Evaluation of oligosaccharide synthesis from lactose and lactulose using β-galactosidases from Kluyveromyces isolated from artisanal cheeses

The β-galactosidase activity of 15 Kluyveromyces strains isolated from cheese belonging to Kluyveromyces lactis and Kluyveromyces marxianus species was tested for the production of oligosaccharides derived from lactose (GOS) and lactulose (OsLu). All Kluyveromyces crude cell extracts (CEEs) produced GOS, such as 6-galactobiose and 3'-, 4'-, and 6'-galactosyl-lactose. At 4 h of reaction, the main trisaccharide formed was 6'-galactosyl-lactose (20 g/100 g of total carbohydrates). The formation of OsLu was also observed by all CEEs tested, with 6-galactobiose, 6'-galactosyl-lactulose, and 1-galactosyl-lactulose being found in all of the reaction mixtures. The synthesis of trisaccharides predominated over other oligosaccharides. K. marxianus strain O3 produced the highest yields of GOS and OsLu after 4 h of reaction, reaching 42 g/100 g of total carbohydrates (corresponding to 80% lactose hydrolysis) and 45 g/100 g of total carbohydrates (corresponding to 87% lactulose hydrolysis), respectively. Therefore, the present study contributes to a better insight into dairy Kluyveromyces β-galactosidases and shows the feasibility of these enzymes to transglycosylate lactose and lactulose, producing high yields of prebiotic oligosaccharides.     

Soil biochemical activities and the geometric mean of enzyme activities after application of sewage sludge and sewage sludge biochar to soil

There is a need to improve the way in which wastes, such as sewage sludges, are managed and a potential way to proceed would be to transform them into biochar. On the other hand, there is a growing interest in the use of soil biochemical properties as indicators of soil quality because they are sensitive to alterations in soil management. Thus, we have studied the effect of a biochar obtained from sewage sludge on soil biochemical properties in an organic soil using two doses of biochar and comparing these results with the control soil and with soils amended with the same two doses of unpyrolyzed sewage sludge. Microbial biomass C, soil respiration, net N mineralization and several enzyme activities (dehydrogenase, β-glucosidase, phosphomoesterase and arylsulphatase) were monitored. The geometric mean of enzyme activities (GMea) was used as a soil quality index. Individual biochemical properties showed a different response to the treatments, while GMea showed an increase in the quality of soils amended with the high biochar dose and a decrease in those amended with a high sewage sludge dose. The geometric mean of enzyme activities was a suitable index to condense the whole set of soil enzyme values in a single numerical value, which was sensitive to management practices.     

The prevalence of Toxocara cati in domestic cats in Mexico City

The faeces of 520 domestic cats, resident in the 16 municipal authorities of Mexico City, were analysed using the method of centrifuging with zinc sulphate (at 33%). Three hundred and ninety-nine animals lived in houses, 121 in apartments. For the purpose of this study they were divided into seven age groups, of 6 months each. Toxocara cati eggs were found in the faeces of 42.5% of the animals. 20.7% of apartment cats and 49.1% of house cats were infected. This difference in the rate of infection is statistically significant (P<0.002.) for the house cats. T. cati was found in all the age groups although most infection was found in animals less than 1 year old.The fact that infection occurred in the whole age range of the cats and in all the municipal authorities means that the risk of being contaminated with this helminth in Mexico City is high. As a consequence, the possibility of developing illness as a result of becoming a host to larva migrans is also high.     

Galápagos land iguana (Conolophus subcristatus) as a seed disperser

The role of the most common land iguana (Conolophus subcristatus) in the Galápagos Islands as an effective seed disperser is explored in this study. A total of 5705 seeds of 32 plant species were identified from 160 scats, 4545 of which (80%) appeared visually undamaged. Germination trials of 849 seeds from 29 species revealed that at least 10 species remained viable after passing through the iguana's gut, although only a small proportion of those seeds (4%) germinated. In any case, we argue that C. subcristatus exerts an important role on the 7 Galapagos islands where it occurs because of its abundance and capacity to ingest and disperse seeds at long distances. Our results strongly suggest that the Galápagos C. subcristatus plays an important role as a seed disperser of not only of native species but also some introduced plants in the Galápagos Islands.     

