Genomic diversity of the locally developed Latvian Darkheaded sheep breed.

The Latvian Darkheaded is the only locally developed sheep breed. The breed was formed at the beginning of the 20th century by crossing local coarse-wooled sheep with the British Shropshire and Oxfordshire breeds. The breed was later improved by adding Ile-de-France, Texel, German blackheads, and Finnsheep to achieve higher prolificacy and better meat quality. Previous studies have reported the Latvian Darkheaded sheep to be closely related to Estonian and Lithuanian Blackface breeds, according to microsatellite data. To expand our knowledge of the genetic resources of the Latvian Darkheaded breed, we conducted a whole-genome resequencing analysis on 40 native sheep. The investigation showed that local sheep harbor genetic diversity levels similar to those observed among other improved breeds of European origin, including Charollais and Suffolk. Genome-wide nucleotide diversity (π) in Latvian Darkheaded sheep was 3.91 × 10-3, whereas the average observed heterozygosity among the 40 animals was 0.267 and 0.438 within the subsample of unrelated individuals. The Ne has rapidly decreased to 200 ten generations ago with a recent drop to Ne 73 four generations ago. However, inbreeding levels based on runs of homozygosity were, on average, low, with FROH ranging between 0.016 and 0.059. The analysis of the genomic composition of the breed confirmed shared ancestry with sheep of British origin, reflecting the history of the breed. Nevertheless, Latvian Darkheaded sheep were genetically separable. The contemporary Latvian Darkheaded sheep population is genetically diverse with a low inbreeding rate. However, further development of breed management programs is necessary to prevent an increase in inbreeding, loss of genetic diversity, and depletion of breed-specific genetic resources, ensuring the preservation of the native Latvian Darkheaded sheep.

Assessment of the inclusion of a feed additive of sodium humate derived from freshwater sapropel in diets for broiler chickens.

Background and Aim: Poultry production is the fastest growing livestock industry in the world, as the rapid growth of and efficient absorption of feed by poultry ensure the production of poultry meat with a relatively low carbon footprint. Seeking new ways to increase livestock productivity as well as poultry product quality, the number of research studies on the use of humic substances of various origins in livestock farming has increased significantly, emphasizing the role of feed additives derived from local resources. The unique capability of humic substances to improve metabolic processes allows the immune protection of the bird body to be strengthened and production efficiency to be increased. This study aimed to identify the effects of sodium humate (NaHum) on the growth performance of broiler chickens and selected blood and ileum microbiota parameters. Materials and Methods: Dietary research was conducted 2 times under production conditions in a poultry facility of a commercial company, with 210 1-day-old, unsorted broiler chickens of both sexes (Ross 308). The broiler chickens were fed with standard commercial feed, the rearing period of 35 days, and slaughtered on day 36. Sodium humate additive was added to drinking water for the research groups of broilers in period from 8th to 35th day of life, 25 mL (Group 1, n = 2 × 35) and 50 mL (Group 2, n = 2 × 35) per liter of drinking water. Sodium humate contained an average of 4.48% dry matter, a kilogram of dry matter containing 104.3 g of crude protein, 3.6 g of crude fiber and 0.9 g of crude fat, 14.3 MJ of metabolic energy, and 5.8 MJ of energy for live weight gain, as well as a very high content of crude ash -759.8 g, including 4.2 g Ca, 4.2 g Na, and 4.81 g Fe, the dry matter digestibility of NaHum was 87.0%, and the absorption capacity of dry matter was 113.2%., the pH level was 13.0, i.e., alkaline. At the end of the dietary research, the productivity and economic efficiency of the research groups of broilers were calculated by live weight gain, carcass weight, feed conversation ratio, and blood and intestinal samples of broilers were analyzed to identify the effects of NaHum on the growth performance and health status of broilers. Results: Dietary research found that adding 25 mL/L and 50 mL/L of NaHum to drinking water for the broiler chickens increased their live weights at the selling age, average live weight gains by 3.06-3.93%, and carcass weights by 5.07-6.06%, while feed conversion increased in terms of both live weight (1.5 and 1.51) and carcass weight (1.84 and 1.86) compared with the control group. The best economic performance in terms of the economic efficiency index and the cost index (CI) was found in Group 1, which was fed with the NaHum additive at an intake rate of 25 mL/L. The NaHum additive modulated the ileal microbiota and metabolic processes in the broiler body. At the same time, a significant decrease in the levels of total protein, alkaline phosphatase and phosphorus (P) in blood was found in the research groups. Conclusion: Considering the positive effects of NaHum derived from freshwater sapropel on the productivity and economic efficiency of broiler chickens, the NaHum feed additive should be further investigated on a larger scale to obtain results that could reasonably be used in practice. This study concluded that a decrease in P levels in the blood was observed when NaHum was added to the drinking water; therefore, it is important to continue the research to draw reasonable conclusions on the effects of NaHum in liquid form on the health performance of farm animals.

