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Topic:Genetics

Genetics in horses encompasses the study of hereditary traits and the genetic makeup that influences various characteristics and health conditions in equine populations. This field involves the analysis of genes and their functions, inheritance patterns, and the impact of genetic variations on traits such as coat color, performance ability, and susceptibility to diseases. Research in equine genetics employs techniques such as genome mapping, sequencing, and genetic testing to identify specific genes and mutations associated with these traits. This page gathers peer-reviewed research studies and scholarly articles that explore the genetic basis of equine traits, the methodologies used in genetic research, and the implications for breeding, health management, and conservation of horse breeds.
Polymer-mineral scaffold augments in vivo equine multipotent stromal cell osteogenesis.
Stem cell research & therapy    March 9, 2018   Volume 9, Issue 1 60 doi: 10.1186/s13287-018-0790-8
Duan W, Chen C, Haque M, Hayes D, Lopez MJ.Use of bioscaffolds to direct osteogenic differentiation of adult multipotent stromal cells (MSCs) without exogenous proteins is a contemporary approach to bone regeneration. Identification of in vivo osteogenic contributions of exogenous MSCs on bioscaffolds after long-term implantation is vital to understanding cell persistence and effect duration. This study was designed to quantify in vivo equine MSC osteogenesis on synthetic polymer scaffolds with distinct mineral combinations 9 weeks after implantation in a murine model. Cryopreserved, passage (P)1, equine bone marrow-derived MSCs (BMSC)...
Differences between horse selection based on two forms of osteochondrosis in fetlock.
Journal of applied genetics    March 9, 2018   Volume 59, Issue 2 225-230 doi: 10.1007/s13353-018-0437-6
Lewczuk D, Bereznowski A, Hecold M, Frąszczak M, Ruść A, Korwin-Kossakowska A, Szyda J, Kamiński S.Horses lose potential opportunities because of health problems. Available breeding strategies are not effective enough, probably also because of the different definition used and its genetic usefulness. The aim of the study was to compare the genetic background estimated by the genome-wide association study (GWAS) for osteochondrosis using two different scaling osteochondrosis (OC)/healthy and osteochondrosis dissecans (OCD)/healthy systems for evaluating the disease status of investigated fetlock joints. Two hundred one Warmblood horses trained for performance tests (87 stallions and 114 mare...
A missense mutation in MYH1 is associated with susceptibility to immune-mediated myositis in Quarter Horses.
Skeletal muscle    March 6, 2018   Volume 8, Issue 1 7 doi: 10.1186/s13395-018-0155-0
Finno CJ, Gianino G, Perumbakkam S, Williams ZJ, Bordbari MH, Gardner KL, Burns E, Peng S, Durward-Akhurst SA, Valberg SJ.The cause of immune-mediated myositis (IMM), characterized by recurrent, rapid-onset muscle atrophy in Quarter Horses (QH), is unknown. The histopathologic hallmark of IMM is lymphocytic infiltration of myofibers. The purpose of this study was to identify putative functional variants associated with equine IMM. A genome-wide association (GWA) study was performed on 36 IMM QHs and 54 breed matched unaffected QHs from the same environment using the Equine SNP50 and SNP70 genotyping arrays. A mixed model analysis identified nine SNPs within a ~ 2.87 Mb region on chr11 that were significantly (...
Blastocyst production after intracytoplasmic sperm injection with semen from a stallion with testicular degeneration.
Reproduction in domestic animals = Zuchthygiene    March 1, 2018   Volume 53, Issue 3 814-817 doi: 10.1111/rda.13153
Roels K, Smits K, Ververs C, Govaere J, D'Herde K, Van Soom A.In horse breeding, intracytoplasmic sperm injection (ICSI) has gained interest to obtain offspring from subfertile individuals. This paper presents a case report of a stallion with severe testicular degeneration. Semen analysis showed very low motility and 83.5% of detached heads. Histology of a testicular biopsy showed severely decreased spermatogenesis, while transmission electron microscopy of the sperm cells revealed no significant abnormalities. A total of 39 oocytes were fertilized by ICSI with frozen-thawed spermatozoa of this stallion: 25 oocytes with intact spermatozoa and 24 with det...
Venom and Antivenom of the Redback Spider (Latrodectus hasseltii) in Japan. Part I. Venom Extraction, Preparation, and Laboratory Testing.
