RESUMEN
Chemical treatments are the main strategy to control gastrointestinal nematodes in sheep, and the emergence of anthelmintic resistance, as consequence, results in control failures and leads to economic losses. Thus, molecular tests may constitute an excellent tool for the early detection of anthelmintic resistance-related mutations. Thus, a polymerase chain reaction (PCR)-based genotyping assay followed by polyacrylamide gel electrophoresis (PAGE) was developed to detect polymorphisms in exon 11 of the acetylcholine receptor monepantel-1 gene (mptl-1) that were previously associated with monepantel resistance through a genome-wide study in Haemonchus contortus. DNA samples recovered from individual and pooled third-stage larvae from two susceptible field-derived isolates and five (three in vivo-derived and two field-derived) resistant populations were used. New polymorphisms, including a 6-bp deletion and a 3-bp insertion, were detected in resistant individuals. These indels, confirmed using sequencing of cloned PCR products, are predicted to result in amino acid changes in transmembrane domain 2 (TMD2) of the MPTL-1 protein. The two susceptible isolates showed only the presence of the wild-type allele (100%), whereas lower frequencies of the wild-type allele were detected in monepantel-resistant populations (11.1 to 66.7%). These findings report new polymorphisms in the mptl-1 gene, validate the results obtained through genomic mapping for monepantel resistance, and provide a PCR-based assay to genotype indels located in exon 11 of mptl-1 in H. contortus.
Asunto(s)
Antihelmínticos , Hemoncosis , Haemonchus , Enfermedades de las Ovejas , Ovinos/genética , Animales , Estudio de Asociación del Genoma Completo , Resistencia a Medicamentos/genética , Antihelmínticos/uso terapéutico , Enfermedades de las Ovejas/tratamiento farmacológico , Exones , Hemoncosis/veterinariaRESUMEN
Single nucleotide polymorphisms (SNPs) are the main type of variation in genome, enabling them to be associated with traits of economic importance in livestock. Genome-wide association studies (GWAS) have led to the discovery of SNPs associated with desirable traits in sheep. However, in these studies, SNPs are genotyped by high-throughput methods in genome scale, which are expensive and require sophisticated equipment and analysis methods. Therefore, the goal of this study was to develop a reliable, rapid, and inexpensive polymerase chain reaction (PCR)-based method to genotype a medium number of animals for a few candidate SNPs previously associated with desirable phenotypes in sheep by GWAS, using markers associated with gastrointestinal nematode resistance as a model. DNA extracted from white-blood cells of 150 sheep was submitted to PCR amplification followed by agarose gel electrophoresis and determination of banding pattern. Tetra-primer ARMS-PCR was successfully optimized after changes in annealing temperature; annealing and extension times; concentration of MgCl2 and DNA; ratios of inner, outer, forward and reverse primer; and addition of adjuvants, for genotyping the OAR2_14765360, OAR6_81718546, OAR11_62887032, and OAR12_69606944 SNPs in sheep. An extensive optimization of tetra-primer ARMS-PCR resulted in a suitable, simple, cost-effective PCR-based method of genotyping four SNP markers previously detected by chip arrays.
