RESUMO
The beef cattle industry has experienced a shift driven by a market demand for healthier meat, cost efficiency and environmental sustainability in recent years. Consequently, there has been a growing focus on the fatty acids content and functions of meat in cattle breeding programmes. Besides, a deeper understanding of the biological mechanisms influencing the expression of different phenotypes related to fatty acid profiles is crucial. In this study, we aimed to identify Single-Nucleotide Variants (SNV) and Insertion/Deletion (InDels) DNA variants in candidate genes related to fatty acid profiles described in genomic, transcriptomic and proteomic studies conducted in beef cattle breeds. Utilizing whole-genome re-sequencing data from Brazilian locally adapted bovine breeds, namely Caracu and Pantaneiro, we identified SNVs and InDels associated with 23,947 genes. From these, we identified 318 candidate genes related to fatty acid profiles that contain variants. Subsequently, we select only genes with SNVs and InDels in their promoter, 5' UTR and coding region. Through the gene-biological process network, approximately 19 genes were highlighted. Furthermore, considering the studied trait and a literature review, we selected the main transcription factors (TF). Functional analysis via gene-TF network allowed us to identify the 30 most likely candidate genes for meat fatty acid profile in cattle. LIPE, MFSD2A and SREBF1 genes were highlighted in networks due to their biological importance. Further dissection of these genes revealed 15 new variants found in promoter regions of Caracu and Pantaneiro sequences. The gene networks facilitated a better functional understanding of genes and TF, enabling the identification of variants potentially related to the expression of candidate genes for meat fatty acid profiles in cattle.
RESUMO
Genome-Wide Association Studies (GWAS) are used for identification of quantitate trait loci (QTL) and genes associated with several traits. We aimed to identify genomic regions, genes, and biological processes associated with number of total and viable oocytes, and number of embryos in Gir dairy cattle. A dataset with 17,526 follicular aspirations, including the following traits: number of viable oocytes (VO), number of total oocytes (TO), and number of embryos (EMBR) from 1641 Gir donors was provided by five different stock farms. A genotype file with 2093 animals and 395,524 SNP markers was used to perform a single-step GWAS analysis for each trait. The top 10 windows with the highest percentage of additive genetic variance explained by 100 adjacent SNPs were selected. The genomic regions identified in our work were overlapped with QTLs from QTL database on chromosomes 1, 2, 5, 6, 7, 8, 9, 13, 17, 18, 20, 21, 22, 24, and 29. These QTLs were classified as External, Health, Meat and carcass, Production or Reproduction traits, and about 38% were related to Reproduction. In total, 117 genes were identified, of which 111 were protein-coding genes. Exclusively associations were observed for 42 genes with EMBR, and 1 with TO. Also, 42 genes were in common between VO and TO, 28 between VO and EMBR, and four genes were in common among all traits. In conclusion, great part of the identified genes plays a functional role in initial embryo development or general cell functions. The protein-coding genes ARNT, EGR1, HIF1A, AHR, and PAX2 are good markers for the production of oocytes and embryos in Gir cattle.
Assuntos
Estudo de Associação Genômica Ampla , Oócitos , Animais , Bovinos/genética , Genótipo , Fenótipo , Locos de Características Quantitativas/genética , Polimorfismo de Nucleotídeo Único/genéticaRESUMO
Runs of homozygosity (ROH) and signatures of selection are the results of selection processes in livestock species that have been shown to affect several traits in cattle. The aim of the current work was to verify the profile of ROH and inbreeding depression in the number of total (TO) and viable oocytes (VO) and the number of embryos (EMBR) in Gir Indicine cattle. In addition, we aim to identify signatures of selection, genes, and enriched regions between Gir subpopulations sorted by breeding value for these traits. The genotype file contained 2093 animals and 420,718 SNP markers. Breeding values used to sort Gir animals were previously obtained. ROH and signature of selection analyses were performed using PLINK software, followed by ROH-based (FROH) and pedigree-based inbreeding (Fped) and a search for genes and their functions. An average of 50 ± 8.59 ROHs were found per animal. ROHs were separated into classes according to size, ranging from 1 to 2 Mb (ROH1-2Mb: 58.17%), representing ancient inbreeding, ROH2-4Mb (22.74%), ROH4-8Mb (11.34%), ROH8-16Mb (5.51%), and ROH>16Mb (2.24%). Combining our results, we conclude that the increase in general FROH and Fped significantly decreases TO and VO; however, in different chromosomes traits can increase or decrease with FROH. In the analysis for signatures of selection, we identified 15 genes from 47 significant genomic regions, indicating differences in populations with high and low breeding value for the three traits.
