Antibacterial potential of commercial and wild lactic acid bacteria strains isolated from ovine and caprine raw milk against Mycoplasma agalactiae

© 2023 Toquet, Bataller, Gomis, Sánchez, Toledo-Perona, De la Fe, Corrales and Gómez-Martín. This is an open-access article distributed under the terms of the Creative Commons CC-BY license, http://creativecommons.org/licenses/by /4.0/ This document is the Published version of a Published Work that appeared in final form in Frontiers in Veterinary Science. To access the final edited and published work see https://doi.org/10.3389/fvets.2023.1197701Introduction: The complexity of fighting contagious agalactia (CA) has raised the necessity of alternative antimicrobial therapies, such as probiotics. Lactic acid bacteria (LAB) are present in the mammary gland of small ruminants and their antimicrobial effect have been previously described against species like Mycoplasma bovis but never against Mycoplasma agalactiae (Ma). This in vitro study aims to evaluate the antimicrobial activity against Ma of ovine and caprine LAB strains and a human commercial probiotic (L2) of Lactobacillus spp. Methods: A total of 63 possible LAB strains were isolated from nine ovine and caprine farms in Spain, three isolates (33B, 248D, and 120B) from the 63 strains were selected, based on their capacity to grow in a specific medium in vitro, for an in vitro experiment to assess their antimicrobial activity against Ma in Ultra High Temperature (UHT) processed goat milk (GM). A women commercial vaginal probiotic was also included in the study. The inoculum of L2 was prepared at a concentration of 3.24 × 108  CFU/mL and the average concentration of the inoculum of the wild LAB varied from 7.9 × 107 to 8.4 × 108  CFU/mL. Results: The commercial probiotic L2 significantly reduced the concentration of Ma to 0.000 log CFU/mL (p < 0.001), strain 33B reduced it from 7.185 to 1.279 log CFU/mL (p < 0.001), and 120B from 6.825 to 6.466 log CFU/mL (p < 0.05). Strain 248D presented a bacteriostatic effect in GM. Moreover, the three wild strains and the commercial probiotic produced a significative reduction of the pH (p < 0.001). Discussion: This is the first in vivo report of the antimicrobial potential of LAB strains against Ma and its interaction. Our results support possible future alternative strategies to antibiotic therapy, previously not contemplated, to fight CA in small ruminants. Further studies are necessary to elucidate the action mechanisms through which these LAB are able to inhibit Ma and to assess the safety of using these strains in possible in vivo studies

The addition of Lactobacillus spp. negatively affects Mycoplasma bovis viability in bovine cervical mucus

© The Author(s). 2020 Open Access This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). This document is the Published version of a Published Work that appeared in final form in BMC Veterinary Research. To access the final edited and published work see https://doi.org/10.1186/s12917-020-02454-9Background Mycoplasma bovis is an important pathogen for the cattle industry worldwide causing significant economic losses. Several transmission routes, including those related to reproduction, have been described. Indeed, the pathogen can colonize the female reproductive tract after artificial insemination (AI) with contaminated semen. Lactobacillus spp.-based probiotics have been used for vaginal dysbiosis treatment in women and cows although their role in controlling cervico-vaginal infections due to M. bovis is unknown. The objective of the present work is to assess the viability of M. bovis (PG45, NCTC 10131) in experimentally contaminated cervical mucus after the addition of Lactobacillus spp. at different concentrations as a competing agent and pH acidifier. Results The addition of probiotic at a concentration higher than 108 colony forming units (CFU/mL had a detrimental effect (P < 0.05) on mycoplasma viability in cervical mucus. This coincided with a significant LAB growth and an important decrease in pH from 8.4 to 5.6 (P < 0.05). However, after the addition of less concentrated probiotic, M. bovis survival was not affected and there was no significant LAB growth despite the drop of pH from 8.4 to 6.73 (P < 0.05). Conclusion The addition of concentrations higher than 108 CFU/mL of Lactobacillus spp. negatively affects M. bovis viability in bovine cervical mucus under in vitro conditions. Although the effect observed on the pathogen viability seems to be related to the pH decrease after LAB proliferation in cervical mucus, further studies are necessary to elucidate if other factors are implicated. Nevertheless, the administration of Lactobacillus spp.-based probiotics might be used in the future to control M. bovis proliferation in the cervico-vaginal tract of cows

The Addition of Lactobacillus spp., Enrofloxacin or Doxycycline Negatively Affects the Viability of Mycoplasma bovis in Diluted Bovine Semen

