Marine Extremophiles: A Source of Hydrolases for Biotechnological Applications

The marine environment covers almost three quarters of the planet and is where evolution took its first steps. Extremophile microorganisms are found in several extreme marine environments, such as hydrothermal vents, hot springs, salty lakes and deep-sea floors. The ability of these microorganisms to support extremes of temperature, salinity and pressure demonstrates their great potential for biotechnological processes. Hydrolases including amylases, cellulases, peptidases and lipases from hyperthermophiles, psychrophiles, halophiles and piezophiles have been investigated for these reasons. Extremozymes are adapted to work in harsh physical-chemical conditions and their use in various industrial applications such as the biofuel, pharmaceutical, fine chemicals and food industries has increased. The understanding of the specific factors that confer the ability to withstand extreme habitats on such enzymes has become a priority for their biotechnological use. The most studied marine extremophiles are prokaryotes and in this review, we present the most studied archaea and bacteria extremophiles and their hydrolases, and discuss their use for industrial applications.     

Sulfated Steroid–Amino Acid Conjugates from the Irish Marine Sponge Polymastia boletiformis

Antifungal bioactivity-guided fractionation of the organic extract of the sponge Polymastia boletiformis, collected from the west coast of Ireland, led to the isolation of two new sulfated steroid-amino acid conjugates (1 and 2). Extensive 1D and 2D NMR analyses in combination with quantum mechanical calculations of the electronic circular dichroism (ECD) spectra, optical rotation, and ¹³C chemical shifts were used to establish the chemical structures of 1 and 2. Both compounds exhibited moderate antifungal activity against Cladosporium cucumerinum, while compound 2 was also active against Candida albicans. Marine natural products containing steroidal and amino acid constituents are extremely rare in nature.     

Dietary fatty acid enrichment during the spawning season increases egg viability and quality in Hippocampus hippocampus

Control of the reproductive cycle is essential to achieve a successful aquaculture industry. Seahorse reared in captivity often shows eggs of poor quality and low offspring survival due to unsuitable rearing and nutritional protocols. This study aimed to evaluate the effect of dietary enrichment, added exclusively during the spawning season, on the egg quality of Hippocampus hippocampus. The effect of enriching mysis throughout the spawning season was analysed in terms of female fecundity, egg viability, egg size and fatty acid profile. The results showed that fecundity was not affected by the enrichment of the diet, but egg viability was increased when females were fed the enriched mysis (84.9 ± 4.1% compared to 62.2 ± 9.4%). Moreover, the content of several PUFA (20:4n‐6, 18:2n‐6, 22:5n‐3, 22:5n‐6, sum of n‐6) decreased throughout the spawning season, especially in the last clutch, in eggs originated from females fed non‐enriched mysis. Eggs from females fed the enriched diet maintained the fatty acid profile throughout the spawning season. In conclusion, enrichment of the diet of H. hippocampus improved egg viability and avoided a decrease in the quality of eggs by maintaining PUFA content in the last clutches of the spawning season.     

Hepatogenous photosensitization associated with liver copper accumulation in buffalos

Four 1.5-year-old, male, Murrah buffalos were maintained during eight months without direct solar exposure during a study of copper toxicosis. Four days after solar exposure, all buffalos presented clinical manifestations consistent with acute photosensitization, including anorexia, apathy, loss of body weight, and generalized cutaneous lesions. Gross lesions were characterized by severe erythema, localized edema, fissures, tissue necrosis, gangrene and crust formation with serous exudation. Liver copper concentration was evaluated, and cutaneous biopsies were taken when clinical signs were evident. The liver copper concentration before solar exposure was increased in all animals. Histopathologic examination of the skin revealed hepatogenous photosensitization characterized by parakeratotic hyperkeratosis, acantholysis, degeneration of squamous epithelial cells, epidermal necrosis with atrophy of sweat glands, and multifocal superficial and deep dermal edema. These findings suggest that asymptomatic accumulation of copper within the liver might have induced hepatic insufficiency thereby resulting in secondary photosensitization when these buffalos were exposed to sunlight.     