Asunto(s)
Técnicas de Genotipaje/métodos , Reacción en Cadena de la Polimerasa/métodos , Ovinos/genética , Animales , ADN/genética , Cartilla de ADN/genética , Estudio de Asociación del Genoma Completo , Genotipo , Fenotipo , Polimorfismo de Nucleótido Simple/genéticaRESUMEN
BACKGROUND: The integration of molecular data from hosts, parasites, and microbiota can enhance our understanding of the complex biological interactions underlying the resistance of hosts to parasites. Haemonchus contortus, the predominant sheep gastrointestinal parasite species in the tropics, causes significant production and economic losses, which are further compounded by the diminishing efficiency of chemical control owing to anthelmintic resistance. Knowledge of how the host responds to infection and how the parasite, in combination with microbiota, modulates host immunity can guide selection decisions to breed animals with improved parasite resistance. This understanding will help refine management practices and advance the development of new therapeutics for long-term helminth control. METHODS: Eggs per gram (EPG) of feces were obtained from Morada Nova sheep subjected to two artificial infections with H. contortus and used as a proxy to select animals with high resistance or susceptibility for transcriptome sequencing (RNA-seq) of the abomasum and 50 K single-nucleotide genotyping. Additionally, RNA-seq data for H. contortus were generated, and amplicon sequence variants (ASV) were obtained using polymerase chain reaction amplification and sequencing of bacterial and archaeal 16S ribosomal RNA genes from sheep feces and rumen content. RESULTS: The heritability estimate for EPG was 0.12. GAST, GNLY, IL13, MGRN1, FGF14, and RORC genes and transcripts were differentially expressed between resistant and susceptible animals. A genome-wide association study identified regions on chromosomes 2 and 11 that harbor candidate genes for resistance, immune response, body weight, and adaptation. Trans-expression quantitative trait loci were found between significant variants and differentially expressed transcripts. Functional co-expression modules based on sheep genes and ASVs correlated with resistance to H. contortus, showing enrichment in pathways of response to bacteria, immune and inflammatory responses, and hub features of the Christensenellaceae, Bacteroides, and Methanobrevibacter genera; Prevotellaceae family; and Verrucomicrobiota phylum. In H. contortus, some mitochondrial, collagen-, and cuticle-related genes were expressed only in parasites isolated from susceptible sheep. CONCLUSIONS: The present study identified chromosome regions, genes, transcripts, and pathways involved in the elaborate interactions between the sheep host, its gastrointestinal microbiota, and the H. contortus parasite. These findings will assist in the development of animal selection strategies for parasite resistance and interdisciplinary approaches to control H. contortus infection in sheep.
Asunto(s)
Hemoncosis , Haemonchus , Microbiota , Parásitos , Enfermedades de las Ovejas , Ovinos/genética , Animales , Parásitos/genética , Estudio de Asociación del Genoma Completo , Multiómica , Heces/parasitología , Enfermedades de las Ovejas/parasitología , Hemoncosis/parasitología , Recuento de Huevos de ParásitosRESUMEN
Visceral toxocariasis is a neglected parasitic zoonosis that occurs through the ingestion of embryonated Toxocara spp. eggs. A wide range of animal species can act as paratenic hosts for this ascarid. The main risk factor for humans is the ingestion of the eggs from contaminated soil; however, infection can also occur through the ingestion of contaminated raw or undercooked infected meat from paratenic hosts. The aim of this study was to verify the presence of Toxocara spp.-specific antibodies in sheep and to determine the risk factors associated with the infection of sheep in Rio Grande do Sul (a major sheep-producing and sheep-consuming state) in southern Brazil. Serum samples collected from 1,642 sheep were tested using an IgG enzyme-linked immunosorbent assay based on the excretory-secretory Toxocara canis antigen. Seroprevalence was 29.0% (477/1,642), and every farm included in the study contained at least one seropositive animal. These results indicate that T. canis infection is widely distributed among sheep herds in Rio Grande do Sul and that it represents a potential risk to human health.
Asunto(s)
Anticuerpos Antihelmínticos/sangre , Antígenos Helmínticos/inmunología , Enfermedades de las Ovejas/epidemiología , Toxocara/inmunología , Toxocariasis/epidemiología , Animales , Ascaris suum/química , Ascaris suum/inmunología , Brasil/epidemiología , Ensayo de Inmunoadsorción Enzimática/veterinaria , Proteínas del Helminto/inmunología , Humanos , Estudios Seroepidemiológicos , Ovinos , Enfermedades de las Ovejas/parasitología , Toxocara/aislamiento & purificación , Toxocara canis/inmunología , Toxocara canis/aislamiento & purificación , Toxocariasis/parasitología , ZoonosisRESUMEN
Haemonchus contortus is a constraint to sheep production. Seeking to reduce the use of hosts and produce parasitic stages in large-scale, a 42-day in vitro culture protocol of H. contortus third-stage larvae was optimized using Dulbecco's modified Eagle's medium (DMEM). In cell-free culture, larvae were maintained at 39.6°C, in acidic media (pH 6.1) for 3 or 6 days with Δ4-dafachronic acid followed by DMEM pH 7.4 supplemented or not with Fildes' reagent. In DMEM pH 7.4 at 37°C, supplementation with Caco-2 cells was compared to Fildes. On Day 14, fourth-stage larvae (L4) development rates in acidic media supplemented (86.8-88.4%) or not (74.4-77.8%) with Fildes and in Caco-2 cell co-culture (92.6%) were similar, and superior to DMEM pH 7.4 with Fildes (0.0%). On Day 21, Caco-2 cell co-culture resulted in higher larvae differentiation (25.0%) and lower degeneration (13.9%) compared to acidic media (1.5-8.1% and 48.6-69.9%, respectively). This is the first report of prolonged in vitro culture of H. contortus larvae using commercial media in co-culture with Caco-2 cells. Although no progression to the adult stage, Caco-2 cell co-culture resulted in morphological differentiation of H. contortus L4 and larval viability for up to 28 days.
