ABSTRACT
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.
ABSTRACT
INTRODUCTION: Crotalaria spectabilis is an important species used as a pre-plant cover for soybean crops to control the proliferation of endoparasitic nematodes. Species from the Crotalaria genus are known for presenting pyrrolizidine alkaloids (PAs) in their composition, however, C. spectabilis is still considered chemically under-explored. OBJECTIVE: The goal of this manuscript is the development and validation of a method for PAs and flavonoids identification and quantification of C. spectabilis seeds and leaves, a toxic plant used for nematode proliferation control in soil, especially in soybean crops. MATERIALS AND METHODS: Seeds and leaves extracts were analysed by high-performance liquid chromatography coupled to tandem mass spectrometry (HPLC-MS/MS) for the identification of the compounds. RESULTS: PAs and phenolic compounds could be identified in both samples based on the MS/MS fragmentation pattern. Molecular formulas of the annotated compounds were confirmed by ultra-high-performace liquid chromatography-quadrupole time-of-flight (UHPLC-QToF), and monocrotaline could also be confirmed by standard comparison. The quantification of monocrotaline was performed by HPLC-MS/MS, resulting in 123 times higher monocrotaline content in seeds than in the leaves, which could explain its efficiency in combating nematode proliferation in soil. CONCLUSION: This was the first report of phenolic compounds in C. spectabilis. The current study highlights the importance of C. spectabilis for nematode control due to the presence of toxic PAs, and the employment of analytical techniques for identification and quantification of compounds present in the extracts.
Subject(s)
Crotalaria , Pyrrolizidine Alkaloids , Chromatography, High Pressure Liquid , Monocrotaline , Plant Extracts , Seeds , Tandem Mass SpectrometryABSTRACT
A study was conducted from December to April 2013, with the aim of evaluating a system of selective antiparasitic treatments using the FAMACHA© color chart compared with a conventional suppressive deworming system every 30 days in Pelibuey ewes during lactation. For the study, 54 ewes were used. They were randomly divided into two groups: FAMACHA and chemical treatments. The ewes in the first group received selective treatment depending on the ocular mucosa coloration (FAMACHA) and body condition score (BCS), while in the second group (chemical) all the animals remained under routine deworming every 30 days. Fecal nematode egg counts, proportion of third-stage trichostrongylid larvae, body condition, coloration of the ocular mucosa, and packed cell volume in the ewes were determined, while in lambs only body weight (BW) was recorded. No significant differences (p > 0.05) were observed in any of the studied variables between groups; however, the use of antiparasitic drugs was reduced during the experimental period in the FAMACHA group and no deaths of lambs or ewes were recorded. The results indicate that during the lactation of ewes, a strategy of selective treatments can be implemented without showing deterioration in major health and productive parameters of these animals.