Ultrasound‐assisted periconal ocular blockade in rabbits

ObjectiveThis study aimed to evaluate the benefit and specifically the feasibility of using ultrasound in ophthalmologic periconal block, and the occurrence of complications.Study designProspective experimental study.AnimalsTen healthy New Zealand White rabbits (6–8 months of age), weighing 2.0–3.5 kg.MethodsRabbits were anesthetized by intramuscular injection of acepromazine (1 mg kg⁻¹), ketamine (30 mg kg⁻¹) and xylazine (3 mg kg⁻¹). Ultrasound-assisted periconal block with lidocaine was performed on 18 eyes. Intraocular pressure was measured by applanation tonometry whereas corneal sensitivity was assessed using an esthesiometer, before and after each periconal anesthesia.ResultsIn all 18 eyes, it was possible to adequately visualize the needle shaft within the periconal space, as well as muscular cone, optic nerve and local anesthetic solution spread. Lidocaine 2% without epinephrine (0.79 ± 0.19 mL) was injected into the periconal space. There was no statistical difference between the intraocular pressure (mean ± SD) measured before (10.9 ± 2.9 mmHg) and after (11.9 ± 3.8 mmHg) the periconal anesthesia (p = 0.38). The effectiveness of the ultrasound-assisted technique was shown according to the values for corneal sensitivity, assessed before and after periconal anesthesia (p < 0.0001). Complications were not observed in this study.ConclusionsEye ultrasonography allowed visualization of all anatomic structures necessary to perform a periconal block, as well as the needle insertion and anesthetic spread in real time. Further studies are required to prove the real potential of ultrasound for reducing the incidence of complications associated with ophthalmic blocks, especially when anatomic disorders of the eye could potentially increase the risk.Clinical relevanceUltrasonography is a painless, noninvasive tool that may improve safety of ophthalmic regional blocks, potentially by reducing the prevalence of globe perforation or penetration of the optic nerve associated with the needle-based techniques.     

Effect of feed restriction on intake of <Emphasis Type="Italic">Moringa oleifera</Emphasis> and <Emphasis Type="Italic">Leucaena leucocephala</Emphasis> and growth performance of rabbits

Two experiments were conducted to evaluate the effect of feed restriction on intake of Moringa oleifera (MO) or Leucaena leucocephala (LL) and growth of rabbits. In experiment one, 45 rabbits (male and female) weighing 1.18 ± 0.15 kg were used. They were randomly distributed to three feed restriction treatments (20, 30, and 40%) with 15 rabbits each (9 females and 6 males) and they were offered M. oleifera (MO) ad libitum. In experiment two, 45 growing male rabbits weighing 0.63 ± 0.113 kg were used. They were randomly assigned to 0, 20, and 30% feed restriction diets, and they have free access to L. leucocephala (LL). Intake of MO increased (P < 0.05) conforming feed restriction increased (40.6, 52.9, and 55.2 g/day of MO for 20, 30, and 40%, respectively). Daily liveweight gain and feed conversion did not differ (P > 0.05), and economic efficiency was similar among treatments. Consumption of LL increased (P < 0.05) in rabbits under the 30% restriction treatment in comparison to that of rabbits restricted 20% (46.0 and 44.4 g/day, respectively). Total feed intake (LL + feed) was highest in 20% restricted rabbits (108.0, 100.8, and 93.2 g/day for 20, 30, and 0%, respectively). Daily liveweight gain and feed conversion were not affected by feed restriction (P > 0.05). Economic efficiency improved twice in feed-restricted rabbits (2.0 and 2.3 for 20 and 30%, respectively) in contrast to that of the control 0% group (1.1). The results suggest that rabbits restricted up to 30% and supplemented with either MO or LL did not affect growth performance and reduced feed cost.     