Assuntos
Endogamia , Polimorfismo de Nucleotídeo Único , Bovinos/genética , Animais , Homozigoto , Genótipo , OócitosRESUMO
Whole genome sequencing of bovine breeds has allowed identification of genetic variants in milk protein genes. However, functional repercussion of such variants at a molecular level has seldom been investigated. Here, the results of a multistep Bioinformatic analysis for functional characterization of recently identified genetic variants in Brazilian Gyr and Guzerat breeds is described, including predicted effects on the following: (i) evolutionary conserved nucleotide positions/regions; (ii) protein function, stability, and interactions; (iii) splicing, branching, and miRNA binding sites; (iv) promoters and transcription factor binding sites; and (v) collocation with QTL. Seventy-one genetic variants were identified in the caseins (CSN1S1, CSN2, CSN1S2, and CSN3), LALBA, LGB, and LTF genes. Eleven potentially regulatory variants and two missense mutations were identified. LALBA Ile60Val was predicted to affect protein stability and flexibility, by reducing the number the disulfide bonds established. LTF Thr546Asn is predicted to generate steric clashes, which could mildly affect iron coordination. In addition, LALBA Ile60Val and LTF Thr546Asn affect exonic splicing enhancers and silencers. Consequently, both mutations have the potential of affecting immune response at individual level, not only in the mammary gland. Although laborious, this multistep procedure for classifying variants allowed the identification of potentially functional variants for milk protein genes.
Assuntos
Caseínas , Proteínas do Leite , Animais , Bovinos/genética , Simulação por Computador , Mutação , Regiões Promotoras GenéticasRESUMO
Bees are important pollinators whose population has declined due to several factors, including infections by parasites and pathogens. Resource sharing may play a role in the dispersal dynamics of pathogens among bees. This study evaluated the occurrence of viruses (DWV, BQCV, ABPV, IAPV, KBV, and CBPV) and microsporidia (Nosema ceranae and Nosema apis) that infect Apis mellifera, as well as pesticide residues in the stingless bees Nannotrigona testaceicornis, Tetragonisca angustula, and Tetragona elongata sharing the same foraging area with A. mellifera. Stingless bees were obtained from 10 nests (two of N. testaceicornis, five of T. angustula, and three of T. elongata) which were kept in the field for 1 year and analyzed for the occurrence of pathogens. Spores of N. ceranae were detected in stingless bees but were not found in their midgut, which indicates that these bees are not affected, but may be vectors of the microsporidium. Viruses were found in 23.4% of stingless bees samples. APBV was the most prevalent virus (10.8%) followed by DWV and BQCV (both in 5.1% of samples). We detected glyphosate and its metabolites in small amounts in all samples. The highest occurrence of N. ceranae spores and viruses was found in autumn-winter and may be related to both the higher frequency of bee defecation into the colony and the low food resources available in the field, which increases the sharing of plant species among the stingless bees and honey bees. This study shows the simultaneous occurrence of viruses and spores of the microsporidium N. ceranae in asymptomatic stingless bees, which suggest that these bees may be vectors of pathogens.
Assuntos
Abelhas , Nosema/fisiologia , Resíduos de Praguicidas/análise , Fenômenos Fisiológicos Virais , Animais , Abelhas/química , Abelhas/microbiologia , Abelhas/virologia , Nosema/isolamento & purificação , Vírus/isolamento & purificaçãoRESUMO
Stingless bees (Apidae: Meliponini) are a group of bees with vestigial stings showing a high level of social organization. They are important pollinators in tropical and subtropical regions, and, in the last decades, stingless beekeeping has increased rapidly in Brazil. Bee-collected pollen and honey of Apis mellifera can be an important source of disease when used as supplements to feed stingless bee colonies, a common and increasing practice adopted by stingless beekeepers. Here, we aimed to investigate the presence of pathogens commonly found in honey bees in diseased colonies of Melipona species in Espírito Santo and São Paulo States, Southeast Brazil. We detected, for the first time, the bacterium Melissococcus plutonius and symptoms of European foulbrood in Melipona spp., associated with brood death and colony losses in some cases. In addition, we tested for the presence of the bacterium Paenibacillus larvae and the fungus Aschosphaera apis, as well as the six more common honey bee viruses in Brazil (BQCV, ABPV, DWV, KBV, IAPV, CBPV) and the microsporidia Nosema apis and Nosema ceranae. However, only one sample of brood was infected with N. ceranae and all other pathogens, with the exception of Melissococcus plutonius, were absent in the analyzed brood. Lastly, we looked for toxic pollen in all food fed to diseased colonies, but none was present.