Elaboration of Nanostructured Levan-Based Colloid System as a Biological Alternative with Antimicrobial Activity for Applications in the Management of Pathogenic Microorganisms.

Considering the documented health benefits of bacterial exopolysaccharides (EPSs), specifically of bacterial levan (BL), including its intrinsic antimicrobial activity against certain pathogenic species, the current study concentrated on the development of active pharmaceutical ingredients (APIs) in the form of colloid systems (CoSs) containing silver nanoparticles (AgNPs) employing in-house biosynthesized BL as a reducing and capping agent. The established protocol of fermentation conditions implicating two species of lactic acid bacteria (LAB), i.e., Streptococcus salivarius K12 and Leuconostoc mesenteroides DSM 20343, ensured a yield of up to 25.7 and 13.7 g L-1 of BL within 72 h, respectively. An analytical approach accomplished by Fourier-transform infrared (FT-IR) spectroscopy allowed for the verification of structural features attributed to biosynthesized BL. Furthermore, scanning electron microscopy (SEM) revealed the crystalline morphology of biosynthesized BL with a smooth and glossy surface and highly porous structure. Molecular weight (Mw) estimated by multi-detector size-exclusion chromatography (SEC) indicated that BL biosynthesized using S. salivarius K12 has an impressively high Mw, corresponding to 15.435 × 104 kilodaltons (kDa). In turn, BL isolated from L. mesenteroides DSM 20343 was found to have an Mw of only 26.6 kDa. Polydispersity index estimation (PD = Mw/Mn) of produced BL displayed a monodispersed molecule isolated from S. salivarius K12, corresponding to 1.08, while this was 2.17 for L. mesenteroides DSM 20343 isolate. The presence of fructose as the main backbone and, to a lesser extent, glucose and galactose as side chain molecules in EPS hydrolysates was supported by HPLC-RID detection. In producing CoS-BL@AgNPs within green biosynthesis, the presence of nanostructured objects with a size distribution from 12.67 ± 5.56 nm to 46.97 ± 20.23 was confirmed by SEM and energy-dispersive X-ray spectroscopy (EDX). The prominent inhibitory potency of elaborated CoS-BL@AgNPs against both reference test cultures, i.e., Pseudomonas aeruginosa, Escherichia coli, Enterobacter aerogenes, and Staphylococcus aureus and those of clinical origin with multi-drug resistance (MDR), was confirmed by disc and well diffusion tests and supported by the values of the minimum inhibitory and bactericidal concentrations. CoS-BL@AgNPs can be treated as APIs suitable for designing new antimicrobial agents and modifying therapies in controlling MDR pathogens.

Genomic Characterization and Initial Insight into Mastitis-Associated SNP Profiles of Local Latvian Bos taurus Breeds.

