Application-Oriented Marine Isomerases in Biocatalysis

The class EC 5.xx, a group of enzymes that interconvert optical, geometric, or positional isomers are interesting biocatalysts for the synthesis of pharmaceuticals and pharmaceutical intermediates. This class, named “isomerases,” can transform cheap biomolecules into expensive isomers with suitable stereochemistry useful in synthetic medicinal chemistry, and interesting cases of production of l-ribose, d-psicose, lactulose, and d-phenylalanine are known. However, in two published reports about potential biocatalysts of marine origin, isomerases are hardly mentioned. Therefore, it is of interest to deepen the knowledge of these biocatalysts from the marine environment with this specialized in-depth analysis conducted using a literature search without time limit constraints. In this review, the focus is dedicated mainly to example applications in biocatalysis that are not numerous confirming the general view previously reported. However, from this overall literature analysis, curiosity-driven scientific interest for marine isomerases seems to have been long-standing. However, the major fields in which application examples are framed are placed at the cutting edge of current biotechnological development. Since these enzymes can offer properties of industrial interest, this will act as a promoter for future studies of marine-originating isomerases in applied biocatalysis.     

Sorption of humic acid on Fe oxides,bacteria, and Fe oxide-bacteria composites

Purpose

Sorption of humic substances on other soil components plays an important role in controlling their function and fate in soil. Sorption of humic substances by individual soil components has been studied extensively. However, few studies reported the sorption characteristic of humic substances on composites of soil components. This study aimed to investigate the sorption characteristics of humic acid on Fe oxide-bacteria composites and improve the understanding on the interaction among humic substance Fe oxide bacteria in soil.

Materials and methods

Humic acid was purchased from Sigma-Aldrich and was purified. Hematite and ferrihydrite were synthesized in the lab. Bacillus subtilis and Pseudomonas putida were cultivated in Luria-Broth medium and harvested at stationary growth phase. Batch sorption experiments were carried out at pH 5.0. Various amounts of humic acid were mixed with 20 mg of Fe oxide, bacteria, or Fe oxide-bacteria composite (oxide to bacteria of 1:1) in 10 mL of KCl (0.02 mol L^?1) to construct sorption isotherms. The effects of phosphate concentration and addition order among humic acid, Fe oxide, bacteria on the sorption of humic acid were also studied. The sorption of humic acid was calculated by the difference between the amount of humic acid added initially and that remained in the supernatant.

Results and discussion

The maximum sorption of humic acid on hematite, ferrihydrite, B. subtilis and P. putida was 73.2, 153.5, 69.1, and 56.7 mg C g^?1, respectively. The maximum sorption of humic acid on examined Fe oxide-bacteria composite was 28.2–57.2 % less than the predicted values, implying that the sorption of humic acid was reduced by the interaction between Fe oxides and bacteria. The presence of phosphate exerted negligible influence on the sorption of humic acid on bacteria while it inhibited the sorption of humic acid on Fe oxides. On Fe oxide-bacteria composites, inhibiting influences followed by promoting or weak inhibiting effects of phosphate with increasing concentration on the sorption of humic acid were found.

Conclusions

The interaction between Fe oxides and bacteria reduced the sorption of humic acid; moreover, the reduction was greater by the interaction of bacteria with ferrihydrite than that with hematite. Phosphate exerted negligible and inhibiting influence on the sorption of humic acid by bacteria and Fe oxides, respectively. On Fe oxide-bacteria composites, humic acid sorption was initially inhibited and then promoted or weakly inhibited by phosphate with increasing concentration.     

Temporary Turnout for Free Exercise in Groups: Effects on the Behavior of Competition Horses Housed in Single Stalls

In Germany, most competition horses are housed in single stalls and free exercise is not permitted in many cases. The reason for not allowing free exercise is mostly the risk of injury. Additionally, opinions exist that the horses’ demand for exercise is fulfilled by training and that the horses’ willingness to perform is negatively influenced by free exercise. In the present study, three turnout practices were investigated with regard to their effect on the behavior of four horses: daily training without free exercise (no turnout), 2-hour turnout (for free exercise) before training, and 2-hour turnout after training. The aim of this study was to determine any differences in the horses’ behavior between the three treatments. The horses’ behavior in the stable was observed through video recordings. The behavior during turnout was observed directly and during training was evaluated by the riders with the aid of a questionnaire. Additionally, the distance covered during turnout was measured by Global Positioning System devices. The behavior within the stall was more restless in the treatment without turnout—which became apparent in significantly more frequent changes between behaviors as compared with the treatments with turnout. The results of Global Positioning System measurement during turnout showed a significantly shorter distance covered when horses were trained before they were turned out compared with turnout before training. If the horses were turned out after training, they also showed less trotting and cantering and more dozing. The horses’ willingness to perform was not significantly different between the three treatments.     