Assuntos
Abelhas/microbiologia , Enterococcaceae/isolamento & purificação , Nosema/isolamento & purificação , Animais , Abelhas/crescimento & desenvolvimento , Brasil , Larva/crescimento & desenvolvimento , Larva/microbiologia , Pupa/crescimento & desenvolvimento , Pupa/microbiologiaRESUMO
Heat stress is an important issue in the global dairy industry. In tropical areas, an alternative to overcome heat stress is the use of crossbred animals or synthetic breeds, such as the Girolando. In this study, we performed a genome-wide association study (GWAS) and post-GWAS analyses for heat stress in an experimental Gir × Holstein F2 population. Rectal temperature (RT) was measured in heat-stressed F2 animals, and the variation between 2 consecutive RT measurements (ΔRT) was used as the dependent variable. Illumina BovineSNP50v1 BeadChip (Illumina Inc., San Diego, CA) and single-SNP approach were used for GWAS. Post-GWAS analyses were performed by gene ontology terms enrichment and gene-transcription factor (TF) networks, generated from enriched TF. The breed origin of marker alleles in the F2 population was assigned using the breed of origin of alleles (BOA) approach. Heritability and repeatability estimates (± standard error) for ΔRT were 0.13 ± 0.08 and 0.29 ± 0.06, respectively. Association analysis revealed 6 SNP significantly associated with ΔRT. Genes involved with biological processes in response to heat stress effects (LIF, OSM, TXNRD2, and DGCR8) were identified as putative candidate genes. After performing the BOA approach, the 10% of F2 animals with the lowest breeding values for ΔRT were classified as low-ΔRT, and the 10% with the highest breeding values for ΔRT were classified as high-ΔRT. On average, 49.4% of low-ΔRT animals had 2 alleles from the Holstein breed (HH), and 39% had both alleles from the Gir breed (GG). In high-ΔRT animals, the average proportion of animals for HH and GG were 1.4 and 50.2%, respectively. This study allowed the identification of candidate genes for ΔRT in Gir × Holstein crossbred animals. According to the BOA approach, Holstein breed alleles could be associated with better response to heat stress effects, which could be explained by the fact that Holstein animals are more affected by heat stress than Gir animals and thus require a genetic architecture to defend the body from the deleterious effects of heat stress. Future studies can provide further knowledge to uncover the genetic architecture underlying heat stress in crossbred cattle.
Assuntos
Bovinos/genética , Redes Reguladoras de Genes , Estudo de Associação Genômica Ampla/veterinária , Resposta ao Choque Térmico/genética , Locos de Características Quantitativas/genética , Alelos , Animais , Cruzamento , Bovinos/fisiologia , Indústria de Laticínios , Feminino , MasculinoRESUMO
Rhipicephalus (Boophilus) microplus is the main cattle ectoparasite in tropical areas. Gir × Holstein crossbred cows are well adapted to different production systems in Brazil. In this context, we performed genome-wide association study (GWAS) and post-GWAS analyses for R. microplus resistance in an experimental Gir × Holstein F2 population. Single nucleotide polymorphisms (SNP) identified in GWAS were used to build gene networks and to investigate the breed of origin for its alleles. Tick artificial infestations were performed during the dry and rainy seasons. Illumina BovineSNP50 BeadChip (Illumina Inc., San Diego, CA) and single-step BLUP procedure was used for GWAS. Post-GWAS analyses were performed by gene ontology terms enrichment and gene transcription factors networks, generated from enriched transcription factors, identified from the promoter sequences of selected gene sets. The genetic origin of marker alleles in the F2 population was assigned using the breed of origin of alleles approach. Heritability estimates for tick counts were 0.40 ± 0.11 in the rainy season and 0.54 ± 0.11 in the dry season. The top ten 0.5-Mbp windows with the highest percentage of genetic variance explained by SNP markers were found in chromosomes 10 and 23 for both the dry and rainy seasons. Gene network analyses allowed the identification of genes involved with biological processes relevant to immune system functions (TREM1, TREM2, and CD83). Gene-transcription factors network allowed the identification of genes involved with immune functions (MYO5A, TREML1, and PRSS16). In resistant animals, the average proportion of animals showing significant SNPs with paternal and maternal alleles originated from Gir breed was 44.8% whereas the proportion of animals with both paternal and maternal alleles originated from Holstein breed was 11.3%. Susceptible animals showing both paternal and maternal alleles originated from Holstein breed represented 44.6% on average, whereas both paternal and maternal alleles originated from Gir breed animals represented 9.3%. This study allowed us to identify candidate genes for tick resistance in Gir × Holstein crossbreds in both rainy and dry seasons. According to the origin of alleles analysis, we found that most animals classified as resistant showed 2 alleles from Gir breed, while the susceptible ones showed alleles from Holstein. Based on these results, the identified genes may be thoroughly investigated in additional experiments aiming to validate their effects on tick resistance phenotype in cattle.