Latvia has two local Bos taurus breeds-Latvian Brown (LBG) and Latvian Blue (LZG)-characterized by a good adaptation to the local climate, longevity, and high fat and protein contents in milk. Since these are desired traits in the dairy industry, this study investigated the genetic background of the LBG and LZG breeds and identified the genetic factors associated with mastitis. Blood and semen samples were acquired, and whole genome sequencing was then performed to acquire a genomic sequence with at least 35× or 10× coverage. The heterozygosity, nucleotide diversity, and LD analysis indicated that LBG and LZG cows have similar levels of genetic diversity compared to those of other breeds. An analysis of the population structure revealed that each breed clustered together, but the overall differentiation between the breeds was small. The highest genetic variance was observed in the LZG breed compared with the LBG breed. Our results show that SNP rs721295390 is associated with mastitis in the LBG breed, and SNPs rs383806754, chr29:43998719CG>C, and rs462030680 are associated with mastitis in the LZG breed. This study shows that local Latvian LBG and LZG breeds have a pronounced genetic differentiation, with each one suggesting its own mastitis-associated SNP profile.

The Evaluation of Roasted Lentils (L. culinaris L.) Quick Meals as An Alternative to Meat Dishes.

Despite the health-promoting benefits, the consumption of lentils in East Europe is low, attracting researchers' interest in solving the problem. The aim of this study was to develop an alternative to animal proteins for nutrient-dense plant-based quick meals using roasted lentils as the primary raw material, performing sensory analysis, and evaluating the content of amino acids, minerals, and vitamins. The consumption of legumes in Latvia is also low, even though most respondents associate the use of legumes with a healthy choice. Roasted lentil quick meals can deliver 15.6% and 26.2% of the reference intake for protein. Furthermore, one-third of the amino acids (AAs) are essential AAs. AA values in prepared quick meals make them promising alternatives to meat products. One portion of ready-roasted lentils with Bolognese sauce provided above 15% of the daily reference intake of thiamin and vitamin B9. One portion of a ready-quick meal of tomato soup with roasted lentils and roasted lentils with Bolognese sauce provided 20.3% and 25.6% of iron, according to daily reference intake. Further studies on the bioavailability of quick meals must be conducted to claim they can replace meat nutritionally.

The Release of Non-Extractable Ferulic Acid from Cereal By-Products by Enzyme-Assisted Hydrolysis for Possible Utilization in Green Synthesis of Silver Nanoparticles.

The present work was undertaken to elucidate the potential contribution of biosynthetically produced ferulic acid (FA) via enzymatic hydrolysis (EH) of rye bran (RB) to the formation of silver nanoparticles (AgNPs) during green synthesis. An analytical approach accomplished by multiple reaction monitoring (MRM) using triple quadrupole mass selective detection (HPLC-ESI-TQ-MS/MS) of the obtained hydrolysate revealed a relative abundance of two isomeric forms of FA, i.e., trans-FA (t-FA) and trans-iso-FA (t-iso-FA). Further analysis utilizing high-performance liquid chromatography with refractive index (HPLC-RID) detection confirmed the effectiveness of RB EH, indicating the presence of cellulose and hemicellulose degradation products in the hydrolysate, i.e., xylose, arabinose, and glucose. The purification process by solid-phase extraction with styrene-divinylbenzene-based reversed-phase sorbent ensured up to 116.02 and 126.21 mg g-1 of t-FA and t-iso-FA in the final eluate fraction, respectively. In the green synthesis of AgNPs using synthetic t-FA, the formation of NPs with an average size of 56.8 nm was confirmed by scanning electron microscopy (SEM) and energy dispersive spectroscopy (EDS) techniques. The inclusion of polyvinylpyrrolidone (PVP-40) in the composition of NPs during synthesis favorably affected the morphological features, i.e., the size and shape of AgNPs, in which as big as 22.4 nm NPs were engineered. Meanwhile, nearly homogeneous round-shaped AgNPs with an average size of 16.5 nm were engineered using biosynthetically produced a mixture of t-FA and t-iso-FA and PVP-40 as a capping agent. The antimicrobial activity of AgNPs against Gram-positive and Gram-negative bacteria, including Pseudomonas aeruginosa, E. coli, Enterococcus faecalis, Bacillus subtilis, and Staphylococcus aureus was confirmed by the disk diffusion method and additionally supported by values of minimum inhibitory (MIC) and bactericidal (MBC) concentrations. Given the need to reduce problems of environmental pollution with cereal processing by-products, this study demonstrated a technological solution of RB rational use in the sustainable production of AgNPs during green synthesis. The AgNPs can be considered as active pharmaceutical ingredients (APIs) to be used for developing new antimicrobial agents and modifying therapies in treating multi-drug resistant (MDR) pathogens.