Asunto(s)
Hemoncosis , Haemonchus , Enfermedades de las Ovejas , Animales , Ovinos , Humanos , Células CACO-2 , Larva , Hemoncosis/veterinaria , Hemoncosis/parasitología , Enfermedades de las Ovejas/parasitologíaRESUMEN
Among the gastrointestinal nematodes affecting sheep, Haemonchus contortus is the most prevalent and virulent, resulting in health problems and production losses. Therefore, selecting sheep resistant to H. contortus is a suitable and sustainable strategy for controlling endoparasites in flocks. Here, 287 lambs of the native Brazilian Morada Nova hair sheep breed were subjected to two consecutive artificial infections with H. contortus and assessed for fecal egg count (FEC), packed cell volume (PCV), and live weight (LW). Forty-four animals ranked as having extreme resistance phenotypes were genotyped using the Illumina OvineSNP50v3 chip. A case−control genome-wide association study (GWAS) detected 37 significant (p < 0.001) markers in 12 ovine chromosomes in regions harboring quantitative trait loci (QTL) for FEC, Trichostrongylus spp. adults and larvae, weight, and fat; and candidate genes for immune responses, mucins, hematological parameters, homeostasis, and growth. Four single-nucleotide polymorphisms (SNP; OAR1_rs427671974, OAR2_rs419988472, OAR5_rs424070217, and OAR17_rs401006318) genotyped by qPCR followed by high-resolution melting (HRM) were associated with FEC and LW. Therefore, molecular markers detected by GWAS for H. contortus resistance in Morada Nova sheep may support animal selection programs aimed at controlling gastrointestinal nematode infections in flocks. Furthermore, genotyping of candidate genes using HRM qPCR may provide a rapid and efficient tool for animal identification.
RESUMEN
A vaccine containing doses ranging from 2 µg to 50 µg of integral membrane proteins from Haemonchus contortus intestinal cells (H11 and H-gal-GP complex) has been shown to be effective for lambs. A vaccine for H. contortus was tested in two-month old grazing Corriedale lambs during an eight-month trial on the outskirts of Bagé, Rio Grande do Sul, Brazil. The animals were kept in a single paddock and allocated to five similar groups according to weight, sex and faecal egg counts (FEC). Vaccinated lambs received 0, 2, 5, 10 or 50 µg of the same antigen diluted in QuilA adjuvant. Vaccine injections were given at days 0, 21, 49, 91, 133, and 175. Lambs were sampled weekly for FEC, packed cell volume (PCV), and plasma ELISA antibody titre. Lambs with PCVs ≤15 % were drenched with levamisole (7.5 mg/kg body weight) as a salvage treatment. During days 77-98 an artificial challenge was administered to increase parasite transmission. Post-artificial challenge (from day 98-217), the FEC of the vaccinated lambs were 59.3 % lower than those of the control lambs. Antigen dose correlated with changes in PCV (r = 0.387 p-value < 0.001). Vaccinated lambs also had higher PCV than controls (p < 0.001; contrast analysis). Salvage treatment was needed in 16.7 % of vaccinated lambs and 88 % of control lambs.