Assuntos
Doenças dos Bovinos/parasitologia , Resistência à Doença/genética , Estudo de Associação Genômica Ampla/veterinária , Rhipicephalus/fisiologia , Infestações por Carrapato/veterinária , Alelos , Animais , Brasil , Cruzamento , Bovinos , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/genética , Feminino , Variação Genética , Masculino , Fenótipo , Polimorfismo de Nucleotídeo Único , Estações do Ano , Especificidade da Espécie , Infestações por Carrapato/epidemiologia , Infestações por Carrapato/genéticaRESUMO
In the present study, lab-on-a-chip electrophoresis (LoaC) was suggested as an alternative method to the conventional polyacrylamide gel electrophoresis under denaturing conditions (SDS-PAGE) to analyze raw cell-free tick hemolymph. Rhipicephalus microplus females were exposed to the entomopathogenic fungus Metarhizium anisopliae senso latu IBCB 116 strain and/or to the entomopathogenic nematode Heterorhabditis indica LPP1 strain. Hemolymph from not exposed or exposed ticks was collected 16 and 24 h after exposure and analyze by SDS-PAGE or LoaC. SDS-PAGE yielded 15 bands and LoaC electrophoresis 17 bands. Despite the differences in the number of bands, when the hemolymph protein profiles of exposed or unexposed ticks were compared in the same method, no suppressing or additional bands were detected among the treatments regardless the method (i.e., SDS-PAGE or chip electrophoresis using the Protein 230 Kit®). The potential of LoaC electrophoresis to detect protein bands from tick hemolymph was considered more efficient in comparison to the detection obtained using the traditional SDS-PAGE method, especially when it comes to protein subunits heavier than 100 KDa. LoaC electrophoresis provided a very good reproducibility, and is much faster than the conventional SDS-PAGE method, which requires several hours for one analysis. Despite both methods can be used to analyze tick hemolymph composition, LoaC was considered more suitable for cell-free hemolymph protein separation and detection. LoaC hemolymph band percent data reported changes in key proteins (i.e., HeLp and vitellogenin) exceptionally important for tick embryogenesis. This study reported, for the first time, tick hemolymph protein profile using LoaC.
Assuntos
Proteínas de Artrópodes/química , Eletroforese em Gel de Poliacrilamida/métodos , Fungos/fisiologia , Rhipicephalus/química , Rhipicephalus/microbiologia , Rhipicephalus/parasitologia , Animais , Proteínas de Artrópodes/metabolismo , Eletroforese em Gel de Poliacrilamida/instrumentação , Feminino , Fungos/classificação , Fungos/genética , Fungos/isolamento & purificação , Hemolinfa/química , Hemolinfa/metabolismo , Dispositivos Lab-On-A-Chip , Nematoides/classificação , Nematoides/genética , Nematoides/isolamento & purificação , Nematoides/fisiologia , Reprodutibilidade dos Testes , Rhipicephalus/metabolismoRESUMO
A total of 32,817 test-day milk yield (TDMY) records of the first lactation of 4,056 Girolando cows daughters of 276 sires, collected from 118 herds between 2000 and 2011 were utilized to estimate the genetic parameters for TDMY via random regression models (RRM) using Legendre's polynomial functions whose orders varied from 3 to 5. In addition, nine measures of persistency in milk yield (PSi) and the genetic trend of 305-day milk yield (305MY) were evaluated. The fit quality criteria used indicated RRM employing the Legendre's polynomial of orders 3 and 5 for fitting the genetic additive and permanent environment effects, respectively, as the best model. The heritability and genetic correlation for TDMY throughout the lactation, obtained with the best model, varied from 0.18 to 0.23 and from -0.03 to 1.00, respectively. The heritability and genetic correlation for persistency and 305MY varied from 0.10 to 0.33 and from -0.98 to 1.00, respectively. The use of PS7 would be the most suitable option for the evaluation of Girolando cattle. The estimated breeding values for 305MY of sires and cows showed significant and positive genetic trends. Thus, the use of selection indices would be indicated in the genetic evaluation of Girolando cattle for both traits.