Analysis of 18 Free Amino Acids in Honeybee and Bumblebee Honey from Eastern and Northern Europe and Central Asia Using HPLC-ESI-TQ-MS/MS Approach Bypassing Derivatization Step.

The profile of amino acids and mono- and disaccharides in conventional polyfloral honey originated from Latvia and Tajikistan and less found in nature bumblebee honey from Russia was investigated. The analysis of free amino acids (FAAs) accomplished by multiple reaction monitoring (MRM) using triple quadrupole mass selective detection (HPLC-ESI-TQ-MS/MS) revealed the presence of 17 FAAs. The concentration of FAAs varied in the range of 0.02-44.41 mg 100 g-1 FW. Proline was the main representative of FAAs, contributing to the total amount of FAAs from 41.7% to 80.52%. The highest concentration of proline was found in bumblebee and buckwheat honey, corresponding to 44.41 and 41.02 mg 100 g-1, respectively. The concentration of essential amino acids (AAs), i.e., leucine, and isoleucine was found to be the highest in buckwheat honey contributing up to 12.5% to the total amount of FAAs. While, the concentration of branched-chain AAs fluctuated within the range of 1.08-31.13 mg 100 g-1 FW, with buckwheat honey having the highest content and polyfloral honey the lowest, respectively. The results of this study confirmed the abundance of FAAs both in honeybee and bumblebee honey. However, the concentration of individual FAAs, such as proline, aspartic acid, leucine, and isoleucine in bumblebee honey was many folds higher than observed in honeybee polyfloral honey.

An Environmentally Friendly Approach for the Release of Essential Fatty Acids from Cereal By-Products Using Cellulose-Degrading Enzymes.

The main intention of the present work was to investigate the ability of cellulose-degrading enzymes (C-DE) to release fatty acids (FAs) from complex matrices of cereal by-products during enzymatic hydrolysis (EH). For this purpose, three types of cereal bran (CB), i.e., wheat, rye, and oat, were used as lignocellulose substrates for three commercially available hydrolytic enzymes, i.e., Viscozyme L, Viscoferm, and Celluclast 1.5 L. The yield and composition of FAs after EH were assessed and compared with those obtained after either conventional Soxhlet extraction or after alkaline-assisted hydrolysis (A-AH) with 10% KOH in 80% MeOH and subsequent liquid-liquid extraction. The experimental results demonstrated that up to 6.3% and 43.7% higher total FA yield can be achieved by EH of rye bran using Celluclast 1.5 L than by A-AH and Soxhlet extraction, respectively. However, the application of Viscoferm for EH of wheat bran ensured up to 7.7% and 13.4% higher total FA yield than A-AH and Soxhlet extraction, respectively. The concentration of essential linolenic acid (C18:3) in lipids extracted after EH of rye bran with Celluclast 1.5 L was up to 24.4% and 57.0% higher than in lipids recovered by A-AH and Soxhlet extraction, respectively. In turn, the highest content of linolenic acid in wheat bran lipids was observed after EH with Viscoferm and Viscozyme L, ensuring 17.0% and 13.6% higher yield than after A-AH, respectively. SEM analysis confirmed substantial degradation of the CB matrix promoted by the ability of C-DE to act specifically on glycosidic bonds in cellulose and on xylosidic bonds in arabinoxylans, arabinans, and other arabinose-containing hemicelluloses. Structural alterations in cell integrity greatly contributed to the release of bound FAs and their better transfer into the extraction solvent. It has been shown that the proposed process of EH can be used for the efficient release of FAs from the CB matrix more sustainably and with a safer profile, thereby conveying greener production of FAs for certain purposes.

