Effects of ultrasound-induced stress on gut microbiota of mice.

Background and Aim: Prolonged stress causes deleterious effects on both the organism and its microbiota. In this study, we examined the effects of exposure to variable frequency ultrasound (US) on the gut microbiota-liver-brain axis of mice. Materials and Methods: This study was conducted on 20 mature clinically healthy sexually naive C57BL/6J male mice (42-45 days old). Group 1 (Normal) consisted of healthy intact mice (n = 10). Group 2 (Stress) consisted of mice subjected to US-induced stress (n = 10) for 20 days with alternating frequencies (20-45 kHz). Stool samples were collected on days 0, 10, and 20, and the corresponding DNA was later subjected to 16SrRNA sequencing. After mice were sacrificed on day 21, the leukocyte count, blood serum biochemical parameters, and liver and brain antioxidant status were measured. Behavioral testing was performed on days 17, 18, and 19. Results: Ultrasound lead to higher stress and anxiety levels; increase in creatinine by 8.29% and gamma-glutamyltransferase activity by 5 times, a decrease in alkaline phosphatase activity by 38.23%, increase of de Ritis coefficient by 21.34%; increased liver and brain superoxide dismutase level by 20.8% and 21.5%, respectively; the stress-related changes in the gut microbiota composition - Bacteroidaceae and Firmicutes. Conclusion: Subjecting mice to 20 days of US-induced stress leads to systemic disorders due to oxidative stress and a decrease in the diversity of the gut microbiota.

Assessment of Antioxidant Stability of Meat Pâté with Allium cepa Husk Extract.

Antioxidants play a very important role in the food industry. Recently, both science and industry have shown substantial preference for natural antioxidants, including searching for antioxidant substances from natural sources without undesirable side effects. The purpose of this study was to evaluate the effect of adding Allium cepa husk extract at a volume of 68 or 34 µL/g of unsalted blanched materials to replace 34% and 17% of the beef broth, respectively, which corresponded to a total antioxidant capacity (TAC) of 44.4 or 22.2 µmol-equiv. Q/100 g meat pté (i.e., 13.42 or 6.71 mg of quercetin/100 g meat pté), on the quality and safety indicators of the developed meat pté. The TAC according to a ferric reducing antioxidant power assay, thiobarbituric acid reactive substances, and physicochemical and microbiological characteristics were determined during the storage of the meat pté. Proximal and UPLC-ESI-Q-TOF-MS analyses were also performed. The addition of yellow onion husk ethanolic extract to the meat pté at both volumes allowed the maintenance of an increased content of antioxidants, which contributed to a decrease in the generation of secondary products of lipid peroxidation for 14 days of storage at 4 ∘C. The results of the microbiological analyses showed that the developed meat ptés were safe according to all indicators of microbial spoilage within 10 days of production. The results supported the use of yellow onion husk extract in the food industry to contribute to improving the functionality of meat products, developing products for a healthy lifestyle, and providing clean-label foods without or with a minimal content of synthetic additives.

Pork Fat and Meat: A Balance between Consumer Expectations and Nutrient Composition of Four Pig Breeds.

Food fat content is one of the most controversial factors from a consumer's point of view. Aim: (1) The trends in consumer attitudes towards pork and the fat and meat compositions in Duroc and Altai meat breeds and Livny and Mangalitsa meat and fat breeds were studied. (2) Methods: Netnographic studies were used to assess Russian consumer purchasing behavior. Protein, moisture, fat, backfat fatty acid content from pigs, longissimus muscles, and backfat from (A) Altai, (L) Livny, and (M) Russian Mangalitsa breeds were compared with those from (D) Russian Duroc. Raman spectroscopy and histology were applied to the backfat analysis. (3) Results: The attitude of Russian consumers to fatty pork is contradictory: consumers note its high fat content as a negative factor, but the presence of fat and intramuscular fat is welcomed because consumers positively associate them with better taste, tenderness, flavor, and juiciness. The fat of the 'lean' D pigs did not show a "healthy" fatty acid ratio, while the n-3 PUFA/n-6 PUFA ratio in the fat of the M pigs was the best, with significant amounts of short-chain fatty acids. The highest UFA content, particularly omega 3 and omega 6 PUFA, was found in the backfat of A pigs with a minimum SFA content. The backfat of L pigs was characterized by a larger size of the adipocytes; the highest monounsaturated and medium chain fatty acid contents and the lowest short-chain fatty acid content; the ratio of omega 3 to omega 6 was 0.07, and the atherogenicity index in L backfat was close to that of D, despite the fact that D pigs are a meat type, while L pigs are a meat and fat type. On the contrary, the thrombogenicity index in L backfat was even lower than the D one. (4) Conclusions: Pork from local breeds can be recommended for functional food production. The requirement to change the promotion strategy for local pork consumption from the position of dietary diversity and health is stated.

