Molecular marker sequences of cattle Cooperia species identify Cooperia spatulata as a morphotype of Cooperia punctata

The genus Cooperia includes important parasites of ruminants and currently contains 34 accepted species. However, even for those species infecting livestock, there is a considerable lack of molecular information and many species are only identifiable using subtle morphological traits. The present study aimed to provide molecular data to allow diagnosis of Cooperia species infecting cattle. Partial sequences of two mitochondrial (cytochrome oxidase 2, 12S rRNA gene) and two nuclear genes (isotype 1 β tubulin gene including two introns, internal transcribed spacers (ITS) were obtained from morphologically identified specimens of Cooperia pectinata, Cooperia punctata and Cooperia spatulata as well as from larvae of pure Cooperia oncophora and C. punctata laboratory isolates. Pairwise identity of ITS-2 sequences was very high and it was the only region able to identify a specimen as Cooperia sp. However, the ITS-2 was unreliable for diagnosis at the species level. All other marker sequences could not unequivocally be allocated to the genus Cooperia but allowed clear species identification with the exception of the pair C. punctata/C. spatulata for which no significant differences were found for any marker sequence. Maximum-likelihood phylogenetic analyses of individual genes as well as a multi-locus analysis covering all four sequences confirmed that specimen identified as C. spatulata were randomly distributed throughout the C. punctata cluster and formed no group of their own. In contrast, the other Cooperia species formed clearly separated and statistically supported clusters. These data indicate that C. spatulata is most likely only a morphotype of C. punctata and the name should be considered a synonym. Combinations of nuclear and mitochondrial markers should be used to identify morphotypes or cryptic species to benefit from excellent barcoding properties of the latter but allowing proper phylogenetic analyses and controlling for lineage sorting that might occur for mitochondrial genotypes within a species

In vitro larvicidal effect of a hydroalcoholic extract from Acacia cochliacantha leaf against ruminant parasitic nematodes

Larvicidal effect was evaluated using acacia cochliacantha leafThe aim of this study was to evaluate the in vitro lethal effect of a hydroalcoholic extract (HAE) from Acacia cochliacantha leaf against three gastrointestinal nematodes species (Haemonchus contortus, H. placei and Cooperiapunctata) of domestic ruminants. The HAE was assessed using five concentrations: 100, 125, 175, 150 and 200 mg/ml; 0.5% Ivermectin was used as a positive control and distilled water, as negative control. The data were normalized using the square root and analysed with a completely randomized design through ANOVA analysis using the general linealmodel (GLM) of the SAS program. The HAE tannin conten was determined through spectrophotometry (UV-visible) and the other major phenols, were identified by chromatographic processes. The results showed an in vitro larvicidal activity of the HAE against the three assessed nematode species with all assessed concentrations. A clear HAE increased concentration dependence effect was observed. The highest activity of the HAE was obtained at the highest concentration (close to 100%, P < 0.05). This result was similar to the one obtained with Ivermectin. On the other hand, the chemical analysis of HAE showed the presence of tannins, caffeoyls and coumaroyl derivates and quercetin as the main compounds. The results suggest that the HAE from this plant species pos- sess in vitro anthelmintic properties. The identified com- pounds in this study would good candidates for further in vivo researches.This study received financial support from CONACYT-Mexico (Project number 251244), INIFAP (Project number 9454133183, through the Researcher Retaining Program)

Implications of between-isolate variation for climate change impact modelling of Haemonchus contortus populations

The impact of climate change on parasites and parasitic diseases is a growing concern and numerous empirical and mechanistic models have been developed to predict climate-driven spatial and temporal changes in the distribution of parasites and disease risk. Variation in parasite phenotype and life-history traits between isolates could undermine the application of such models at broad spatial scales. Seasonal variation in the transmission of the haematophagous gastrointestinal nematode Haemonchus contortus, one of the most pathogenic helminth species infecting sheep and goats worldwide, is primarily determined by the impact of environmental conditions on the free-living stages. To evaluate variability in the development success and mortality of the free-living stages of H. contortus and the impact of this variability on future climate impact modelling, three isolates of diverse origin were cultured at a range of temperatures between 15°C and 37°C to determine their development success compared with simulations using the GLOWORM-FL H. contortus model. No significant difference was observed in the developmental success of the three isolates of H. contortus tested, nor between isolates and model simulations. However, development success of all isolates at 37°C was lower than predicted by the model, suggesting the potential for overestimation of transmission risk at higher temperatures, such as those predicted under some scenarios of climate change. Recommendations are made for future climate impact modelling of gastrointestinal nematodes

Evaluación in vitro e in vivo de plantas bioactivas sobre el control de cooperia punctata en bovinos

Ciencias de la Producción y de la Salud Anima

Scatter plot of spicule length of <i>Cooperia pectinata</i>, <i>Cooperia punctata</i> and <i>Cooperia spatulata</i> collected from a calf in Brazil.

<p>Mean spicule length of 30 <i>C</i>. <i>pectinata</i>, 43 <i>C</i>. <i>punctata</i> and 12 <i>C</i>. <i>spatulata</i> was plotted and compared using One-Way-ANOVA and Bonferroni post-hoc tests. All groups were significantly different with p<0.0001 but it must be kept in mind that the <i>C</i>. <i>punctata</i> worms were preselected according to a smaller over-all length which clearly introduces a systematical bias. Individual values are shown in black while mean ± SD are indicated in red.</p

Ventral view of spicules of <i>Cooperia</i> spp. from a Brazilian calf.

<p>(A) <i>Cooperia pectinata</i> males have large spicules with corrugated edges in the middle third (arrow). (B) <i>Cooperia punctata</i> spicules show a large concavity near the middle, which has a distinct border and a lateral narrow projection. (C) <i>Cooperia spatulata</i> spicules have a small concavity (small arrow) and a large ventral flange (large arrow).</p

Recent Advances in the Control of Endoparasites in Ruminants from a Sustainable Perspective

Consumer awareness of animal welfare and environmental health has led to a plateau level of global consumption putting serious pressure on the livestock industry [...

Pairwise sequence identities for intra- and inter-species comparisons at the mitochondrial <i>cox-2</i> locus.

<p>Pairwise identities are plotted as box plots for a partial cytochrome oxidase 2 (<i>cox-2</i>) sequence (A) and the third codon position of the same fragment (B). Whiskers indicate 5–95% quantiles with outliers represented by dots. The mean of each dataset is marked by a cross. Intra-species comparisons and inter-species comparisons within the genus <i>Cooperia</i> are shown in black and blue, respectively. Comparisons between <i>C</i>. <i>punctata</i> and <i>C</i>. <i>spatulata</i> are drawn in red. All comparisons between a <i>Cooperia</i> sequence and a representative <i>H</i>. <i>contortus</i> sequence are plotted in green. All accession numbers for sequences used in the analysis are given in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0200390#pone.0200390.s002" target="_blank">S2 Table</a>. Index letters (a-d in panel A and a-f in panel B) are used to indicate significant differences between groups as revealed by Kruskal-Wallis test followed by a Dunn’s post hoc test between all <i>Cooperia</i> groups. Only groups sharing no index letter are significantly different (p<0.05).</p