Effect of sodium diacetate on fermentation, aerobic stability, and microbial diversity of alfalfa silage.

Alfalfa (Medicago sativa L.) is a vital source of forage protein for ruminants, yet its ensiling poses challenges due to high buffering capacity and low water-soluble carbohydrates (WSC). This study investigated the impact of sodium diacetate (SDA) on alfalfa silage quality and aerobic stability. SDA was applied at four different rates to wilted alfalfa on a fresh basis: 0 g/kg, 3 g/kg, 5 g/kg, and 7 g/kg, and silages were ensiled in laboratory-scale silos for 45 days, followed by 7 days of aerobic exposure. A 16S rRNA gene sequencing assay using GenomeLab™ GeXP was performed to determine the relationship between dominant isolated lactic acid bacteria species and fermentation characteristics and aerobic stability on silage. The results showed that Lentilolactobacillus brevis, Pediococcus pentosaceus and Enterococcus faecium were the most prevalent bacteria when silos were opened, whereas Weissella paramesenteroides, Bacillus cereus, B. megaterium and Bacillus spp. were most prevalent bacteria after 7 days of aerobic exposure. Dry matter, pH, and WSC content were not affected by SDA, but doses above 5 g/kg induced a homofermentative process, which increased lactic acid concentration and lactic acid to acetic acid ratio, decreased yeast count during aerobic exposure, and improved aerobic stability. These findings offer useful information for optimizing SDA usage in silage, assuring improved quality and longer storage, and thereby improving animal husbandry and sustainable feed practices.

Characterisation of Single Nucleotide Polymorphisms and Haplotypes of MSTN Associated with Growth Traits in European Sea Bass (Dicentrarchus labrax).

The myostatin (MSTN) gene, known as growth differentiation factor-8 (GDF-8), is a member of the transforming growth factor-ß (TGF-ß) superfamily and plays a specific inhibitory role during the critical phases of skeletal muscle mass development in vertebrates. This study was conducted to investigate MSTN polymorphisms in harvest size European sea bass reared in Turkey. Nine single nucleotide polymorphisms (SNPs) and two indels were identified in exons 1-3 of MSTN in the European sea bass population The associations between the g.16612A indel located in intron 1 and standard length were significant. The MSTN g.15252 T > A locus in intron 2 was significantly related to the total weight, fillet weight and standard length (P < 0.05). The relationship between the g.14873C > T locus in exon 3 of MSTN and standard height, head length, body length, pre-anal length, abdominal length, post-anal length and head width was significant (P < 0.05). According to the results of the haplotype analysis, two haplogroup and eight haplotype combinations were detected in the population. The haplogroup 2 had significant associations with all measured growth traits (P < 0.05). Thus, SNPs and haplotypes identified in this study could be useful for European sea bass breeding and marker-assisted selection.

Polymorphism detection of DGAT1 and Lep genes in Anatolian water buffalo (Bubalus bubalis) populations in Turkey.

Acyl-CoA: diacylglycerol-acyltransferase 1 (DGAT1) enzyme plays a key role in controlling the synthesis rate triglyceride from diacylglycerol. Leptin (LP, OB, obese) is an important hormone that synthesizes mostly from adipose tissue and regulates glucose metabolism and homeostasis. DGAT1 and Lep genes are closely related to reproduction, growth, milk yield and composition in water buffalo breeds. This study aimed to identify genetic variation in the DGAT1 and Lep gene regions in 150 water buffalo individuals from five different provinces of Turkey using DNA sequencing. A total of 38 nucleotide variations and indels have identified 761 bp long partial intron 2 and exon 3 and 5 ' UTR regions of the Lep gene in Anatolian water buffalo populations; 422 bp long partial exon 7-9 and exon 8 regions of DGAT1 gene were amplified and two mutations were defined in the point of 155 and 275 nucleotide that is three genotypes for S allele and Y allele of DGAT1 gene in intron 7 in Anatolian buffalo populations, respectively. These SNPs may have an effect on reproduction, growth, milk yield and composition in water buffalo populations and may prove to be useful for water buffalo breeding.

Effect of Different Kefir Source on Fermentation, Aerobic Stability, and Microbial Community of Alfalfa Silage.

The present study has been one of the first attempts to thoroughly examine the effects of different kefir sources on fermentation characteristics, aerobic stability, and microbial communities of alfalfa silages. The effects of commercial kefir (CK) and homemade kefir culture (HK) applied with untreated a common control (CON) and three different application doses (5.0, 5.7, and 6.0 log cfu g-1) on wilted alfalfa and stored at an ambient temperature of 25-30 °C are studied. After 45 days of ensiling, fermentation characteristics and aerobic stability of silages were measured, and bacterial diversity was investigated by 16S ribosomal RNA gene sequencing using the GenomeLab™ GeXP platform. Both CK and HK accelerate more lactic acid production and reduced ammonia nitrogen concentration. Factor analysis of kefir sources suggests that the addition of kefir improves the aerobic stability of silages, even the initial water-soluble carbohydrate (WSC) content is inadequate via its antimicrobial effect on yeast and mold formation. Enterococcus faecium, Pediococcus pentosaceous and Lactobacillus brevis were dominant bacterial species among the treated groups at silo opening, while Lactobacillus plantarum and Lactobacillus brevis became dominant bacterial species after 7 days of aerobic exposure. In conclusion, the application of kefir on alfalfa silages improves fermentation quality and aerobic stability even with low WSC content.