竹子处理后的化学品可追朔性

盐溶液如CCA和CCB是用来防治竹子腐烂、虫蛀和白蚁的主要化学品。通过测定竹秆中化学品的总量,评价了这些方法的有效性。将大耳竹劈成条,浸泡在10%的CCB溶液中(31.75的铬,13%的铜和5.25%的硼),并评价了从竹条到浸入溶液之间的时间的影响。利用电子显微镜确定了化学品的可追朔性。结果证实,铬和铜氧化随着竹解剖分子不均匀分布。     

Morphological and photosynthetic alterations in the Yellow-ipe, <Emphasis Type="Italic">Tabebuia chrysotricha</Emphasis> (Mart. Ex DC.) Standl., under nursery shading and gas exchange after being transferred to full sunlight

The objective of this study was to evaluate the effect of nursery shading on the Yellow-ipe seedling (Tabebuia chrysotricha) growth, photosynthesis, and photosynthetic acclimation after being transferred into direct sunlight. The Yellow-ipe seedlings were grown under 0, 50, 70 and 95% shade. At the 134th day of sowing, leaf gas exchange and chlorophyll were measured under current growth shading, after exposure to 15 min and two day full sunlight. With the increase of shading, the Yellow-ipe seedlings allocated more biomass to the stem and leaves and less to the roots, and there was an increase in the leaf area ratio and specific leaf area. In relation to 0% of shading there was a increase of 211% in stem, 116% in leaf, and a reduction of 200% in roots biomass when seedling were grown under 95% of shading. The total biomass accumulation had a high correlation with collar diameter (r = 0.96). More than 70% of the shading reduced the photosynthesis, and after transferring the seedlings into full sunlight, more than 50% of the shading induced a reduction in chlorophyll, stomatal conductance, photosynthesis and instantaneous carboxylation efficiency, suggesting the presence of a photoinhibition process. The Yellow-ipe seedling growth under nursery conditions should not be done under more than 50% shading, which may result in the lower seedling quality and poorer acclimation to transplantation, particularly to severe degraded areas with direct sunlight. The species can be used for recovering from totally devastated forest areas to initial recovery when full canopy are forming.     

Influence of leaf position that correspond to whole plant severity and diagrammatic scale for white spot of corn

The main objective of this research is to determine the influence of leaf position on corn plants with white spot caused by Pantoea ananatis, which better represents the infection on the whole plant. A diagrammatic scale to quantify the severity of the disease was elaborated and validated. For scale elaboration, the minimal and maximal limits of the disease severity observed in the field were considered, and intermediate levels followed logarithmic increments according to the Weber–Fechner stimulation law. The scale has nine classes: 0.1, 1, 2, 4, 8, 16, 24, 32 and 64%. For scale evaluation, a severity evaluation for white spot was performed by 10 raters with no experience in disease evaluation. Initially, severity estimation was performed without a scale for 41 leaves with different levels of severity. Afterward, the same raters used the proposed diagrammatic scale. Through linear regression to compare the actual and estimate severity values, the raters’ accuracy and precision were analyzed. Satisfactory accuracy and precision were achieved when estimation was performed with a diagrammatic scale. To determine the best leaf disease severity evaluation, correlation and regression analyses were performed with 25 plants of five genotypes, for a total of 284 leaves analyzed. Results analysis leads us to conclude that the severity of white spot on corn plants significantly correlates with the disease mean severity of leaves 0 and -1, i.e., a leaf of the corn ear and the one immediately below it. This scale provided good levels of accuracy and precision (a mean R² of 94%), with errors concentrating around 10%. Raters presented increased reproducibility (R² > 90% in 82% of cases) of severity estimates. The proposed diagrammatic scale is considered adequate to estimate the severity of white spot in corn for germplasm evaluations, for epidemiological studies and for evaluation of control strategies for this disease.     

Ralstonia solanacearum decreases volumetric growth of trees and yield of kraft cellulose of Eucalyptus spp.

