Compliance with Nutritional Recommendations and Gut Microbiota Profile in Galician Overweight/Obese and Normal-Weight Individuals.

Different research studies have identified specific groups or certain dietary compounds as the onset and progression of obesity and suggested that gut microbiota is a mediator between these compounds and the inflammation associated with pathology. In this study, the objective was to evaluate the dietary intake of 108 overweight (OW), obese (OB), and normal-weight (NW) individuals and to analyze their gut microbiota profile to determine changes and associations with Body Mass Index (BMI) and diet. When individuals were compared by BMI, significant differences in fiber and monounsaturated fatty acids (MUFAs) intake were observed, showing higher adequacy for the NW group. The analysis of gut microbiota showed statistical differences for 18 ASVs; Anaerostipes and Faecalibacterium decreased in the OW/OB group, whereas the genus Oscillospira increased; the genus was also found in the LEFSe analysis as a biomarker for OW/OB. Roseburia faecis was found in a significantly higher proportion of NW individuals and identified as a biomarker for the NW group. Correlation analysis showed that adequation to nutritional recommendation for fiber indicated a higher abundance of Prevotella copri, linearly correlated with F. prausnitzii, Bacteroides caccae, and R. faecis. The same correlation was found for the adequation for MUFAs, with these bacteria being more abundant when the intake was adjusted to or below the recommendations.

Hábitos alimentarios en la población gallega durante el confinamiento por la COVID-19 / Food habits in the Galician population during confinement by COVID-19

INTRODUCCIÓN: en plena crisis por la COVID-19, el consumo alimentario ha pasado a llevarse a cabo en su totalidad en el hogar y las visitas a los centros de distribución de alimentos se han visto reducidas. Asimismo, otros hábitos, como la práctica de ejercicio físico o las horas de sueño, se han visto también modificadas. OBJETIVOS: el objetivo de este trabajo ha sido evaluar los cambios producidos en los hábitos alimentarios y en el estilo de vida de la población gallega durante el período de confinamiento por la COVID-19. MÉTODOS: se elaboró una encuesta en Microsoft Forms para adultos residentes en Galicia y se distribuyó, de manera aleatoria, a través de un servicio de mensajería instantánea para móviles a través de Internet. RESULTADOS: el número de participantes fue de 1350 personas, mayores de edad, siendo el promedio de edad de 63,2 ± 8,1 años y estando la muestra compuesta por un 30 % de hombres y un 70 % de mujeres. Los datos muestran una tendencia hacia el consumo de alimentos saludables, como frutas y verduras, y un descenso de otros menos saludables, como son los alimentos procesados. Un 46 % de la población indicó estar comiendo más que antes del confinamiento, aumentando el picoteo entre horas (54 %) y la ansiedad por la comida, debido a situaciones de estrés o nervios, en un 40 % de los encuestados. Esto se ha traducido en un incremento del peso corporal, indicado por un 44 % de la muestra, siendo la media de kilos ganada de 2,8 ± 1,5. CONCLUSIONES: durante el confinamiento, los hábitos alimentarios y el estilo de vida de la población gallega se han visto modificados. El incremento de peso experimentado en una población que ya partía de un elevado porcentaje de sobrepeso y obesidad al inicio del confinamiento podría tener un impacto negativo en el desarrollo de patologías crónicas no transmisibles

INTRODUCTION: during the COVID-19 crisis food habits have changed among the population as food consumption is now carried out totally at home, and visits to groceries or food centers have decreased considerably. Likewise, other habits, such as physical activity or sleeping hours have also been modified. OBJECTIVES: the objective of this work was to evaluate the changes occurred in eating and lifestyle habits among the adult Galician population during the confinement period ordered in Spain during the COVID-19 pandemic. METHODS: a survey was conducted using Microsoft Forms, and spread among the population using an instant messaging service for mobiles through the internet. RESULTS: a total of 1350 Galician adults completed the questionnaire; mean of age was 63.2 ± 8.1 years, 30 % were men, and 70 % were women. The obtained data show a trend towards consumption of healthy foods such as fruits and vegetables, and a decrease in less healthy ones such as processed foods. In total, 46 % of the population reported they were eating more than before the confinement, with an increase in snacking between meals (54 %) and in anxiety about food due to situations of stress or nervousness bieng reported by 40 % of respondents. This has resulted in an increase in body weight, reported by 44 % of participants, with the average gain being 2.8 ± 1.5 kg. CONCLUSIONS: during confinement, the eating and lifestyle habits of the Galician population have been modified. The weight gain observed in a population that already had a high level of overweight and obesity at confinement onset aggravates the present situation and may have a negative impact on the development of chronic, non-communicable diseases

Natural Compounds as Antimicrobial Agents.

