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Introdução: A caseína constitui cerca de 80% das proteínas presentes no leite de vaca. Durante a digestão é liberado o peptídeo opióide β-casomorfina-7 (BCM-7), que afeta os mecanismos de propulsão gastrointestinal, a motilidade, além de inibir as secreções gástricas e pancreáticas. A β-caseína bovina possui duas variantes, β-caseína A1 e A2. A diferença é a alteração de um nucleotídeo (SNP), que acarreta a troca de um aminoácido. Com genótipo A2, em homozigose, durante a digestão a liberação de BCM-7 não ocorre ou ocorre em quantidades pequenas. Devido ao alto custo da genotipagem por sequenciamento ou ship de SNP, a ARMS-PCR é uma metodologia de genotipagem de SNPs envolvendo somente um PCR seguido de eletroforese em gel, que torna a genotipagem simples e econômica. Objetivo: O objetivo foi avaliar a ARMS-PCR para genotipagem do SNP A1/A2 no gene da β-caseína. Material e Método: Foram coletadas amostras de sangue de 20 vacas da raça Girolando produtoras de leite. Foi extraído o DNA das amostras de sangue e realizada a amplificação do loco do gene da β-caseína pela técnica ARMS-PCR com quatro primers, dois internos e dois externos. Resultados: Após a amplificação as amostras foram analisadas por eletroforese em gel de agarose a 2%, corada com GelRed e visualizadas em luz UV. As mesmas amostras foram amplificadas pela PCR convencional e sequenciadas em 3 repetições cada. Após a identificação do genótipo, as amostras foram classificadas como positivas (A2/A2) e negativas (A1/A1 e A1/A2) para as amostras da ARMS-PCR em relação ao genótipo apresentado no sequenciamento. Foram analisadas a sensibilidade e a especificidade. Todas as amostras amplificaram e foram genotipadas pela técnica de ARMS-PCR. A sensibilidade e especificidade da ARMS-PCR foi de 100%, em todas as amostras a identificação do genótipo foi idêntica a observada pelo sequenciamento do loco de DNA. Dos 20 animais avaliados, 40% eram A2/A2, 25% A1/A1 e 35% A1/A2. Concluímos que a ARMS-PCR é eficiente para genotipagem do SNP da β-caseína (leite A1 e A2), e pode ser aplicada para a genotipagem dos animais.

Background: Current reproductive management of bovine elite populations involves the use of assisted reproductive technologies (ARTs), aiming to obtain the greatest genetic gain. However, inadequate use of ARTs may lead to loss of genetic diversity in the offspring. Objective: To assess the genetic diversity in elite female cattle populations used in commercial in vitro embryo production. Methods: Using genetic and ecological approaches for the study of populations based on microsatellite markers, we assessed the genetic diversity between and within populations of cows used in commercial in vitro embryo production programs in Brazil. Results: Endogamy within populations varied from zero to 9.1%, while heterozygosity between populations (FST) was

The aim of this study was to analyze the effects of genetic group and sex on residual feed intake, performance, testicular size, carcass and morphometric traits in Santa Ines purebred and Dorper ´ Santa Ines crossbred lambs. A total of 32 lambs, with initial body weight of 18±3.7 kg were used. Analysis of variance, considering the interaction between sex and genetic group of all the traits were performed. Crossbred females (-0.018±0.06) and purebred males (-0.018±0.05) were more efficient (p

The objective of this study was to evaluate the variation of surface temperature through infrared thermography (IRT) in different anatomical regions during the estrous cycle of hair sheep. The estrous cycle of 20 Santa Ines hair breed ewes was synchronized and, during this synchronization period (15 days), environmental conditions (temperature, humidity and wet-bulb globe temperature) and body temperatures were evaluated. The temperatures of the rectum and vagina were measured with a digital thermometer and the surface temperature of the anus, vulva, muzzle, left eye and left ear were measured with an infrared thermal camera. The mean, standard deviation, regression and correlation of each temperature of each anatomical region were evaluated regarding the estrous cycle. The eye was not a good anatomical region to measure the temperature variation during the estrous cycle (R2 = 0.011, p > 0.05). The vulva was the region with greatest potential for evaluation of surface temperature variation with IRT during estrus synchronization (β = 0.0805; R2= 0.767, p

