Analyze Diet

Topic:DNA

DNA in horses refers to the genetic material that carries the hereditary information necessary for the growth, development, functioning, and reproduction of equine species. It consists of sequences of nucleotides that encode the genetic instructions used in the development and functioning of horses. DNA analysis in horses can provide insights into genetic diversity, lineage, and breed characteristics. It is also utilized in identifying genetic disorders, understanding hereditary traits, and assisting in selective breeding programs. This page compiles peer-reviewed research studies and scholarly articles that explore the structure, function, and applications of DNA analysis in equine genetics and breeding.
Ancient Patrilineal Lines and Relatively High ECAY Diversity Preserved in Indigenous Horses Revealed With Novel Y-Chromosome Markers.
Frontiers in genetics    May 21, 2020   Volume 11 467 doi: 10.3389/fgene.2020.00467
Liu S, Yang Y, Pan Q, Sun Y, Ma H, Liu Y, Wang M, Zhao C, Wu C.Extremely low nucleotide diversity of modern horse Y-chromosome has been reported, and only poor phylogenetic resolution could be resulted from limited Y-chromosome markers. In this study, three types of horse Y-chromosome markers, including Single-nucleotide polymorphisms (SNPs), copy number variants (CNVs), and allele-specific CNVs, were developed by screening more than 300 male horses from 23 indigenous Chinese horse populations and 4 imported horse breeds. Fourteen segregating sites including a novel SNP in the AMELY gene were found in approximately 53 kb of male-specific Y-chromosome sequ...
Mitochondrial DNA variation in the Italian Heavy Draught Horse.
PeerJ    May 15, 2020   Volume 8 e8996 doi: 10.7717/peerj.8996
Lancioni H, Cardinali I, Giontella A, Antognoni MT, Miglio A.In the last decades, Italy as well as other developed countries have registered a decrease in the population size of many local horse breeds. The continuous crossbreeding has determined the dilution of genetic heritage of several native breeds. The Italian Heavy Draught Horse (IHD) is the only autochthonous Italian coldblooded horse among these breeds; therefore, it represents a resource to be preserved. In 1927, the first generation of this breed was officially created by crossing different Heavy Draught horses with local mares and recorded in a Studbook. Methods: To provide the first compreh...
Signatures of selection analysis using whole-genome sequence data reveals novel candidate genes for pony and light horse types.
Genome    May 14, 2020   Volume 63, Issue 8 387-396 doi: 10.1139/gen-2020-0001
Salek Ardestani S, Aminafshar M, Zandi Baghche Maryam MB, Banabazi MH, Sargolzaei M, Miar Y.Natural selection and domestication have shaped modern horse populations, resulting in a vast range of phenotypically diverse breeds. Horse breeds are classified into three types (pony, light, and draft) generally based on their body type. Understanding the genetic basis of horse type variation and selective pressures related to the evolutionary trend can be particularly important for current selection strategies. Whole-genome sequences were generated for 14 pony and 32 light horses to investigate the genetic signatures of selection of the horse type in pony and light horses. In the overlappin...
Molecular typing of equine papillomavirus and autovaccination to treat horses with cutaneous papillomatosis.
Australian veterinary journal    May 10, 2020   Volume 98, Issue 8 405-410 doi: 10.1111/avj.12954
Onen EA.The aim of this study was to evaluate formalin-inactivated autovaccination to treat cutaneous papillomatosis and to perform molecular typing of the papillomavirus in four horses (two foals, one 3-year-old filly and a 5-year-old stallion). Methods: Histopathological slides of lesions were prepared and stained with haematoxylin and eosin (H&E) to establish a diagnosis that was based on observation koilocytosis, which is a pathognomonic cytopathic change that is associated with papillomatosis, using light microscopy. Polymerase chain reaction (PCR) and DNA sequencing were performed using the ...
Phylogenetic Characteristics of Echinococcus granulosus Sensu Lato in Uzbekistan.