Effect of Essential Oils Supplemented with Caprylic Acid and Sodium Chloride against Faecal ESBL-Producing Escherichia coli Isolated from Pigs.

The purpose of the present investigation was to compare the antibacterial activity of six commercial and lab-scale extracted essential oils (EOs) alone or in combination with caprylic acid (CA) and sodium chloride (NaCl) against faecal Escherichia coli with and without extended-spectrum beta-lactamase (ESBL) encoding genes, and of isolates classified as multidrug-resistant (MDR). Gas chromatography−mass spectrometry (GC−MS) was used for the analysis of chemical composition of EOs, while the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) assays were carried out to elucidate the antibacterial activity of non-supplemented and supplemented EOs against different resistance levels of E. coli strains. The main compounds in commercial EOs were aromatic monoterpenoids (30−56%) and p-cymene (8−35%), while the main compounds in the lab-scale EOs were aromatic monoterpenoids (12−37%) and γ-terpinene (18−22%). Commercial EOs exhibited superior inhibitory activity of E. coli in comparison to lab-scale produced EOs. Antibacterial activity of EOs was significantly enhanced by enrichment of the EOs with NaCl (p < 0.001) or CA (p = 0.012). Most of the non-supplemented EOs exhibited lower activity against MDR and ESBL producing E. coli. In contrast, EOs supplemented with CA and especially NaCl was equally effective against ESBL and non-ESBL as well as MDR and non-MDR E. coli. It was found that supplementation of EOs with NaCl could enhance the antibacterial activity towards ESBL and MDR E. coli isolates. However, additional studies are needed to clarify the potential risks of developing resistance.

AK-2011 strain for the development of a vaccine against equine rhinopneumonitis.

Equine rhinopneumonitis is an acute, highly contagious disease found virtually worldwide. The purpose of the studies presented in this paper is to develop a technology for the manufacture of a cell-derived equine rhinopneumonitis vaccine, as well as to assess the safety and immunogenicity of the newly developed vaccine in laboratory animals model. The object of the studies was the AK-2011 strain isolated from the horses suffering from rhinopneumonitis during an outbreak of abortions. The viability of the AK-2011 strain was assessed using a continuous line of calf trachea cells, a continuous line of calf kidney cells, a continuous line of sheep kidney cells, a continuous line of bovine kidney cells, a continuous line of green monkey kidney cells, a continuous line of Syrian hamster kidney cells, a primary trypsinized culture of horse kidney cells grown in tubes and flasks and the AK-2011 laboratory strain of equine rhinopneumonitis virus with biological activity of 6.0 lg TCID50/cm 3 . Sequencing and polymerase chain reaction analysis were performed. The virus isolated from the ORF68 gene in Kazakhstan appeared to be the most similar to the T-953 and 2222-03 strains isolated in the USA and Australia, respectively, in terms of phylogenetics. As to primary infections, cytopathic effects (CPEs) induced by the AK-2011 virus stain (dilution 101 ) in calf trachea and horse kidney cell cultures were stable from the first to tenth passages, with biological activity of 5.75-6.00 lg TCID50/cm 3 . CPEs caused by the virus were apparent on days 2-3, further developed intensively and extended to 60-80% of the cell monolayer on days 5-7. The vaccine results can be used to immunize horses on farms against rhinopneumonia, and horses should be immunized twice with an interval of 2-3 months.

Lignocellulose-Degrading Enzymes: A Biotechnology Platform for Ferulic Acid Production from Agro-Industrial Side Streams.