The use of infrared thermography has not been used previously to assist sheep reproduction. The aim of this study was evaluate the viability of infrared thermography to identify body surface temperature patterns during ewes' estrous cycle. The estrous cycles of 20 Santa Ines ewes were synchronized through hormone treatment (progesterone implant). The ewes were submitted to measurements of body temperature (rectal and vaginal) with a digital thermometer and surface temperature of the anus, vulva, muzzle, left ear and left eye by infrared thermography. Temperature and humidity of the environment and wet-bulb globe temperature (WBGT) were measured. After the progesterone implant's removal, the follicular dynamic was evaluated daily during five days with ultrasound. The estrous cycle was separated into six phases to facilitate interpretation of the differences in body temperatures in each period. Phase 1 was the period before estrous synchronization, phase 2 was the beginning of estrous synchronization, phase 3 was the intermediate period and end of synchronization, phase 4 was the period of estrus manifestations, phase 5 was the ovulation period, and phase 6 was the post-ovulatory period. There was a difference among phases for all body regions evaluated (P

The LIM and SH3 protein 1 (LASP1) and the ligand dependent nuclear receptor corepressor-like (LCORL) genes were selected as candidates in this study of growth and height of equines of the Brazilian Pony breed. Until now, no genomic studies on the morphometry of Brazilian ponies have been reported in the literature. Both genes are correlated with osteogenesis and transcription factor TFIID of genes involved in skeletal growth, respectively. The objective of this study was to analyze two height associated mutations in LASP1 and LCORL genes and their associations with equine morphometry. This is the first study that genotyped these known variants in the Brazilian Pony breed. The DNA was extracted from hair follicles and the two variants were amplified by PCR. The genotyping was performed by RFLP markers through restriction enzyme cleavage; the samples were submitted to capillary electrophoresis for allelic discrimination by size; and the polymorphism was identified and associated with the morphometric measures of the animals. Regarding the LASP1 gene, the ponies with genotypes G/G and G/A presented the smallest measures for head length, neck length, withers height, croup height and body length. Mutation analysis [T > C] of the LCORL gene identified the T/T genotype in all ponies except one individual of the control group, with shoulder height of 1.69 m, which presented the C/C genotype, confirming the association of this gene with the height of horses of the Brazilian Pony breed. The stature and morphometry of horses of this breed were influenced by variants of the LASP1 gene.

Uterine illnesses are the major problem in horse reproductive programs. The diagnosis of these pathologies is often neglected or does not provide enough information about the cause. The aim of this study was to evaluate different diagnoses to provide more accurate information about endometritis and the real endometrial profile at the beginning of a breeding season. Fifty-one mares with reproductive failure were evaluated. Two different uterine cytology diagnoses, culture of uterine flush material and biopsy assay, were performed. The data were analyzed by the Chi-squared test to compare different diagnostic methods for the same sample from each mare, and sensitivity and specificity tests were performed. Most of evaluations showed no association between cytology and other diagnostic methods, except with bacterial culture. The most common bacterium found was Escherichia colli, but it was not associated with neutrophil presence in cytology. The presence of β-hemolitic streptococcus was well characterized by positive cytology and cloudy lavage fluid aspect. Endometrial profile was affected by age and whether the mare was a donor or recipient. The most sensitive diagnostic method was gross aspect of lavage fluid. Endometritis should be diagnosed by at least two methods to increase accuracy. Endometritis is not the only uterine disease that causes embryo loss, since endometrial fibrosis can also be a cause.