The Korean journal of parasitology    April 30, 2020   Volume 58, Issue 2 205-210 doi: 10.3347/kjp.2020.58.2.205
Kim HJ, Yong TS, Shin MH, Lee KJ, Park GM, Suvonkulov U, Kovalenko D, Yu HS.Echinococcosis occurs mainly in areas with heavy livestock farming, such as Central Asia, America, and Australia. Echinococcus granulosus sensu lato (s.l.) infection causes echinococcosis in intermediate hosts, such as sheep, cattle, goats, camels, and horses. Numerous cases of echinococcosis occur in Uzbekistan as stock farming is a primary industry. Epidemiological and genetic studies of E. granulosus s.l. are very important for mitigating its impact on public health and the economy; however, there are no such studies on E. granulosus s.l. in Uzbekistan. In the present study, to determine wh...
Equine Hoof Canker: Bovine Papillomavirus Infection Is Not Associated With Impaired Keratinocyte Differentiation.
Veterinary pathology    April 29, 2020   Volume 57, Issue 4 525-534 doi: 10.1177/0300985820921820
Apprich V, Licka T, Freiler S, Gabriel C.Impaired keratinocyte differentiation has recently been suggested as a key event in equine hoof canker development. Koilocytotic appearance of keratinocytes, one of the most characteristic morphological alterations in hoof canker tissue, is also a common marker for papillomavirus (PV) infection, and bovine PV-1 and/or -2 (BPV-1/2) has previously been detected in equine canker patients. Therefore, the present study aimed to correlate the frequency and severity of koilocytotic keratinocytes with BPV detection in hoof canker samples. Hoof tissue of 5/18 canker-affected horses and 2/6 control hors...
Detection of Neorickettsia risticii, the agent of Potomac horse fever, in horses from Rio de Janeiro, Brazil.
Scientific reports    April 29, 2020   Volume 10, Issue 1 7208 doi: 10.1038/s41598-020-64328-2
Paulino PG, Almosny N, Oliveira R, Viscardi V, Müller A, Guimarães A, Baldani C, da Silva C, Peckle M, Massard C, Santos H.This study aims to report the presence of Neorickettsia risticii DNA in blood samples from naturally infected horses in Rio de Janeiro, provide clinicopathological findings related to the infection, and report the phylogenetic diversity of the 16S rDNA of N. risticii in order to evaluate its heterogeneity. Real-time quantitative polymerase chain reaction (qPCR) was performed to investigate the presence of N. risticii in samples collected from horses (n = 187). Five positive samples were found in the molecular screening. Hypoalbuminemia and high levels of creatine kinase and lactate dehydro...
Genes encoding equine β-lactoglobulin (LGB1 and LGB2): Polymorphism, expression, and impact on milk composition.
PloS one    April 22, 2020   Volume 15, Issue 4 e0232066 doi: 10.1371/journal.pone.0232066
Wodas L, Mackowski M, Borowska A, Puppel K, Kuczynska B, Cieslak J.β-lactoglobulin is one of the most abundant milk whey proteins in many mammal species, including the domestic horse. The aim of this study was to screen for polymorphism in the equine LGB1 and LGB2 gene sequences (all exons, introns, and 5'-flanking region) and to assess potential relationship of particular genotypes with gene expression levels (measured in milk somatic cells) and milk composition traits (protein, fat, lactose, and total β-lactoglobulin content). Direct DNA sequencing analysis was performed for twelve horse breeds: Polish Primitive Horse (PPH), Polish Coldblood Horse (PCH), ...
A missense mutation in ASIP is associated with light point variation in donkeys.
Animal genetics    April 20, 2020   Volume 51, Issue 4 629 doi: 10.1111/age.12940
Yu Y, Shang S, Zhang X, Wang Z, Dang W, Zhang J, Zhu Y, Dang R, Irwin DM, Zhang S.No abstract available
Insertion of Telomeric Repeats in the Human and Horse Genomes: An Evolutionary Perspective.
International journal of molecular sciences    April 18, 2020   Volume 21, Issue 8 2838 doi: 10.3390/ijms21082838
Santagostino M, Piras FM, Cappelletti E, Del Giudice S, Semino O, Nergadze SG, Giulotto E.Interstitial telomeric sequences (ITSs) are short stretches of telomeric-like repeats (TTAGGG)n at nonterminal chromosomal sites. We previously demonstrated that, in the genomes of primates and rodents, ITSs were inserted during the repair of DNA double-strand breaks. These conclusions were derived from sequence comparisons of ITS-containing loci and ITS-less orthologous loci in different species. To our knowledge, insertion polymorphism of ITSs, i.e., the presence of an ITS-containing allele and an ITS-less allele in the same species, has not been described. In this work, we carried out a gen...