Asunto(s)
Hemoncosis/veterinaria , Glicoproteínas de Membrana/inmunología , Enfermedades de las Ovejas/prevención & control , Vacunas/inmunología , Animales , Antihelmínticos/administración & dosificación , Antihelmínticos/uso terapéutico , Relación Dosis-Respuesta Inmunológica , Heces/parasitología , Hemoncosis/prevención & control , Haemonchus , Levamisol/administración & dosificación , Levamisol/uso terapéutico , Recuento de Huevos de Parásitos , Ovinos , Vacunas/administración & dosificaciónRESUMEN
Gastrointestinal nematodes are a major constraint in sheep production. Breeding for resistance has proven to be an effective and feasible approach to address this problem. The use and investigation of genetic markers for resistance traits could accelerate genetic progress and lead to a better understanding of underlying molecular mechanisms. Thus, the aim of this study was to evaluate if five single nucleotide polymorphisms SNPs OAR2_14765360, OAR6_81718546, OAR11_62887032, OAR12_69606944 and OAR15_59871543 are associated with resistance and resilience traits in a flock of the Morada Nova sheep breed. Lambs were submitted to two consecutive parasite challenges by oral infection with 4000 infective larvae L3) of Haemonchus contortus. Fecal egg counts (FEC), packed cell volume (PVC) and body weight were measured every one or two weeks for 42 days in each trial. DNA samples from 287 lambs, 131 ewes and 4 rams were amplified by ARMS-PCR or PCR-RFLP and genotypes were determined. Analysis of variance (ANOVA) was used for association analyses between genotypes and phenotypes. In case of significant association, the allele substitution effect was calculated based on a linear model. OAR2_14765360 and OAR12_69606944 were associated with FEC, and OAR12_69606944 also had significant effects on PCV and weight gain, showing favourable associations of the CC genotype with all evaluated traits. Both OAR6_81718546 and OAR11_62887032 were associated with weight gain, and OAR6_81718546 had an additional effect on PCV. OAR15_59871543 was not polymorphic in the population. OAR6_81718546 and OAR12_69606944 presented significant allele substitution effects of -1.06⯱â¯0.52â¯kg for the T allele on final body weight and 0.74⯱â¯0.32 for the C allele in PCV of the same sampling date, respectively. This is the first report of SNPs associated with gastrointestinal nematode resistance in this sheep breed. Our findings support the existence of quantitatice trait loci (QTL) for resistance and resilience in linkage disequilibrium with the polymorphic SNPs and suggest their future use for explorations of these traits in Morada Nova sheep.
Asunto(s)
Resistencia a la Enfermedad/genética , Hemoncosis/veterinaria , Haemonchus/genética , Polimorfismo de Nucleótido Simple , Enfermedades de las Ovejas/genética , Animales , Brasil , Hemoncosis/genética , Hemoncosis/parasitología , Ovinos , Enfermedades de las Ovejas/parasitologíaRESUMEN
Cells of the nucleus pulposus (NP) in the intervertebral disc are derived directly from the embryonic notochord. In humans, a shift in NP cell population coincides with the beginning of age-related changes in the extracellular matrix that can lead to spinal disorders. To begin identifying the bases of these changes, the manner by which relevant environmental factors impact cell function must be understood. This study investigated the roles of biochemical, nutritional, and physical factors in regulating immature NP cells. Specifically, we examined cell morphology, attachment, proliferation, and expression of genes associated with the notochord and immature NP (Sox9, CD24, and type IIA procollagen). Primary cells isolated from rat caudal discs were exposed to different media formulations and physical culture configurations either in 21% (ambient) or 2% (hypoxic) O2. As expected, cells in alginate beads retained a vacuolated morphology similar to chordocytes, with little change in gene expression. Interestingly, NP tissues not enzymatically digested were more profoundly influenced by oxygen. In monolayer, alpha-MEM preserved vacuolated morphology, produced the highest efficiency of attachment, and best maintained gene expression. DMEM and Opti-MEM cultures resulted in high levels of proliferation, but these appeared to involve small non-vacuolated cells. Gene expression patterns for cells in DMEM monolayer cultures were consistent with chondrocyte de-differentiation, with the response being delayed by hypoxia. Overall, results indicate that certain environmental conditions induce cellular changes that compromise the notochordal phenotype in immature NP. These results form the foundation on which the mechanisms of such changes can be elucidated.