Background: Assessing breeding soundness is important to predict the potential fertility of stallions. Objective: To investigate the association of testis traits, total number of spermatozoa, and testosterone with fertility of Mangalarga Marchador breed stallions. Methods: The traits measured were testicular length, width and height, right and left testicular volume and total testicular volume. We also evaluated the total number of spermatozoa (TNS), serum testosterone concentration and fertility of stallions (by embryo recovery from donor mares) in the breeding and non-breeding seasons during 1 year. Analysis of variance was performed to verify the effects of age category (AC; young/adult) and reproductive season on the traits. Correlations and repeatability of the traits were also calculated. Results: The AC did not influence testis traits (p > 0.05). Reproduction season affected left testicular length, left testicular height, right testicular width and height (p 0.05). We observed strong correlations between all testicular measurements and testicular volume (0.53 to 0.75), medium correlation between TNS and testis volume (0.32), and low correlation between testosterone and testis traits (0.20). Repeatability of the characteristics of left and right testis was medium to high (0.22 to 0.78). Embryo recovery rate was 60% and its correlation with TNS was 0.44. Conclusion: Testis size evaluation is recommended over the evaluation of serum testosterone concentration to predict sperm output. Sperm output (TNS) is a good predictor of stallion fertility. Keywords: equine, hormone, male, reproduction, testis size. Resumen Antecedentes: Es importante realizar examen andrologico de sementales para predecir su potencial reproductivo. Objetivo: Evaluar la asociacion entre caracteristicas testiculares, numero total de espermatozoides y testosterona con la fertilidad de sementales de raza Mangalarga Marchador. Metodos: Las caracteristicas edidas fueron: longitud testicular, ancho y altura del testiculo. Tambien fue calculado el volumen testicular derecho, izquierdo y total, asi como el numero total de espermatozoides (TNS), concentracion serica de testosterona y fertilidad (por recuperacion embrionaria de las yeguas) de los sementales dentro y fuera de la temporada reproductiva durante un ano. Se realizo analisis de varianza para evaluar el efecto de categoria de edad (AC) y estacion reproductiva sobre las caracteristicas. Resultados: La AC no influencio las caracteristicas testiculares (p > 0,05). La estacion reproductiva influencio la longitud del testiculo izquierdo, la altura del testiculo izquierdo, y el ancho y altura del testiculo derecho (p 0,05). Se observo una alta correlacion entre todas las caracteristicas testiculares y el volumen testicular (0,53 a 0,75), mediana correlacion entre TNS y el volumen testicular (0,32), y una baja correlacion entre las medidas testiculares y la testosterona (0,20). La repetitividad de las medidas testiculares fue media a alta (0,22 a 0,78). La tasa de recuperacion embrionaria fue de 60% y la correlacion con TNS fue 0,44. Conclusion: Para predecir la produccion espermatica se recomienda evaluar las caracteristicas testiculares, sin necesidad de medir testosterona. La produccion espermatica es un buen predictor de la fertilidad de los sementales. Palabras clave: equino, hormona, macho, reproduccion, tamano testicular. Resumo Antecedentes: Realizar o exame andrologico no garanhao, e importante para predizer seu potencial reprodutivo. Objetivo: Avaliar a associacao das caracteristicas testiculares, numero total de espermatozoides, testosterona e fertilidade de garanhoes Mangalarga Marchador. Metodos: As caracteristicas mensuradas foram: comprimento do testiculo, largura do testiculo e altura do testiculo. Tambem foi calculado o volume testicular, direito, esquerdo e total, assim como calculado o numero total de espermatozoides (TNS), concentracao serica de testosterona e fertilidade dos garanhoes (pela recuperacao embrionaria das eguas doadoras) durante e fora da estacao reprodutiva durante um ano. Foi feita analise de variância para verificar efeito da categoria de idade (AC) e estacao reprodutiva sobre as caracteristicas. Resultados: A AC nao influenciou nas caracteristicas testiculares (p > 0,05). Estacao reprodutiva influenciou no comprimento testicular esquerdo, altura testicular esquerda, largura e altura testicular direita (p 0,05). Foi observado alta correlacao entre todas caracteristicas testiculares e volume testicular (0,53 a 0,75), media correlacao entre TNS e volume testicular (0,32), e baixa correlacao entre medidas testiculares e testosterona (0,20). A repetitividade das medidas testiculares foi meia a altas (0,22-0,78). A taxa de recuperacao embrionaria foi 60% e a correlacao com TNS foi de 0,44. Conclusao: E recomendada a avaliacao das caracteristicas testiculares para predizer a producao espermatica sem necessidade de mensurar testosterona. A producao espermatica e um bom preditor da fertilidade dos garanhoes. Palavras chave: equino, hormonio, macho, reproducao, tamanho testicular.

In sheep, there is a lack of information on the behaviour of hemodynamic indices and parameters of blood flow velocity of the uterine artery during pregnancy, making it impossible to determine the real meaning of the values found and their probable relevance in normal physiological or pathological states. The objective of the present study was to evaluate the blood flow velocity parameters and hemodynamic indices of the uterine artery in ewes (18) during pregnancy (33). Based on non-invasive colour Doppler sonography, we evaluated the peak systolic velocity (PS), end diastolic velocity (ED), time-averaged maximum and minimum velocity in a cardiac cycle (TAMAX and TAMEAN), pulsatility index (PI), resistance index (RI), systolic/diastolic ratio (S/D), heart rate (HR), arterial diameter (AD) and the blood flow volume (BFV). Examinations started on day 28 and continued at two-week intervals until parturition. The Doppler parameters and the diameter of the uterine artery underwent significant changes during pregnancy. In the evaluated animals, indices related to resistance of the uterine artery, namely the impedance of blood flow, decreased throughout the initial, middle and late stages of gestation (PI: 1.15, 1.04, 0.97; RI: 0.61, 0.59, 0.57; S/D: 2.68, 2.52, 2.39, respectively) (p < 0.05). In contrast, the contents related to higher uterine irrigation increased during gestation (PS, ED, TAMAX, TAMEAN, HR, AD and BFV) (p < 0.05). No differences were found between the means of the variables in relation to the right and left uterine arteries and between single and multiple pregnancies in the hemodynamic indices. The present study is the first to demonstrate changes in uterine hemodynamics throughout pregnancy in sheep.