Pyrethroids levels in paddy field water under Mediterranean conditions: measurements and distribution modelling

The cultivation of rice (Oriza sativa L.) under Mediterranean conditions regularly requires the use of treated wastewater due to shortage of freshwater. As a consequence, the intensification of rice production to supply the uprising demand of grain could break the stability between agriculture and environment. In this work, we studied the occurrence and distribution of pyrethroids in surface water and groundwater collected during two periods (flooding and dry soil conditions) in paddy fields located in the Spanish Mediterranean coast. Pyrethroids were detected at concentrations ranging from 14 to 1450 ng L^?1 in surface water and from 6 to 833 ng L^?1 in groundwater. The results obtained were evaluated statistically using principal component analysis, and differences between both sampling campaigns were found, with lower concentrations of the target compounds during the flooding sampling event. Moreover, a geographic information system program was used to represent a model distribution of the obtained results, showing wastewater treatment plants as the main sources of contamination and the decrease of pyrethroids during flooding condition when water flows over the paddy fields. The impact of these compounds on water quality was discussed.     

Occurrence of Lipophilic Marine Toxins in Shellfish from Galicia (NW of Spain) and Synergies among Them

Lipophilic marine toxins pose a serious threat for consumers and an enormous economic problem for shellfish producers. Synergistic interaction among toxins may play an important role in the toxicity of shellfish and consequently in human intoxications. In order to study the toxic profile of molluscs, sampled during toxic episodes occurring in different locations in Galicia in 2014, shellfish were analyzed by liquid chromatography tandem mass spectrometry (LC–MS/MS), the official method for the detection of lipophilic toxins. The performance of this procedure was demonstrated to be fit for purpose and was validated in house following European guidelines. The vast majority of toxins present in shellfish belonged to the okadaic acid (OA) group and some samples from a particular area contained yessotoxin (YTX). Since these toxins occur very often with other lipophilic toxins, we evaluated the potential interactions among them. A human neuroblastoma cell line was used to study the possible synergies of OA with other lipophilic toxins. Results show that combination of OA with dinophysistoxin 2 (DTX2) or YTX enhances the toxicity triggered by OA, decreasing cell viability and cell proliferation, depending on the toxin concentration and incubation time. The effects of other lipophilic toxins as 13-desmethyl Spirolide C were also evaluated in vitro.     

Genetic connectivity of the South American fur seal (Arctocephalus australis) across Atlantic and Pacific oceans revealed by mitochondrial genes

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Sugars Profiles of Different Chestnut (Castanea sativa Mill.) and Almond (Prunus dulcis) Cultivars by HPLC-RI

Sugar profiles of different almond and chestnut cultivars were obtained by high-performance liquid chromatography (HPLC), by means of a refractive index (RI) detector. A solid-liquid extraction procedure was used in defatted and dried samples. The chromatographic separation was achieved using a Eurospher 100-5 NH₂ column using an isocratic elution with acetonitrile/water (70:30, v/v) at a flow rate of 1.0 ml/min. All the compounds were separated in 16 min. The method was optimized and proved to be reproducible and accurate. Generally, more than 95% of sugars were identified for both matrixes. Sugars profiles were quite homogeneous for almond cultivars; sucrose was the main sugar (11.46 ± 0.14 in Marcona to 22.23 ± 0.59 in Ferragnes g/100 g of dried weight), followed by raffinose (0.71 ± 0.05 in Ferraduel to 2.11 ± 0.29 in Duro Italiano), glucose (0.42 ± 0.12 in Pegarinhos two seeded to 1.47 ± 0.19 in Ferragnes) and fructose (0.11 ± 0.02 in Pegarinhos two seeded to 0.59 ± 0.05 in Gloriette). Commercial cultivars proved to have higher sucrose contents, except in the case of Marcona. Nevertheless, chestnut cultivars revealed a high heterogeneity. Sucrose was the main sugar in Aveleira (22.05 ± 1.48), Judia (23.30 ± 0.83) and Longal (9.56 ± 0.91), while glucose was slightly prevalent in Boa Ventura (6.63 ± 0.49). The observed variance could serve for inter-cultivar discrimination.