Japanese journal of infectious diseases    February 28, 2018   Volume 71, Issue 2 116-121 doi: 10.7883/yoken.JJID.2017.291
Matsumura T, Mashiko R, Sato T, Itokawa K, Maekawa Y, Ogawa K, Isawa H, Yamamoto A, Mori S, Horita A, Ginnaga A, Miyatsu Y, Takahashi M, Taki H....The redback spider (Latrodectus hasseltii Thorell) reportedly invaded Japan in September 1995. To date, 84 redback spider bite cases have been reported; 7 of these cases employed the antivenom. Antivenom has been imported from Australia in the past, but because of restrictions on exportation it was evident that nearly all of the antivenom present in Japan would expire during 2014. In 2014, a plan was proposed to experimentally manufacture and stockpile a horse antiserum for ourselves, using redback spiders indigenous to Japan. A total of 11,403 female spiders were captured alive: 1,217 from th...
Progenitor cells in auricular cartilage demonstrate cartilage-forming capacity in 3D hydrogel culture.
European cells & materials    February 27, 2018   Volume 35 132-150 doi: 10.22203/eCM.v035a10
Otto IA, Levato R, Webb WR, Khan IM, Breugem CC, Malda J.Paramount for the generation of auricular structures of clinically-relevant size is the acquisition of a large number of cells maintaining an elastic cartilage phenotype, which is the key in producing a tissue capable of withstanding forces subjected to the auricle. Current regenerative medicine strategies utilize chondrocytes from various locations or mesenchymal stromal cells (MSCs). However, the quality of neo-tissues resulting from these cell types is inadequate due to inefficient chondrogenic differentiation and endochondral ossification, respectively. Recently, a subpopulation of stem/pr...
Concentrations of follicle stimulating hormone associated with follicle selection, number of follicles, and ipsilateral vs contralateral relationships in mares.
Theriogenology    February 24, 2018   Volume 113 159-165 doi: 10.1016/j.theriogenology.2018.02.017
Ginther OJ, Domingues RR, Dangudubiyyam SV, Araujo ER.Diameter deviation or follicle selection during a follicular wave begins with continuous growth rate of F1 (dominant or selected follicle; DF) and decreasing growth rate of F2 (largest subordinate). Intraovarian patterns based on presence or absence of the DF and CL are DF-CL, DF, CL, and devoid (neither DF nor CL). The DF and CL relationships in a pair of ovaries are ipsilateral (DF-CL pattern and devoid pattern) and contralateral (DF pattern and CL pattern). The effects of patterns and relationships on F1, F2, and FSH during deviation were determined in 21 mares. Diameters of F1 and F2 at ex...
Ancient genomes revisit the ancestry of domestic and Przewalski’s horses.
Science (New York, N.Y.)    February 22, 2018   Volume 360, Issue 6384 111-114 doi: 10.1126/science.aao3297
Gaunitz C, Fages A, Hanghøj K, Albrechtsen A, Khan N, Schubert M, Seguin-Orlando A, Owens IJ, Felkel S, Bignon-Lau O, de Barros Damgaard P....The Eneolithic Botai culture of the Central Asian steppes provides the earliest archaeological evidence for horse husbandry, ~5500 years ago, but the exact nature of early horse domestication remains controversial. We generated 42 ancient-horse genomes, including 20 from Botai. Compared to 46 published ancient- and modern-horse genomes, our data indicate that Przewalski's horses are the feral descendants of horses herded at Botai and not truly wild horses. All domestic horses dated from ~4000 years ago to present only show ~2.7% of Botai-related ancestry. This indicates that a massive genomic ...
Molecular Genetic Manipulation of Sarcocystis neurona.
Current protocols in microbiology    February 22, 2018   Volume 48 20D.2.1-20D.2.14 doi: 10.1002/cpmc.48
Howe DK, Yeargan M, Simpson L, Dangoudoubiyam S.Sarcocystis neurona is a member of the important phylum Apicomplexa and the primary cause of equine protozoal myeloencephalitis (EPM). Moreover, S. neurona is the best-studied species in the genus Sarcocystis, one of the most successful parasite taxa, as virtually all vertebrate animals may be infected by at least one species. Consequently, scientific investigation of S. neurona will aid in the control of EPM and neurologic disease in sea mammals, while also improving our understanding of a prominent branch on the apicomplexan phylogenetic tree. These protocols describe methods that expand the...
