Beneficial microorganisms for corals (BMCs) ameliorate environmental stress, but whether they can prevent mortality and the underlying host response mechanisms remains elusive. Here, we conducted omics analyses on the coral Mussismilia hispida exposed to bleaching conditions in a long-term mesocosm experiment and inoculated with a selected BMC consortium or a saline solution placebo. All corals were affected by heat stress, but the observed “post-heat stress disorder” was mitigated by BMCs, signified by patterns of dimethylsulfoniopropionate degradation, lipid maintenance, and coral host transcriptional reprogramming of cellular restructuration, repair, stress protection, and immune genes, concomitant with a 40% survival rate increase and stable photosynthetic performance by the endosymbiotic algae. This study provides insights into the responses that underlie probiotic host manipulation. We demonstrate that BMCs trigger a dynamic microbiome restructuring process that instigates genetic and metabolic alterations in the coral host that eventually mitigate coral bleaching and mortality.
The use of morphine, the standard opioid drug, is limited by its undesirable effects, such as tolerance, physical dependence, and hyperalgesia (increased pain sensitivity). Clinical and preclinical studies have reported development of hyperalgesia after prolonged opioid administration or after a single dose of intrathecal (i.t.) morphine in uninjured rats. However, whether a single standard systemic morphine dose is sufficient to decrease the nociceptive threshold in rats is unknown. Here, we showed that a single morphine subcutaneous injection induces analgesia followed by a long-lasting delayed hyperalgesia in uninjured and PGE2 sensitized rats. The i.t injection of extracellular signal-regulated kinase (ERK) inhibitor blocked morphine-induced analgesia, without interfering with the morphine-induced hyperalgesia. However, i.t. injection of SB20358, a p38 inhibitor and SP660125, a JNK inhibitor, decreased the morphine-induced hyperalgesia. Consistently with the behavioral data, Western Blot analysis showed that ERK is more phosphorylated 1 h after morphine, i.e., when the analgesia is detected. Moreover, phospho-p38 and phospho-JNK levels are upregulated 96 h after morphine injection, time that coincides with the hyperalgesic effect. Intrathecal (i.t.) oligodeoxynucleotide (ODN) antisense to cAMP-responsive element binding protein (CREB) attenuated morphine-induced hyperalgesia. Real-time polymerase chain reaction (RT-PCR) analysis showed that CREB downstream genes expressions were significantly up-regulated 96 h after morphine injection in spinal cord. Together, our data suggest that central ERK is involved in the analgesic and hyperalgesic effects of morphine while JNK, p38, and CREB are involved in the morphine-induced delayed hyperalgesia.
Crotalphine (CRP) is a structural analogue to a peptide that was first identified in the crude venom from the South American rattlesnake Crotalus durissus terrificus. This peptide induces a potent and long-lasting antinociceptive effect that is mediated by the activation of peripheral opioid receptors. The opioid receptor activation regulates a variety of intracellular signaling, including the mitogen-activated protein kinase (MAPK) pathway. Using primary cultures of sensory neurons, it was demonstrated that crotalphine increases the level of activated ERK1/2 and JNK-MAPKs and this increase is dependent on the activation of protein kinase Cζ (PKCζ). However, whether PKCζ-MAPK signaling is critical for crotalphine-induced antinociception is unknown. Here, we biochemically demonstrated that the systemic crotalphine activates ERK1/2 and JNK and decreases the phosphorylation of p38 in the lumbar spinal cord. The in vivo pharmacological inhibition of spinal ERK1/2 and JNK, but not of p38, blocks the antinociceptive effect of crotalphine. Of interest, the administration of a PKCζ pseudosubstrate (PKCζ inhibitor) prevents crotalphine-induced ERK activation in the spinal cord, followed by the abolishment of crotalphine-induced analgesia. Together, our results demonstrate that the PKCζ-ERK signaling pathway is involved in crotalphine-induced analgesia. Our study opens a perspective for the PKCζ-MAPK axis as a target for pain control.
Deep subsurface microbial communities are more abundant in coarse-grained sedimentary rocks such as sandstones than in fine-grained mudstones. The low porosity and low permeability of mudstones are believed to restrict microbial life. Then, it is expected that distinct, isolated microbial communities may form in sandstones separated by mudstones. In this context, the connectivity between microbial communities in different sandstone units can be investigated to infer evolutionary patterns of diversification in space-time, which may potentially contribute with relevant data for analyses of hydraulic connectivity and stratigraphic correlation. In this work, we used high throughput DNA sequencing of a ribosomal 16S gene fragment to characterize the prokaryotic communities found in Permian sandstone samples of the same core that are separated by one mudstone interval, in the Charqueadas coal field, Parana Basin (Southern Brazil). Our samples were collected at ~300 m deep, in porous sandstones separated by a thick mudstone package. Differences in the bacterial community structure between samples were observed for the classified OTUs, from phylum to genus. Molecular biology might be further applied as a possible tool to help to understand the spatial and temporal distribution of depositional facies, and the efficiency of low permeability rocks to compartmentalize reservoirs. Ongoing studies aim to extend the present investigation into further analyses regarding lateral changes in microbial communities present in the same sandstone units.
Anaerobic digestion (AD) is a process resulting from the anaerobic metabolism of specific microorganisms that produce an eco-friendly type of energy and a stabilized soil fertilizer. We described the microbial communities and their changes in three depths of BioKöhler® biodigester, fed with cattle manure for 18 days, under anaerobic incubation at the psychrophilic temperature range (~20°C). During the experiment, the maximum methane content in the raw biogas was 79.9%. Non-metric multidimensional scaling (MDS) showed significant differences among microbial communities in the bottom, medium, and upper depths. Considering all the periods of incubation, the microbial communities changed until the eighth day, and they remained stable from eighth to seventeenth days. Bacteroidetes, Firmicutes, and Synergistetes were the most abundant phyla in samples, representing approximately 41% of the total OTUs. The relative abundance of the phyla Euryarchaeota, Actinobacteria, Firmicutes, and Verrucomicrobia changed from bottom to medium sampling points. Moreover, Crenarchaeota differed in frequencies from medium to upper, and Acidobacteria from bottom to upper samples. Lentisphaerae, Chloroflexi, and LD1 were different solely at the bottom, whereas OP9 and Tenericutes only in the medium. Psychrophilic AD performed in this work removed pathogens like Salmonella and Escherichia, as observed at the digestate analyzed. This type of treatment of raw manure besides producing eco-friendly energy efficiently also generates a stabilized and safe biomass that can be used as fertilizer in soils.
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