Asunto(s)
Antígeno CD24/genética , Colágeno Tipo II/genética , Disco Intervertebral/metabolismo , Notocorda/metabolismo , Factor de Transcripción SOX9/genética , Alginatos/metabolismo , Animales , Adhesión Celular/genética , Hipoxia de la Célula , Proliferación Celular , Forma de la Célula/genética , Células Cultivadas , Medios de Cultivo/metabolismo , Regulación de la Expresión Génica , Ácido Glucurónico/metabolismo , Ácidos Hexurónicos/metabolismo , Disco Intervertebral/citología , Masculino , Notocorda/citología , Fenotipo , Ratas , Ratas Sprague-Dawley , Factores de TiempoRESUMEN
BACKGROUND: Haemonchus contortus, a gastrointestinal nematode parasite of sheep, is mainly controlled by anthelmintics; the occurrence of anthelmintic resistance leads to treatment failures and increases economic burden. Because molecular mechanisms involved in drug resistance can be elucidated by genomic studies, an extreme quantitative trait locus (X-QTL) mapping approach was used to identify co-segregation of the resistance phenotype with genetic markers to detect the genome-wide variants associated with monepantel resistance in H. contortus. METHODS: A cross between H. contortus isolates using parental susceptible (Par-S) males and monepantel resistant (Par-R) females resulted in SR progeny, while reciprocal cross resulted in RS progeny. Pools (n = 30,000) of infective larvae (L3) recovered from Par-R, and from SR and RS populations in the F3 generation, collected both before (unselected group) and 7 days after (selected group) selection with monepantel treatment in sheep hosts, were subjected to genome sequencing (Pool-Seq). Pairwise comparisons of allele frequencies between unselected and selected groups were performed for each population by Fisher's exact test (FET) and for both populations combined by a Cochran-Mantel-Haenszel (CMH) test. RESULTS: Mapping rates varied from 80.29 to 81.77% at a 90.4X mean coverage of aligned reads. After correction for multiple testing, significant (P < 0.05) changes in allele frequencies were detected by FET for 6 and 57 single nucleotide polymorphisms (SNPs) in the SR and RS populations, respectively, and by the CMH test for 124 SNPs in both populations. The significant variants located on chromosome 2 generated a selection signal in a genomic region harboring the mptl-1, deg-3 and des-2 genes, previously reported as candidates for monepantel resistance. In addition, three new variants were identified in the mptl-1 gene. CONCLUSIONS: This study expands knowledge on genome-wide molecular events underlying H. contortus resistance to monepantel. The identification of a genome region harboring major genes previously associated with monepantel resistance supports the results of the employed X-QTL approach. In addition, a deletion in exon 11 of the mptl-1 gene should be further investigated as the putative causal mutation leading to monepantel resistance.
Asunto(s)
Aminoacetonitrilo/análogos & derivados , Antihelmínticos/farmacología , Resistencia a Medicamentos/genética , Haemonchus/efectos de los fármacos , Haemonchus/genética , Sitios de Carácter Cuantitativo , Aminoacetonitrilo/farmacología , Animales , Femenino , Variación Genética , Masculino , Mutación , FenotipoRESUMEN
Bovine babesiosis is a tick-borne disease caused by Babesia spp. haemoprotozoans. The disease is of great importance at tick enzootic unstable areas and hampers cattle production in several developing countries. The available immunisation alternatives are pre-immunition and attenuated vaccines. Despite being efficient and protective, they are unsafe as they use cattle blood cells as inoculum and may potentially spread other diseases. Another alternative to help in babesiosis control would be the identification of genetically resistant cattle to Babesia bovis infection. The objective of this work was to phenotype cattle based on primary response against B. bovis infection. Two-hundred and forty half-sib Hereford and Aberdeen Angus heifers (120 animals from each breed), 12-18-month-old naïve cattle, originated from a tick-free area in Southern Brazil, were used in the experiment. Animals were monitored following an inoculation with 1x10(7)B. bovis parasitised erythrocytes. Results showed three different phenotypes: 1-'susceptible', animals with babesiosis clinical signs that received treatment to avoid death; 2-'intermediate', animals with clinical signs: parasitaemia, >or=21.5% reduction in packed cell volume (PCV) and increase in body temperature when compared to their pre-challenge physiological parameters, no specific treatment was needed as animals self recovered from the disease, and 3-'resistant', animals without clinical signs that showed B. bovis presence in blood smears, <21.5% PCV reductions, with little or no increase in body temperature and no need for babesiosis treatment. The frequencies of each phenotype were: 45.4, 26.7, and 27.9%, respectively, demonstrating the existence of phenotypic variation for B. bovis in Bos taurus cattle.