Genetic Diversity and Structure of Iranian Horses’ Population Based on Mitochondrial Markers.
Journal of equine veterinary science    February 22, 2018   Volume 64 107-111 doi: 10.1016/j.jevs.2018.02.011
Hedayat Evrigh N, Omri M, Boustan A, Seyedsharifi R, Vahedi V.The source and history of horse domestication have been studied for decades in biology and archeology. The aim of this study was to evaluate the genetic structure and the genetic relationship between six breeds of native horses using mitochondrial D-loop region. Blood samples were collected from 96 horses. Total DNA was extracted, and 430 bp of D-Loop region (hyper variable) was amplified and sequenced using Sanger sequencing methods. The analysis of data led to identify 48 polymorphic sites that create 52 haplotypes. The plotted phylogenic tree for haplotypes of Iranian native horses is plac...
Pedigree analysis and inbreeding effects over morphological traits in Campolina horse population.
Animal : an international journal of animal bioscience    February 22, 2018   1-10 doi: 10.1017/S175173111800023X
Bussiman FO, Perez BC, Ventura RV, Peixoto MGCD, Curi RA, Balieiro JCC.Genetic improvement, without control of inbreeding, can go to loss of genetic variability, reducing the potential for genetic gains in the domestic populations. The aim of this study was to analyze the population structure and the inbreeding depression in Campolina horses. Phenotype information from 43 465 individuals was analyzed, data provided by the Campolina Breeders Association. A pedigree file containing 107 951 horses was used to connected the phenotyped individuals. The inbreeding coefficient was performed by use of the diagonal of the relationship matrix and the genealogical parameter...
Lack of support for adaptation of post-glacial horses to woodlands.
Nature ecology & evolution    February 21, 2018   Volume 2, Issue 4 582-583 doi: 10.1038/s41559-018-0491-9
Sommer RS, Hegge C, Schmölcke U.No abstract available
How is the Morphometry of Stallions and Mares Show-Winning and Nonwinning Campolina Brazilian Breed with Batida and Picada Gaits?
Journal of equine veterinary science    February 21, 2018   Volume 64 34-40 doi: 10.1016/j.jevs.2018.02.012
Sousa AS, Jesus IIC, Araújo Oliveira CA, Costa RB, de Godoi FN.The aim of this study was to evaluate the morphometric measurements of stallion and mares show-winning and nonwinning Campolina breed with batida and picada gaits. A total of 167 horses (74 stallions and 93 mares), consisting of 102 batida and 65 picada gait animals, were used. Among these horses, 96 were awarded and 71 were not. Twenty linear, seven angular, and 11 girth measurements were taken. Results from the judging sessions were collected, and awards were observed in both sexes and both gaits. A descriptive statistical analysis and an analysis of variance were performed using the MEANS s...
Molecular Characterization of the Horn Fly Haematobia irritans Infesting Horses in Central Anatolia Region in Turkey.
Journal of equine veterinary science    February 21, 2018   Volume 64 49-54 doi: 10.1016/j.jevs.2018.02.004
Onder Z, Duzlu O, Yildirim A, Ciloglu A, Okur M, Inci A.This study reports intense horn fly infestations of horses raised in an important wetland ecosystem, Sultan Marshes in Central Anatolia, Turkey. In total, seven horses raised together were found to be harbored over 500 flies per site of each animal. Totally, 376 fly specimens were collected from the horses by using the nets and were subjected to the laboratory for species identification. All flies were morphologically identified as the adults of Haematobia irritans. Partial fragments of mitochondrial cytochrome oxidase I (mt-COI) gene from totally 50 isolates were amplified for sequence and ph...
Hereditary Multiple Exostoses-A Comprehensive Examination of a 4-Year-Old Dutch Warmblood Male With Maternal Links to HME in First and Second Generations.
Journal of equine veterinary science    February 21, 2018   Volume 64 55-58 doi: 10.1016/j.jevs.2018.02.009
May-Davis S, Doveren A, Sweere R, Smak J, Vermeulen Z.Hereditary multiple exostoses (HME) is a condition rarely diagnosed in horses, which has not been previously reported in Dutch Warmblood horses. Its presentation resembles that diagnosed in humans, and it is predominantly active during periods of skeletal maturation. This case study reports a link between a granddam with no obvious HME lesions and its first and second generations presenting with HME, albeit with different sires. The severity in affected horses (n = 5) varied from one foreleg to the other with extensive multiple lesions of 123 of 205 bones in a half sibling in the first genera...
