RESUMO

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.

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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.

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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.(AU)

Haemonchus contortus provoca grandes prejuízos à ovinocultura. Visando reduzir a utilização de ovinos hospedeiros e produzir estágios parasitários em larga escala, um protocolo para o cultivo in vitro por 42 dias de larvas de terceiro estádio de H. contortus foi realizado em meio Eagle, modificado por Dulbecco (DMEM). No cultivo sem células, as larvas foram mantidas a 39,6°C e incubadas em DMEM ácido (pH 6,1) por 3 ou 6 dias com 4Δ-ácido dafacrônico seguido por DMEM pH 7,4 suplementado ou não com reagente de Fildes. Em DMEM pH 7,4 a 37°C, a suplementação com células Caco-2 foi comparada à suplementação com Fildes. No Dia 14, as taxas de desenvolvimento até o quarto estádio larvar (L4) foram similares em meio ácido sem células suplementado (86,8-88,4%) ou não (74,4-77,8%) com Fildes e em co-cultura com células Caco-2 (92,6%), e superiores ao desenvolvimento em DMEM pH 7,4 com Fildes (0,0%). No Dia 21, o co-cultivo com células Caco-2 resultou em maior diferenciação (25,0%) e menor degeneração (13,9%) das larvas em comparação ao meio ácido (1,5-8,1% e 48,6-69,9%, respectivamente). Este é o primeiro relato de cultivo in vitro prolongado de H. contortus em meio comercial em co-cultivo com células Caco-2. Apesar da ausência de progressão até o estágio adulto, o co-cultivo com células Caco-2 resultou em diferenciação de L4 e manutenção da viabilidade das larvas de H. contortus por até 28 dias in vitro.(AU)

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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.

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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.

RESUMO

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.

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The record of occurrence of acanthocephalan parasites on tambaqui farms in Northern Brazil has increased in recent years along with reports of associated economic losses, justifying further epidemiological studies of this parasite. As such, this study evaluated the occurrence of the acanthocephalan Neoechinorhynchus buttnerae and the parasite-host relationship in tambaqui (Colossoma macropomum) from three fish farms in the Amazonian town of Rio Preto da Eva, Brazil. Fish from the farms (N = 18 per farm) were collected for parasitological examination, determination of the relative condition factor and correlation tests between the parameters assessed. Of the fish examined, 37 were parasitized with N. buttnerae (68.5% prevalence), with intensity of infection variation ranging from 1 to 1,219 specimens per fish (12,279 specimens counted). In two farms was detected the high mean intensity of infection, 230.8 and 451.3 parasites per fish, with a total count of 4,155 and 8,123 parasites, respectively, whereas the third farm had a single fish infected with only one specimen. The weight-length ratio equation indicated that the fish were growing at the same rate in weight and length. It was observed significant correlation between parasite intensity and fish length, as well as significant negative correlation with parasite intensity and condition factor. The gastrointestinal tract contained ostracod invertebrates, which are intermediate N. buttnerae hosts. The high N. buttnerae intensity affect their physiological condition as suggested by the negative correlation between parasitic intensity and condition factor, emphasizing the need to implement suitable prophylactic measures to prevent infection by the parasite.(AU)

Nos últimos anos tem crescido o registro de ocorrências de parasitos acantocéfalos em pisciculturas de tambaqui da região Norte do Brasil, com relatos de perdas econômicas, justificando assim a necessidade de aprofundar dados epidemiológicos desta doença parasitária. Assim, o objetivo deste estudo foi avaliar a ocorrência do acantocéfalo Neoechinorhynchus buttnerae e a relação parasito-hospedeiro em tambaqui (Colossoma macropomum) de três pisciculturas do município de Rio Preto da Eva (AM), Brasil. Foram amostradas três pisciculturas no município alvo, coletando 18 peixes em cada uma, para análises parasitológicas e cálculo do fator de condição relativo e correlações associadas. Foram quantificados 12.279 espécimes de N. buttnerae em 37 peixes parasitados (prevalência 68,5%), com intensidade de infecção variando de 1 a 1.219 parasitos por peixe. Duas propriedades apresentaram as maiores intensidades médias de infecção, 230,8 e 451,3 parasitas por peixe; com 4.155 e 8.123 parasitos quantificados, respectivamente, enquanto na terceira foi encontrado apenas 1 parasito em 1 peixe infectado. A equação da relação peso-comprimento indicou que os peixes estavam crescendo na mesma proporção em peso e comprimento. Foi observada correlação significativa entre intensidade parasitária e comprimento dos peixes, assim como correlação negativa significativa entre intensidade parasitária e fator de condição. No trato gastrointestinal dos tambaquis foi observada a presença de invertebrados ostracodas; hospedeiros intermediários desta espécie de acantocéfalo. Portanto, a alta carga parasitária de N. buttnerae pode ter comprometido o desempenho dos peixes e afetado a sua condição fisiológica como sugerido pela correlação negativa entre intensidade parasitária e fator de condição, e enfatiza-se a importância do emprego de medidas adequadas de profilaxia para prevenção de infecções por este parasito.(AU)