Differences in Antioxidant Potential of Allium cepa Husk of Red, Yellow, and White Varieties.

The effective management of agro-industry organic waste for developing high-commercial-value products is a promising facet of the circular economy. Annually, more than 550,000 tons of waste that is potentially rich in biologically active substances is generated worldwide while processing onions (Allium cepa L.). The antioxidant potential of red, yellow, and white onion husks was studied using FRAP, ORAC, chemiluminescence, and UPLC-ESI-Q-TOF-MS analysis methods. The extraction of phenolic compounds from onion husks was more effective when using an aqueous solution of 70% ethanol as compared with water. Ethanolic extract from red onion husks exhibited the highest TACORAC and TACFRAP values, averaging 2017.34 µmol-equiv. Trolox/g raw material and 2050.23 µmol-equiv. DQ/g raw material, respectively, while the white onion exhibited much lower levels of antioxidants. According to the chemiluminescence results, it was determined that the red and yellow onion husks contained antioxidants of three types of power, while white onion husks only contained medium and weak types. The highest content of flavonoids was found in red onion husks, averaging 1915.90 ± 9.92 µg-eq. DQ/g of raw material and 321.42 ± 2.61 µg-eq. DQ/g of raw material for ethanol and water, respectively, while yellow onion husks exhibited 544.06 ± 2.73 µg-eq. DQ/g of raw material and 89.41 ± 2.08 for ethanol and water, respectively. Quercetin and its glycosides were the most representative flavonoids, and a number of substances with different pharmacological and biological properties were also identified.

Bioactive Compounds of Porcine Hearts and Aortas May Improve Cardiovascular Disorders in Humans.

Functional foods promote health benefits in human metabolism, with bioactive compounds acting as therapeutic agents. The aim was to investigate the biological effects of a pâté made of pork hearts and aortas, minced, sterilised and packed in tins. Adults (61-66 years old) with a body mass index of 26.4-60.7 kg/m2 (n = 36) were randomly divided into two groups: one group consumed a low-calorie diet (LCD), while the other consumed an LCD with the developed meat product (MP) for 28-30 days. Serum biochemical parameters, anthropometry and blood pressure were measured. Consumption of an LCD + MP by experimental group participants helped to maintain reduced cholesterol levels. The difference in total cholesterol was significantly different (p = 0.018) from that of the control group, mainly due to the difference in low-density lipoprotein cholesterol (p = 0.005). Six peptides with potential cholesterol-binding properties and four peptides with potential antioxidant activity were identified in the MP, while elevation of the content of two peptides with potential angiotensin-converting enzyme-inhibitory activity was detected in patients' plasma. Intervention with the MP can be considered as a supportive therapy to the main treatment for medical cardiovascular diseases due to a positive effect on serum cholesterol.

Local Experimental Intracerebral Hemorrhage in Rats.

(1) Background: Hemorrhagic stroke is a lethal disease, accounting for 15% of all stroke cases. However, there are very few models of stroke with a hemorrhagic etiology. Research work is devoted to studying the development of cerebrovascular disorders in rats with an intracerebral hematoma model. The aim of this study was to conduct a comprehensive short-term study, including neurological tests, biochemical blood tests, and histomorphological studies of brain structures. (2) Methods: The model was reproduced surgically by traumatizing the brain in the capsula interna area and then injecting autologous blood. Neurological deficit was assessed according to the McGrow stroke-index scale, motor activity, orientation-exploratory behavior, emotionality, and motor functions. On Day 15, after the operation, hematological and biochemical blood tests as well as histological studies of the brain were performed. (3) Results: The overall lethality of the model was 43.7%. Acute intracerebral hematoma in rats causes marked disorders of motor activity and functional impairment, as well as inflammatory processes in the nervous tissue, which persist for at least 14 days. (4) Conclusions: This model reflects the situation observed in the clinic and reproduces the main diagnostic criteria for acute disorders of cerebral circulation.

Antioxidant effect of ethanolic onion (Allium cepa) husk extract in ageing rats.