Biorefining by enzymatic hydrolysis (EH) of lignocellulosic waste material due to low costs and affordability has received enormous interest amongst scientists as a potential strategy suitable for the production of bioactive ingredients and chemicals. In this study, a sustainable and eco-friendly approach to extracting bound ferulic acid (FA) was demonstrated using single-step EH by a mixture of lignocellulose-degrading enzymes. For comparative purposes of the efficiency of EH, an online extraction and analysis technique using supercritical fluid extraction-supercritical fluid chromatography-mass spectrometry (SFE-SFC-MS) was performed. The experimental results demonstrated up to 369.3 mg 100 g-1 FA release from rye bran after 48 h EH with Viscozyme L. The EH of wheat and oat bran with Viscoferm for 48 h resulted in 255.1 and 33.5 mg 100 g-1 of FA, respectively. The release of FA from bran matrix using supercritical fluid extraction with carbon dioxide and ethanol as a co-solvent (SFE-CO2-EtOH) delivered up to 464.3 mg 100 g-1 of FA, though the extractability varied depending on the parameters used. The 10-fold and 30-fold scale-up experiments confirmed the applicability of EH as a bioprocessing method valid for the industrial scale. The highest yield of FA in both scale-up experiments was obtained from rye bran after 48 h of EH with Viscozyme L. In purified extracts, the absence of xylose, arabinose, and glucose as the final degradation products of lignocellulose was proven by high-performance liquid chromatography with refractive index detection (HPLC-RID). Up to 94.0% purity of FA was achieved by solid-phase extraction (SPE) using the polymeric reversed-phase Strata X column and 50% EtOH as the eluent.

The Prevalence and Characterization of Fecal Extended-Spectrum-Beta-Lactamase-Producing Escherichia coli Isolated from Pigs on Farms of Different Sizes in Latvia.

The aim of this study was to determine the prevalence of fecal ESBL-producing Escherichia coli (E. coli) in pigs on large and small farms in Latvia, to characterize beta-lactamase genes and establish an antimicrobial resistance profile. Fecal samples (n = 615) were collected from 4-week, 5-week, 6-week, 8-week, 12-week and 20-week-old piglets, pigs and sows on four large farms (L1, L2, L3, L4) and three small farms (S1, S2, S3) in Latvia. ChromArt ESBL agar and combination disc tests were used for the screening and confirmation of ESBL-producing E. coli. The antimicrobial resistance was determined by the disc diffusion method and ESBL genes were determined by polymerase chain reaction (PCR). Subsequently, ESBL-producing E. coli was confirmed on three large farms, L1 (64.3%), L2 (29.9%), L3 (10.7%) and one small farm, S1 (47.5%); n = 144 (23.4%). The prevalence of ESBL-producing E. coli differed considerably between the large and small farm groups (26.9% vs. 12.7%). Of ESBL E. coli isolates, 96% were multidrug-resistant (MDR), demonstrating there were more extensive MDR phenotypes on large farms. The distribution of ESBL genes was blaTEM (94%), blaCTX-M (86%) and blaSHV (48%). On the small farm, blaSHV dominated, thus demonstrating a positive association with resistance to amoxicillin-clavulanic acid, ceftazidime and cefixime, while on the large farms, blaCTX-M with a positive association to cephalexin and several non-beta lactam antibiotics dominated. The results indicated the prevalence of a broad variety of ESBL-producing E. coli among the small and large farms, putting the larger farms at a higher risk. Individual monitoring of ESBL and their antimicrobial resistance could be an important step in revealing hazardous MDR ESBL-producing E. coli strains and reviewing the management of antibiotic use.

Highly-Efficient Release of Ferulic Acid from Agro-Industrial By-Products via Enzymatic Hydrolysis with Cellulose-Degrading Enzymes: Part I-The Superiority of Hydrolytic Enzymes Versus Conventional Hydrolysis.

Historically Triticum aestívum L. and Secale cereále L. are widely used in the production of bakery products. From the total volume of grain cultivated, roughly 85% is used for the manufacturing of flour, while the remaining part is discarded or utilized rather inefficiently. The limited value attached to bran is associated with their structural complexity, i.e., the presence of cellulose, hemicellulose, and lignin, which makes this material suitable mostly as a feed supplement, while in food production its use presents a challenge. To valorize these materials to food and pharmaceutical applications, additional pre-treatment is required. In the present study, an effective, sustainable, and eco-friendly approach to ferulic acid (FA) production was demonstrated through the biorefining process accomplished by non-starch polysaccharides degrading enzymes. Up to 11.3 and 8.6 g kg-1 of FA was released from rye and wheat bran upon 24 h enzymatic hydrolysis with multi-enzyme complex Viscozyme® L, respectively.