© The Authors. 2020. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). This document is the Published version of a Published Work that appeared in final form in Animals. To access the final edited and published work see https://doi.org/10.3390/ani10050837Mycoplasma bovis is an important etiologic agent of bovine mycoplasmosis in cattle. Different transmission routes have been described, including those related to reproduction. The presence of mycoplasma in semen has led to its appearance in infection-free areas through artificial insemination (AI). Semen was recently reported to be the initial source of two M. bovis mastitis outbreaks in two closed dairy herds in Finland. This questions the effectiveness of the antimicrobials currently used in semen extenders to control the pathogens in contaminated semen. They should be re-evaluated, or alternative measures to antimicrobials should be tested to obtain M. bovis-free semen. This in vitro study aimed to assess different strategies to reduce the risk of transmission of M. bovis through AI technologies. The viability of M. bovis (PG45, NCTC 10131) in bull semen diluted (DS) in a Tris-citrate-fructose solution was tested, after the addition of enrofloxacin, doxycycline or a Lactobacillus spp.-based probiotic. The data show the susceptibility of the pathogen to the addition of 0.125 μg/mL of enrofloxacin or 0.0625 μg/mL of doxycycline and to the addition of the probiotic at a concentration of 3.24 × 106 colony forming units (CFU)/mL or 3.24 × 108 CFU/mL in DS. The Tris-citrate-fructose medium negatively affected the viability of M. bovis, although this effect was lower than that observed after the addition of the probiotic and antimicrobials (p < 0.05). Our results may support new strategies for reducing the risk of M. bovis transmission through AI

Avances en criopreservación y transferencia no quirúrgica de embriones porcinos = Advances in cryopreservation and non-surgical transfer of porcine embryos / Jesús Gomis Almendro.

Texto en inglés, resumen en español.Tesis-Universidad de Murcia.Consulte la tesis en: BCA. GENERAL. ARCHIVO UNIVERSITARIO. TM 4607

Avances en criopreservación y transferencia no quirúrgica de embriones porcinos = Advances in cryopreservation and non-surgical transfer of porcine embryos.

La industria porcina tiene un considerable interés en el uso de la transferencia de embriones, incluyendo la criopreservación de los mismos. Sin embargo, la utilización comercial de la transferencia de embriones en esta especie ha sido muy limitada debido principalmente a la necesidad de utilizar procedimientos quirúrgicos para la obtención y transferencia de los embriones, y a las dificultades encontradas para la criopreservación de dichos embriones mediante los protocolos tradicionales. Últimamente, se han abierto nuevas expectativas para la utilización comercial de esta tecnología gracias al desarrollo de un procedimiento para transferir embriones por vía no quirúrgica y al desarrollo de la vitrificación como método alternativo a los procedimientos tradicionales de criopreservación. El objetivo de esta tesis es mejorar la transferencia vía no quirúrgica de embriones vitrificados-calentados y, desarrollar un protocolo de vitrificación eficiente para los estadios tempranos de desarrollo de embriones porcinos obtenidos, pudiendo aumentar su viabilidad post-calentamiento. Palabras claves: Transferencia de embriones; porcino; Vitrificación; Blastocistos; MEM vitaminas; Forskolin; Delipidación; Desarrollo embrionario; Criopreservación; Cigoto; Embriones de 2 células; Embriones de 4 células; In vivo; Cerdo. Summary The swine industry has a considerable interest for the use of embryo transfer and cryopreservation since these technologies may allow the transport and storage of valuable genetic material with minimal health risks and cost. However, the commercial application of embryo transfer in pig has been traditionally limited due to the need of surgical transfer procedures and the difficulties for long-term storage of pig embryos by using traditional cryopreservation methods. However, new perspectives for embryo transfer and embryo cryopreservation have occurred with the development of a new device, for non-surgical embryo transfer and with the development of vitrification as an alternative to traditional cryopreservation methods. This thesis was designed to improve the non-surgical deep intrauterine embryo transfer procedure with vitrified embryos and develop an efficient and practical vitrification protocol for porcine embryos at very early developmental stages. Keywords: Embryo transfer; Porcine; embryos; Vitrification; Blastocysts; MEM vitamins; Forskolin; Delipidation; Embryo development; Cryopreservation; zygote; 2-Cell embryos; 4-Cell embryos; In vivo; Pi

