic nematode species and will facilitate their study in laboratory and field settings.

Factors affecting distribution and mobility of trace elements (Cu, Pb, Zn) in a perennial grapevine (Vitis vinifera L.) in the Champagne region of France.

Soil and Vitis vinifera L. (coarse and fine roots, leaves, berries) concentration and geochemical partitioning of Cu, Pb and Zn were determined in a contaminated calcareous Champagne plot to assess their mobility and transfer. Accumulation ratios in roots remained low (0.1-0.4 for Cu and Zn, <0.05 for Pb). Differences between elements resulted from vegetation uptake strategy and soil partitioning. Copper, significantly associated with the oxidisable fraction (27.8%), and Zn with the acid soluble fraction (33.3%), could be mobilised by rhizosphere acidification and oxidisation, unlike Pb, essentially contained in the reducible fraction (72.4%). Roots should not be considered as a whole since the more reactive fine roots showed higher accumulation ratios than coarse ones. More sensitive response of fine roots, lack of correlation between chemical extraction results and vegetation concentrations, and very limited translocation to aerial parts showed that fine root concentrations should be used when assessing bioavailability.

Pulmonary vagal innervation is required to establish adequate alveolar ventilation in the newborn lamb.

To investigate the effects of bilateral intrathoracic vagotomy on the establishment of continuous breathing and effective gas exchange at birth, we studied 8 chronically instrumented, unanesthetized, sham-operated and 14 vagotomized newborn lambs after a spontaneous, unassisted vaginal delivery. Fetal lambs were instrumented in utero to record sleep states, diaphragmatic electromyogram, blood pressure, arterial pH, and blood-gas tensions. Six of eight sham-operated lambs established effective gas exchange within 10 min of birth, whereas 12 of 14 vagotomized animals developed respiratory acidosis and hypoxemia (P = 0.008). Breathing frequency in vagotomized newborns was significantly lower during the entire postnatal period compared with sham-operated newborns. Vagotomized subjects also remained hypothermic during the entire postnatal period (P < 0.05). Bronchoalveolar lavage indicated an increased minimum surface tension, whereas lung histology showed perivascular edema and partial atelectasis in the vagotomized group. We conclude that stimulation of breathing and effective gas exchange are critically dependent on intact vagal nerves during the transition from fetal to neonatal life.

Umbilical arterial blood flow and plasma prostaglandin E2 concentrations during arousal and breathing movements in fetal sheep.

To investigate the effects of lung distension and oxygenation on umbilical blood flow (UBF) and plasma prostaglandin E2 (PGE2) in relation to arousal and stimulation of breathing movements, we studied eight chronically instrumented, unanesthetized fetal sheep between 137 and 143 d of gestation. Electrocorticogram, electro-oculogram, nuchal and diaphragmatic electromyograms, arterial pH and blood gas tensions, Hb oxygen saturation, body temperature, and UBF were recorded in each fetus. Electrocorticogram, electro-oculogram, and nuchal electromyograms were used to define sleep states. No sooner than 4 d after surgery, fetal lungs were distended with 100% O2 or N2 in a randomized order via an in situ Y-endotracheal tube. PGE2 concentrations were analyzed by RIA. A significant increase in fetal arousal and stimulation of breathing during nonrapid eye movement sleep was observed during lung distension with O2 as compared with control periods and lung distension with nitrogen. In all sleep states, UBF significantly decreased during oxygenation as compared with the control values. However, no significant correlation was observed between the time of the onset of arousal and the decrease in UBF. Lung distension with N2 resulted in increased plasma PGE2 concentrations, whereas, no change was observed during oxygenation. Our data suggest that an increase in fetal partial pressure of arterial O2 leads to a decrease in UBF. However, the onset of arousal and stimulation of breathing during lung distension and oxygenation were not dependent on a decrease in plasma PGE2 concentrations.

Effect of bilateral vagotomy on oxygenation, arousal, and breathing movements in fetal sheep.

To investigate the effects of bilateral cervical vagotomy on arousal and breathing responses, we studied eight sham-operated and eight chronically instrumented unanesthetized vagotomized sheep fetuses between 136 and 144 days of gestation (term approximately 147 days). Each fetus was instrumented to record sleep states, diaphragmatic electromyogram, blood pressure, pH, and blood gas tensions. In a randomized order, fetal lungs were distended with four different O2 concentrations: 0 (100% N2), 21, 50, and 100% at a continuous positive airway pressure of 30 cmH2O via an in situ Y-endotracheal tube. Under control conditions, inspiratory time and the duration of the single longest breathing episode decreased from 598 +/- 99 (SD) ms and 24 +/- 10 min in sham group to 393 +/- 162 ms and 11.0 +/- 3.0 min in vagotomized group (P = 0.04 and 0.033), respectively. In response to lung distension with 100% N2, breathing time decreased from 44 +/- 17 to 20 +/- 18% (P = 0.045) in sham-operated fetuses, whereas it remained unchanged in the vagotomized group. In response to 100% O2, fetal arterial PO2 increased in five of eight fetuses sham-operated from 18.2 +/- 5.1 to 227 +/- 45 Torr (P = 0.0001) and in six of eight vagotomized fetuses from 18.5 +/- 4.4 to 172 +/- 39 Torr (P < 0.001). Although arousal was observed in all oxygenated fetuses at the onset of breathing, the duration of arousal was markedly attenuated in vagotomized fetuses (14 +/- 10 vs. 46 +/- 29 min in sham group; P = 0.024). Frequency and amplitude of breathing and respiratory output (frequency x amplitude) increased only in sham group (P = 0.02, 0.004, and 0.0002, respectively). We conclude that in response to lung distension and oxygenation, arousal and stimulation of breathing during active and quite sleep are critically dependent on intact vagal nerves.