The role of natural antioxidants in preventing of age-relating diseases is evident. The vegetable industry generates a large amount of waste, which is a good source of antioxidants. The aim of the study was the investigation of the antioxidant effect of long-term consumption of ethanolic yellow onion husk extract in ageing laboratory rodents. Twenty male Wistar albino rats were divided randomly into two groups (n = 10): a control group and an experimental group that received ethanolic yellow onion husk extract (2 mL/rat diluted with distilled water; activity of 4.44 µmol-equiv. quercetin) for 188 days. Oxygen radical absorbance capacity and ferric reducing antioxidant power assays were used to determine the total antioxidant capacity of the extract, which amounted to 941.4 ± 32.7 µmol equiv. Trolox/g raw material and 167.4 ± 16.4 µmol-equiv. quercetin/g raw material, respectively. Oral intake of the onion husk extract affected the indicators of the antioxidant system of the liver and the brain but not of the blood and plasma, mainly due to elevations in the activity of catalase and superoxide dismutase in the liver by 44.4% and 79.1%, respectively, and in the brain by three-fold and 79.1%, respectively. The availability, cheapness and high antioxidant potential of onion waste qualifies it a good source of functional ingredients and bioactive substances applicable in the food and pharmaceutical industries.

Effects of tissue-specific biomolecules on piglets after-weaning period.

BACKGROUND AND AIM: Now-a-days antibiotics are the main tool for correcting the pathological conditions of pigs; unfortunately, antibiotics are a potential threat to the environment, as they lead to the spread of antibiotic-resistant infections. This study aimed to study the immunomodulatory encapsulated biomolecules on piglets in the post-weaning period. MATERIALS AND METHODS: An immunomodulator based on biomolecules obtained from animal raw materials included in alginate capsules to improve absorption has been developed. The study presents the results of a study on 25 weaned piglets (25-30 days old) which received biomolecules at a dose of 200 mg/piglet for 14 days, followed by 400 mg/piglet from days 15 to 28. Blood was taken from animals for analysis (biochemical, hematological, cytometric, and enzyme immunoassay) and the integral index of blood serum antimicrobial activity was determined. RESULTS: Experimental animals, whose initial weight was 1.6 times less than that of the control animals, were able to bridge this gap and, on the 28th day, there were no differences in weight. Stimulation of the production of cytokines interleukin (IL)-2 and IL-4 was observed and the antimicrobial resistance of blood serum to Escherichia coli also increased. A positive effect on the metabolism of piglets was noted, which helped them adapt to a change in diet (from colostrum to solid food). CONCLUSION: The results show that the immunomodulation at the dose of 150 mg/kg body weight has a great potential for improving weaned pigs.

Lateral flow immunoassay for sensitive detection of undeclared chicken meat in meat products.

This study presents the development of an immunochromatographic test system aimed at the detection of chicken additives in meat products. It is based on sandwich-format lateral flow immunoassay (LFIA) of immunoglobulins as a biomarker for species identification. The LFIA based on gold nanoparticles as a label for anti-species antibodies was used to determine chicken immunoglobulins and, accordingly, chicken meat in food products. Absence of cross-reactivity with mammalian species tested in the study confirmed high specificity of the determination. The test system showed good sensitivity and rapidity, allowing for the detection of as low as 0.063% (w/w) chicken meat in raw meat mixtures within 20 min. As a result of the testing of raw and cooked meat products, it was shown that the test system can reliably recognize specific immunoglobulins even after heat processing. The proposed assay can be recommended for rapid on-site screening control of the composition and quality of meat products.

Development of Immunohistochemical Methods for Casein Detection in Meat Products.

To increase production efficiency of meat products, milk protein additives are often used. Despite a number of advantages, use of dairy ingredients involves a certain risk, namely the allergenic potential of milk proteins. A number of methods have been developed to detect milk-origin raw materials in foodstuffs, including immunological reference methods. This study presents newly developed immunohistochemical (IHC) methods for casein detection in meat products. Casein was successfully detected directly in meat products where sensitivity was determined at 1.21 and specificity at 0.28. The results obtained from the IHC were compared with the Enzyme-Linked Immuno Sorbent Assay (ELISA) and there was no statistically significant difference between the IHC and ELISA methods (p > 0.05). The correspondence between the methods was 72% in total. The highest correspondence was reached in frankfurters (90%), the lowest in canned pâté (44%).

Lateral Flow Immunoassay to Detect the Addition of Beef, Pork, Lamb, and Horse Muscles in Raw Meat Mixtures and Finished Meat Products.

A lateral flow immunoassay for sensitive detection of skeletal troponin I (TnI) as a specific, thermostable marker of muscle tissue was developed. Due to the antibodies' choice, the assay specifically detects mammalian TnI (in beef, pork, lamb, and horse) but does not detect bird TnI (in chicken or turkey), thus enabling differentiation of these types of raw meat materials. The assay is based on a sandwich format of the analysis using gold nanoparticles as labels. The time of the assay is 15 min, and the TnI detection limit is 25 ng/mL. A buffer solution is proposed for efficient extraction of TnI from muscle tissues and from finished meat products that have undergone technological processing (smoking-cooking-smoking, cooking and smoking). The possibility of detecting beef addition in minced chicken down to 1% was demonstrated.