Effect of Probiotics and Herbal Products on Intestinal Histomorphological and Immunological Development in Piglets.

The aim of the study was to evaluate the effect of probiotics and herbal products on the intestinal histomorphological and immunological development in piglets. Accordingly, 2-week-old piglets were allocated in 4 groups: C (basal diet), Pro (basal diet + probiotics), Pro+B (basal diet + probiotics + buckwheat bran), and H (powder of herbs). After 6 weeks of the experiment, 4 piglets from each experimental group were randomly selected and slaughtered at a slaughterhouse. Samples of tissue and digestive content from the jejunum and colon were collected for bacteriological, histological, and immunohistochemical examination. The results showed that probiotics increased the number of Lactobacillus spp. in the small (p < 0.05) and large intestines. The intestinal histomorphology was improved (p < 0.05) in all experimental groups by an increased villus height, VH : CD ration, colon crypt depth, and number of Ki-67+ epithelial cells. A higher number (p < 0.05) of goblet cells and their acidification were observed in group Pro, while the density of goblet cells was decreased by the herbs. Probiotics increased (p < 0.05) the number of intraepithelial lymphocytes (IELs), density of CD3+ cells in Peyer's patches (PPs), and lamina propria (LP). In group H, a dual effect on the CD3+ cell distribution was observed. The herbs reduced (p < 0.05) the number of IELs and CD3+ in LP but increased the distribution of CD3+ cells in PPs. In the colon, herbs increased CD3+ cells in LP as well. It suggests that probiotics and herbs had influence on the intestinal histomorphology and the ability to modulate the mucosal immune system; however, the combination of probiotics and buckwheat bran was not so convincing, probably due to the inhibitory effect of the buckwheat bran on the probiotics used.

Cells and Cytokines in Milk of Subclinically Infected Bovine Mammary Glands after the Use of Immunomodulatory Composition GLP 810.

The aim of this study was to investigate the effect of intramammary infusions of natural composition GLP 810 with immunomodulating properties on the local nonspecific cellular and humoral immune response in cows with subclinical mastitis. The composition GLP 810 consists of lactic acid, lysozyme, glycopeptides, and 0.9% solution of NaCl. The following parameters were studied: (1) leukocyte differential distribution in milk, (2) expression of cytokines in milk leukocytes, (3) antibacterial activity, and (4) milk quality. Nineteen mammary glands in five lactating cows were infused with 10 mL of GLP 810, and nineteen other glands from five control cows were treated with 10 mL 0.9% NaCl. The results showed that after intramammary administration of the composition GLP 810 three times with 48 h intervals, the following effects on leukocyte populations in milk were observed: (1) an increase in the number of polymorphonuclear leukocytes and lymphocytes and (2) a decrease in the number of macrophages. A reduction in the number of pathogenic bacteria was also detected. The analyses of tumour necrosis factor-alpha, interleukin-10, and beta-defensin-2 revealed that the production of the aforementioned cytokines significantly increased, whereas no significant effects on interleukin-1 and caspase-6 expression in milk leukocytes were recorded. However, there were significant differences between mammary glands with high and low milk somatic cell count. The results suggest that the composition GLP 810 has an immunomodulatory effect on mammary glands and it could be used for improving the immune response in cows with subclinical mastitis during lactation.

Valorization of Wild Apple (Malus spp.) By-Products as a Source of Essential Fatty Acids, Tocopherols and Phytosterols with Antimicrobial Activity.

