RESUMO
Approximately 80% of the world's cattle are raised in regions with a high risk of tick-borne diseases, resulting in significant economic losses due to parasitism by Rhipicephalus (Boophilus) microplus. However, the lack of a systemic biology approach hampers a comprehensive understanding of tick-host interactions that mediate tick resistance phenotypes. Here, we conducted a genome-wide association study (GWAS) of 2933 Braford cattle and found 340 single-nucleotide polymorphisms (SNPs) associated with tick counts. Gene expression analyses were performed on skin samples obtained from previously tick-exposed heifers with extremely high or low estimated breeding values for R. microplus counts. Evaluations were performed both before and after artificial infestation with ticks. Differentially expressed genes were found within 1-Mb windows centered at significant SNPs from GWAS. A total of 330 genes were related to the breakdown of homeostasis that was induced by larval attachment to bovine skin. Enrichment analysis pointed to a key role of proteolysis and signal transduction via JAK/STAT, NFKB and WNT/beta catenin signaling pathways. Integrative analysis on matrixEQTL revealed two cis-eQTLs and four significant SNPs in the genes peptidyl arginine deiminase type IV (PADI4) and LOC11449251. The integration of genomic data from QTL maps and transcriptome analyses has identified a set of twelve key genes that show significant associations with tick loads. These genes could be key candidates to improve the accuracy of genomic predictions for tick resistance in Braford cattle.
Assuntos
Resistência à Doença , Estudo de Associação Genômica Ampla , Polimorfismo de Nucleotídeo Único , Rhipicephalus , Infestações por Carrapato , Animais , Bovinos , Rhipicephalus/genética , Rhipicephalus/fisiologia , Infestações por Carrapato/veterinária , Infestações por Carrapato/genética , Infestações por Carrapato/parasitologia , Infestações por Carrapato/imunologia , Resistência à Doença/genética , Biologia de Sistemas , Doenças dos Bovinos/genética , Doenças dos Bovinos/imunologia , Doenças dos Bovinos/parasitologia , Locos de Características Quantitativas , Feminino , Interações Hospedeiro-Parasita/genética , Interações Hospedeiro-Parasita/imunologiaRESUMO
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
This study aimed to evaluate the transcriptional changes occurring in isolated perfused mammary alveolar tissue in response to inoculation with S. agalactiae and to identify the most affected biological functions and pathways after 3 h. Four udders taken at slaughter from cows with healthy mammary gland were perfused ex situ with warmed and gassed Tyrode's solution. Mammary alveolar tissue samples were taken from the left fore and rear quarters (IQ-inoculated quarters) before inoculation (hour 0) and at 3 h post inoculation (hpi) and at the same times from control right fore and rear quarters (not inoculated: NIQ). A total of 1756 differentially expressed genes (DEGs) were identified between IQ and NIQ at 3 hpi using edgeR package. Within this set of DEGs, 952 were up regulated and mainly involved with innate immune response and inflammatory response, e.g., CD14, CCL5, TLR2, IL-8, SAA3, as well as in transcriptional regulation such as FOS, STAT3 and NFKBIA. Genes down-regulated (804) included those involved with lipid synthesis e.g., APOC2, SCD, FABP3 and FABP4. The most affected pathways were chemokine signaling, Wnt signaling and complement and coagulation cascades, which likely reflects the early stage response of mammary tissue to S. agalactiae infection. No significant gene expression changes were detected by RNA-Seq in the others contrasts. Real time-PCR confirmed the increase in mRNA abundance of immune-related genes: TLR2, TLR4, IL-1ß, and IL-10 at 3 hpi between IQ and NIQ. The expression profiles of Casp1 and Bax for any contrasts were unaffected whereas Bcl2 was increased in IQ, which suggests no induction of apoptosis during the first hours after infection. Results provided novel information regarding the early functional pathways and gene network that orchestrate innate immune responses to S. agalactiae infection. This knowledge could contribute to new strategies to enhance resistance to this disease, such as genomic selection.
