Search Results (15)

Background/Objectives: Advances in nanopore sequencing have opened new avenues for studying DNA methylation at single-base resolution, yet their application in epigenetic ageing research remains underdeveloped. Methods: We present a novel framework that leverages the unique capabilities of nanopore sequencing to profile and interpret age-associated methylation patterns in native DNA. Results: Unlike conventional array-based approaches, long reads sequencing captures full CpG context, accommodates diverse and repetitive genomic regions, removes bisulfite conversion steps, and is compatible to the latest reference genome. Conclusions: This work establishes nanopore sequencing as a powerful tool for next-generation epigenetic ageing studies, offering a scalable and biologically rich platform for anti-ageing interventions monitoring and longitudinal ageing studies. Full article

This study aimed to determine the allele frequencies and genetic diversity of 21 autosomal short tandem repeat (STR) loci from the Expanded U.S. Core Loci and European Standard Set in the Romanian population. A random sample of 928 unrelated men from all Romanian counties was analyzed using the Investigator 24plex QS and Investigator 24plex GO! Kits (Qiagen). The genotypes were determined, and the allele frequencies were calculated using the STRidER tool. The results provide updated population genetic data for the Romanian population, which is essential for accurate calculation of DNA evidence weight in forensic casework. Full article

Contamination of agricultural ecosystems with antibiotics including sulfamethoxazole (SMX) can create favorable conditions to increase bacterial abundance in soil with antibiotic-resistant genes and can also affect plants. The aim of this research was to assess the phytotoxicity of tomato after irrigation with SMX degraded in 20 min using the non-thermal plasma-ozonation technique (T20). To achieve this, two experiments were performed at the scales of Petri dishes and pots using Solanum lycopersicum L. species, cultivar Zaraza, subjected to irrigation treatments that were compared to a distilled water control. In plates, T20 solution improved root length and also seedling vigor indexes, but the germination index, germination speed, and biomass were slightly decreased. In soil, although T20 reduced the seedling root length, their growth was not inhibited (15.3%), while in plates they exhibited a growth promotion effect with 90% more than the control. The physical–chemical and geochemical variables measured in the soil were suitable for crop characteristics and plant growth and showed statistically significant variations after harvesting. In T20-treated shoots, compared to SMX, better results were obtained for their length, assimilatory pigments, and biomass, thus selectively reducing the tomato seedling phytotoxicity depending on the endpoints, type of control, and growth methods tested. Full article

Sturgeons are facing critical endangerment due to overfishing, habitat destruction, pollution and climate change. Their roe, highly prized as caviar, has driven the overexploitation, severely depleting wild populations. In recent years sturgeon aquaculture has experienced significant growth, primarily aimed at providing high-quality caviar and secondarily meat. This sector generates significant quantities of by-products, which are mainly treated as waste, being mostly discarded, impacting the environment, even though they are a source of bioactive molecules and potential applications in various sectors. This article presents a review of the proximate composition and nutritional value of sturgeon caviar and meat, also exploring the potential of the by-products, with an emphasis on the processing of these components, the chemical composition and the functional and bioactive properties. Although sturgeon caviar, meat, and by-products are highly valuable both nutritionally and economically, adopting sustainable practices and innovative approaches is crucial to ensuring the industry’s future growth and maintaining ecological balance. Despite some limitations, like the deficient standardization of the methods for extracting and processing, sturgeon by-products have a tremendous potential to increase the overall value of sturgeon aquaculture and to promote a zero-waste approach, contributing to achieving the Sustainable Development Goals adopted by all United Nations Member States in 2015. Full article

Fourteen lactic acid bacteria (LAB) strains were isolated from the intestinal tract and feces of piglets. Among these isolates, only twelve were biochemically (API 50 CHL and ABIS online for bacterial identification) and genetically (16S rRNA sequencing) confirmed as Limosilactobacillus fermentum, Lactobacillus acidophilus, and Lactiplantibacillus plantarum. Experiments to evaluate the probiotic potential of the isolates including pH tolerance (pH 2.0 and 3.0), bile salts (0.3% ox gall) resistance, hemolysis activity, antibiotic susceptibility, and high-temperature resistance were tested. Only two isolates from identified strains exhibited high survival rates when exposed to low pH and bile salts, these were L. acidophilus IBNA 76 and L. plantarum IBNA 84. The antibiotic test presented 100% resistance of both strains to gentamicin, kanamycin, lincomycin, colistin sulfate, erythromycin, amikacin, oxytetracycline, enrofloxacin, streptomycin, and tilmicosin, lower than the 0.6 mm inhibition zone diameter. Promising isolates (L. acidophilus IBNA 76 and L. plantarum IBNA 84) were exposed to the spray-drying technique based on visible probiotic potential and survival rates. Carrier matrix material was used as a maltodextrin-glucose solution. The encapsulation probiotic isolated survived both over 67% and 77%, corresponding to a decrease in strain viability from 10⁹ to 10⁷ CFU/g. After further in vitro evaluations, the findings of this study showed that, from all LAB strains, L. acidophilus IBNA 76 and L. plantarum IBNA 84 may be considered probiotic candidates for animal nutrition and may have promising performance in piglet feed due to their origin of isolation. Full article