Microsatellite markers for evaluating the diversity of the natural killer complex and major histocompatibility complex genomic regions in domestic horses.
HLA    February 21, 2018   Volume 91, Issue 4 271-279 doi: 10.1111/tan.13211
Horecky C, Horecka E, Futas J, Janova E, Horin P, Knoll A.Genotyping microsatellite markers represents a standard, relatively easy, and inexpensive method of assessing genetic diversity of complex genomic regions in various animal species, such as the major histocompatibility complex (MHC) and/or natural killer cell receptor (NKR) genes. MHC-linked microsatellite markers have been identified and some of them were used for characterizing MHC polymorphism in various species, including horses. However, most of those were MHC class II markers, while MHC class I and III sub-regions were less well covered. No tools for studying genetic diversity of NKR com...
Analysis of Genetic Diversity and Structure of Guanzhong Horse Using Microsatellite Markers.
Animal biotechnology    February 20, 2018   Volume 30, Issue 1 95-98 doi: 10.1080/10495398.2017.1416392
Zeng L, Chen N, Yao Y, Dang R, Chen H, Lei C.To determine the genetic diversity and validate the pedigree record of Chinese Guanzhong horse, 67 individuals were genotyped with eight microsatellite markers. In our study, the mean observed and expected heterozygosities were 0.51 and 0.66, respectively. The mean observed number of alleles for the Guanzhong horse was 3.88. Nonetheless, the total value of F multiloci clearly indicates that about 0.5% of overall genetic variation is due to line founder differences, while differences among individuals are responsible for the remaining 99.5%. In addition, the polymorphic information content (PIC...
Stallion semen quality depends on major histocompatibility complex matching to teaser mare.
Molecular ecology    February 19, 2018   Volume 27, Issue 4 1025-1035 doi: 10.1111/mec.14490
Jeannerat E, Marti E, Berney C, Janett F, Bollwein H, Sieme H, Burger D, Wedekind C.The major histocompatibility complex (MHC) has repeatedly been found to influence mate choice of vertebrates, with MHC-dissimilar mates typically being preferred over MHC-similar mates. We used horses (Equus caballus) to test whether MHC matching also affects male investment into ejaculates after short exposure to a female. Semen characteristics varied much among stallions. Controlling for this variance with a full-factorial within-subject experimental design, we found that a short exposure to an MHC-dissimilar mare enhanced male plasma testosterone and led to ejaculates with elevated sperm nu...
Review: The potential of seminal fluid mediated paternal-maternal communication to optimise pregnancy success.
Animal : an international journal of animal bioscience    February 19, 2018   Volume 12, Issue s1 s104-s109 doi: 10.1017/S1751731118000083
Bromfield JJ.Artificial insemination has been a landmark procedure in improving animal agriculture over the past 150 years. The utility of artificial insemination has facilitated a rapid improvement in animal genetics across agricultural species, leading to improvements of growth, health and productivity in poultry, swine, equine and cattle species. The utility of artificial insemination, as with all assisted reproductive technologies side-steps thousands of years of evolution that has led to the development of physiological systems to ensure the transmission of genetics from generation to generation. The ...
Species-specific regulation of angiogenesis by glucocorticoids reveals contrasting effects on inflammatory and angiogenic pathways.
PloS one    February 15, 2018   Volume 13, Issue 2 e0192746 doi: 10.1371/journal.pone.0192746
Morgan R, Keen J, Halligan D, O'Callaghan A, Andrew R, Livingstone D, Abernethie A, Maltese G, Walker B, Hadoke P.Glucocorticoids are potent inhibitors of angiogenesis in the rodent in vivo and in vitro but the mechanism by which this occurs has not been determined. Administration of glucocorticoids is used to treat a number of conditions in horses but the angiogenic response of equine vessels to glucocorticoids and, therefore, the potential role of glucocorticoids in pathogenesis and treatment of equine disease, is unknown. This study addressed the hypothesis that glucocorticoids would be angiostatic both in equine and murine blood vessels.The mouse aortic ring model of angiogenesis was adapted to assess...