Asunto(s)
Babesia bovis , Babesiosis/veterinaria , Enfermedades de los Bovinos/parasitología , Animales , Anticuerpos Antiprotozoarios , Antiprotozoarios/uso terapéutico , Babesiosis/tratamiento farmacológico , Babesiosis/genética , Babesiosis/inmunología , Temperatura Corporal , Bovinos , Enfermedades de los Bovinos/tratamiento farmacológico , Enfermedades de los Bovinos/genética , Enfermedades de los Bovinos/inmunología , Femenino , Predisposición Genética a la Enfermedad , Inmunoglobulina G/sangre , Parasitemia , Fenotipo , Factores de TiempoRESUMEN
An experiment was conducted with the objective of evaluating whether a 5000 L3 larvae Haemonchus placei primary infection, a less pathogenic parasite species for sheep, could attenuate highly pathogenic Haemonchus contortus infections in lambs. Thirty nine 6-month-old lambs were divided in three primary-infection groups: (HcPI) H. contortus-primary infected lambs, (HpPI) H. placei-primary infected lambs, and (CTRL) non-infected control lambs. Later, these same individuals, then aged 10-month old, were cross challenged with Haemonchus parasite species, creating four groups: HcPI challenged with H. placei (HcPI-HpCH), HpPI challenged with H. contortus (HpPI-HcCH), HcPI challenged with H. contortus (HcPI-HcCH), and CTRL. After a 60-day challenge period, all animals were necropsied for gastrointestinal worm counts. HcPI faecal egg count average was found to be twice the HpPI FEC group (p<0.0001). The HcPI also showed lower packed cell volume averages compared to the other groups (p<0.0001). Both H. contortus- and H. placei-primary infections displayed immune responses with similar IgG levels. For the challenge trial, the larval doses used were not enough to trigger clinic infection signs in all treated groups, compared to controls, and H. placei primary infection was not able to maintain anti-H. contortus IgG levels in a subsequent H. contortus infection.
Asunto(s)
Hemoncosis/veterinaria , Haemonchus/clasificación , Enfermedades de las Ovejas/parasitología , Animales , Heces/parasitología , Hemoncosis/inmunología , Hemoncosis/parasitología , Haemonchus/inmunología , Recuento de Huevos de Parásitos/veterinaria , Ovinos , Enfermedades de las Ovejas/inmunologíaRESUMEN
Genetic markers for sheep resistance to gastrointestinal parasites have long been sought by the livestock industry as a way to select more resistant individuals and to help farmers reduce parasite transmission by identifying and removing high egg shedders from the flock. Polymorphisms related to the major histocompatibility complex and interferon (IFN)-γ genes have been the most frequently reported markers associated with infection. Recently, a new picture is emerging from genome-wide studies, showing that not only immune mechanisms are important determinants of host resistance but that gastrointestinal mucus production and hemostasis pathways may also play a role.
Asunto(s)
Resistencia a la Enfermedad/genética , Genoma/genética , Interacciones Huésped-Parásitos/genética , Parasitosis Intestinales/veterinaria , Infecciones por Nematodos/veterinaria , Enfermedades de las Ovejas/genética , Animales , Cruzamiento , Marcadores Genéticos/genética , Parasitosis Intestinales/genética , Nematodos/fisiología , Infecciones por Nematodos/genética , Ovinos/genéticaRESUMEN
Gastrointestinal (GI) parasitic infection is the main health constraint for small ruminant production, causing loss of weight and/or death. Red Maasai sheep have adapted to a tropical environment where extreme parasite exposure is a constant, especially with highly pathogenic Haemonchus contortus. This breed has been reported to be resistant to gastrointestinal parasite infection, hence it is considered an invaluable resource to study associations between host genetics and resistance. The aim of this study was to identify polymorphisms strongly associated with host resistance in a double backcross population derived from Red Maasai and Dorper sheep using a SNP-based GWAS analysis. The animals that were genotyped represented the most resistant and susceptible individuals based on the tails of phenotypic distribution (10% each) for average faecal egg counts (AVFEC). AVFEC, packed cell volume (AVPCV), and live weight (AVLWT) were adjusted for fixed effects and co-variables, and an association analysis was run using EMMAX. Revised significance levels were calculated using 100,000 permutation tests. The top five significant SNP markers with - log10 p-values >3.794 were observed on five different chromosomes for AVFEC, and BLUPPf90/PostGSf90 results confirmed EMMAX significant regions for this trait. One of these regions included a cluster of significant SNP on chromosome (Chr) 6 not in linkage disequilibrium to each other. This genomic location contains annotated genes involved in cytokine signalling, haemostasis and mucus biosynthesis. Only one association detected on Chr 7 was significant for both AVPCV and AVLWT. The results generated here reveal candidate immune variants for genes involved in differential response to infection and provide additional SNP marker information that has potential to aid selection of resistance to gastrointestinal parasites in sheep of a similar genetic background to the double backcross population.