Arterial oxygen tension threshold range for the onset of arousal and breathing in fetal sheep.

Mechanisms for the control of episodic fetal breathing movements or the onset of continuous breathing at birth remain unknown. Lung distension with 100% O2 at a continuous positive airway pressure of 30 cm H2O may induce arousal and continuous breathing. To investigate 1) the threshold range of arterial oxygen tension (PaO2) for the onset of arousal and breathing and 2) the graded response of breathing to various levels of PaO2, we studied 10 fetal sheep between 135 and 142 d of gestation (term = 147 +/- 2 d). Each fetus was instrumented to record sleep states, diaphragmatic electromyogram, arterial pH, and blood gas tensions. PaO2 threshold was determined through an indwelling O2 sensor catheter. Fetal lungs were distended at a continuous positive airway pressure of 40 cm H2O with 100% N2 or with O2 ranging from 40 to 100% via an in situ endotracheal tube. At the onset of arousal (n = 10), PaO2, arterial carbon dioxide tension, and Hb O2 saturation increased from control values of 21.7 +/- 0.75 torr (2.9 +/- 0.09 kPa), 41.8 +/- 1.1 torr (5.47 +/- 0.15 kPa), and 52.9 +/- 2.6% to 65.6 +/- 9.6 torr (8.74 +/- 1.28 kPa), 46.9 +/- 1.3 torr (6.25 +/- 0.17 kPa), and 92.9 +/- 2.06%, respectively, whereas the pH decreased from 7.31 +/- 0.006 to 7.27 +/- 0.009 (mean +/- SEM; p = 0.001, 0.04, 0.002, and 0.001, respectively). Seven of 10 fetuses breathed continuously. In these fetuses, PaO2 and arterial carbon dioxide tension further increased and pH decreased; however, no further significant increase in Hb O2 saturation was observed.(ABSTRACT TRUNCATED AT 250 WORDS)

The effects of lung distension, oxygenation, and gestational age on fetal behavior and breathing movements in sheep.

Lung distension with 100% O2 at a continuous positive airway pressure of 30 cm H2O may induce continuous fetal breathing movements (FBM) in sheep. The objectives of this study were 1) to investigate the relative roles of lung distension and oxygenation and 2) to test the hypothesis that FBM can be induced during labor, when normally they are greatly reduced or absent. We studied 13 chronically instrumented, unanesthetized fetal sheep between 128 and 144 d of gestation (term = 147 +/- 2 d). Each fetus was instrumented to record sleep states, diaphragm electromyogram, blood pressure, arterial pH, and blood gas tensions. The fetal lungs were distended via an in situ endotracheal tube with four different concentrations of O2 (0, 21, 50 and 100%) at a continuous positive airway pressure of 10, 20 and 30 cm H2O in a randomized order. No change in any recorded physiologic variable was observed at 129 +/- 1 or 132 +/- 1 d of gestation. At 135 +/- 1 and 138 +/- 1 d, in response to a continuous positive airway pressure of 30 cm H2O and 100% O2, pH decreased (p = 0.0004 and 0.005, respectively) and arterial O2 tension increased (p = 0.004 and 0.02, respectively). However, increases in 1) breathing time, 2) breathing time/low-voltage electrocortical activity ratio, 3) duration of arousal, and 4) length of single breathing epochs were observed only at 138 +/- 1 d. Lung distension with N2 resulted in a decrease in FBM. Six fetuses were studied during labor.(ABSTRACT TRUNCATED AT 250 WORDS)

The tricho-rhino-phalangeal syndrome revisited.

The tricho-rhino-phalangeal syndrome (TRPS) is a rare congenital disorder, characterized by (1) a peculiar and somewhat pear-shaped nose, (2) sparse and brittle scalp hair, and (3) radiographic evidence of cone-shaped epiphyses of the hands. On the basis of clinical, radiographic and genetic criteria, two subtypes (type I and II) are discerned. We describe an intermediate "hybrid" variant of the TRPS in a patient with clinical and radiographic features of TRPS type I, but with a clearly abnormal karyotype, consistent with TRPS type II. The radiographic findings of the syndrome are reviewed, with particular emphasis on the cone-shaped epiphyses in the hands, the changes in the coxo-femoral joints and the atypical appearance of the pubic symphysis.