Potential Anti-Obesity,Anti-Steatosis,and Anti-Inflammatory Properties of Extracts from the Microalgae Chlorella vulgaris and Chlorococcum amblystomatis under Different Growth Conditions

Microalgae are known as a producer of proteins and lipids, but also of valuable compounds for human health benefits (e.g., polyunsaturated fatty acids (PUFAs); minerals, vitamins, or other compounds). The overall objective of this research was to prospect novel products, such as nutraceuticals from microalgae, for application in human health, particularly for metabolic diseases. Chlorella vulgaris and Chlorococcum amblystomatis were grown autotrophically, and C. vulgaris was additionally grown heterotrophically. Microalgae biomass was extracted using organic solvents (dichloromethane, ethanol, ethanol with ultrasound-assisted extraction). Those extracts were evaluated for their bioactivities, toxicity, and metabolite profile. Some of the extracts reduced the neutral lipid content using the zebrafish larvae fat metabolism assay, reduced lipid accumulation in fatty-acid-overloaded HepG2 liver cells, or decreased the LPS-induced inflammation reaction in RAW264.7 macrophages. Toxicity was not observed in the MTT assay in vitro or by the appearance of lethality or malformations in zebrafish larvae in vivo. Differences in metabolite profiles of microalgae extracts obtained by UPLC-LC-MS/MS and GNPS analyses revealed unique compounds in the active extracts, whose majority did not have a match in mass spectrometry databases and could be potentially novel compounds. In conclusion, microalgae extracts demonstrated anti-obesity, anti-steatosis, and anti-inflammatory activities and could be valuable resources for developing future nutraceuticals. In particular, the ultrasound-assisted ethanolic extract of the heterotrophic C. vulgaris significantly enhanced the anti-obesity activity and demonstrated that the alteration of culture conditions is a valuable approach to increase the production of high-value compounds.     

A modified agglutination test for the diagnosis of Besnoitia besnoiti infection

Bovine besnoitiosis is caused by Besnoitia besnoiti, an obligate intracellular apicomplexan parasite. Affected animals present cutaneous and systemic manifestations and the disease may lead to considerable economic losses. Although generally associated to tropical and subtropical areas, bovine besnoitiosis is now considered an emergent disease in Europe, due to the increasing number of new cases and apparent geographical expansion. In this study we evaluated the performance of a modified agglutination test (B-MAT) in the serodiagnosis of bovine besnoitiosis in comparison to the indirect immunofluorescent-antibody test (IFAT). To establish optimal protocol conditions we used bovine sera with a known infection status for B. besnoiti infection. Positive animals (n=36) presented B. besnoiti dermal cysts and anti-B. besnoiti specific antibodies, as determined by the indirect immunofluorescence test (IFAT). Negative animals (n=103) were from non-endemic areas in Portugal and negative by the IFAT. From here, we evaluated the sensitivity and specificity of the B-MAT relative to the IFAT with a panel of sera from herds with history of bovine besnoitiosis in Portugal, Spain and France (n=402), using three serum dilutions (1:80, 1:160, 1:320). Considering the positive cut-off at 1:160 serum dilution, the B-MAT showed an almost perfect test agreement with the IFAT; (κ=0.968; 95% CI: 0.941-0.996) with a relative sensitivity of 97.2% (95% CI: 94.1-100%) and a relative specificity of 99.3% (95% CI: 98.4-100%). As a simple and inexpensive technique the B-MAT represents a valuable tool for the diagnosis and study of the epidemiology of bovine besnoitiosis.     

Epidemiological Analysis of Leptospira spp. Infection in Equids from the Brejo Paraibano Microregion of Brazil

Leptospirosis is a zoonotic disease that occurs in humans and several animal species, including cattle, pigs, and horses, and it has a worldwide distribution. In equids, leptospirosis manifests through recurrent uveitis and abortion and other reproductive disorders. The objective of the present study was to investigate the epidemiological situation of Leptospira spp. infection in equids from the Brejo Paraibano microregion in the northeast of Brazil, that is, to assess the prevalence of and identify the major risk factors for infection. For this purpose, blood samples were taken from 257 equids (204 horses, 46 mules, and 7 donkeys), and a microscopic agglutination test (MAT) was used as serological analysis, with 24 serovars of Leptospira spp. as antigens. Antibodies were observed in 16.2% of horses, 13.0% of mules, and 28.6% of donkeys. Animals between the ages of 2.5 and 11 years had a risk of infection 0.4 times lower than those younger than 2.5 years old (odds ratio, 0.4; 95% confidence interval, 0.18-0.90). Further studies are needed to identify potential wild reservoirs and the main routes of infections.     

Sulfated Glucan from the Green Seaweed Caulerpa sertularioides Inhibits Adipogenesis through Suppression of Adipogenic and Lipogenic Key Factors

Sulfated polysaccharides (SPS) from seaweeds have great biochemical and biotechnological potential. This study aimed to investigate the effect of SPS isolated from the seaweed Caulerpa sertularioides on adipogenic differentiation as a possible alternative treatment for obesity. The SPS-rich extract from the seaweed C. sertularioides was fractioned into three SPS-rich fractions (F0.5; F0.9; and F1.8) chemically characterized. Among these four samples, only F0.9 showed a significant inhibitory effect on adipogenesis of 3T3-L1 preadipocytes. Ten SPS-rich fractions were isolated from F0.9 through ion-exchange chromatography. However, only the fraction (CS0.2) containing a sulfated glucan was able to inhibit adipogenesis. CS0.2 reduces lipid accumulation and inhibits the expression of key adipogenic (PPARγ, C/EBPβ, and C/EBPα) and lipogenic markers (SREBP-1c, Fabp4, and CD36). The data points to the potential of sulfated glucan from C. sertularioides for the development of functional approaches in obesity management.     