The objective of this study was evaluate the efficiency in water and food use in sheep production, using two new traits (residual water intake - RWI; residual feed and residual water intake - RFRWI). For this purpose, we evaluated the relationships between the traits total water intake, RWI and RFRWI with water measures, productive performance and carcass traits (ultrasound). We used 32 lambs with age of 3 months: 16 Santa Ines breed (8 males and 8 females) and 16 crossbred 7/8 Dorper x Santa Ines (8 males and 8 females). They were fed at an automated feed and water station (Intergado®), where the food and water intake by each animal was measured automatically. Residual water intake (RWI), residual feed intake (RFI) and residual body weight gain (RWG) were calculated using the MIXED procedure in SAS®. Correlation coefficients between total water intake (TWI), RWI and all traits (water measures, performance and carcass traits) were calculated using the CORR procedure (P ≤ 0.05). Total water intake showed mean 3.15 L/day during the study and the RWI ranged from 0.66 L to -0.89 L, which represents a difference of 1.55 L between the most and least efficient animal. There was positive correlation between less efficient animals in water consumption (positive RWI) and water intake (r = 0.31); total water intake in relation to dry matter intake (r = 0.36); total water intake in relation to mid-trial metabolic body weight (r = 0.26); residual feed and residual water intake (r = 0.99) and gain/feed ratio (r = 0.44). There was negative correlation between less efficient animal and dry matter intake in relation to body weight (r= -0.33) and feed conversion ratio (r= -0.44). The evaluation of the new variables, RWI and RFRWI, proved effective in identifying the most efficient animals in water use. Moreover, these findings open the possibility of considering the traits RWI and RFRWI in sheep selection, because performance and carcass traits are not adversely affected by selecting for these new traits.

The aim of this study was to evaluate seminal and spermatozoa quality of stallions from North of Rio de Janeiro State in the breeding season and non-breeding...

The objective of this study was to identify polymorphisms in the myostatin and leptin genes in Santa Inês (SI) and crossbreed (SI x Dorper) sheep, to verify the effect of these polymorphisms on carcass traits. We evaluated seventy sheep of 8-month-old at the Federal Institute of Espírito Santo, Brazil. Data collected were slaughter weight (SW), hot carcass weight (HCW), cold carcass weight (CCW), loin weight, tenderloin weight and fat thickness (FT). The hot carcass yield (HCY) was calculated by the formula (HCW/SW) x 100. We collected hairs from each animal for DNA extraction by the alkaline protocol. The animals were genotyped for the G>A mutation in nucleotide 9827 of the myostatin gene and for three polymorphisms in exon 3 of the leptin gene, by the PCR-RFLP technique. The amplicons the myostatin and leptin gene were cleaved with restriction enzyme for allelic discrimination. The alleles were recorded for each animal and analysis of variance was performed to check the influence of the mutations on the carcass traits. The mutant allele of the myostatin gene showed association with increased measures of CCW, FT and with reduced HCY. Among the three alleles of the leptin gene, only one showed an effect (increased CCW). The other alleles were not associated with any traits.

PURPOSE: This study aimed to associate DNA variants in promoter and exon flanking regions of the CYP19A1 gene with in vitro embryo production traits in cattle. The role of transcription factor binding sites created or lost due to DNA sequence variation and their possible effect on gene expression was also evaluated. METHODS: We collected date from Gyr dairy oocyte donor cows (Bos taurus indicus) at a commercial in vitro embryo production farm and analyzed the genotype–phenotype association with in vitro production traits. Using Sanger sequencing and web-based software, we assessed important CYP19A1 gene regions in oocyte donor cows and analyzed the effects of variants on the transcription factor binding sites. RESULTS: Two SNP mutations significantly associated with oocyte production, oocyte viability, embryo development, and pregnancies were found (T > C in the untranslated exon 1 flanking region ([GenBank: AJ250379.1]: rs718446508 T > C), and a T > C in the 5′-upstream region (1.1 promoter) ([GenBank: AC_000167.1]: rs41651668 T > C). Six new transcription factor binding sites were created. A binding site for transcription factors associated with the development of the placenta and embryo implantation was eliminated due to variations in the DNA sequence identified. CONCLUSIONS: The CYP19A1 gene contributes to genetic variation of in vitro embryo production traits in cattle. The complexity of the physiological phenomena related to estrogen pathways and their influence on reproduction in cattle allow indication of the mutations evaluated here as possible genetic markers for embryo production traits, which should be validated in the next steps of marker-assisted selection.