Genetic and subunit vaccines based on the stem domain of the equine influenza hemagglutinin provide homosubtypic protection against heterologous strains.
Vaccine    February 15, 2018   Volume 36, Issue 12 1592-1598 doi: 10.1016/j.vaccine.2018.02.019
Ibañez LI, Caldevilla CA, Paredes Rojas Y, Mattion N.H3N8 influenza virus strains have been associated with infectious disease in equine populations throughout the world. Although current vaccines for equine influenza stimulate a protective humoral immune response against the surface glycoproteins, disease in vaccinated horses has been frequently reported, probably due to poor induction of cross-reactive antibodies against non-matching strains. This work describes the performance of a recombinant protein vaccine expressed in prokaryotic cells (ΔHAp) and of a genetic vaccine (ΔHAe), both based on the conserved stem region of influenza hemagglut...
Abnormal locomotor muscle recruitment activity is present in horses with shivering and Purkinje cell distal axonopathy.
Equine veterinary journal    February 12, 2018   Volume 50, Issue 5 636-643 doi: 10.1111/evj.12813
Aman JE, Valberg SJ, Elangovan N, Nicholson A, Lewis SS, Konczak J.Cerebellar Purkinje cell axonal degeneration has been identified in horses with shivering but its relationship with abnormal hindlimb movement has not been elucidated. Objective: To characterise surface electromyographic (sEMG) hindlimb muscle activity in horses with shivering, correlate with clinical scores and examine horses for Purkinje axonal degeneration. Methods: Descriptive controlled clinical study. Methods: The hindlimb of seven shivering and six control draught horses were clinically scored. Biceps femoris (BF), vastus lateralis (VL), tensor fasciae latae and extensor digitorum longu...
Chondrogenic Differentiation of Defined Equine Mesenchymal Stem Cells Derived from Umbilical Cord Blood for Use in Cartilage Repair Therapy.
International journal of molecular sciences    February 10, 2018   Volume 19, Issue 2 537 doi: 10.3390/ijms19020537
Desancé M, Contentin R, Bertoni L, Gomez-Leduc T, Branly T, Jacquet S, Betsch JM, Batho A, Legendre F, Audigié F, Galéra P, Demoor M.Cartilage engineering is a new strategy for the treatment of cartilage damage due to osteoarthritis or trauma in humans. Racehorses are exposed to the same type of cartilage damage and the anatomical, cellular, and biochemical properties of their cartilage are comparable to those of human cartilage, making the horse an excellent model for the development of cartilage engineering. Human mesenchymal stem cells (MSCs) differentiated into chondrocytes with chondrogenic factors in a biomaterial appears to be a promising therapeutic approach for direct implantation and cartilage repair. Here, we cha...
A 2-Year-Old Filly With Hereditary Equine Regional Dermal Asthenia: The First Case Report From China.
Journal of equine veterinary science    February 9, 2018   Volume 64 1-4 doi: 10.1016/j.jevs.2018.01.010
Li 李靖 J, Liu B, Yu F, Liu T, Peng Y, Fu Y.Hereditary equine regional dermal asthenia (HERDA) is an autosomal recessive inheritable disorder described in the Quarter Horses and related breeds. In this case report, a 2-year-old Quarter Horse filly was diagnosed with HERDA based on clinical findings and genetic testing. The observed clinical signs were stretchy, loose and thin skin, and open wounds on the upper body. Skin biopsy results were consistent with the common findings previously described in the literature. This is the first HERDA case report in China (and in Asia). Genetic testing protocols should be implemented for breeding fa...
Genetic diversity of equine herpesvirus 1 isolated from neurological, abortigenic and respiratory disease outbreaks.
Transboundary and emerging diseases    February 9, 2018   Volume 65, Issue 3 817-832 doi: 10.1111/tbed.12809
Bryant NA, Wilkie GS, Russell CA, Compston L, Grafham D, Clissold L, McLay K, Medcalf L, Newton R, Davison AJ, Elton DM.Equine herpesvirus 1 (EHV-1) causes respiratory disease, abortion, neonatal death and neurological disease in equines and is endemic in most countries. The viral factors that influence EHV-1 disease severity are poorly understood, and this has hampered vaccine development. However, the N752D substitution in the viral DNA polymerase catalytic subunit has been shown statistically to be associated with neurological disease. This has given rise to the term "neuropathic strain," even though strains lacking the polymorphism have been recovered from cases of neurological disease. To broaden understan...