Feeding farm animals with aflatoxin-contaminated feed can cause various severe toxic effects, leading to increased susceptibility to infectious diseases and increased mortality, weight loss, poor performance and reduced reproductive capability. Following ingestion of contaminated foodstuffs, aflatoxins are metabolized and biotransformed differently in animals. Swine metabolism is not effective in detoxifying and excreting aflatoxins, meaning the risk of aflatoxicosis is increased. Thus, it is of great importance to elucidate the metabolism and all metabolic pathways associated with this mycotoxin. The damage induced by AFB1 in cells and tissues consists of inhibition of cell proliferation, carcinogenicity, immunosuppression, mutagenicity, oxidative stress, lipid peroxidation and DNA damage, leading to pathological lesions in the liver, spleen, lymph node, kidney, uterus, heart, and lungs of swine. At present, it is a challenging task and of serious concern to completely remove aflatoxins and their metabolites from feedstuff; thus, the aim of this study was a literature review on the deleterious effects of aflatoxins on swine metabolism, as well as alternatives that contribute to the detoxification or amelioration of aflatoxin-induced effects in farm animal feed. Full article

The sturgeons (order Acipenseriformes) are caviar producers and some of the most valuable fish species worldwide. Due to different reasons, wild populations are now at the brink of extinction. The high demand for caviar has led to the development of aquaculture for restocking and caviar production. Since the caviar from different species has different prices depending on the quality and attempts of commercial fraud based on species substitution have been found, correct species identification is more than necessary. We report a new multistep methodology for an accurate species identification based on both nuclear and mitochondrial markers. Our test integrates data from the analysis of microsatellites (Afu19, Afu34, Afu39, Afu54, Aox27, AoxD234, AnacC11 and AnacE4), nuclear gene markers (RPS7, vimentin and rhodopsin) and mtDNA barcoding to give a reliable molecular diagnostic for five sturgeon species (Huso huso, Acipenser stellatus, Acipenser ruthenus, Acipenser gueldenstaedtii and Acipenser baerii). In addition to species identification, our methodology allows the identification of bester, sterbe and best beluga hybrids, but also the identification of hybrids of unknown origin. This methodology has a good potential to contribute to the conservation of highly threatened sturgeon populations and also to the traceability of their products. Full article

A total of 15 strains of lactic acid bacteria (LAB) were isolated from the broiler chicken’s gastrointestinal tract. All isolates were phenotypical and genetically identified. Among these isolates, only six were biochemical (API 50 CHL and ABIS soft) and genetically (16S rRNA sequencing) confirmed as Lactobacillus acidophilus, Limosilactobacillus fermentum, Levilactobacillus brevis, and Ligilactobacillus salivarius. Probiotic properties, including tolerance to pH (pH 2.0 and 3.0), bile salts (0.3% oxgall), hemolysis activity, and antibiotic susceptibility, were evaluated. Three isolates of the latter isolates showed high resistance at low pH values (73.74% to 98.20%) and bile salt concentrations (77.89% to 99.49%). The antibiotic test presented 100% resistance of LAB to gentamicin, lincomycin, enrofloxacin, and streptomycin lower than the 0.5 mm inhibition zone diameter. Selected strains (L. acidophilus IBNA 64, L. salivarius IBNA 33, and L. salivarius IBNA 41) were exposed to the spray-drying process based on observable probiotic potential. A maltodextrin-glucose solution was used as a thermoprotectant. After spray drying, a reduction in strain viability was noted (10⁸ to 10⁴ CFU/g). In conclusion, only L. salivarius (IBNA 33 and IBNA 41) could be used as a possible probiotic, and further studies are needed to ensure their safe application in the animal nutrition field with beneficial effects for improving performance and pathogen microorganism control from intestines equilibrating the microbiota composition. Full article