During the first two decades of this century, conventional antimicrobial compounds have been found out to have more bacterial resistance. What has also been worrying is the rediscovery of the so-called "natural compounds", which in turn have a good name among the average citizen because of the former's plant or animal origin. However, they do not form a well-classified group of substances. This Special Issue consists of five reviews focusing on clinical bacteria applications in food and their specific effects upon virulent bacterial factors. You will also find a research on much needed, new antimicrobials sourced in extreme environments, and secondary metabolites of Burkholderia. This issue includes 12 original research papers which will provide you with an in-depth coverage of the protein extract activity, as well as the activity of other plant extracts, on fighting bacteria, fungi or diarrhea. Their use in broilers or laying eggs for production purposes has also been focused on in order to improve gut microbiota. Last but not least, we should not forget about honey and its effect; Allium sativum-fermented extracts, as well as other "natural" compounds, have been studied in their fight against biofilms. Furthermore, we have also examined the use of essential oils, which are currently used in edibles such as fresh sausages. The present work also deals with other applications such as natural compound derivatives as well as compound mixtures.

Confirmation of ß-agonist residues in bovine retina and liver using HPLC-MS/MS and evaluation of matrix-dependent problems.

ß-Agonists or ß-adrenergic agonists are synthetic drugs sometimes used for therapeutic purposes in veterinary medicine. However, these compounds also belong to a class of illegal growth promoters that decrease the fat content in farm animals. The purpose of this study was to develop an analytical LC-MS/MS confirmatory method for the detection of nine ß-agonists in bovine retina and liver tissues. The obtained CCα values ranged from 0.29 to 1.54 µg kg-1 in retina and from 0.03 to 0.18 µg kg-1 in liver, and CCß values from 0.50 to 2.63 µg kg-1 and from 0.05 to 0.30 µg kg-1, respectively. Additionally, the importance of a matrix effect in these two tissues for the achievement of accurate detection of non-compliant samples was studied and discussed. It was concluded that the main characteristic of this phenomena is its considerable variability from one animal to another. Also, the total amount of matrix in the final extract can provide differences in the matrix-matched calibration line.

Assessment of Tetracyclines Residues and Tetracycline Resistant Bacteria in Conventional and Organic Baby Foods.

Children are very vulnerable to bacterial infections and they are sometimes subject to antimicrobials for healing. The presence of resistance genes may counteract effects of antimicrobials. This work has thereby compared the amount of tetracycline resistance genes, tet(A) and tet(B), between conventional and organic meat-based or vegetable-based baby foods and used the quantification of these genes to assess the presence of tetracycline residues in these samples. Counts of bacteria harboring the tet(A) gene were higher than those containing tet(B), and there was no difference between the organic and the conventional samples. Samples with detectable amounts of tetracycline residues were also positive for the presence of tet genes, and when the presence of the genes was not detected, the samples were also negative for the presence of residues. The percentages of tetracycline residues were higher in organic samples than in conventional ones. It cannot be concluded that organic formulas are safer than conventional ones for the studied parameters.

Monitoring the presence of 13 active compounds in surface water collected from rural areas in northwestern Spain.

Drug residues are considered environmental contaminants, and their occurrence has recently become a matter of concern. Analytical methods and monitoring systems are therefore required to control the continuous input of these drug residues into the environment. This article presents a suitable HPLC-ESI-MS/MS method for the simultaneous extraction, detection and quantification of residues of 13 drugs (antimicrobials, glucocorticosteroids, anti-inflammatories, anti-hypertensives, anti-cancer drugs and triphenylmethane dyes) in surface water. A monitoring study with 549 water samples was carried out in northwestern Spain to detect the presence of drug residues over two sampling periods during 2010, 2011 and 2012. Samples were collected from rural areas with and without farming activity and from urban areas. The 13 analytes were detected, and 18% of the samples collected showed positive results for the presence of at least one analyte. More collection sites were located in rural areas than in urban areas. However, more positive samples with higher concentrations and a larger number of analytes were detected in samples collected from sites located after the discharge of a WWTP. Results indicated that the WWTPs seems to act as a concentration point. Positive samples were also detected at a site located near a drinking water treatment plant.