Evaluation of Genetic Diversity and Structure of Turkish Water Buffalo Population by Using 20 Microsatellite Markers.

The present study was aimed to investigate the genetic diversity among 17 Turkish water buffalo populations. A total of 837 individuals from 17 provincial populations were genotyped, using 20 microsatellites markers. The microsatellite markers analyzed were highly polymorphic with a mean number of alleles of (7.28) ranging from 6 (ILSTS005) to 17 (ETH003). The mean observed and expected heterozygosity values across all polymorphic loci in all studied buffalo populations were 0.61 and 0.70, respectively. Observed heterozygosity varied from 0.55 (Bursa (BUR)) to 0.70 (Mus (MUS)). It was lower than expected heterozygosity in most of the populations indicating a deviation from Hardy-Weinberg equilibrium. The overall value for the polymorphic information content of noted microsatellite loci was 0.655, indicating their suitability for genetic diversity analysis in buffalo. The mean FIS value was 0.091 and all loci were observed significantly deviated from Hardy-Weinberg Equilibrium (HWE), most likely based on non-random breeding. The 17 buffalo populations were genetically less diverse as indicated by a small mean FST value (0.032 ± 0.018). The analysis of molecular variance (AMOVA) analysis indicated that about 2% of the total genetic diversity was clarified by population distinctions and 88 percent corresponded to differences among individuals. The information produced by this study can be used to establish a base of national conservation and breeding strategy of water buffalo population in Turkey.

Associations between genetic variants of the insulin-like growth factor I (IGF-I) gene and growth traits in European sea bass (Dicentrarchus labrax, L.).

The IGF-I is a significant peptide hormone to cell growth, development, reproduction, and metabolism of teleost fish. This study was conducted to investigate the IGF-I gene polymorphism and to reveal the association between the genotypes of IGF-I gene and growth traits in 80 harvest size European sea bass (Dicentrarchus labrax, L.) reared in Izmir region of Turkey. The 821 bp long exon 1 and 5' UTR regions of IGF-I gene were amplified, and PCR products were analyzed via DNA sequencing method. Two novel single nucleotide polymorphisms (SNPs) such as g.46672A > G and g.46749C > T in the 5' UTR region of IGF-I gene in the European sea bass were detected for the first time with this study. Associations between the genotypes of IGF-I g.46749C > T locus and body weight were found statistically significant (p < 0.05). The genotypes of IGF-I g.46749C > T and g.46672A > G loci were found significantly associated with total length (p < 0.05). However, both loci did not show significant associations with body width. The studied gene region sequences were entered to NCBI GenBank database with the accession numbers: MK621680-MK621682. Thus, these polymorphisms have a significant effect on the body weight and total length which could be useful for sea bass selection and breeding for marker-assisted selection (MAS) program.

Leptin Receptor Gene Polymorphisms in Some Turkish Donkey Populations.

Leptin receptor is a fundamental regulator in physiological functions of the regulation of food intake, energy homeostasis, immune function, and reproduction as well as on ovarian follicular cells on the placenta and lactating mammary glands. The aim of this study was to investigate the LEPR gene polymorphism in 60 donkeys reared in Thrace region of Turkey. A 585 bp long partial intron 6, exon 7, intron 7, and exon 8 regions of LEPR gene were amplified, and polymerase chain reaction products analyzed via DNA sequencing. A novel single-nucleotide polymorphism (SNP) was identified as g.713668A>G in the seventh exon region of LEPR gene. This novel SNP was first identified, and the partial DNA sequence of LEPR gene in donkeys was reported for the first time in this study, and these sequences were deposited to NCBI Genbank database with the accession number: MK807114-MK807115. The A>G transition revealed a silent mutation (CAA-CAG) in glutamine amino acid. This nucleotide mutation could cause the changes of secondary structure of protein and expression level of LEPR hormone. For this reason, additional studies are needed to reveal new SNPs and in the LEPR gene that may affect economic traits and structure of protein in donkey breeds.

Genetic diversity of κ-casein (CSN3) and lactoferrin (LTF) genes in the endangered Turkish donkey (Equus asinus) populations.