Asunto(s)
Cruzamientos Genéticos , Tracto Gastrointestinal/parasitología , Predisposición Genética a la Enfermedad , Ovinos/genética , Animales , Mapeo Cromosómico , Estudio de Asociación del Genoma Completo , Recuento de Huevos de Parásitos , Polimorfismo de Nucleótido Simple , Ovinos/parasitologíaRESUMEN
Galectins and collectins are proteins classified in the lectin family that have the ability to recognize molecular patterns associated with pathogens. Studies on cattle have demonstrated high expression of these proteins during infection with gastrointestinal nematodes. The aim of this study was to investigate whether the level of Haemonchus contortus infection would alter the expression of galectins (Gal11 and Gal14) and collectins (SPA and CGN) in sheep. Twelve Corriedale sheep exposed to natural infection with nematodes were divided into two groups: group 1 (G1, n = 7) and group 2 (G2, n = 5), with low and high parasite burdens, respectively, based on fecal egg counts and abomasal parasite counts. The fecal egg counts and abomasal parasite counts were significantly different (p < 0.05) between the groups. Galectin and collectin gene expression was observed in all sheep abomasal samples. However, animals with lower infection levels showed lower expression of the genes Gal14, SPA and CGN (p < 0.05). Expression of lectins was associated with the abomasal H. contortus burden, thus suggesting that these proteins may have a role in controlling of this infection.
Asunto(s)
Colectinas/biosíntesis , Galectinas/biosíntesis , Hemoncosis/veterinaria , Enfermedades de las Ovejas/metabolismo , Animales , Colectinas/genética , Femenino , Galectinas/genética , Expresión Génica , Hemoncosis/genética , Hemoncosis/metabolismo , Haemonchus , Masculino , OvinosRESUMEN
Toll-like receptor 4 (TLR4) is a key factor in the innate immune recognition of lipopolysaccharide (LPS) from Gram-negative bacteria. Previous studies from our group identified differences in the expression profile of TLR4 and genes affected by the TLR4 signaling pathway among pigs that shed varying levels of Salmonella, a Gram-negative bacterium. Therefore, genetic variation in this gene may be involved with the host's immune response to bacterial infections. The current study screened for single nucleotide polymorphisms (SNPs) in the TLR4 gene and tested their association with Salmonella fecal shedding. Pigs (n = 117) were intranasally challenged at 7 weeks of age with 1 × 10(9) CFU of S. Typhimurium χ4232 and were classified as low or persistent Salmonella shedders based on the levels of Salmonella being excreted in fecal material. Salmonella fecal shedding was determined by quantitative bacteriology on days 2, 7, 14, and 20/21 post exposure, and the cumulative levels of Salmonella were calculated to identify the low (n = 20) and persistent (n = 20) Salmonella shedder pigs. From those 40 animals, the TLR4 region was sequenced, and 18 single nucleotide polymorphisms (SNPs) in TLR4 were identified. Twelve SNPs have been previously described and six are novel SNPs of which five are in the 5' untranslated region and one is in intron 2. Single marker association test identified 13 SNPs associated with the qualitative trait of Salmonella fecal shedding, and seven of those SNPs were also associated with a quantitative measurement of fecal shedding (P < 0.05). Using a stepwise regression process, a haplotype composed of SNPs rs80787918 and rs80907449 (P ≤ 4.0 × 10(-3)) spanning a region of 4.9 Kb was identified, thereby providing additional information of the influence of those SNPs on Salmonella fecal shedding in pigs.
Asunto(s)
Derrame de Bacterias/genética , Polimorfismo de Nucleótido Simple/genética , Salmonella/fisiología , Sus scrofa/genética , Sus scrofa/microbiología , Receptor Toll-Like 4/genética , Animales , Área Bajo la Curva , Recuento de Colonia Microbiana , Haplotipos/genética , Fenotipo , Salmonella/crecimiento & desarrolloRESUMEN
Galectins and collectins are proteins classified in the lectin family that have the ability to recognize molecular patterns associated with pathogens. Studies on cattle have demonstrated high expression of these proteins during infection with gastrointestinal nematodes. The aim of this study was to investigate whether the level of Haemonchus contortus infection would alter the expression of galectins (Gal11 and Gal14) and collectins (SPA and CGN) in sheep. Twelve Corriedale sheep exposed to natural infection with nematodes were divided into two groups: group 1 (G1, n = 7) and group 2 (G2, n = 5), with low and high parasite burdens, respectively, based on fecal egg counts and abomasal parasite counts. The fecal egg counts and abomasal parasite counts were significantly different (p < 0.05) between the groups. Galectin and collectin gene expression was observed in all sheep abomasal samples. However, animals with lower infection levels showed lower expression of the genes Gal14, SPA and CGN (p < 0.05). Expression of lectins was associated with the abomasal H. contortus burden, thus suggesting that these proteins may have a role in controlling of this infection.