Detection of veterinary drug residues in surface waters collected nearby farming areas in Galicia, North of Spain.

The occurrence of pharmaceuticals in the aquatic environment has become a matter of concern in the last decade due to potential risks posed to non-target organisms and the potential for unintended human exposure via food chain. This concern has been driven by a high detection frequency for drugs in environmental samples; these substances are produced in large quantities and are used in both veterinary and human medicine, leading to deposition and potential effects in the environment. However, few studies have focused on the presence of pharmaceuticals in rural areas associated with farming activities in comparison to urban areas. The aim of this study is to investigate the occurrence of pharmaceutically active compounds in surface waters collected from urban and rural areas in northwestern Spain. A monitoring study was conducted with 312 river water samples analysed by high-performance liquid chromatography coupled to tandem mass spectrometry. Positive detection of pharmaceuticals was made for 51 % of the samples. Decoquinate, sulfamethazine, sulfamethoxypyridazine and trimethoprim were the drugs most frequently detected, being present in more than 10 % of the samples. The sampling sites located downstream of the discharge points for wastewater treatment plants yielded the highest number of positive samples, 13 % of the positive samples were detected in these sites and 38 % of the samples collected near the collection point of a drinking water treatment plant were positive.

Determination of the Presence of Three Antimicrobials in Surface Water Collected from Urban and Rural Areas.

Due to the continuous release of antimicrobials into the environment, the aim of this study was to compare the frequency of detection of sulfamethazine, sulfamethoxypyridazine and trimethoprim in surface water collected from urban and rural areas in Northwestern Spain. A monitoring study was conducted with 314 river water samples analyzed by high-performance liquid chromatography coupled to tandem mass spectrometry. The results indicated that 37% of the samples contained residues of at least one of the investigated antimicrobials, and every sampling site yielded positive samples. At sites located near the discharge points of wastewater treatment plants and near the collection point of a drinking-water treatment plant, more than 6% of the samples were positive for the presence of antimicrobial residues.

Direct quantification and distribution of tetracycline-resistant genes in meat samples by real-time polymerase chain reaction.

The evolution of antimicrobial-resistant bacteria has become a threat to food safety and methods to control them are necessary. Counts of tetracycline-resistant (TR) bacteria by microbiological methods were compared with those obtained by quantitative PCR (qPCR) in 80 meat samples. TR Enterobacteriaceae counts were similar between the count plate method and qPCR (P= 0.24), whereas TR aerobic mesophilic bacteria counts were significantly higher by the microbiological method (P < 0.001). The distribution of tetA and tetB genes was investigated in different types of meat. tetA was detected in chicken meat (40%), turkey meat (100%), pork (20%), and beef (40%) samples, whereas tetB was detected in chicken meat (45%), turkey meat (70%), pork (30%), and beef (35%) samples. The presence of tetracycline residues was also investigated by a receptor assay. This study offers an alternative and rapid method for monitoring the presence of TR bacteria in meat and furthers the understanding of the distribution of tetA and tetB genes.

Detection and quantitative analysis of 21 veterinary drugs in river water using high-pressure liquid chromatography coupled to tandem mass spectrometry.