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RESUMO

The record of occurrence of acanthocephalan parasites on tambaqui farms in Northern Brazil has increased in recent years along with reports of associated economic losses, justifying further epidemiological studies of this parasite. As such, this study evaluated the occurrence of the acanthocephalan Neoechinorhynchus buttnerae and the parasite-host relationship in tambaqui (Colossoma macropomum) from three fish farms in the Amazonian town of Rio Preto da Eva, Brazil. Fish from the farms (N = 18 per farm) were collected for parasitological examination, determination of the relative condition factor and correlation tests between the parameters assessed. Of the fish examined, 37 were parasitized with N. buttnerae (68.5% prevalence), with intensity of infection variation ranging from 1 to 1,219 specimens per fish (12,279 specimens counted). In two farms was detected the high mean intensity of infection, 230.8 and 451.3 parasites per fish, with a total count of 4,155 and 8,123 parasites, respectively, whereas the third farm had a single fish infected with only one specimen. The weight-length ratio equation indicated that the fish were growing at the same rate in weight and length. It was observed significant correlation between parasite intensity and fish length, as well as significant negative correlation with parasite intensity and condition factor. The gastrointestinal tract contained ostracod invertebrates, which are intermediate N. buttnerae hosts. The high N. buttnerae intensity affect their physiological condition as suggested by the negative correlation between parasitic intensity and condition factor, emphasizing the need to implement suitable prophylactic measures to prevent infection by the parasite.

Nos últimos anos tem crescido o registro de ocorrências de parasitos acantocéfalos em pisciculturas de tambaqui da região Norte do Brasil, com relatos de perdas econômicas, justificando assim a necessidade de aprofundar dados epidemiológicos desta doença parasitária. Assim, o objetivo deste estudo foi avaliar a ocorrência do acantocéfalo Neoechinorhynchus buttnerae e a relação parasito-hospedeiro em tambaqui (Colossoma macropomum) de três pisciculturas do município de Rio Preto da Eva (AM), Brasil. Foram amostradas três pisciculturas no município alvo, coletando 18 peixes em cada uma, para análises parasitológicas e cálculo do fator de condição relativo e correlações associadas. Foram quantificados 12.279 espécimes de N. buttnerae em 37 peixes parasitados (prevalência 68,5%), com intensidade de infecção variando de 1 a 1.219 parasitos por peixe. Duas propriedades apresentaram as maiores intensidades médias de infecção, 230,8 e 451,3 parasitas por peixe; com 4.155 e 8.123 parasitos quantificados, respectivamente, enquanto na terceira foi encontrado apenas 1 parasito em 1 peixe infectado. A equação da relação peso-comprimento indicou que os peixes estavam crescendo na mesma proporção em peso e comprimento. Foi observada correlação significativa entre intensidade parasitária e comprimento dos peixes, assim como correlação negativa significativa entre intensidade parasitária e fator de condição. No trato gastrointestinal dos tambaquis foi observada a presença de invertebrados ostracodas; hospedeiros intermediários desta espécie de acantocéfalo. Portanto, a alta carga parasitária de N. buttnerae pode ter comprometido o desempenho dos peixes e afetado a sua condição fisiológica como sugerido pela correlação negativa entre intensidade parasitária e fator de condição, e enfatiza-se a importância do emprego de medidas adequadas de profilaxia para prevenção de infecções por este parasito.

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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.

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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.

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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.

Assuntos

RESUMO

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.(AU)

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.(AU)

Assuntos

RESUMO

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.

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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.

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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.

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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.

Assuntos

RESUMO

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.