Metabolic, Molecular, and Behavioral Effects of Western Diet in Serotonin Transporter-Deficient Mice: Rescue by Heterozygosity?

Reduced function of the serotonin transporter (SERT) is associated with increased susceptibility to anxiety and depression and with type-2 diabetes, which is especially true in older women. Preference for a "Western diet" (WD), enriched with saturated fat, cholesterol, and sugars, may aggravate these conditions. In previous studies, decreased glucose tolerance, central and peripheral inflammation, dyslipidemia, emotional, cognitive, and social abnormalities were reported in WD-fed young female mice. We investigated the metabolic, molecular, and behavioral changes associated with a 3-week-long dietary regime of either the WD or control diet in 12-month-old female mice with three different Sert genotypes: homozygous (Slc6a4) gene knockout (Sert -/-: KO), heterozygous (Sert +/-: HET), or wild-type mice (Sert +/+: WT). In the WT-WD and KO-WD groups, but not in HET-WD-fed mice, most of changes induced by the WD paralleled those found in the younger mice, including brain overexpression of inflammatory marker Toll-like receptor 4 (Tlr4) and impaired hippocampus-dependent performance in the marble test. However, the 12-month-old female mice became obese. Control diet KO mice exhibited impaired hippocampal-dependent behaviors, increased brain expression of the serotonin receptors Htr2c and Htr1b, as well as increased Tlr4 and mitochondrial regulator, peroxisome proliferator-activated receptor gamma-coactivator-1a (Ppargc1a). Paradoxically, these, and other changes, were reversed in KO-WD mutants, suggesting a complex interplay between Sert deficiency and metabolic factors as well as potential compensatory molecular mechanisms that might be disrupted by the WD exposure. Most, but not all, of the changes in gene expression in the brain and liver of KO mice were not exhibited by the HET mice fed with either diet. Some of the WD-induced changes were similar in the KO-WD and HET-WD-fed mice, but the latter displayed a "rescued" phenotype in terms of diet-induced abnormalities in glucose tolerance, neuroinflammation, and hippocampus-dependent performance. Thus, complete versus partial Sert inactivation in aged mice results in distinct metabolic, molecular, and behavioral consequences in response to the WD. Our findings show that Sert +/- mice are resilient to certain environmental challenges and support the concept of heterosis as evolutionary adaptive mechanism.

Hypolipidemic and anti-inflammatory effects of aorta and heart tissues of cattle and pigs in the atherosclerosis rat model.

The aim of this study was to investigate the effects of aorta and heart tissues obtained from cattle and pigs on atherosclerosis disorders. Atherosclerosis model rats were provided with the respective diets consisting of aorta and heart tissues. Administration of each tissue suppressed body weight gain as compared to that of the control. In particular, the aorta tissues of pigs and cattle demonstrated significant suppressions in body weight gain in the model rats. The aorta tissues of pigs and cattle showed a significant increase and decrease in the serum high-density lipoproteins and atherogenic index, respectively, which was correlated with the increase in apolipoprotein A1. Hematological analysis revealed that aorta tissues of pigs and cattle clearly reduced the ratio of granulocytes/lymphocytes in the atherosclerosis rats. Serum vascular cellular adhesion molecule-1 levels in the atherosclerosis rats, which were administered these aorta tissues, were also significantly reduced. Additionally, there was an increase in von Willebrand factor in the rat serum. Based on the results obtained, the aorta tissues of pigs and cattle, in particular, demonstrated positive effects in the atherosclerosis rats due to the alteration of lipid metabolism and reduction in inflammation related to atherosclerosis.

Enzyme immunoassay and proteomic characterization of troponin I as a marker of mammalian muscle compounds in raw meat and some meat products.

The skeletal muscle protein troponin I (TnI) has been characterized as a potential thermally stable and species-specific biomarker of mammalian muscle tissues in raw meat and meat products. This study proposed a technique for the quantification of TnI comprising protein extraction and sandwich enzyme-linked immunosorbent assay (ELISA). The technique is characterized by a TnI detection limit of 4.8 ng/ml with quantifiable concentrations ranging from 8.7 to 52 ng/ml. The method was shown to be suitable for detection of TnI in mammalian (beef, pork, lamb, and horse) meat but not in poultry (chicken, turkey, and duck) meat. In particular, the TnI content in beef was 0.40 3 ± 0.058 mg/g of wet tissue. The TnI estimations obtained for the pork and beef samples using ELISA were comparable to the proteomic analysis results. Thus, the quantitative study of TnI can be a convenient way to assess the mammalian muscle tissue content of various meat products.