In this study, the κ -casein (CSN3) and lactoferrin (LTF) genes which were found in association with milk production traits in different animal species were studied firstly in Turkish donkey populations. A total of 108 donkeys from different regions of Turkey were used in order to reveal the different genotypes of CSN3 and LTF genes by using polymerase chain reaction - restriction fragment length polymorphism (PCR-RFLP) and DNA sequencing methods. To determine the genetic polymorphism, we attempted to digest a fragment of 235 bp of the CSN3 gene and a fragment of 751 bp of the LTF gene using PstI, and DraII, EagI and MboI restriction enzymes, respectively. Neither the CSN3 gene nor the LTF gene had enzyme recognition sites with the PstI, DraII and MboI restriction enzymes in all of the studied samples. However, the LTF gene was only distinguished with the EagI restriction enzyme. Three genotypes were identified in the LTF gene with the EagI restriction enzyme: GG homozygotes (667, 84 bp), AG heterozygotes (751; 667, 84 bp) and AA homozygotes (751 bp). The transition from guanine to adenine in 89 bp of the LTF gene lacks the restriction site and different genotypes are obtained. This novel single nucleotide polymorphism (SNP) has been firstly detected in donkeys. According to the results, the G allele was predominant in the LTF-EagI gene in the studied Turkish donkey populations. In this study, all the genotype distributions of LTF-EagI were not found in Hardy-Weinberg equilibrium ( P < 0.05 ). The CSN3 and LTF genes have not been studied before in donkeys, so the results are the preliminary results of these gene regions in donkeys.

Associations between genetic variants of the POU1F1 gene and production traits in Saanen goats.

This study was conducted to determine the polymorphisms of the POU1F1 gene and their relationships with milk yield and components, litter size, birth weight, and weaning weight in goats. For this purpose, a total of 108 Saanen goats from two different farms (Bornova and Manisa) were used as animal materials. Polymorphisms at the exon 6 and the 3 ' flanking region of the POU1F1 gene were determined by using PCR-RFLP with PstI and AluI restriction enzymes and DNA sequencing analyses. Two alleles and three genotypes were identified by AluI or PstI digestions of the POU1F1 gene. The genotypes frequencies of TT, TC, and CC were 64.8 %, 31.5 % and 3.7 % for the PstI locus; 54.6 %, 31.5 % and 13.9 % for the AluI locus, respectively. T allele frequencies (0.56 and 0.88 for the AluI locus, 0.80 and 0.81 for the PstI locus, respectively) were predominant in both loci at the Bornova and Manisa farms. In terms of POU1F1-AluI and POU1F1-PstI loci, two populations were found to be in Hardy-Weinberg equilibrium. In the POU1F1-AluI locus, significant associations were found between genotypes and lactation milk yield and litter size. Similarly, a significant relationship between genotypes and birth weight in the POU1F1-PstI locus ( p < 0.05 ) was determined. The TC and CC genotypes were observed to be higher than the TT genotype for lactation milk yield and litter size at the POU1F1-AluI locus. Birth weight was found to be higher in animals that have the CC genotype at the POU1F1-PstI locus. In conclusion, the POU1F1 gene can be used as a molecular marker for economic features like reproduction, growth, milk content and yield in Saanen goats.

The Identification of Novel Single-Nucleotide Polymorphisms of Equine Beta-Lactoglobulin and Lactotransferrin Genes.

Lactoferrin (LTF), also called lactotransferrin, is an iron-binding protein and member of transferrin family, whereas ß-LG is an important milk protein and belongs to the ligand-binding protein family of lipocalins and binds retinol. Beta-lactoglobulin (ß-LG) is a major whey protein and it is a member of lipocalins family. ß-LG is also known as a major allergen reason of cow milk protein allergy. Yet, no information has been reported about ß-LG and LTF genes in donkeys. In this study, the genetic variation of ß-LG and LTF genes were analyzed via DNA sequencing in 77 donkeys, which were individuals reared in Thrace region of Turkey. Four novel SNPs are identified as g.12887G>A, g.12973G>A, g.12654G>A, and g.13057T>C in the third intron region of ß-LG gene and a genetic variant g.272719G>A in the 14th intron of LTF gene. The DNA sequences of ß-LG and LTF genes in donkeys are reported for the first time in this study and these sequences were deposited to NCBI GenBank database, with the accession number MK125015 and MK211165, respectively. These variations may have an effect on milk yield and content in donkeys, so further investigations are needed to prove this hypothesis and these may be candidate SNPs for donkey breeding.

Genetic characterization of some Turkish sheep breeds based on the sequencing of the Ovar-DRB1 gene in the major histocompatibility complex (MHC) gene region.

In this research, Ovar-DRB1 gene in the major histocompatibility complex (MHC) gene region was surveyed by DNA sequencing in some of the native sheep breeds that are reared in Turkey. A total of 80 samples were collected from eight different Turkish native sheep breeds, and these samples were used for DNA sequencing. The exon 2 region of Ovar-DRB1 in the MHC gene region was polymerase chain reaction (PCR) amplified and sequenced. A total of 25 new alleles were revealed in the Ovar-DRB1 gene in Turkish native sheep breeds with 24 variable sites; only 13 sites were parsimony informative. The average pairwise genetic distance was 0.029 % for the Ovar-DRB1 gene exon 2 region. The sequence variations at eight different positions (7026, 7036, 7040, 7053, 7059, 7069, 7131 and 7214) are found in all of the studied samples. G → C transversion at position 7081 is only seen in Akkaraman sheep breed, whereas T → C transition at position 7097 is only seen in one sample from the Akkaraman breed. Overall, two main groups were detected among the 25 alleles from Turkish native sheep breeds. All Daǧliç and Kivircik alleles and one allele from Karayaka, Malya and Sakiz are grouped together while all the other breeds are grouped in the other branch.