Colectinas e galectinas são proteínas da família das lectinas que possuem a capacidade de reconhecer padrões moleculares associados aos patógenos. Estudos em bovinos têm demonstrado a alta expressão dessas proteínas durante a infecção por nematoides gastrintestinais. O objetivo deste estudo foi investigar se o nível de infecção de Haemonchus contortus altera a expressão de colectinas (SPA e CGN) e galectinas (Gal11 e Gal14) de ovinos. Doze ovinos da raça Corriedale expostos a infecção natural com nematoides foram separados em dois grupos: grupo 1 (G1, n=7) com menor grau de parasitismo; e grupo 2 (G2, n=5) com maior grau, a partir da contagem do número de parasitos recuperados do abomaso e OPG. A contagem de OPG e de parasitos recuperados do abomaso dos grupos G1 e G2 apresentaram diferença estatística (p<0,05). A expressão dos genes de colectinas e galectina foi observada em todas as amostras de abomaso dos ovinos, porém animais com menor grau de infecção apresentaram menor expressão dos genes de Gal14, SPA e CGN (p<0,05). A expressão de lectinas foi associada ao número de H. contortus encontrados no abomaso de ovinos, indicando um possível papel dessas proteínas no controle da infecção.
Asunto(s)
Animales , Masculino , Enfermedades de las Ovejas/metabolismo , Colectinas/biosíntesis , Galectinas/biosíntesis , Hemoncosis/veterinaria , Ovinos , Expresión Génica , Colectinas/genética , Galectinas/genética , Hemoncosis/genética , Hemoncosis/metabolismo , HaemonchusRESUMEN
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.
Asunto(s)
Enfermedades de los Bovinos/inmunología , Inmunoglobulina G/biosíntesis , Ixodidae/inmunología , Infestaciones por Garrapatas/veterinaria , Animales , Bovinos , Enfermedades de los Bovinos/parasitología , Susceptibilidad a Enfermedades/inmunología , Susceptibilidad a Enfermedades/veterinaria , Femenino , Inmunidad Innata/inmunología , Inmunoglobulina A/biosíntesis , Inmunoglobulina A/sangre , Inmunoglobulina E/biosíntesis , Inmunoglobulina E/sangre , Inmunoglobulina G/sangre , Masculino , Saliva/inmunología , Infestaciones por Garrapatas/inmunología , Infestaciones por Garrapatas/parasitologíaRESUMEN
A influência da herança citoplasmática sobre características reprodutivas foi investigada em bovinos Brangus-Ibagé (3/8 Nelore x 5/8 Aberdeen Angus). Os efeitos genéticos aditivos foram responsáveis por 12% ± 11% da variação fenotípica observada no primeiro intervalo entre partos, mas esta contribuição decresceu para zero quando todos os intervalos entre partos (IEP) foram considerados. A herdabilidade da idade para o primeiro parto (IPP, em dias) foi estimada em 0,19 ± 0,09. A linhagem mitocondrial teve um efeito negligenciável na variância fenotípica do intervalo entre partos (0,0 ± 0,02), peso do terneiro ao nascer (0,0 ± 0,01) e peso da vaca ao parto (0,0 ± 0,01). No entanto, para a idade ao primeiro parto, a linhagem mitocondrial contribuiu com 0,15 ± 0,07 da variação total. O peso da vaca ao parto teve efeito linear significante em IEP e IPP. Três mutações na região D-loop do mtDNA afetaram significantemente o IEP (T®C nos sítios 16.113 e 16.119) ou o peso do terneiro ao nascer (T®C no sítio 16.113). As variantes C diminuíram o intervalo entre partos (29 e 32 dias, respectivamente) e aumentaram o peso do terneiro (0,6kg). Embora os efeitos sejam pequenos, a seleção de fêmeas portadoras dessas mutações poderia melhorar o desempenho reprodutivo e o desenvolvimento deste rebanho.