RESUMO
The monitoring of resistance of cattle tick populations in Brazil to the chemical bases in use is largely limited to investigation of the phenotypic profile. There are few studies investigating the role played by the genotypic profile in acaricide resistance in the country. Therefore, the aim of the present study was to carry out molecular characterization and trace out the genetic profile of populations of Rhipicephalus microplus with respect to resistance to the organophosphate and pyrethroid chemical groups. For that purpose, larvae were genotyped belonging to 587 populations for pyrethroids and 306 for organophosphates, using the polymerase chain reaction technique. It was found that 75.49% and 97.44% of the larvae studied showed resistance to the organophosphates and pyrethroids, respectively. Among the populations resistant to pyrethroids, 91.9% were heterozygotes, showing that most of the resistant populations have only one allele responsible for resistance. Therefore, it is possible to conclude that the genotyped populations have high resistance to organophosphates, and even more so to pyrethroids. This information is fundamental for understanding the mechanisms of resistance of R. microplus to acaricides, to enable improvement of control techniques.
Assuntos
Resistência a Inseticidas/genética , Inseticidas , Organofosfatos , Piretrinas , Rhipicephalus/genética , Animais , Brasil , Bovinos , Doenças dos Bovinos/parasitologia , Doenças dos Bovinos/prevenção & controle , Genótipo , Larva , Rhipicephalus/classificação , Infestações por Carrapato/parasitologia , Infestações por Carrapato/prevenção & controle , Infestações por Carrapato/veterináriaRESUMO
Until the mid-1990s, the only microsporidium known to infect bees of the genus Apis was Nosema apis. A second species, Nosema ceranae, was first identified in 1996 from Asian honey bees; it is postulated that this parasite was transmitted from the Asian honey bee, Apis cerana, to the European honey bee, Apis mellifera. Currently, N. ceranae is found on all continents and has often been associated with honey bee colony collapse and other reports of high bee losses. Samples of Africanized drones collected in 1979, preserved in alcohol, were analyzed by light microscopy to count spores and were subjected to DNA extraction, after which duplex PCR was conducted. All molecular analyses (triplicate) indicated that the drones were infected with both N. ceranae and N. apis. PCR products were sequenced and matched to sequences reported in the GenBank (Acc. Nos. JQ639316.1 and JQ639301.1). The venation pattern of the wings of these males was compared to those of the current population living in the same area and with the pattern of drones collected in 1968 from Ribeirão Preto, SP, Brazil, from a location close to where African swarms first escaped in 1956. The morphometric results indicated that the population collected in 1979 was significantly different from the current living population, confirming its antiquity. Considering that the use of molecular tools for identifying Nosema species is relatively recent, it is possible that previous reports of infections (which used only light microscopy, without ultrastructural analysis) wrongly identified N. ceranae as N. apis. Although we can conclude that N. ceranae has been affecting Africanized honeybees in Brazil for at least 34 years, the impact of this pathogen remains unclear.
Assuntos
Abelhas/microbiologia , Nosema/classificação , África , Distribuição Animal , Animais , Abelhas/anatomia & histologia , Colapso da Colônia/história , Colapso da Colônia/microbiologia , Contagem de Colônia Microbiana , História do Século XX , Masculino , Dados de Sequência Molecular , Nosema/genética , Nosema/isolamento & purificação , Reação em Cadeia da Polimerase , Dinâmica Populacional , Análise de Sequência de DNA , Asas de Animais/anatomia & histologiaRESUMO
Reactive dairy cows are more susceptible to stress, and this may result in negative effects on milk yield and quality. The aims of this study were to investigate the relationships between temperament traits and concentration of milk cortisol and oxytocin, milk yield, milkability, and milk quality in Holstein-Gyr cows. Temperament traits were assessed in 76 Holstein-Gyr cows in the milking parlor (by scoring milking reactivity and recording the numbers of steps and kicks during pre-milking udder preparation and when fitting the milking cluster) and during handling in the corral (by measuring the time to enter in the squeeze chute, ET and flight speed, FS). Milk samples were collected for milk quality (% fat, % protein, % lactose, and somatic cell count, SCC), and milk cortisol and oxytocin. Milk yield, milking time, and average flow were also measured. The calmer cows during milking management (class 'low') produced milk with higher protein (p = 0.028) content and tendencies for lower fat (p = 0.056) and higher lactose (p = 0.055) contents. Regarding the hormones, the most reactive cows (class 'high') in the milking and handling corral produced milk with higher concentrations of cortisol (p<0.001) and oxytocin (p = 0.023). In addition, the temperament of the animals affected some of the productive measures evaluated. Cows with reactive temperament had lower milk flow and longer milking time than the intermediate ones and had higher fat and a tendency for lower protein percentage in milk compared to cows with intermediate temperaments. Calm and intermediate cows in the handling corral produced more milk and presented better milkability parameters, such as a shorter milking time and greater average milk flow. Our results suggest that the cows' behavioral reactivity can be related to the intensity of their response to stress during handling.