The amplified production of fruit as well as burgeoning demand for plant-made food products have resulted in a sharp increase of waste. Currently, millions of tons of by-products are either being discarded or utilized rather ineffectively. However, these by-products may be processed and further incorporated as functional ingredients in making high-value food products with many physiological and biochemical effects. The chemical analysis of pomace oils using gas chromatography-mass spectrometry (GC/MS) and reversed-phase-liquid chromatography coupled with fluorescence detector (RP-HPLC/FLD) systems led to the identification and quantification of 56 individual lipophilic compounds including unsaturated, polyunsaturated and saturated fatty acids, as well as phytosterols and four homologs of tocopherol. The oils recovered from by-products of Malus spp. (particularly cv. "Ola") are rich in fatty acids such as linolenic (57.8%), α-linolenic (54.3%), and oleic (25.5%). The concentration of total tocopherols varied among the Malus species and dessert apples investigated, representing the range of 16.8⁻30.9 mg mL-1. The highest content of total tocopherols was found in M. Bernu prieks, followed by M. cv. "Ola", and M. × Soulardii pomace oils. A significantly higher amount of δ-tocopherol was established in the oil of M. Bernu prieks, indicating that this species could be utilized as a natural and cheap source of bioactive molecules. ß-Sitosterol was the prevalent compound determined in all tested pomace oils with a percentage distribution of 10.3⁻94.5%. The main triterpene identified in the oils was lupeol, which varied in the range of 0.1⁻66.3%. A targeted utilization of apple pomace would facilitate management of tons of by-products and benefit the environment and industry.

Presence of Methicillin-resistant Staphylococcus Aureus in Slaughterhouse Environment, Pigs, Carcasses, and Workers.

INTRODUCTION: Methicillin-resistant Staphylococcus aureus (MRSA) is a highly resistant and difficult to cure zoonotic microorganism, which makes up a large part of food toxic infections and has shown high prevalence among pig population all over the world. The aim of the study was to establish the occurrence of MRSA in slaughterhouses, evaluate its antimicrobial resistance, and verify whether there are any differences or similarities with reference to other European countries. MATERIAL AND METHODS: A total of 100 pigs, 105 carcasses, 19 workers, and 24 samples from the environment of several slaughterhouses were examined by conventional microbial and molecular methods. RESULTS: In total, 78 MRSA isolates were found. MRSA prevalence in slaughtered pigs varied from 8.0% to 88.6% depending on the slaughterhouse, reaching higher prevalence in slaughterhouses with higher slaughter capacity. In total, 21.1% of all workers were carriers of MRSA and 6.7% of carcasses were contaminated with MRSA. The 98.2% of MRSA isolates were resistant to penicillin, 89.1% to tetracycline, 60.1% to erythromycin, 65.5% to gentamycin, and 15 different spa types were found, among which spa type t01333 was most widespread. CONCLUSION: The study indicated that MRSA prevalence and spa types differed according to slaughterhouse slaughter capacity and good hygiene practices. Quite high MRSA occurrence among slaughterhouse workers is one of the main factors which increase pork contamination risk.

MRSA in Pig Population.

Methicillin resistant Staphylococcus aureus (MRSA) is widespread worldwide in different types of animal species and as a zoonosis takes a great risk for human health not only as a food toxicoinfection, but also as a highly resistant pathogen causing serious soft tissue infectious, septicaemia and even death. One of the most affected food-producing animal species is swine in the production of which new antibiotics in big amounts are used more and more continuously, increasing antimicrobial resistance. In this study several commercial pig farms and pigs with different age groups as well as farm workers and samples from environment were examined with the purpose of detecting MRSA prevalence and evaluating antimicrobial resistance. A total of 85 isolated MRSA strains were characterised by conventional microbial and molecular methods. MRSA was found in all farms. MRSA prevalence in different pig age groups and farms varied from none to 79.2% reaching higher values among 3-3.5 (26.6%) and 4-4.5 (31.9%) old pigs. The 98.7% of 74 further investigated MRSA isolates were resistant to penicillin, 94.9% to tetracycline, 45.6% to cephalexin and 10 different spa types were found among which spa type t011 was the most widespread. To the best of our knowledge, this is the first time MRSA was researched in sow milk and the first description of the presence of MRSA in several age groups of pigs in Latvia.