Assessing the sexual behavior of stallions is extremely important because sexual behavior is linked to reproductive efficiency. The objective of this study was to evaluate the sexual behavior, testosterone concentration and libido of Mangalarga Marchador stallions from northern Rio de Janeiro State, Brazil. The sexual behavior of 10 stallions was evaluated during semen collection while mounting a mare in estrus. Blood samples from each animal were collected for the evaluation of serum testosterone. The study was conducted during the breeding and non-breeding seasons with animals separated into two age categories: young ( 5 years). The libido of the stallions was evaluated using the observations of reaction time (RT), mount time (MT) and mount and ejaculation time (MEjT). Analysis of variance and analysis of data frequency were performed. The stallion behavior and testosterone concentrations were not affected by the breeding season or stallion age ( p > 0.05). The overall mean of RT was 14.0 ± 2.4 s, MT was 32.7 ± 3.5 s and MEjT was 24.3 ± 0.8 s. We observed a few instances of a mount without an erection and a mount without ejaculation a maximum of 4 and 1 times, respectively. The percentage of stallions that did not perform Flehmen, biting, kicking, sniffing and vocalization were 72, 93, 93, 34 and 45% respectively. The average testosterone concentration was 0.64 ± 0.06 ng/mL. We concluded that the animals exhibited good libido overall, thereby indicating that they were well adapted to the environmental conditions of northern Rio de Janeiro state, Brazil.

This work aimed to investigate the relationship between echogenicity, testicular biometry, and accessory gland biometry and evaluate the echogenicity pattern of these structures in Santa Ines rams. Fifty-four healthy Santa Ines sheep were classified into three age groups: group 1 - 3 to 5 months (pre-pubertal); group 2 - 7 to 11 months (pubertal); and group 3 - 13 to 28 months (sexually mature). The averages of testicular biometry and bulbourethral gland biometry were different among age groups. The testicular parenchyma was homogeneous with low echogenicity (pre-pubertal group) and medium echogenicity (pubertal and sexually mature groups), suggesting that testicular echogenicity can be a helpful tool to identify the start of puberty. The accessory glands were heterogeneous with low echogenicity in all age groups. Correlations were high between weight, biometry of testis and of bulbourethral gland and echogenicity of the accessory glands, indicating that sexually mature rams with higher body development have larger testicles and larger glands. We recommend the use of testicular biometry as a complementary evaluation of the reproductive efficiency of Santa Ines rams. The average values of echogenicity and the biometry of accessory glands are important parameters to assist the selection of breeding rams.

