Assembly and Analysis of Haemonchus contortus Transcriptome as a Tool for the Knowledge of Ivermectin Resistance Mechanisms.

Haemonchus contortus (Hc) is an important parasitic nematode of small ruminants. In this study we assembled the transcriptome of Hc as a model to contribute to the knowledge about the profile of the differential gene expression between two Mexican Hc strains under different anthelmintic resistance statuses, one susceptible and the other resistant to ivermectin (IVMs and IVMr, respectively), in order to improve and/or to have new strategies of control and diagnosis. The transcript sequence reads were assembled and annotated. Overall, ~127 Mbp were assembled and distributed into 77,422 transcript sequences, and 4394 transcripts of the de novo transcriptome were matched base on at least one of the following criteria: (1) Phylum Nemathelminthes and Platyhelminthes, important for animal health care, and (2) ≥55% of sequence identity with other organisms. The gene ontology (GO) enrichment analysis (GOEA) was performed to study the level of gene regulation to IVMr and IVMs strains using Log Fold Change (LFC) filtering values ≥ 1 and ≥ 2. The upregulated-displayed genes obtained via GOEA were: 1993 (for LFC ≥ 1) and 1241 (for LFC ≥ 2) in IVMr and 1929 (for LFC ≥ 1) and 835 (for LFC ≥ 2) in IVMs. The enriched GO terms upregulated per category identified the intracellular structure, intracellular membrane-bounded organelle and integral component of the cell membrane as some principal cellular components. Meanwhile, efflux transmembrane transporter activity, ABC-type xenobiotic transporter activity and ATPase-coupled transmembrane transporter activity were associated with molecular function. Responses to nematicide activity, pharyngeal pumping and positive regulation of synaptic assembly were classified as biological processes that might be involved in events related to the anthelmintic resistance (AR) and nematode biology. The filtering analysis of both LFC values showed similar genes related to AR. This study deepens our knowledge about the mechanisms behind the processes of H. contortus in order to help in tool production and to facilitate the reduction of AR and promote the development of other control strategies, such as anthelmintic drug targets and vaccines.

Detection and genetic characterization of feline retroviruses in domestic cats with different clinical signs and hematological alterations.

Feline leukemia virus (FeLV) and feline immunodeficiency virus (FIV) are globally distributed retroviruses that infect domestic cats and cause various syndromes that can lead to death. The aim of this study was to detect and genotype feline retroviruses in Mexican domestic cats. We used PCR assays to identify proviral DNA and viral RNA in 50 domestic cats with different clinical signs and hematological alterations. Endogenous FeLV (enFeLV) was identified in the genomic DNA of all cats in the study, and we detected transcripts of the LTR region of enFeLV in 48 individuals. Exogenous FeLV (exFeLV) was found in 13 cats. Furthermore, we detected FIV proviral DNA in 10 cats. The enFeLV sequences were shown to be the most variable, while the exFeLV sequences were highly conserved and related to previously reported subgroup A sequences. Sequencing of the FIV gag gene revealed the presence of subtype B in the infected cats.

Effect of entomopathogenic native fungi from paddock soils against Rhipicephalus microplus larvae with different toxicological behaviors to acaricides.

The objectives of this study are to: (1) evaluate the in vitro acaricidal effect of 54 Metarhizium anisopliae strains, six Beauveria bassiana strains and one Purpureocilium lilacinum strain, against the larvae of two populations of Rhipicephalus microplus (multi-resistant and susceptible to chemical acaricides); and (2) determine the lethal concentrations required to eliminate the 50% (LC50) and 99% (LC99) of larvae through the use of entomopathogenic fungi (EF) with high acaricidal effects. The mortality percentage was evaluated by larval immersion tests at a dose of 1 × 108 conidia/mL for each fungal strain. For calculating LC50 and LC99, four doses (1 × 108, 1 × 107, 1 × 106 and 1 × 105) were used. Nine strains of M. anisopliae and the P. lilacinum strain showed a high mortality percentage in the R. microplus larvae of both populations. The best strains that showed the lowest values of LC50 and LC99 for tick elimination were MaV50 and PlV01. In conclusion, several strains of entomopathogenic fungi showed a high acaricidal effect against the R. microplus larvae of both populations, suggesting that these fungi might be a promissory adjuvant in the control of R. microplus, including those who are resistant. Finally, the discovery of a P. lilacinum strain with a high acaricidal effect is also reported.

Molecular characterization of the ORF2 of Torque teno sus virus 1a and Torque teno sus virus 1b detected in cases of postweaning multisystemic wasting syndrome in Mexico.