Evaluation of dietary histidine supplementation on meagre,Argyrosomus regius,juvenile growth,haematological profile,stress and muscle cellularity

Histidine plays an important role in haemoglobin synthesis, antioxidant defence system and stress resistance, and it is thought to be a limiting amino acid in most meagre diets. In this study, two different diets, a control and a diet supplemented with 3% histidine, were tested in 60‐day‐old meagre (initial weight—6.2 g) for 82 days. Survival varied between 92% and 94%, and neither SGR nor whole fish composition seemed to be affected by the diet. When fish were submitted to a stress test (outside water for 30 s), it was observed that the levels of glucose and lactate significantly increased in both treatments. The concentration of cortisol increased after the stress test only in the His 30 g/kg group. Muscle cellularity was affected by the diets, where the control group showed a higher mean fibre area and lower fibre density. These results, associated with a lower frequency of smaller fibres, suggest a decrease in fibre recruitment and a higher fibre hypertrophy. In conclusion, the supplementation of diets with 3% led to a more pronounced response to the stress test in terms of cortisol levels and increased muscle fibre recruitment, decreasing muscle fibre hypertrophy contribution to muscle growth.     

Slow-release N fertilisers are not an alternative to urea for fertilisation of autumn-grown tall cabbage

Tall cabbage (Brassica oleracea var. costata, cv. Penca de Mirandela) is grown in Portugal during autumn and winter months when heavy rains can be expected. In this agrosystem the management of N is a considerable challenge due to the risk of nitrate leaching. Field experiments with tall cabbage and rye were carried out during the growing seasons of 2007/2008 and 2008/2009. The experimental design included three fertilisers that delay N availability by different mechanisms [Floranid permanent, containing isobutylidene diurea (IBDU); Basacote, a polymer-coated fertiliser; and Entec 26, containing 3,4-dimethylpyrazole phosphate (DMPP) as nitrification inhibitor], two urea treatments (total N applied basally; and divided into two equal rates in preplant and topdress application), and a zero N control. The fertilisers were applied at a rate of 120 kg N ha^?1. A pot experiment with tall cabbage followed by ryegrass grown in the same pots was carried out in 2007/2008. The fertiliser treatments were essentially the same and a rate of 2.94 g N/pot was applied. Dry matter yield, plant N concentration, plant N recovery and indices of N-use efficiency were compared among the fertiliser treatments. In the pot experiment, soil nitrate concentration was also monitored by using anion exchange membranes inserted directly into the soil. Urea produced mean DM yields statistically higher than control in all crops and in both field and pot experiments and years. No significant differences in DM yield were usually found among fertilised treatments. Basacote produced DM yields often not significantly different from that of control. In the field experiment of tall cabbage, plants recovered 20 and 50% of the amount of N applied, respectively in Basacote and in the other fertilised plots. In the pot experiment, tall cabbage + ryegrass recovered 49.6 and ≈100% of N applied, respectively in Basacote and in the other fertilised plots. From the strictly agronomic point of view, none of the slow-release N materials provided advantages over urea to be advised for this agrosystem, considering also their high cost.     

Pyogenic bacterial infections in humans with MyD88 deficiency

MyD88 is a key downstream adapter for most Toll-like receptors (TLRs) and interleukin-1 receptors (IL-1Rs). MyD88 deficiency in mice leads to susceptibility to a broad range of pathogens in experimental settings of infection. We describe a distinct situation in a natural setting of human infection. Nine children with autosomal recessive MyD88 deficiency suffered from life-threatening, often recurrent pyogenic bacterial infections, including invasive pneumococcal disease. However, these patients were otherwise healthy, with normal resistance to other microbes. Their clinical status improved with age, but not due to any cellular leakiness in MyD88 deficiency. The MyD88-dependent TLRs and IL-1Rs are therefore essential for protective immunity to a small number of pyogenic bacteria, but redundant for host defense to most natural infections.     

Quince (Cydonia oblonga miller) fruit characterization using principal component analysis

This paper presents a large amount of data on the composition of quince fruit with regard to phenolic compounds, organic acids, and free amino acids. Subsequently, principal component analysis (PCA) is carried out to characterize this fruit. The main purposes of this study were (i) the clarification of the interactions among three factors-quince fruit part, geographical origin of the fruits, and harvesting year-and the phenolic, organic acid, and free amino acid profiles; (ii) the classification of the possible differences; and (iii) the possible correlation among the contents of phenolics, organic acids, and free amino acids in quince fruit. With these aims, quince pulp and peel from nine geographical origins of Portugal, harvested in three consecutive years, for a total of 48 samples, were studied. PCA was performed to assess the relationship among the different components of quince fruit phenolics, organic acids, and free amino acids. Phenolics determination was the most interesting. The difference between pulp and peel phenolic profiles was more apparent during PCA. Two PCs accounted for 81.29% of the total variability, PC1 (74.14%) and PC2 (7.15%). PC1 described the difference between the contents of caffeoylquinic acids (3-O-, 4-O-, and 5-O-caffeoylquinic acids and 3,5-O-dicaffeoylquinic acid) and flavonoids (quercetin 3-galactoside, rutin, kaempferol glycoside, kaempferol 3-glucoside, kaempferol 3-rutinoside, quercetin glycosides acylated with p-coumaric acid, and kaempferol glycosides acylated with p-coumaric acid). PC2 related the content of 4-O-caffeoylquinic acid with the contents of 5-O-caffeoylquinic and 3,5-O-dicaffeoylquinic acids. PCA of phenolic compounds enables a clear distinction between the two parts of the fruit. The data presented herein may serve as a database for the detection of adulteration in quince derivatives.