Assuntos
Perfilação da Expressão Gênica/veterinária , Glândulas Mamárias Animais/metabolismo , Infecções Estreptocócicas/veterinária , Streptococcus agalactiae , Animais , Bovinos , Feminino , Perfilação da Expressão Gênica/métodos , Regulação da Expressão Gênica , Imunidade/genética , Inflamação/genética , Mastite Bovina/genética , Mastite Bovina/imunologia , Mastite Bovina/microbiologia , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Infecções Estreptocócicas/genética , Infecções Estreptocócicas/imunologiaRESUMO
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
Diarrhea and respiratory diseases pose significant challenges in the rearing of pre-weaned calves, motivating the investigation of tools to improve gastrointestinal tract development, health, and overall performance in young calves. Consequently, the primary objective of this study was to assess the effectiveness of an additive incorporated into milk replacer to promote the development and health of the animals. Forty-six dairy calves were randomly assigned into two treatments: control (CON, n = 23; with 15 females and 8 males), and sodium butyrate (SB, n = 23; with 15 females and 8 males). The calves in the SB treatment group were supplemented with 4 g/d of unprotected sodium butyrate (Adimix, Adisseo, China), added to the milk replacer from 4 to 60 days of age. Water and starter were fed ad libitum. The study evaluated several parameters, including feed intake, nutrient digestibility, ruminal pH, ammonia and volatile fatty acids, blood metabolites (glucose, insulin-like growth factor type 1, urea, ß-hydroxybutyrate), hemogram, health scores, performance, and feed efficiency. Bull calves were euthanized at 60 days of age for organ comparison, while heifer calves were assessed for carryover effects up to 90 days of age. Data were analyzed independently using linear mixed models using the nlme package in R, and the Artools package for non-parametric categorical outcomes. Although the feed intake and performance variables exhibited differences within weeks, no divergence was observed between treatment groups. Notably, a positive treatment-by-week interaction was identified for starter feed intake (p = 0.02) and total dry matter intake (p = 0.04) during pre-weaning for CON animals. Ruminal parameters, blood metabolites, and hemogram values such as glucose, urea, insulin-like growth factor type 1, mean corpuscular value, lymphocytes, and neutrophils displayed differences within weeks during the pre-weaning stage, but similar results within groups. No differences between supplemented and non-supplemented calves were found across nutrient digestibility, organ development, and histology. Regarding health scores, differences were noted within weeks for fecal and respiratory scores during the pre-weaning stage, and only the respiratory score during the post-weaning stage. Consequently, butyrate supplementation did not elicit improvements or negative effects in the body development or health status of dairy calves.
RESUMO
The objective of this study was to evaluate blood cells and metabolites, insulin-like growth factor-1 (IGF-1), digestibility, internal organs weight and histology, gene expression, and spleen cell proliferation of pre-weaned bull calves supplemented with a blend of essential oils in milk replacer (MR). Sixteen newborn Holstein × Gyr crossbred dairy bull calves, with body weight at birth of 33.3 ± 3.7 kg, were housed in individual sand bedded pens, blocked by genetic composition, and randomly assigned to 1 of 2 treatments in a randomized complete block design: Control (CON, n = 8) and blend of essential oils supplementation (BEO, n = 8, 1 g/day/calf, Apex Calf, Adisseo, China). The commercial blend was composed by plant extracts derived from anise, cinnamon, garlic, rosemary, and thyme. Animals were fed 5 L of MR/day reconstituted at 15% (dry matter basis), divided into two equal meals. Water and starter were provided ad libitum. ß-hydroxybutyrate, urea, and glucose were evaluated weekly, IGF-1 was evaluated biweekly, and total blood cell count was performed every four weeks until the end of the trial at eight weeks of age. Feed samples were collected three times a week and polled for weekly analysis. Apparent total nutrient digestibility was determined from d 56 to 60 of age. On d 60 ± 1, animals were euthanized for organ weight, histology, spleen cell proliferation, and intestinal gene expression analysis. Data were analyzed independently using linear mixed models using the REML method in the nlme package in R for continuous outcomes. A non-parametric test was used for ordered categorical outcomes using the Artools package in R. There were no differences between groups for blood evaluations, digestibility, gene expression, and a spleen cell proliferation assay. However, BEO calves presented a heavier pancreas, heavier intestines, bigger ileum villi, and higher cecum butyrate levels (P < 0.05), demonstrating that the EO supplementation helped intestinal development and symbiotic bacteria. It was also observed in CON animals' heavier respiratory tract and a higher eosinophil count (P < 0.05). Therefore, the organs where eosinophils are more active had a better response for BEO animals. No differences were found in the intestinal gene expression in the immune context. These results demonstrate that supplementing essential oils in MR could contribute to gut development and immune function. However, more research is needed to understand its impact on body development and define the best dosage and route of administration.