Assuntos
Leite , Ocitocina , Feminino , Bovinos , Animais , Leite/metabolismo , Ocitocina/metabolismo , Lactação/fisiologia , Hidrocortisona/metabolismo , Temperamento , Lactose/metabolismo , Indústria de Laticínios/métodos , Glândulas Mamárias Animais/fisiologiaRESUMO
Macrophages are classified upon activation as classical activated M1 and M2 anti-inflammatory regulatory populations. This macrophage polarization is well characterized in humans and mice, but M1/M2 profile in cattle has been far less explored. Bos primigenius taurus (taurine) and Bos primigenius indicus (indicine) cattle display contrasting levels of resistance to infection and parasitic diseases such as C57BL/6J and Balb/c murine experimental models of parasite infection outcomes based on genetic background. Thus, we investigated the differential gene expression profile of unstimulated and LPS stimulated monocyte-derived macrophages (MDMs) from Holstein (taurine) and Gir (indicine) breeds using RNA sequencing methodology. For unstimulated MDMs, the contrast between Holstein and Gir breeds identified 163 Differentially Expressed Genes (DEGs) highlighting the higher expression of C-C chemokine receptor type five (CCR5) and BOLA-DQ genes in Gir animals. LPS-stimulated MDMs from Gir and Holstein animals displayed 1,257 DEGs enriched for cell adhesion and inflammatory responses. Gir MDMs cells displayed a higher expression of M1 related genes like Nitric Oxide Synthase 2 (NOS2), Toll like receptor 4 (TLR4), Nuclear factor NF-kappa-B 2 (NFKB2) in addition to higher levels of transcripts for proinflammatory cytokines, chemokines, complement factors and the acute phase protein Serum Amyloid A (SAA). We also showed that gene expression of inflammatory M1 population markers, complement and SAA genes was higher in Gir in buffy coat peripheral cells in addition to nitric oxide concentration in MDMs supernatant and animal serum. Co-expression analyses revealed that Holstein and Gir animals showed different transcriptional signatures in the MDMs response to LPS that impact on cell cycle regulation, leukocyte migration and extracellular matrix organization biological processes. Overall, the results suggest that Gir animals show a natural propensity to generate a more pronounced M1 inflammatory response than Holstein, which might account for a faster immune response favouring resistance to many infection diseases.
Assuntos
Cruzamento , Bovinos , Perfilação da Expressão Gênica/veterinária , Redes Reguladoras de Genes/efeitos dos fármacos , Lipopolissacarídeos/farmacologia , Macrófagos/química , Animais , Diferenciação Celular/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Lipopolissacarídeos/efeitos adversos , Ativação de Macrófagos , Macrófagos/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Análise de Sequência de RNA/veterinária , Especificidade da EspécieRESUMO
Whole-genome re-sequencing, alignment and annotation analyses were undertaken for 12 sires representing four important cattle breeds in Brazil: Guzerat (multi-purpose), Gyr, Girolando and Holstein (dairy production). A total of approximately 4.3 billion reads from an Illumina HiSeq 2000 sequencer generated for each animal 10.7 to 16.4-fold genome coverage. A total of 27,441,279 single nucleotide variations (SNVs) and 3,828,041 insertions/deletions (InDels) were detected in the samples, of which 2,557,670 SNVs and 883,219 InDels were novel. The submission of these genetic variants to the dbSNP database significantly increased the number of known variants, particularly for the indicine genome. The concordance rate between genotypes obtained using the Bovine HD BeadChip array and the same variants identified by sequencing was about 99.05%. The annotation of variants identified numerous non-synonymous SNVs and frameshift InDels which could affect phenotypic variation. Functional enrichment analysis was performed and revealed that variants in the olfactory transduction pathway was over represented in all four cattle breeds, while the ECM-receptor interaction pathway was over represented in Girolando and Guzerat breeds, the ABC transporters pathway was over represented only in Holstein breed, and the metabolic pathways was over represented only in Gyr breed. The genetic variants discovered here provide a rich resource to help identify potential genomic markers and their associated molecular mechanisms that impact economically important traits for Gyr, Girolando, Guzerat and Holstein breeding programs.