Understanding the Intestinal Microbiome in Health and Disease.
The Veterinary clinics of North America. Equine practice    February 7, 2018   Volume 34, Issue 1 1-12 doi: 10.1016/j.cveq.2017.11.005
Costa MC, Weese JS.This article provides readers with the basic concepts necessary to understand studies using recent molecular methods performed in intestinal microbiome assessment, with special emphasis on the high throughput sequencing. This review also summarizes the current knowledge on this topic and discusses future insights on the interaction between the intestinal microbiome and equine health.
Determinants of gestation length in Thoroughbred mares on German stud farms.
Animal reproduction science    February 7, 2018   Volume 191 22-33 doi: 10.1016/j.anireprosci.2018.02.001
Ewert M, Lüders I, Böröcz J, Uphaus H, Distl O, Sieme H.The aim of the present study was to analyze the effects of stallion and mare, their ages, and maternal lineage on the gestation length (GL) in Thoroughbreds. In addition, additive genetic effects of the dam, stallion and fetus were analyzed. Data were taken from 1993 through 2009, and included 16,226 pregnancies from 5959 Th oroughbred mares mated with 290 different stallions. All analyses were performed using linear mixed models. The GL ranged from 306 to 390 days, with a mean length of 347.0 ± 14.4 days. Mating of mares with stallions aged 17 years and older resulted in a significantly ...
Equid herpesvirus 8: Complete genome sequence and association with abortion in mares.
PloS one    February 7, 2018   Volume 13, Issue 2 e0192301 doi: 10.1371/journal.pone.0192301
Garvey M, Suárez NM, Kerr K, Hector R, Moloney-Quinn L, Arkins S, Davison AJ, Cullinane A.Equid herpesvirus 8 (EHV-8), formerly known as asinine herpesvirus 3, is an alphaherpesvirus that is closely related to equid herpesviruses 1 and 9 (EHV-1 and EHV-9). The pathogenesis of EHV-8 is relatively little studied and to date has only been associated with respiratory disease in donkeys in Australia and horses in China. A single EHV-8 genome sequence has been generated for strain Wh in China, but is apparently incomplete and contains frameshifts in two genes. In this study, the complete genome sequences of four EHV-8 strains isolated in Ireland between 2003 and 2015 were determined by I...
Constituents of neutrophil extracellular traps induce in vitro collagen formation in mare endometrium.
Theriogenology    February 7, 2018   Volume 113 8-18 doi: 10.1016/j.theriogenology.2018.02.001
Rebordão MR, Amaral A, Lukasik K, Szóstek-Mioduchowska A, Pinto-Bravo P, Galvão A, Skarzynski DJ, Ferreira-Dias G.Neutrophil extracellular traps (NETs) are DNA complexes carrying nuclear and cytoplasmic proteins, such as elastase (ELA), cathepsin-G (CAT) and myeloperoxidase (MPO). Mare endometrosis is a chronic degenerative process characterized by excessive collagen in endometrium. While NETs fight bacteria that cause endometritis, they may trigger endometrial fibrogenesis. The aim was to evaluate the in vitro effect of some NETs components on mare endometrial fibrogenesis and determine its relationship with histopathology or estrous cycle. Endometrial explants were incubated with NETs components (ELA, ...
Morphofunctional diversity of equine of varied genetic compositions raised in the Pantanal biome of Brazil.
Tropical animal health and production    February 7, 2018   Volume 50, Issue 5 1033-1040 doi: 10.1007/s11250-018-1527-5
de Rezende MPG, de Souza JC, Carneiro PLS, Bozzi R, Jardim RJD, Malhado CHM.Evaluating phenotypic diversity makes it possible to identify discrepancies in aptitudes among animals of different genetic bases, which is an indicator of adaptive or selective differences between populations. The objective of this work was to evaluate the morphofunctional diversity of 452 male and female adult equines (Arabian, Quarter Mile, Pantaneiro, and Criollo breeds, and undefined crossbreeds of horses and mules) raised in the Pantanal biome (Brazil). Linear measurements were performed to estimate conformation indexes. Initially, a discriminant analysis was performed, regardless of the...
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