Worldwide Torque teno sus virus (TTSuV, genus Iotatorquevirus) species have been regarded as possible agents associated with porcine circovirus-associated disease. Iotatorquevirus species possess high genomic variability, suggesting that diverse genotypes are widely geographically distributed. In this study, we validated the genomic variability of Iotaroquevirus species in pigs with postweaned multisystemic wasting syndrome. Genomic DNA from nine TTSuV1a-positive tissues and 15 TTSuV1b-positive tissues was used to amplify the complete ORF2 of each species by nested PCR to perform a molecular characterization. It was found that Mexican TTSuV1a sequences belong to genotype B, sharing phylogenetic origin, high nucleic acid and amino acid sequence similarity and dominant epitope conformation with commercially linked countries, such as the United States, Canada and China, whereas the Mexican TTSuV1b sequences belong to genotype A, being more divergent among each other and displaying low nucleotide identity with worldwide genotype A sequences. In both Iotatorquevirus species, a PTPase-like signature motif was identified in the predicted amino acid sequence, being more conserved for Mexican TTSuV1b sequences than for Mexican TTSuV1a sequences, in which several substitutions were observed. These changes may influence the conformation of dominant epitopes as different arrays were determined among TTSuV1a genotypes. ORF2 variability may account for pathogenic differences by modifying viral replication and immune response, as depicted for human TTV.

Genetic polymorphism of the alpha s1-casein locus in five populations of goats from Mexico

With the objective of estimating allele frequencies, and testing for population divergence for the CSN1S1 locus, genotypes of animals from five goat populations; Saanen (n = 97), Alpine (n = 81) Toggenburg (n = 92), local goats with external appearance similar to the Murciana-Granadina breed from Central Mexico (n = 26) and heterogeneous local animals denominated Mosaico Lagunero (n = 30), from Northern Mexico, were identified using PCR and Xmn1 PCR-RFLP methodology. For Saanen, Alpine and Toggenburg, the sum of E and F alleles had the largest frequencies (from 0.468 to 0.789), while for the groups local Murciana-Granadina and Mosaico Lagunero the sum of the most frequent allelic groups (A and B), were 0.385 and 0.533 respectively. Both local Murciana-Granadina and Mosaico Lagunero populations showed heterozygote excess (P < 0.08). The percentage of the total genetic variation (F ST) explained by population differences was 5.16. There was genetic differentiation for most pair comparisons between populations (P < 0.05), excepting for Alpine versus Toggenburg, and Toggenburg versus Mosaico Lagunero (P > 0.05). For Saanen and Alpine the frequencies of alleles E and F were similar to the same breeds previously analyzed in Europe. Therefore there are opportunities of increasing the frequency of the strong alleles for protein content Gene Assisted Selection (GAS) in these two breeds. For Toggenburg the most frequent allelic groups were F (0.32) and B (0.21). Results indicate differentiation between most populations for this locus. Moreover, heterozygote excess in local populations indicated breed admixture.

Hacia un proyecto nacional e investigación en genomas de animales domésticos / Towards a national research genomic project on domestic animals

La investigación científica orientada a identificar, estudiar y aprovechar genes, así como a entender la organización y evolución del genoma, ha llevado a proyectos genómicos de gran escala como el Proyecto del Genoma Humano y de otros organismos de interés científico y económico para el hombre. En México es posible conformar este tipo de proyectos de forma original, con base en la extensa diversidad genética presente en poblaciones biológicas nativas que componen el patrimonio genómico nacional y utilizando diversos recursos disponibles en el dominio público. Los animales domésticos nativos en México contituyen un recurso genético valioso y único para identificar genes importantes para la industria pecuaria y la biotecnología. Sin embargo, estas variedades están amenazadas debido a las políticas de absorción y reemplazo por variedades comerciales mejoradas provenientes de países más desarrollados. La investigación genómica en animales domésticos puede ser importante para desarrollar y mantener una ganadería competitiva en una economía de mercado mundial. Por ello, se propone un proyecto genómico en animales domésticos en México cuyas metas serían: la preservación, estudio y la explotación de recursos genéticos autóctonos; la identificación, clonación y estudio de genes importantes desde el punto de vista comercial y médico, y la transferencia de tecnología a la industria animal local. Este proyecto puede ser el fundamento para estimular el desarrollo de una industria pecuaria nacional más moderna, competitiva e independiente, más adecuada a condiciones propias tanto ecológicas y económicas, como culturales y de mercado