Assuntos
Bovinos/genética , Mutação INDEL , Polimorfismo de Nucleotídeo Único , Animais , Brasil , Cruzamento , Bovinos/classificação , Feminino , Genótipo , Sequenciamento de Nucleotídeos em Larga Escala/veterinária , Masculino , Anotação de Sequência Molecular , Análise de Sequência com Séries de Oligonucleotídeos/veterinária , Análise de Sequência de DNA/veterinária , Especificidade da EspécieRESUMO
Two species of Spiroplasma (Mollicutes) bacteria were isolated from and described as pathogens of the European honey bee, Apis mellifera, ~30 years ago but recent information on them is lacking despite global concern to understand bee population declines. Here we provide a comprehensive survey for the prevalence of these two Spiroplasma species in current populations of honey bees using improved molecular diagnostic techniques to assay multiyear colony samples from North America (U.S.A.) and South America (Brazil). Significant annual and seasonal fluctuations of Spiroplasma apis and Spiroplasma melliferum prevalence in colonies from the U.S.A. (n = 616) and Brazil (n = 139) occurred during surveys from 2011 through 2013. Overall, 33% of U.S.A. colonies and 54% of Brazil colonies were infected by Spiroplasma spp., where S. melliferum predominated over S. apis in both countries (25% vs. 14% and 44% vs. 38% frequency, respectively). Colonies were co-infected by both species more frequently than expected in both countries and at a much higher rate in Brazil (52%) compared to the U.S.A. (16.5%). U.S.A. samples showed that both species were prevalent not only during spring, as expected from prior research, but also during other seasons. These findings demonstrate that the model of honey bee spiroplasmas as springtime-restricted pathogens needs to be broadened and their role as occasional pathogens considered in current contexts.
Assuntos
Abelhas/microbiologia , Spiroplasma/isolamento & purificação , Animais , Carga Bacteriana , Brasil , Estações do Ano , Spiroplasma/classificação , Spiroplasma/genética , Estados UnidosRESUMO
In dairy farm animals, one the most important goal of the selection is the improvement of milk yield and composition. Several studies have demonstrated that the candidate genes of the kappa-casein (CSN3) and β - lactoglobulin (β-LG) are associated with milk yield, milk quality and health traits in dairy animals. Therefore the aim of this study was to detect polymorphisms in CSN3 and β-LG genes and its association with milk yield in up to 305 days (305MY) and predicted transmission capacity (PTA) for 305MY in Girolando cattle. Totally, 138 bulls and 729 cows (n=867) were sampled. The genotypes of both genes were obtained by the PCR-RFLP method using HinfI and HaeIII enzymes for CSN3 and β-LG genes, respectively. Statistical results revealed two alleles A and B for both genes. The genotypes and alleles more frequents for CSN3 and β-LG genes were respectively: AA (0.7324) and A (0.8558), and AB (0.4827) and A (0.5017). The x2 test revealed that the two loci were at Hardy-Weinberg equilibrium (p0.05). The allele variants of β-LG and CSN3 might be more investigated before include them into future breeding schemes designed for Girolando dairy cattle with objective of improving milk traits as milk yield in up to 305 days (305MY) and predicted transmission capacity (PTA) for 305MY.
Em rebanhos leiteiros, um dos objetivos mais importantes da seleção é a melhoria da produção e composição do leite. Vários estudos demonstraram que os genes candidatos da kappa-caseína (CSN3) e da β-lactoglobulina (β-LG) estão associados a produção de leite, qualidade do leite e características de saúde em animais leiteiros. Portanto, o objetivo deste estudo foi detectar polimorfismos nos genes CSN3 e β-LG e avaliar possíveis associaçoes desses polimorfismos com a produção de leite em até 305 dias (305MY) e a capacidade de transmissão prevista (PTA) de leite em bovinos da raça Girolando. No total, 138 touros e 729 vacas (n = 867) foram amostrados. A genotipagem foi realizada pelo método PCR-RFLP utilizando as enzimas HinfI e HaeIII para os genes CSN3 e β-LG, respectivamente. Os resultados estatísticos revelaram dois alelos A e B para ambos os genes. Os genotipos e alelos mais frequentes para os genes CSN3 e β-LG foram respectivamente: AA (0,7324) e A (0,8558) e AB (0,4827) e A (0,5017). O teste x2 revelou que os dois loci estavam em equilibrio de Hardy-Weinberg (p 0,05). Portanto, as variantes alélicas identificadas nos genes β-LG e CSN3 devem ser mais investigadas antes de serem incluidas nos programas de melhoramento desenhados para bovinos leiteiros Girolando objetivando melhorar as características do leite analisadas no presente estudo.
