Rhipicephalus (Boophilus) microplus is responsible for reducing animal welfare, causing a drop in productive performance and transmitting hemoparasites. The main strategy of tick control is application of synthetic acaricides. However, parasite resistance to these compounds is a major concern. Therefore, the acaricidal and repellent in vitro effect of the Ocotea elegans essential oil on larvae and adult females of R. (B.) microplus were evaluated. The larval packet test (LPT), larval repellency test (RT) and adult immersion test (AIT) were performed. The essential oil was analyzed by gas chromatography (GC/FID) and the structure of the oil's major constituent (92.2% sesquirosefuran) was elucidated by nuclear magnetic resonance. In the AIT, efficacy higher than 90% was detected from the concentration 25 mg/mL upward. In both LPTs performed after 48 h, only the 100 mg/mL concentration resulted in mortalities above 70%. On the other hand, the essential oil caused an average of 95.8% repellency from 0.78 to 100 mg/mL. The LC in the two LPT (48 h) tests were 59.68 and 25.59 mg/mL, respectively. The LC and LC in the AIT were 4.96 and 17.37 mg/mL, and in the RT they were 0.04 and 1.24 mg/mL respectively. We conclude that the essential oil of O. elegans leaves has a significant acaricidal effect on engorged females and on larval repellency of R. (B.) microplus ticks, and can be a promising alternative for the control of this ectoparasite.
Due to the great economic impact of Haemonchus contortus on sheep farming, there is an increasing number of studies addressing host resistance against this nematode, including identification of directly related immune mechanisms. In this context, relative gene expression by RT-qPCR have been largely used, due to its rapidity, high sensitivity, specificity, and reproducibility. Although, appropriate reference gene selection is crucial for accurate interpretation of results. In this study, five reference genes (GAPDH, G6PDH, YWHAZ, ACTB, and B2M) were tested for expression stability in abomasum (fundic and pyloric regions) and abomasal lymph nodes of Morada Nova sheep classified as resistant (n = 5) or susceptible (n = 5) to H. contortus infection in a flock of 151 animals. GAPDH combined with YWHAZ were selected as reference genes for abomasal fundic region and abomasal lymph nodes, whereas YWHAZ was the most stable gene for abomasal pyloric region. These genes presented the lowest intra- and inter-group variations and, consequently, highest stability. In contrast, expression of G6PDH was the least stable in all tissues. The impact of reference gene selection was demonstrated by relative quantification of a pro-inflammatory cytokine (TNFα) in abomasal fundic region. Significant differences in TNFα expression levels between resistant and susceptible groups were only observed when the most stable genes (GAPDH combined with YWHAZ) or GAPDH were used as reference genes, whereas no significant differences were observed when other tested reference genes were used. It was demonstrated that normalization of expression data using inappropriate reference genes may significantly influence interpretation results.
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