INTRODUCTION: The use of veterinary drugs in food production focuses on the control and improvement of animal health. The disadvantage of this practice is that pharmaceuticals and their metabolites are released into the environment, finding their way to natural water systems and becoming a potential risk to non-target organism. METHODS: This paper reports the development and validation of a quantitative method, based on high-performance liquid chromatography coupled to tandem mass spectrometry, for the simultaneous analysis of 21 veterinary drugs, antimicrobials, corticosteroids, coccidiostats and antifungal agents, in surface water. RESULTS: The precision of the method was established by calculating the mean recoveries, which were in the range of 94-101%. The developed method was employed to conduct the first monitoring study on the presence of veterinary drugs in the Galicia region, Northwest of Spain and was applied to 235 surface water samples. Eleven veterinary drugs were detected at concentrations from below the limit of quantification to 2,978.6 ng L(-1). Limits of detection and quantification were in the range of 6.2 (betamethasone, cortisone, decoquinate, dexamethasone, maduramycin, monensin, narasin, salinomycin, sulfachloropyridazine, sulfamethoxypyridazine and trimethoprim) to 12.5 ng L(-1) (for the rest of the selected drugs) and 12.5 (betamethasone, cortisone, decoquinate, dexamethasone, maduramycin, monensin, narasin, salinomycin, sulfachloropyridazine, sulfamethoxypyridazine and trimethoprim) to 25.0 ng L(-1) (for the remaining pharmaceuticals), respectively. CONCLUSION: Sulfonamides were the group most frequently found, which are widely used in veterinary medicine.

Determination of naturally occurring progestogens in bovine milk as their oxime derivatives using high performance liquid chromatography-electrospray ionization-tandem mass spectrometry.

BACKGROUND: Hormones and hormone-like substances which are present in the environment have been repeatedly accused of being the cause of most endocrine disruption. However, the possible role of endogenous hormones in food of animal origin deserves to be discussed as well. The relation between steroid hormones and several human health problems has been previously reported, such as prostate and breast cancer, perturbation of human reproduction and endocrine disruption on humans and wildlife. This research is particularly concerned with cow's milk, which contains a considerable amount of sex hormones. RESULTS: A liquid chromatography-tandem mass spectrometry method has been developed for the simultaneous detection and quantification of four naturally occurring steroid hormones in commercial bovine milk (pregnenolone (P5), progesterone (P4), 17-hydroxypregnenolone (17-OHP5), 17-hydroxyprogesterone (P4)). Oxime derivatives of steroids were analyzed in positive ionization and multiple reaction monitoring mode. Methodology has been validated according to Decision 2002/657/EC criteria. CONCLUSION: This method has been successfully used in real samples. It is fast and easy-handling and provides a useful tool for the assessment of progestogens in bovine milk.

Development and validation of an LC-MS/MS confirmatory method for residue analysis of cyproheptadine in urine of food-producing animals.

The possible off-label and illegal use of cyproheptadine (CYP) as an appetite stimulant for food-producing animals creates the need for methods capable of detecting it. A high-performance liquid chromatography tandem mass spectrometry method (LC-MS/MS) was developed to identify CYP in bovine urine, according to Commission Decision 2002/657/EC. Two multiple reaction monitoring (MRM) transitions for each analyte were monitored: 288.1/96.1 and 288.1/191.2 for CYP and 282.1/167.2 and 282.1/116.3 for diphenylpyraline hydrochloride (DPP), which was used as an internal standard. The solid phase extraction technique without a liquid-liquid step gives good results in urine samples from treated animals. The analytical method was successfully validated for linearity (0.15-10 ng/mL), with intraday precision of 9.4%, interday precision of 20.4%, and accuracy of 96.7%. The decision limit (CCalpha) and detection capability (CCbeta) were 0.48 and 0.82 ng/mL, respectively.

Development of a rapid and confirmatory procedure to detect 17beta-estradiol 3-benzoate treatments in bovine hair.

A high-performance liquid chromatography-tandem mass spectrometry method (LC-MS/MS) was developed for efficient and confirmatory surveillance of illegal use of estradiol benzoate, even when this substance is used in reproductive control. After cryogenic grinding, estradiol benzoate was extracted from hair with acetonitrile for 24 h on a rocking table. The validation of the method was based on Commission Decision 2002/657/EC using the deuterated analogue of estradiol benzoate as internal standard. Decision limit (0.81 ng/g), detection capability (1.38 ng/g), repeatability CV% (13.7), within in laboratory reproducibility CV% (15.6%), and trueness (99.3%) were calculated. Using the proposed methodology the presence of estradiol benzoate in samples obtained from animals treated to synchronize their estrous cycles can be confirmed.

Cryogenic grinding pre-treatment improves extraction efficiency of fluoroquinolones for HPLC-MS/MS determination in animal tissue.