Whole genome detection of sequence and structural polymorphism in six diverse horses.
PloS one    April 9, 2020   Volume 15, Issue 4 e0230899 doi: 10.1371/journal.pone.0230899
Al Abri MA, Holl HM, Kalla SE, Sutter NB, Brooks SA.The domesticated horse has played a unique role in human history, serving not just as a source of animal protein, but also as a catalyst for long-distance migration and military conquest. As a result, the horse developed unique physiological adaptations to meet the demands of both their climatic environment and their relationship with man. Completed in 2009, the first domesticated horse reference genome assembly (EquCab 2.0) produced most of the publicly available genetic variations annotations in this species. Yet, there are around 400 geographically and physiologically diverse breeds of hors...
Gallop Racing Shifts Mature mRNA towards Introns: Does Exercise-Induced Stress Enhance Genome Plasticity?
Genes    April 9, 2020   Volume 11, Issue 4 410 doi: 10.3390/genes11040410
Cappelli K, Mecocci S, Gioiosa S, Giontella A, Silvestrelli M, Cherchi R, Valentini A, Chillemi G, Capomaccio S.Physical exercise is universally recognized as stressful. Among the "sport species", the horse is probably the most appropriate model for investigating the genomic response to stress due to the homogeneity of its genetic background. The aim of this work is to dissect the whole transcription modulation in Peripheral Blood Mononuclear Cells (PBMCs) after exercise with a time course framework focusing on unexplored regions related to introns and intergenic portions. PBMCs NGS from five 3 year old Sardinian Anglo-Arab racehorses collected at rest and after a 2000 m race was performed. Apart from d...
Animal domestication in the era of ancient genomics.
Nature reviews. Genetics    April 7, 2020   Volume 21, Issue 8 449-460 doi: 10.1038/s41576-020-0225-0
Frantz LAF, Bradley DG, Larson G, Orlando L.The domestication of animals led to a major shift in human subsistence patterns, from a hunter-gatherer to a sedentary agricultural lifestyle, which ultimately resulted in the development of complex societies. Over the past 15,000 years, the phenotype and genotype of multiple animal species, such as dogs, pigs, sheep, goats, cattle and horses, have been substantially altered during their adaptation to the human niche. Recent methodological innovations, such as improved ancient DNA extraction methods and next-generation sequencing, have enabled the sequencing of whole ancient genomes. These gen...
Genome-wide association study for host genetic factors associated with equine herpesvirus type-1 induced myeloencephalopathy.
Equine veterinary journal    April 6, 2020   Volume 52, Issue 6 794-798 doi: 10.1111/evj.13261
Dunuwille WMB, YousefiMashouf N, Balasuriya UBR, Pusterla N, Bailey E.Equid herpesvirus (EHV-1) infections in horses can lead to equine herpesvirus myeloencephalopathy (EHM), characterised by neurological clinical signs. The sporadic occurrence of the disease in horse herds suggests a host genetic component. A recent study reported an association between the occurrence of EHM and genetic markers on horse chromosome 6 (ECA6). Objective: To investigate the association of EHM with genetic host factors, especially with reference to the association reported for ECA6. Methods: Genome-wide association study (GWAS) was conducted based on 94 horses that had EHV-1 infecti...
A De Novo MITF Deletion Explains a Novel Splashed White Phenotype in an American Paint Horse.
The Journal of heredity    April 4, 2020   Volume 111, Issue 3 287-293 doi: 10.1093/jhered/esaa009
Magdesian KG, Tanaka J, Bellone RR.Splashed white is a coat color pattern in horses characterized by extensive white patterning on the legs, belly, and face often accompanied by blue eyes and deafness. Three mutations in microphthalmia-associated transcription factor (MITF) and two mutations in Paired Box 3 (PAX3) have been identified that explain splashed white patterns (SW1-SW5). An American Paint Horse stallion with a splashed white phenotype and blue eyes, whose parents were not white patterned, was negative for the 5 known splashed white variants and other known white spotting alleles. This novel splashed white phenotype (...
Characterization and phylogenetic analysis of the first complete mitochondrial genome of Cylicocyclus radiatus.