SUMMARY Evaluation of the testis size is very important in stallions’ selection, since it is an indirect determinant of their reproductive potential. The aim of this study was to evaluate the biometric testicular traits, calculate the volume testicular and analyze the serum testosterone concentration of the stallion. During six months were measured the tests of adults Brazilian Ponies stallions from northern of Rio de Janeiro State, Brazil. The length, the width and the height of the testicles (TL, TW and TH) were measured. From these measurements, the total testicular volume (TTV) was calculated. The serum testosterone concentration was also evaluated. The data were studied by analysis of variance and were calculated the correlations among the traits. The average of the testis biometry were TL 5.8 ± 0.6 cm; TW 3.8 ± 0.5 cm; TH 4.7 ± 0.4 cm and 111.2 ± 27.7 for TTV. Mean of serum testosterone concentration was 0.89 ± 0.52 ng/dl. High correlations between biometry testicular traits with testicular volume were observed and low to moderate correlations were observed for testicular traits, testosterone and sperm output. The testis size could be an important trait that assists to the prediction of stallion's reproductive potential of the Brazilian ponies stallions. It is suggested that there would be no fluctuation in serum testosterone concentration in Brazilian ponies stallions that coult afect on its fertility. RESUMO Avaliação do tamanho dos testículos é muito importante na seleção de garanhões, uma vez que é um determinante indireto do seu potencial reprodutivo. O objetivo deste estudo foi avaliar as características de biometria testiculares, calcular o volume testicular e analisar a concentração sérica de testosterona de garanhões. Durante seis meses, foram medidos os testículos de garanhões Pôneis da raça Brasileiro adultos, criados no Norte do Estado do Rio de Janeiro, Brasil. O comprimento a largura e a altura dos testículo (TL, TW e TH) foram mensurados. A partir dessas medidas, calculou-se o volume testicular total (TTV). Os dados foram avaliados por análise de variância e foram calculadas as correlações entre as características. A média geral das características de biometria testicular e volume testicular foram: TL 5,8 ± 0,6 cm; TW 3,8 ± 0,5 cm; TH 4,7 ± 0,4 cm e TTV 111.2 ± 27,7. A média de concentração sérica de testosterona foi de 0,89 ± 0,52 ng/dl. Altas correlações foram observadas entre as características de biometria testicular e volume testicular. Baixas a médias correlações entre as características testiculares com produção espermática e com concentração de testosterona foram observadas. O tamanho testicular pode ser uma importante característica que auxilia na predição do potencial reprodutivo do garanhão pôneis da raça Brasileira. Sugere-se que não há variações na concentração sérica de testosterona nos pôneis da raça Brasileira que poderia afetar em sua fertilidade.

The aim of this study was to evaluate the breeding behavior of Brazilian Pony stallions during semen collection to examine the effect of age category (AC) and month of evaluation on the characteristics of breeding behavior. Nine stallions were evaluated during the breeding and non-breeding season, 5-8 (AC1) and 9-13 years (AC2) of age. The variables evaluated were reaction time (RT), mount time one (MT1) and two (MT2), mount and ejaculation time (MEjT), mounting without erection (MWEr), mounting without ejaculation (MWEj), flehmen, kicks, bites, smells, vocalizations and serum testosterone concentration. Analyses of variance and correlation were performed. The average scores were RT: 56.2±92.9s, MT1: 115.0±122.4s, MT2: 196.4±160.7s, MEjT: 21.8±3.2s, MWEr: 0.8±1.3s, MWEj: 2.1±3.5s, flehmen: 3.8±3.0s, kicks: 0.3±1.2s, bites: 1.0±2.0s, smells: 5.7±3.5s, vocalizations: 6.4±6.3s and testosterone: 0.89±0.52ng/mL. The AC affected the RT (P0.05) but not the other traits. Month of evaluation did not affect most variables (P0.05). The correlations between age and RT and between age and MT1 and MT2 were significant. The correlation between testosterone and MEjT was moderate and negative. The breeding behavior of Brazilian Pony stallions was similar across different months of evaluation and different age categories. The month of evaluation and age category also did not affect testosterone concentrations, meaning that semen collection is feasible throughout the year.

Este estudo objetivou investigar a variabilidade genética em dois loci do gene MC1R associados à cor de pelagem em ovinos Santa Inês. Pelos foram coletados da cauda de 121 animais com diferentes pigmentações. A genotipagem foi realizada por PCR RFLP com enzimas de restrição (NlaIII e BmgBI). As frequências alélicas e genotípicas esperadas no equilíbrio de Hardy-Weinberg foram verificadas no programa GENEPOP, e a diversidade genética, no programa ARLEQUIN. Os fragmentos observados no locus Nla1-MC1R apresentaram 46, 110 e 134 pb, frequência genotípica de 37% no genótipo homozigoto selvagem (AA), 23% no genótipo homozigoto mutante (TT) e 40% no genótipo heterozigoto (AT), com frequência de 42,8% para o alelo mutante. O alelo (A) foi associado à pelagem preta, sendo que o genótipo (T/T) aumenta em 4,688 vezes a probabilidade de serem multicoloridos (p < 0,001). No locus Bmg3-MC1R, os fragmentos apresentaram 107, 260 e 372 pb, 20% do genótipo homozigoto selvagem (GG), 12% do genótipo homozigoto mutante (AA) e 68% do genótipo heterozigoto (AG), com frequência alélica de 45,8% para o alelo mutante. O alelo (A) foi associado com a pelagem preta, sendo que o genótipo (A/G) acresce em 19,92 vezes a probabilidade de serem multicoloridos (p < 0,0001). Mutações no gene MC1R exibiram elevada variabilidade genética (0,49).