Single Amino Acid Polymorphisms in the Fasciola hepatica Carboxylesterase Type B Gene and Their Potential Role in Anthelmintic Resistance.

The expression of the Fasciola hepatica carboxylesterase type B (CestB) gene is known to be induced upon exposure to the anthelmintic triclabendazole (TCBZ), leading to a substantial rise in enzyme-specific activity. Furthermore, the nucleotide sequence of the CestB gene displays variations that can potentially result in radical amino acid substitutions at the ligand binding site. These substitutions hold the potential to impact both the ligand-protein interaction and the catalytic properties of the enzyme. Thus, the objective of our study was to identify novel CestB polymorphisms in TCBZ-resistant parasites and field isolates obtained from a highly endemic region in Central Mexico. Additionally, we aimed to assess these amino acid polymorphisms using 3D modeling against the metabolically oxidized form of the anthelmintic TCBZSOX. Our goal was to observe the formation of TCBZSOX-specific binding pockets that might provide insights into the role of CestB in the mechanism of anthelmintic resistance. We identified polymorphisms in TCBZ-resistant parasites that exhibited three radical amino acid substitutions at positions 147, 215, and 263. These substitutions resulted in the formation of a TCBZSOX-affinity pocket with the potential to bind the anthelmintic drug. Furthermore, our 3D modeling analysis revealed that these amino acid substitutions also influenced the configuration of the CestB catalytic site, leading to alterations in the enzyme's interaction with chromogenic carboxylic ester substrates and potentially affecting its catalytic properties. However, it is important to note that the TCBZSOX-binding pocket, while significant for drug binding, was located separate from the enzyme's catalytic site, rendering enzymatic hydrolysis of TCBZSOX impossible. Nonetheless, the observed increased affinity for the anthelmintic may provide an explanation for a drug sequestration type of anthelmintic resistance. These findings lay the groundwork for the future development of a molecular diagnostic tool to identify anthelmintic resistance in F. hepatica.

Morphological characterization and first molecular identification of the eleven Eimeria species that infect sheep from Mexico.

Within livestock production, enteric diseases play an important role, since they cause severe economic losses due to mortality, growth depression, and reduction in the conversion rate. Coccidiosis caused by Eimeria spp. is a parasitic disease of high morbidity that affects various animal species, including sheep. In sheep, eleven species of Eimeria have been identified mainly through microscopical identification of the oocysts; however, this technique has certain limitations that make it difficult to identify the different Eimeria species. The objective of the present study was to morphologically identify the eleven species of Eimeria that infect sheep in the southeastern region of the State of Mexico, as well as obtain the partial sequence of the ITS-1 rRNA region of each species and analyze it phylogenetically. A total of 412 samples were collected from the 13 municipalities that comprise the region I of the State of Mexico, out of which, 40 had approximately 80% of a single Eimeria species. Among these, the eleven Eimeria species reported in sheep were identified. The phylogenetic analysis showed that the species reported in this study are associated with those reported in rabbits, bovines, and birds. It is suggested that the phylogenetic division of sheep in two clades may be associated with the presence or absence of the residual body. It is proposed that the present methodology can be used effectively for diagnosis and to obtain information about the epidemiology of ovine coccidial infection. The results obtained in this study constitute the first report of the ITS-1 region of the eleven Eimeria species that infect sheep worldwide.

Influence of levamisole and Freund's adjuvant on mouse immunisation with antigens of adults of the liver fluke Fasciola hepatica Linnaeus, 1758.

We have studied the influence of both levamisole (AL) and Freund's adjuvant (AF) on the immunisation of mice with the secretory antigens of adults of the liver fluke Fasciola hepatica Linnaeus, 1758. Total IgG antibodies were detected in all groups where the F. hepatica antigen was administered, been levels of IgG1 increased respect to IgG2a antibodies. During immunisation, IL-4 and IFN-γ were only detected in AL and AF groups, but after infection, IL-4 boosted in all groups. IFN-γ increased two fold in AF and AL groups compared to the saline solution (AS) group. Worm recovering was of 32-35% in groups administered without antigen whereas in AS, AL and AF groups recovering was of 25%, 12% and 8%, respectively. Macroscopical lesions in the liver were scarce in AL and AF groups. Our data suggest that immunisation of mice with antigens of F. hepatica enhances the immune response avoiding both liver damage and worm establishment after challenge infection. The murine model of fasciolosis has appeared to be useful to elucidate the mechanism by which the parasite modulates immune responses toward a Th2 type but also the development of Th1 type-inducing vaccines.

Comparación del efecto protector de una vacuna y un ionóforo contra coccidias en pollos / Comparison between vaccine protector effect and a ionofore against coccidiosis in chickens

Se realizó un estudio con pollos de engorda alojados en piso, con el objeto de comparar la eficacia de una vacuna con Eimeria tenella, E. necatrix, E. acervulina y E. máxima contra el ionóforo narazina en el alimento en la prevención de coccidiosis. Se emplearon 320 pollos, divididos en cuatro lotes, cada uno con 4 repeticiones de 20 aves. El lote a recibió alimento sin coccidiostato. A las tres semanas de iniciado el experimento, los tres fueron confrontados con un inóculo formado con una mezcla de E. tenella, E. necatrix, E. acervulina y E. máxima. El lote D fue testigo negativo. Las variables registradas fueron peso, consumo de alimentos, índice de ooquistes en heces y cama, lesiones, mortalidad y pigmentación de los tarsos. Los resultados mostraron diferencias significativas entre las 0-4 semanas en la ganancia de peso y conversión alimenticia, la mejor ganancia de peso correspondió al grupo D, seguida del A, B y C, respectivamente. La conversión alimenticia fue más eficiente para el grupo D. Los datos de 4 a 7 y de 0 a 7 semanas no mostraron diferencias estadísticas en ninguno de los parámetros analizados. El índice anticoccidial fue el mejor para el grupo A seguido del B y C, respectivamente, lo cual indica que el inmunógeno proporcionó una adecuada protección contra la coccidiosis aviar