Veterinary parasitology    April 3, 2020   Volume 281 109097 doi: 10.1016/j.vetpar.2020.109097
Hu L, Zhang M, Sun Y, Bu Y.Cylicocyclus spp. are common parasites in the intestine of equine animals. Hosts infected by these strongylid nematodes might face emaciation, anemia, and even death, all of which lead to significant economic losses. In the present study, the complete mitochondrial genome (mitogenome) of Cylicocyclus radiatus, the type species of the genus Cylicocyclus, was sequenced and characterized for the first time using next-generation sequencing technology. Results show that the mitogenome of C. radiatus is a double-stranded 13,836 bp circular molecule, containing 12 protein-coding genes, 22 tRNA genes,...
Effect of relaxin on semen quality variables of cryopreserved stallion semen.
Animal reproduction science    April 2, 2020   Volume 216 106351 doi: 10.1016/j.anireprosci.2020.106351
Elkhawagah AR, Nervo T, Poletto M, Martino NA, Gallo D, Bertero A, Vincenti L.The aim of the study was to ascertain effects of different concentrations of relaxin added to extender medium during the pre-freezing incubation periods on quality variables of stallion frozen-thawed spermatozoa. Semen samples collected from three stallions were filtered, diluted with skim milk, and centrifuged at 600g for 10 min. Sperm pellets were suspended in BotuCrio freezing medium to a final concentration of 50 × 10 sperm/mL. The diluted semen was divided into five experimental groups supplemented with 0 (control), 12.5, 25, 50, or 100 ng/mL of relaxin. The semen samples were transferre...
Comparative analysis of CpG islands in equine infectious anemia virus strains.
Virus genes    April 1, 2020   Volume 56, Issue 3 339-346 doi: 10.1007/s11262-020-01749-1
Liu Q, Yu YY, Wang HY.Increasing evidence suggests that DNA methylation has key roles in the replication of retroviruses, including lentiviruses, and pathogenesis of diseases. However, the precise characteristics of CpG islands are not known for many retroviruses. In this study, we compared the distribution of CpG islands among strains of equine infectious anemia virus (EIAV), a lentivirus in the family Retroviridae and a model for HIV research. We identified CpG islands in 32 full-length EIAV genomic sequences obtained from the GenBank database using MethPrimer. Only one CpG island, from 100 to 120 bp, was identi...
Genetic Differentiation of the Two Types of Polish Cold-blooded Horses Included in the National Conservation Program.
Animals : an open access journal from MDPI    March 24, 2020   Volume 10, Issue 3 542 doi: 10.3390/ani10030542
Gurgul A, Jasielczuk I, Semik-Gurgul E, Pawlina-Tyszko K, Szmatoła T, Polak G, Bugno-Poniewierska M.The current role of the horse as a companion animal resulted in a decrease of interest in breeding and usage of draft horses. This meant that the population of cold-blooded horses in Poland has been dramatically reduced during the last decades. To avoid impoverishment of the gene pool of the local horse population, a conservation program was established which involves draft horses and other local horse breeds. The draft horses bred in Poland can be subdivided in a few horse types of which the most widespread and consolidated are Sztumski and Sokólski horses. These two subpopulations are pheno...
First case of fatal equine meningoencephalitis caused by Halicephalobus gingivalis in Mexico.
Veterinary parasitology, regional studies and reports    March 18, 2020   Volume 20 100399 doi: 10.1016/j.vprsr.2020.100399
Avila VA, López-García Y, Hernández-Castro R, Salas-Garrido CG, Ramírez-Lezama J, Calderón-Villa R, Martínez-Chavarría LC.Aberrant nematode larval migration in the CNS of horses is rare but frequently fatal; one of the main etiological agents involved in this illness is Halicephalobus gingivalis. This soil nematode has been associated with several fatal equine meningoencephalitis reports worldwide; however, it had never been diagnosed in horses of Mexico. A 10 year-old Andalusian horse presented dysphagia, fever, weakness, prostration and ataxia; the patient expired during the medical attention. Post mortem examination was performed and no gross alterations were found. Histopathology revealed meningoencephalitis...
A Subset of Equine Gastric Squamous Cell Carcinomas Is Associated With Equus Caballus Papillomavirus-2 Infection.