An efficiency extraction of fluoroquinolones in chicken muscle was achieved by pulverizing it in a freezer mill before treatment with NaOH (10mM)/MeCN (1:1). The improvement of cryogenic grinding in the extraction was demonstrated for the same piece (whole leg) of four chickens treated with enrofloxacin in equal doses. A confirmatory method based on high-performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) was used to analyze the extracts. The chromatographic separation was achieved in 5 min with a Synergi Fusion-RP 80A (50 x 2 mm, 4 microm) column filled with a hybrid polymer. The HPLC was coupled with a detector based in a quadrupole-linear ion trap Q-TRAP that allows a confirmatory detection according to the European legislation. The specificity of the method was assessed by testing a number of representative blank muscle samples (n = 10) to verify the absence of potential interfering compounds. The limits of detection and quantitation were 2 and 5 ng g(-1) of quinolones in muscle samples, respectively. The chromatographic method was demonstrated to be linear for the range studied (5-500 ng g(-1)) with the P value for lack-of-fit in the ANOVA table greater or equal to 0.10 (calibration coefficient 0.9998 and 0.9996 for ciprofloxacin and enrofloxacin, respectively). The mean intra-day relative standard deviation (RSD) (n = 6, c = 50 ng g(-1)) was 6%; inter-day assay gave a RSD of 12%. The extraction and clean-up were carried out in one step with very satisfactory recovery data (between 65 and 101%).

Microbiological and physicochemical properties of fresh retail cuts of beef packaged under an advanced vacuum skin system and stored at 4 degrees C.

The effect of an advanced vacuum skin packaging system on the microbiological and physicochemical properties of fresh retail cuts of beef (including meat portions from six different anatomical regions) stored at 4 degrees C was compared with the effect of traditional vacuum packaging. The vacuum skin packaging system whose effect on meat quality was evaluated in this work displayed two remarkable features: (i) the instantaneous heating of the lower surface of the upper film of the package before the film descended over the meat surface and (ii) the tight disposition of the plastic film on the meat surface, which avoided wrinkles and purges. Throughout storage at 4 degrees C, rates of bacterial growth were statistically significantly slower in beef portions processed with the vacuum skin packaging system than in those processed with traditional vacuum packaging, with average differences of 2.07, 1.60, and 1.25 log CFU/g in counts of aerobic mesophiles, anaerobes, and lactic acid bacteria, respectively. pH values were statistically significantly lower for beef portions packaged with the vacuum skin system than for those that were vacuum packaged in the traditional manner, probably because of the relative predominance of lactic acid bacteria observed in such samples, which coincided with both higher meat firmness values and a slower meat tenderization process. The vacuum skin system prevented the appearance of undesirable coloration on the meat surface and also significantly improved the commercial color of the meat as determined on the basis of luminosity (L*) and the redness (a*). Overall, the quality (as determined by microbiological and physicochemical analyses and by visual examination) of fresh retail cuts of beef packaged with the vacuum skin system and stored at 4 degrees C was higher than that of meat samples processed with the traditional vacuum-packaging system.

Comparison of extraction methods for the recovery, amplification and species-specific analysis of DNA from bone and bone meals.

We report the effect of several parameters on the efficiency of recovery of DNA from animal bones. The effects of preheating the samples (at either 60 degrees C or 100 degrees C) at different intervals (from 1 h to overnight) in different media (water, 0.5 M ethylenediaminetetraacetic acid (EDTA), or 0.5 M EDTA + 0.05% sodium dodecyl sulfate (SDS) were investigated. The effect of slight (5 min) or intense (30 min) pretreatments with ultrasound was also evaluated. Several different treatments with proteinase K (ranging from 200 to 800 microg, and lasting from 1 to 3 h) at 65 degrees C were also considered. Additionally, two different DNA extraction methods (based on silica resins and purification columns, respectively) were evaluated. The recovery of DNA from the samples was 40% higher when the bones were preheated in 0.5 M EDTA at 60 degrees C for 1 h, this being followed by treatment with 800 microg of proteinase K for 3 h. The DNA thus obtained was successfully amplified by polymerase chain reaction (PCR) using a set of primers specific to a 359 bp region of the mitochondrial cytochrome b gene, and the species of origin were identified by visualizing the restriction fragment length polymorphism (RFLP) with the endonucleases PalI and MboI.