Ralstonia solanacearum is currently one of the most important plant pathogenic bacteria worldwide, with a wide geographical distribution and host diversity. The pathogen infects more than 200 plant species belonging to approximately 50 plant families, including Eucalyptus spp. Although, high losses have been reported in nurseries, little is known on the negative impact of the disease in the field. In this study, we evaluated the incidence of R. solanacearum and its effect on volumetric growth and cellulose yield of discoloured wood chips obtained from infected trees of one clone of Eucalyptus urophylla and two hybrid clones of Eucalyptus urophylla × E. grandis. The average incidence of bacterial wilt ranged between 60.6% and 72.4%. Volumetric growth of infected trees disease decreased 78.6% and 81.7% at 18 and 30 months, respectively. The pulp screen yield of three clones decreased between 3.2 and 6.4%, with an average 4.3%. The results of this work provide useful information on the losses of volumetric growth and pulp yield of eucalypt caused by R. solanacearum.     

Sufficiency ranges and crop nutrient removals for peppermint (Mentha X piperita L.) established from field and pot fertilizer experiments

Peppermint is an important aromatic and medicinal plant used across the world in pharmaceutical, cosmetic and food industries. However, there is a lack of agronomic research on this crop which hinders the implementation of best agricultural practice at farm level. Plant analysis, for instance, cannot be used as a tool to implement a suitable fertilizer recommendation program, since sufficiency ranges and crop nutrient removals have not yet been established. Thus, the main objectives of the present work were to assess the response of peppermint to varying nitrogen (N), phosphorus (P), potassium (K) and boron (B) rates, to establish sufficiency ranges from macro, micronutrients and SPAD-readings and to estimate crop nutrient removals in the aboveground biomass. Field trials and pot experiments were conducted from 2013 to 2015 in a wide range of conditions involving 12 N, P, K or B fertilizer trials and a total of 48 cuts of biomass. Nitrogen fertilization increased dry matter yield of peppermint on the vast majority of sampling dates. In contrast, P, K, or B did not produce a significant effect on dry matter yield in any of the experiments. The sufficiency ranges set for macronutrients N, P, K, Ca and Mg are respectively 32.0 – 42.0, 1.2 – 4.5, 10.0 – 30.0, 7.0 – 23.0, and 4.0 – 10.0 g kg^?1. Those for micronutrients B, copper (Cu), iron (Fe), zinc (Zn), and manganese (Mn) are respectively 20 – 200, 5 – 25, 100 – 600, 25 – 300, and 30 – 200 mg kg^?1. Sufficiency range for SPAD-readings is 45 – 50 SPAD units. All these ranges were established for the commercial harvesting date. The amounts of N, P, K, calcium (Ca), and magnesium (Mg) removed in aboveground biomass are respectively 22.7, 1.6, 26.4, 16.4 and 4.8 kg Mg^?1 of dry biomass.     

Influence of Sugarcane Straw on Aminocyclopyrachlor Leaching in a Green-Cane Harvesting System

The straw on the soil in the green cane harvesting system acts as a physical barrier to the action of the pre-emergence herbicides. However, some herbicides have physico-chemical characteristics to pass through the straw in water solution. The aim was to evaluate the influence of different densities of sugarcane straw on the leaching of aminocyclopyrachlor in a tropical soil (Oxisol—Typic Hapludox) in a system of green cane harvesting. The experiment was carried out using glass columns filled with soil, with and without sugarcane straw (0, 5, 10, and 20 t ha^?1) on the top of the soil. ¹⁴C-aminocyclopyrachlor was applied followed by 200 mm of simulated rain for 48 h. The herbicide was detected in all soil layers (0–30 cm), and in the leachate (>?30 cm), in small trace amounts (≤?0.21%) in all treatments, showing that aminocyclopyrachlor undergoes a high degree of leaching in the clayey soil. In the absence of straw, 39% of the herbicide remained in the upper layer (0–5 cm). In the presence of straw, the herbicide was distributed through the soil profile, mainly from 0 to 25 cm. Forty percent more herbicide was retained in the straw with 20 t ha^?1 of sugarcane straw. Thus, it is concluded that the straw in high amounts retains aminocyclopyrachlor, but does not prevent it from reaching the soil; the high solubility of the herbicide and intense rainfall in a short period of time are factors that contribute to the herbicide passing through this barrier.