Veterinary pathology    March 17, 2020   Volume 57, Issue 3 427-431 doi: 10.1177/0300985820908797
Alloway E, Linder K, May S, Rose T, DeLay J, Bender S, Tucker A, Luff J.Squamous cell carcinoma (SCC) is the most common neoplasm of the equine stomach. However, the mechanisms underlying malignant transformation are unknown. As papillomavirus-2 (EcPV-2) is a likely cause of some genital SCCs, we hypothesized that EcPV-2 is associated with a subset of equine gastric SCCs. To this aim, we performed polymerase chain reaction (PCR) and in situ hybridization (ISH) for EcPV-2 E6/ E7 oncogenes on 11 gastric SCCs and on gastric samples from 15 control horses with no SCC. PCR for EcPV-2 was positive in 7/11 (64%) gastric SCCs; non-SCC gastric samples were all negative. I...
Detection of intra-articular gene therapy in horses using quantitative real time PCR in synovial fluid and plasma.
Drug testing and analysis    March 16, 2020   Volume 12, Issue 6 743-751 doi: 10.1002/dta.2785
Haughan J, Jiang Z, Stefanovski D, Moss KL, Ortved KF, Robinson MA.Gene therapy promotes the expression of missing or defective genes and can be curative following administration of a single dose. Gene therapy is prohibited in equine athletes by regulatory bodies due to the high potential for abuse and novel analytical methods are needed for detection. The goal of this study was to detect the administration of an experimental gene therapy: a recombinant adeno-associated viral vector (rAAV) carrying a transgene for the anti-inflammatory cytokine IL-10 (rAAV-IL10). Twelve horses were randomly assigned to receive an intra-articular injection of rAAV-IL10 or phos...
Transporting and Exercising Unconditioned Horses: Effects on Microflora Populations.
Journal of equine veterinary science    March 13, 2020   Volume 90 102988 doi: 10.1016/j.jevs.2020.102988
Szemplinski KL, Thompson A, Cherry N, Guay K, Smith WB, Brady J, Jones T.The objective of this study was to determine if transportation and exercise stress in horses affect the microflora populations in the equine hindgut. Four horses were subjected to three transport periods (0, 3, and 6 hours) with a 7-d rest period between each transport. Horses were fed 0.91 kg/day of Purina Impact All Stages 12% and had ad libitum access to Cynodon dactylon (Coastal Bermudagrass) hay. Fecal samples were collected before (0 hours) and after (48 hours) transport. In addition, three horses underwent a different standardized exercise test with a 7-d rest period between each ex...
Gut Microbiomes of Endangered Przewalski’s Horse Populations in Short- and Long-Term Captivity: Implication for Species Reintroduction Based on the Soft-Release Strategy.
Frontiers in microbiology    March 12, 2020   Volume 11 363 doi: 10.3389/fmicb.2020.00363
Tang L, Li Y, Srivathsan A, Gao Y, Li K, Hu D, Zhang D.Captivity maybe the only choice for survival of many endangered vertebrates, and understanding its broad effects is important for animal management and conservation, including breeding endangered species for subsequent release. Extreme environmental changes during captivity may influence survival ability in the wild. Captivity decreases gut bacterial diversity in a wide range of animals. However, most studies directly compare animals living in captivity with those in the wild, and there is a lack of understanding of effects of gradient shift in lifestyle during species reintroduction based on ...
Development of a nested PCR assay for detection of Streptococcus equi subspecies equi in clinical equine specimens and comparison with a qPCR assay.
Journal of microbiological methods    March 9, 2020   Volume 172 105887 doi: 10.1016/j.mimet.2020.105887
Noll LW, Stoy CPA, Wang Y, Porter EG, Lu N, Liu X, Burklund A, Peddireddi L, Hanzlicek G, Henningson J, Chengappa MM, Bai J.Streptococcus equi subsp. equi is a Gram positive bacterial pathogen commonly associated with strangles in horses, a respiratory disease characterized by abscessation of submandibular and retropharyngeal lymph nodes which can lead to obstruction of the airway. Several real-time PCR (qPCR) assays have been developed for detection of S. equi from horses with many targeting conserved regions of the S. equi cell wall-associated M-protein (SeM), a major virulence factor and immunogen of S. equi. Our objective was to develop a nested PCR (nPCR) targeting SeM and an 18S rRNA internal control gene for...
Characterization and comparison of the bacterial microbiota in different gastrointestinal tract compartments of Mongolian horses.