Assuntos
Caseínas/genética , Lactoglobulinas/genética , Leite/química , Polimorfismo de Nucleotídeo Único , Proteínas do Leite/análise , Reação em Cadeia da Polimerase , Técnicas de GenotipagemRESUMO
In dairy farm animals, one the most important goal of the selection is the improvement of milk yield and composition. Several studies have demonstrated that the candidate genes of the kappa-casein (CSN3) and β - lactoglobulin (β-LG) are associated with milk yield, milk quality and health traits in dairy animals. Therefore the aim of this study was to detect polymorphisms in CSN3 and β-LG genes and its association with milk yield in up to 305 days (305MY) and predicted transmission capacity (PTA) for 305MY in Girolando cattle. Totally, 138 bulls and 729 cows (n=867) were sampled. The genotypes of both genes were obtained by the PCR-RFLP method using HinfI and HaeIII enzymes for CSN3 and β-LG genes, respectively. Statistical results revealed two alleles A and B for both genes. The genotypes and alleles more frequents for CSN3 and β-LG genes were respectively: AA (0.7324) and A (0.8558), and AB (0.4827) and A (0.5017). The x2 test revealed that the two loci were at Hardy-Weinberg equilibrium (p<0.001). The allele substitution effects for the variants were not significant on 305MY and PTA for 305MY (p>0.05). The allele variants of β-LG and CSN3 might be more investigated before include them into future breeding schemes designed for Girolando dairy cattle with objective of improving milk traits as milk yield in up to 305 days (305MY) and predicted transmission capacity (PTA) for 305MY.(AU)
Em rebanhos leiteiros, um dos objetivos mais importantes da seleção é a melhoria da produção e composição do leite. Vários estudos demonstraram que os genes candidatos da kappa-caseína (CSN3) e da β-lactoglobulina (β-LG) estão associados a produção de leite, qualidade do leite e características de saúde em animais leiteiros. Portanto, o objetivo deste estudo foi detectar polimorfismos nos genes CSN3 e β-LG e avaliar possíveis associaçoes desses polimorfismos com a produção de leite em até 305 dias (305MY) e a capacidade de transmissão prevista (PTA) de leite em bovinos da raça Girolando. No total, 138 touros e 729 vacas (n = 867) foram amostrados. A genotipagem foi realizada pelo método PCR-RFLP utilizando as enzimas HinfI e HaeIII para os genes CSN3 e β-LG, respectivamente. Os resultados estatísticos revelaram dois alelos A e B para ambos os genes. Os genotipos e alelos mais frequentes para os genes CSN3 e β-LG foram respectivamente: AA (0,7324) e A (0,8558) e AB (0,4827) e A (0,5017). O teste x2 revelou que os dois loci estavam em equilibrio de Hardy-Weinberg (p <0,001). Os efeitos de substituiçao alélica para as variantes nao foram significativos para as características 305 MY e PTA para 305MY (p> 0,05). Portanto, as variantes alélicas identificadas nos genes β-LG e CSN3 devem ser mais investigadas antes de serem incluidas nos programas de melhoramento desenhados para bovinos leiteiros Girolando objetivando melhorar as características do leite analisadas no presente estudo.(AU)
Assuntos
Caseínas/genética , Lactoglobulinas/genética , Leite/química , Polimorfismo de Nucleotídeo Único , Proteínas do Leite/análise , Técnicas de Genotipagem , Reação em Cadeia da PolimeraseRESUMO
ABSTRACT Due to their ecological and economic importance, honey bees have attracted much scientific attention, which has intensified due to the recent population decline of these insects in the several parts of the world. Among the factors related to these patterns, infection by pathogens are the most relevant, mainly because of the easy dissemination of these microorganisms. Although no zoonotic diseases are associated with these insects, the presence of infectious agents in bee products should still be considered because they play a role as disease dispersers, increasing the risk to animal health. Because of the possibility of dispersion of pathogens via bee products, this work aimed to identify the presence of spores of the pathogens Paenibacillus larvae, Ascosphaera apis and Nosema spp. in samples of honey, pollen and royal jelly that are registered with Brazil's Federal Inspection Service (S.I.F.) and commercially available in the state of São Paulo. Of the 41 samples of bee products analyzed, only one showed no contamination by any of these pathogens. N. ceranae and P. larvae had the highest prevalence considering all the samples analyzed (present in 87.80% and 85.37% of the total, respectively), with N. apis present in 26.83% and A. apis present in 73.17% of the samples. These results provide support for the formulation of government regulations for sanitary control of exotic diseases by preventing dispersion of pathogens, including through illegal importation, since local and international trade and the transfer of colonies between regions play important roles in the dispersion of these microorganisms.