The purpose of this study was to investigate the combined effects of aflatoxin B1 and ochratoxin A on protein expression and catalytic activities of CYP1A2, CYP2E1, CYP3A29 and GSTA1 and the preventive effect of dietary byproduct antioxidants administration against these mycotoxin damage. Three experimental groups (E1, E2, E3) and one control group (C) of piglets after weaning (TOPIGS-40 hybrid) were fed with experimental diets for 30 days. A basal diet containing normal compound feed for starter piglets was used as a control treatment and free of mycotoxin. The experimental groups were fed as follows: E1—basal diet plus a mixture (1:1) of two byproducts (grapeseed and sea buckthorn meal), E2—the basal diet experimentally contaminated with mycotoxins (479 ppb OTA and 62ppb AFB1) and E3—basal diet containing 5% of the mixture (1:1) of grapeseed and sea buckthorn meal and contaminated with the mix of OTA and AFB1. After 4 weeks, the animals were slaughtered, and tissue samples were taken from liver and kidney in order to perform microsomal fraction isolation, followed by protein expression and enzymatic analyses. The protein expressions of CYP2E1 and CYP3A29 were up-regulated in an insignificant manner in liver, whereas in kidney, those of CYP1A2, CYP2E1 and CYP3A29 were down-regulated. The enzymatic activities of CYP1A2, CYP2E1 and CYP3A29 decreased in liver, in a significant manner, whereas in kidney, these increased significantly. The co-presence of the two mycotoxins and the mixture of grape seed and sea buckthorn meal generated a tendency to return to the control values, which suggest that grapeseed and sea buckthorn meal waste represent a promising source in counteracting the harmful effect of ochratoxin A and aflatoxin B. Full article

The use of natural feed additives could be a beneficial approach to maintaining the health of chickens and a way to improve food digestion. Flaxseed is a rich source of omega-3 fatty acid, alpha linolenic acid, oleic acid, and fiber. The purpose of this study was to evaluate the effects of dietary inclusion of 4% flaxseed on the intestinal morphology, specific enzymatic activity, and cecal microbiome in broiler chickens. The 4-week feeding trial was conducted on 100 Cobb 500 (14 days of age) unsexed broiler chickens divided into two groups: a control group (C) and an experimental group (E). The broilers were housed in boxes of size 3 m² (each group was housed in a single box with 10 replicates, 5 chickens per replicate) and reared on permanent wood shaves litter (10–12 cm thick). At the end of the experiment, chickens (n = 10) were sacrificed and tissue samples were harvested from the duodenum, jejunum, and cecum for histological, enzymatic, and microbiome analyses. In group E, histological analysis revealed a significant increase in villus height (p < 0.001) possibly leading to enhanced intestinal nutrient absorption. An increase in the specific activities of α-amylase (p < 0.05), invertase (p < 0.01), and endo-β-1,4-glucanase (p < 0.001) was noticed in the E group for the duodenum and jejunum compared to the control group. In contrast, maltase activity decreased in the duodenum and increased in the jejunum in the E group. The trypsin and lipase specific activities did not vary in a significant way. In addition, the cecal microbiome of the E group was characterized by an increase in Lactobacilli (p < 0.01) and Clostridium coccoides and a decrease in Bacteroides, Ruminoccocus, Enterobacteriaceae, and Clostridium leptum. In conclusion, our results suggest that dietary supplementation of flaxseed meal may boost intestinal health status in poultry. Full article

The purpose of this study was to investigate the potential of a byproduct mixture derived from grapeseed and sea buckthorn oil industry to mitigate the harmful damage produced by ochratoxin A and aflatoxin B1 at hepatic and renal level in piglets after weaning. Forty cross-bred TOPIGS-40 hybrid piglets after weaning were assigned to three experimental groups (E1, E2, E3) and one control group (C), and fed with experimental diets for 30 days. The basal diet was served as a control and contained normal compound feed for starter piglets without mycotoxins. The experimental groups were fed as follows: E1—basal diet plus a mixture (1:1) of two byproducts (grapeseed and sea buckthorn meal); E2—the basal diet experimentally contaminated with mycotoxins (479 ppb OTA and 62ppb AFB1); and E3—basal diet containing 5% of the mixture (1:1) of grapeseed and sea buckthorn meal and contaminated with the mix of OTA and AFB1. After 4 weeks, the animals were slaughtered, and tissue samples were taken from liver and kidney in order to perform gene expression and histological analysis. The gene expression analysis showed that when weaned piglets were fed with contaminated diet, the expression of most analyzed genes was downregulated. Among the CYP450 family, CYP1A2 was the gene with the highest downregulation. According to these results, in liver, we found that mycotoxins induced histomorphological alterations in liver and kidney and had an effect on the expression level of CYP1A2, CYP2A19, CYP2E1, and CYP3A29, but we did not detect important changes in the expression level of CY4A24, MRP2 and GSTA1 genes. Full article