Assuntos
Alho , Fator de Crescimento Insulin-Like I , Óleos de Plantas , Animais , Bovinos , Masculino , Antioxidantes , Expressão Gênica , Fator de Crescimento Insulin-Like I/genética , Baço , Óleos de Plantas/farmacologiaRESUMO
The aim of this study was to identify possible effects of different vaccination strategies (concomitantly or not) against brucellosis and clostridia on intake, performance, feeding behavior, blood parameters, and immune responses of dairy heifers calves. Fifty heifers calves were enrolled [38 Gyr (Zebu, Bos taurus indicus) and 12 5/8 Holstein × Gyr]. At 120 d of age, animals were randomly distributed among 3 groups: B (n = 18), vaccinated against brucellosis; C (n = 14), vaccinated against clostridia and CB (n = 18), vaccinated concomitantly for both. Rectal and thermographic temperatures were evaluated on days -1, 0, 1, 2, 3, 5, 7,10, 14, and 28 relatives to the vaccination day. Feed and water intake, body weight (BW), and feeding behavior were monitored daily by an electronic feeding system. Blood was sampled on days 0, 3, 7, 14, and 28, relative to the vaccination day for determination of glucose and ß -hydroxybutyrate (BHBA) concentrations. Blood sampled on day 0 (prevaccination) and on days 28 and 42 were used to evaluate the immune response against Brucella abortus and clostridia. There was an increase in rectal temperature between the first and the third day postvaccination in the 3 groups. The thermography revealed an increase of local temperature for 7 d on groups B and CB. Group C had increased local temperature for a longer period, lasting for up to 14 d. Dry mater intake was reduced for groups B and CB, but no alteration was observed for group C. No alterations regarding initial BW, final BW, average daily weight gain, and feed efficiency were observed. No differences were observed for the 3 vaccination groups for blood parameters throughout the evaluation period. The concomitant vaccination against brucellosis and clostridia led to lower neutralizing antibody titers against epsilon toxin of Clostridium perfringens and botulinum toxin type C of C. botulinum (C > CB > B). When cellular proliferation assay and serological tests to B. abortus were evaluated, no differences were observed between groups B and CB. The present results indicate that the concomitant vaccination against brucellosis and clostridia has no relevant impact on the intake, performance, and feeding behavior of dairy calves. However, the concomitant vaccination of vaccines against these 2 pathogens impacts animal immunity against clostridial infections.
Assuntos
Brucelose , Doenças dos Bovinos , Ração Animal/análise , Animais , Peso Corporal , Brucelose/veterinária , Bovinos , Doenças dos Bovinos/prevenção & controle , Comportamento Alimentar , Feminino , Imunidade , Vacinação/veterináriaRESUMO
Ticks (Acari: Ixodidae) are bloodsucking ectoparasitic arthropods of human and veterinary medical importance. Tick saliva has been shown to contain a wide range of bioactive molecules with vasodilatory, antihemostatic, and immunomodulatory activities. We have previously demonstrated that saliva from Rhipicephalus sanguineus ticks inhibits the maturation of dendritic cells (DCs) stimulated with LPS. Here we examined the mechanism of this immune subversion, evaluating the effect of tick saliva on Toll-like receptor (TLR)-4 signalling pathway in bone marrow-derived DCs. We demonstrated that R. sanguineus tick saliva impairs maturation of DCs stimulated with LPS, a TLR-4 ligand, leading to increased production of interleukin (IL)-10 and reduced synthesis of IL-12p70 and TNF-alpha. The immunomodulatory effect of the tick saliva on the production of pro-inflammatory cytokines by DCs stimulated with LPS was associated with the observation that tick saliva inhibits the activation of the ERK 1/2 and p38 MAP kinases. These effects were independent of the expression of TLR-4 on the surface of DCs. Additionally, saliva-treated DCs also presented a similar pattern of cytokine modulation in response to other TLR ligands. Since the recent literature reports that several parasites evade immune responses through TLR-2-mediated production of IL-10, we evaluated the effect of tick saliva on the percentage of TLR-2(+) DCs stimulated with the TLR-2 ligand lipoteicoic acid (LTA). The data showed that the population of DCs expressing TLR-2 was significantly increased in DCs treated with LTA plus saliva. In addition, tick saliva alone increased the expression of TLR-2 in a dose- and time-dependent manner. Our data suggest that tick saliva induces regulatory DCs, which secrete IL-10 and low levels of IL-12 and TNF-alpha when stimulated by TLR ligands. Such regulatory DCs are associated with expression of TLR-2 and inhibition of ERK and p38, which promotes the production of IL-10 and thus down-modulates the host's immune response, possibly favouring susceptibility to tick infestations.
Assuntos
Quinases de Proteína Quinase Ativadas por Mitógeno/metabolismo , Rhipicephalus/fisiologia , Saliva/metabolismo , Receptor 2 Toll-Like/metabolismo , Animais , Células Cultivadas , Quimiocinas/metabolismo , Células Dendríticas , Feminino , Regulação da Expressão Gênica , Camundongos , Ratos , Receptor 2 Toll-Like/genéticaRESUMO
Cattle present variable levels of resistance to ticks and the immune correlates of these heritable phenotypes must be known in order to develop effective vaccines. The antibody responses to tick salivary antigens were examined in cattle of tick-susceptible (Holstein) and tick-resistant (Nelore) breeds. After heavy infestations, levels of IgG1 and IgG2 antibodies decreased in Holsteins and remained the same in Nelores. Conversely, levels of IgE antibodies increased in Holsteins. Different sizes of tick burdens modulated the IgG1 antibody response in a susceptible breed (Aberdeen): levels were higher than in controls in heavily infested animals, but not in those undergoing intermediary or minimal infestations. The three experimental groups presented similar levels of IgG2 antibodies. Levels of IgE antibodies were higher only in animals undergoing intermediate infestations. These results indicate that tick infestations suppress the IgG antibody response in susceptible breeds, that IgE antibodies are not protective, and that the dose of tick saliva modulates the isotype of host antibody responses.