Seasonal and Diurnal Changes in Starch Content and Sugar Profiles of Bermudagrass in the Piedmont Region of the United States

Seasonal and diurnal patterns of sugar accumulation in bermudagrass (Cynodon dactylon) pastures were monitored to evaluate risk factors for pasture-associated laminitis of ponies and horses. Bermudagrass was collected from four plots in the morning and afternoon on a weekly basis, from mid-July until late August. Tissue was air-dried to simulate hay, or frozen to retain the sugar profiles of fresh pasture. Samples were analyzed colorimetrically for total water-soluble and ethanol-soluble carbohydrates, and electrochemically for starch. In addition, sugars were separated and quantified by high-performance liquid chromatography coupled to pulsed amperometric detection. The dominant sugars in extracts were glucose, fructose, and sucrose. Some minor peaks, corresponding to tri- and tetrasaccharides, were also detected in some extracts. Starch increased over time in fresh and dried tissue, and concentrations varied diurnally in fresh, but not in dried tissue (P = .021). Sucrose in dried tissue decreased and then increased, with higher concentrations than in fresh tissue on all sampling dates (P = .024). Glucose and fructose exhibited diurnal variation on one and two dates, respectively (P = .034 and .0028, respectively). These results reveal trends in carbohydrate concentrations and profiles that may help to evaluate the likelihood of equine laminitis outbreaks on bermudagrass.     

Why Are Mares with Pneumovagina Susceptible to Bacterial Endometritis? A Personal Opinion

Pneumovagina is the abnormal presence of air in the vagina, which may lead to pneumometra and endometritis. Predisposing factors are abnormal perineal conformation and weakness of the vestibulovaginal sphincter. This report examines why ponies and some other breeds are less susceptible to pneumovagina than Thoroughbreds and consequently less susceptible to endometritis. Observations on vaginal ballooning during vaginoscopic examination led to the conclusion that there is a fundamental difference in intra-abdominal pressures between horses and ponies, and between mares with high or low body condition scores. This difference in the extreme cases, for example, a small fat pony versus a large thin Thoroughbred, means that the pressure gradient in the former is from the uterus through the vagina to the exterior, whereas the case is reversed in the latter. Although the uterus is at a negative pressure relative to the exterior, evacuation of fluids is seriously disadvantaged, leading to an inability to resolve uterine infections.     

Role of wild ruminants in the epidemiology of bluetongue virus serotypes 1, 4 and 8 in Spain

ABSTRACT: Although the importance of wild ruminants as potential reservoirs of bluetongue virus (BTV) has been suggested, the role played by these species in the epidemiology of BT in Europe is still unclear. We carried out a serologic and virologic survey to assess the role of wild ruminants in the transmission and maintenance of BTV in Andalusia (southern Spain) between 2006 and 2010.A total of 473 out of 1339 (35.3%) wild ruminants analyzed showed antibodies against BTV by both ELISA and serum neutralization test (SNT). The presence of neutralizing antibodies to BTV-1 and BTV-4 were detected in the four species analyzed (red deer, roe deer, fallow deer and mouflon), while seropositivity against BTV-8 was found in red deer, fallow deer and mouflon but not in roe deer. Statistically significant differences were found among species, ages and sampling regions. BTV RNA was detected in twenty-one out of 1013 wild ruminants (2.1%) tested. BTV-1 and BTV-4 RNA were confirmed in red deer and mouflon by specific rRT-PCR.BTV-1 and BTV-4 seropositive and RNA positive wild ruminants, including juveniles and sub-adults, were detected years after the last outbreak was reported in livestock. In addition, between the 2008/2009 and the 2010/2011 hunting seasons, the seroprevalence against BTV-1, BTV-4 and BTV-8 increased in the majority of provinces, and these serotypes were detected in many areas where BTV outbreaks were not reported in domestic ruminants. The results indicate that wild ruminants seem to be implicated in the dissemination and persistence of BTV in Spain.     

Evaluation of nonpolar metabolites in plant extracts by 13C NMR spectroscopy

(13)C nuclear magnetic resonance (NMR) spectroscopy was explored as a simple and efficient technique for the quantitative analysis of nonpolar metabolites in plants. The method was first optimized with a mixture of known metabolites and then applied to the nonpolar leaf extracts of plants harvested in the Valencian community (eastern Spain) belonging to three different genera: Euphorbia (Euphorbiaceae), Araujia (Apocynaceae), and Morus (Moraceae). Furthermore, an exhaustive analysis of Euphorbia characias leaf and stem extracts from different geographic locations allowed that quantitative (13)C NMR spectroscopy is a suitable tool for metabolic profiling purpose.     

Simultaneous determination of phenolic compounds and saponins in quinoa (Chenopodium quinoa Willd) by a liquid chromatography-diode array detection-electrospray ionization-time-of-flight mass spectrometry methodology

A new liquid chromatography methodology coupled to a diode array detector and a time-of-flight mass spectrometer has been developed for the simultaneous determination of phenolic compounds and saponins in quinoa (Chenopodium quinoa Willd). This method has allowed the simultaneous determination of these two families of compounds with the same analytical method for the first time. A fused-core column C18 has been used, and the analysis has been performed in less than 27 min. Both chromatographic and electrospray ionization time-of-flight mass spectrometry parameters have been optimized to improve the sensitivity and to maximize the number of compounds detected. A validation of the method has also been carried out, and free and bound polar fractions of quinoa have been studied. Twenty-five compounds have been tentatively identified and quantified in the free polar fraction, while five compounds have been tentatively identified and quantified in the bound polar fraction. It is important to highlight that 1-O-galloyl-β-D-glucoside, acacetin, protocatechuic acid 4-O-glucoside, penstebioside, ethyl-m-digallate, (epi)-gallocatechin, and canthoside have been tentatively identified for the first time in quinoa. Free phenolic compounds have been found to be in the range of 2.746-3.803 g/kg of quinoa, while bound phenolic compounds were present in a concentration that varies from 0.139 and 0.164 g/kg. Indeed, saponins have been found to be in a concentration that ranged from 5.6 to 7.5% of the total composition of whole quinoa flour.