Acipenser stellatus is a critically endangered species due to the anthropic influence. It has been intensively captured for decades because of its high economic value, its roe being used in the caviar industry. Therefore, Acipenser stellatus is intensively raised in fish farms for both conservation and economical purposes. Aquaculture aims to optimize the feeding regime of juveniles in order to improve its profitability. The purpose of this study was to investigate if Acipenser stellatus can adapt to a starvation/refeeding regime by assessing the effects of this regime on oxidative stress biomarkers and antioxidant defense mechanisms in juveniles raised under aquaculture conditions. The juveniles were subjected to two regimes: a 7-day starvation period followed by 21 days of refeeding, respectively a 14-day starvation period followed by 21 days of refeeding. The results showed that both starvation/refeeding regimes induced an enhancement of antioxidant enzymes activities in the intestine of the juveniles. The oxidative damage was counteracted at the protein level. However, lipid peroxidation was significantly induced in the intestine of the juveniles subjected to 14/21-day starvation/refeeding regime. The 7/21-day starvation/refeeding regime proved to be more suitable for Acipenser stellatus and therefore, it could be useful to optimize the feeding practice in aquaculture production. Full article

The purpose of this study was to examine the effects of dietary inclusion of two additives at the final concentration of 0.5% bilberry (E1) and 1% walnut (E2) leaves powder in the basal diet on digestive health of hens. A total number of 90 Tetra SL hens were divided into two experimental groups (E1 and E2) and one control group (C) consisting of 30 hens each. After four weeks, 10 hens of each group were sacrificed and tissue samples and intestinal content were taken from the duodenum, jejunum, and cecum in order to perform histological, enzymatic, and microbiota analyses. In groups E1 and E2, the histological analysis showed a significant increase of villus height, resulting probably in increased absorption of nutrients in duodenum and jejunum. A decrease in the specific activity of alpha-amylase and trypsin in E1 and E2 for both duodenum and jejunum compared to the control one was also recorded. In addition, the maltase and invertase specific activity in duodenum increased, a tendency that was kept for maltase but not for invertase in jejunum. The cecal microbiota of E1 and E2 individuals was characterized by an increase of Firmicutes and Lactobacilli and a decrease of Enterobacteriaceae. In conclusion, our results indicate that bilberry and walnut leaves additives in feed may improve the health status of the poultry gastrointestinal tract. Full article

The Mangalitsa pig, a swine breed belonging to the protected gene fund of original and primitive animal breeds of the FAO (Food and Agriculture Organization), has been known to inhabit Romanian territories since the 19th century. The aim of this study was to compare the Mangalitsa breed with several European and Asiatic swine breeds in order to emphasize its uniqueness and to elucidate its origin. For this purpose, we analyzed a 613 bp mitochondrial DNA D-loop fragment and 1140 bp of the cytochrome b gene in a population of Mangalitsa pigs and the polymorphic sites were compared with sequences from GenBank originating from other swine breeds. Taking into account the total of 24 breeds and 5 different Wild Boar populations analyzed, 86 polymorphic sites representing 32 haplotypes were observed, with an average percentage of polymorphic sites of 4.9%. Three Neighbor-Joining phylogenetic trees were constructed based on Kimura 2-parameter distances, using D-loop, cytochrome b and mitochondrial reunited sequences. For the analyzed Mangalitsa population, four distinct haplotypes were identified, including one that was common to other breeds. Our study suggests that the Mangalitsa swine originate from primitive breeds which might be directly derived from the Wild Boar. Full article

Acipenseriformes are composed of 25 sturgeon species and two paddlefish species distributed exclusively in the northern hemisphere. The Danube River and the Black Sea were originally inhabited by six sturgeon species but two are extinct and only four are still reproducing currently in the Lower Danube: Huso huso, Acipenser stellatus, A. gueldenstaedtii and A. ruthenus. Sturgeon species hybridize more easily than other fish and the determination of pure species or hybrid status is important for conservation and for breeding in fish farms. This survey demonstrated that morphological determination of this status is not reliable and a molecular tool, based on eight microsatellites genotypes is proposed. This method, based on three successive statistical analyses including Factorial Correspondence Analysis (FCA), Structure assignation and NewHybrids status determination, showed a high efficiency in discriminating pure species specimens from F1, F2 and two kinds of backcross individuals involving three of the four reproducing Lower Danube sturgeon species. Full article