MicrobiologyOpen    March 9, 2020   Volume 9, Issue 6 1085-1101 doi: 10.1002/mbo3.1020
Su S, Zhao Y, Liu Z, Liu G, Du M, Wu J, Bai D, Li B, Bou G, Zhang X, Dugarjaviin M.The intestinal microbiota plays an important role in the health and metabolism of the host. Next-generation sequencing technology has enabled the characterization of the gut microbiota of several animal species. We analyzed the intestinal microbiota in six different parts of the gastrointestinal tracts (GITs) of five Mongolian horses by sequencing the 16S rRNA gene V3-V4 hypervariable region. All horses were kept in the natural habitat of the Inner Mongolia grassland. Significant differences were observed among the microbiota compositions of the distinct GIT regions. In addition, while the mic...
Rapid isothermal duplex real-time recombinase polymerase amplification (RPA) assay for the diagnosis of equine piroplasmosis.
Scientific reports    March 5, 2020   Volume 10, Issue 1 4096 doi: 10.1038/s41598-020-60997-1
Lei R, Wang X, Zhang D, Liu Y, Chen Q, Jiang N.Equine piroplasmosis (EP) is a severe disease of horses caused by the tick-borne protozoa Theileria equi (T. equi) and Babesia caballi (B. caballi). Infectious carriers are not always symptomatic, meaning there is a risk to non-enzootic areas. Regulatory tests for EP include sero-epidemiological methods for equine babesiosis, but these lack specificity due to cross-reactivity with other Babesia species. In this study, we present a real-time quantitative recombinase polymerase amplification (qRPA) method for fast simultaneous detection of both T. equi and B. caballi. In this method, primers and...
Molecular sexing and preliminary assessment of population sex ratio of the endangered Malayan tapir (Tapirus indicus) in Peninsular Malaysia.
Scientific reports    March 4, 2020   Volume 10, Issue 1 3973 doi: 10.1038/s41598-020-60552-y
Lim QL, Tan YL, Ng WL, Yong CSY, Ismail A, Rovie-Ryan JJ, Rosli N, Annavi G.A molecular sexing method by polymerase chain reaction (PCR) amplification of a portion of the sex-determining region Y (SRY) and the zinc finger (ZF) gene, as well as six equine Y-chromosome-specific microsatellite markers, were tested in the Malayan tapir (Tapirus indicus). While the microsatellite markers did not yield any male-specific amplicons for sex-typing, the SRY/ZF marker system produced reliable molecular sexing results by accurately sex-typing 31 reference Malayan tapirs, using whole blood, dried blood spot (DBS), or tissue samples as materials for DNA extraction. The marker syste...
Potential for residual contamination by Streptococcus equi subspp equi of endoscopes and twitches used in diagnosis of carriers of strangles.
Equine veterinary journal    March 2, 2020   Volume 52, Issue 6 884-890 doi: 10.1111/evj.13248
Svonni E, Andreasson M, Fernström LL, Rydén A, Pringle J, Riihimäki M.Endoscopic examinations are essential for diagnosis and treatment of strangles (S equi infection) in horses. However, even after disinfection, endoscopes may retain viable bacteria or bacterial DNA. Twitches are commonly used during endoscopic examinations and can thus also potentially transmit the organism to other horses. Objective: To evaluate the efficacy of different disinfectant methods to eliminate S equi from experimentally contaminated endoscopes and twitches and the effectiveness of field disinfection of endoscopes used in sampling carriers of S equi. Methods: Experimental contami...
Comparison of different mathematical models to assess seasonal variations in the longevity of DNA integrity of cooled-stored stallion sperm.
Andrologia    February 28, 2020   Volume 52, Issue 4 e13545 doi: 10.1111/and.13545
Ortiz I, Quiñones-Pérez C, Hidalgo M, Consuegra C, Diaz-Jimenez M, Dorado J, Vega-Pla JL, Crespo F.Dynamic assessment of sperm DNA fragmentation (SDF) has shown to give fuller understanding of stallion semen quality; however, there have been limited attempts to use this parameter to investigate seasonal changes in productive functions. The aims of this study were to: (a) establish a reliable mathematical model to describe the longevity of cooled-stored sperm DNA integrity; (b) to examine the effect of seasonal variations on SDF. Ejaculates were cooled to 5°C, and SDF was analysed after 0, 6 and 24 hr of storage. The coefficient of determination (R ) was calculated after fine-tuning linear...
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