PRESENTE Y FUTURO DEL DIAGNÓSTICO DE GESTACIÓN EN EL GANADO BOVINO

To diagnose pregnancy in cattle an accurate, safe, economical and early method must be used. Today, several methods are available in the cattle industry, while others are still under development. The objective of this study is to review bibliographically the methods for the diagnosis of pregnancy in bovines and their use in current and future livestock. On the one hand, direct methods such as transrectal palpation and transrectal ultrasound of the reproductive tract are considered the most widely used methods for the diagnosis of pregnancy in bovines today. These methods work well and are economically interesting. However, they are also invasive and require some experience to perform properly. In general, these methods are used three to four weeks after artificial insemination. Second, indirect methods for the diagnosis of pregnancy in bovines can be used for the early detection of pregnancy and therefore can save time. The presence of the embryo must be determined without directly visualizing the embryonic structures. Due to technological advances, indirect biochemical methods are being developed to detect certain molecules associated with pregnancy and predict the presence of an embryo at an even earlier stage, by quantifying hormones such as progesterone (P4) or glycoproteins associated with it. pregnancy (PAG) using rapid test kits. The detection of molecules such as interferon tau (IFNt) or microRNA (miRNA) and early pregnancy factors (EPF) are examples of these promising new methods still under development.Para llevar a cabo el diagnóstico de gestación en el ganado bovino, se debe utilizar un método preciso, seguro, económico y que se pueda realizar de manera temprana. Aunque varias técnicas están actualmente disponibles en el mercado, otras siguen todavía en desarrollo, siendo posibles herramientas diagnósticas a tener en cuenta en un futuro. Por ello, el presente trabajo tiene como objetivo hacer una revisión sobre las diversas técnicas de diagnóstico de gestación (DG) y su potencial uso a nivel comercial, tanto en el presente como en el futuro, de la ganadería bovina. Los métodos directos para el DG, como son la palpación directa o la ecografía del tracto reproductor vía transrectal, siguen siendo los más empleados en la actualidad durante el control gestacional en el ganado vacuno. Son técnicas diagnósticas con buenos resultados y son interesantes a nivel económico, pero son herramientas invasivas y requieren una cierta experiencia por parte del técnico que las realiza. En general, se aplican a partir de la 3ª-4ª semana posterior a la inseminación artificial. Por otro lado, los métodos indirectos de DG son menos invasivos que los directos. Mediante este tipo de diagnósticos vía indirecta se puede detectar la presencia o ausencia de un embrión, sin visualizar directamente estructuras gestacionales. Existen métodos indirectos físicos, como la vigilancia del retorno al estro, así como técnicas indirectas bioquímicas, que permiten evaluar mediante el uso de kits rápidos ciertas sustancias como la progesterona (P4) o las Glicoproteínas Asociadas a la Gestación (GPAG), producidas durante la gestación de forma temprana. Otras técnicas novedosas y prometedoras, pero que todavía se encuentran en desarrollo, son la evaluación de moléculas como el interferón tau (IFNt), los micro-ARN (miARN) y/o los Factores de Gestación Temprana (FGT)

Present and future of pregnancy diagnosis in cattle

Para llevar a cabo el diagnóstico de gestación en el ganado bovino, se debe utilizar un método preciso, seguro, económico y que se pueda realizar de manera temprana. Aunque varias técnicas están actualmente disponibles en el mercado, otras siguen todavía en desarrollo, siendo posibles herramientas diagnósticas a tener en cuenta en un futuro. Por ello, el presente trabajo tiene como objetivo hacer una revisión sobre las diversas técnicas de diagnóstico de gestación (DG) y su potencial uso a nivel comercial, tanto en el presente como en el futuro, de la ganadería bovina. Los métodos directos para el DG, como son la palpación directa o la ecografía del tracto reproductor vía transrectal, siguen siendo los más empleados en la actualidad durante el control gestacional en el ganado vacuno. Son técnicas diagnósticas con buenos resultados y son interesantes a nivel económico, pero son herramientas invasivas y requieren una cierta experiencia por parte del técnico que las realiza. En general, se aplican a partir de la 3ª-4ª semana posterior a la inseminación artificial. Por otro lado, los métodos indirectos de DG son menos invasivos que los directos. Mediante este tipo de diagnósticos vía indirecta se puede detectar la presencia o ausencia de un embrión, sin visualizar directamente estructuras gestacionales. Existen métodos indirectos basados en signos clínicos, como la vigilancia del retorno al estro, así como técnicas indirectas bioquímicas, que permiten evaluar mediante el uso de kits rápidos ciertas sustancias como la progesterona (P4) o las Glicoproteínas Asociadas a la Gestación (GPAG), producidas durante la gestación de forma temprana. Otras técnicas novedosas y prometedoras, pero que todavía se encuentran en desarrollo, sonla evaluación de moléculas como el interferón tau (IFNτ), los micro-ARN (miARN) y/o los Factores de Gestación Temprana (FGT).To diagnose pregnancy in cattle an accurate, safe, economical and early method must be used. Today, several methods are available in the cattle industry, while others are still under development. The objective of this study is to review bibliographically the methods for the diagnosis of pregnancy in bovines and their use in current and future livestock. On the one hand, direct methods such as transrectal palpation and transrectal ul-trasound of the reproductive tract are considered the most widely used methods for the diagnosis of pregnancy in bovines today. These methods work well and are economically interesting. However, they are also invasive and require some experience to perform properly. In general, these methods are used three to four weeks after artificial insemination. Second, indirect methods for the diagnosis of pregnancy in bovines can be used for the early detection of pregnancy and therefore can save time. The presence of the embryo must be determined without directly visualizing the embryonic structures. Due to technological advances, indirect biochemical methods are being developed to detect certain molecules associated with pregnancy and predict the presence of an embryo at an even earlier stage, by quantifying hormones such as progesterone (P4) or glycoproteins asso-ciated with it. pregnancy (PAG) using rapid test kits. The detection of molecules such as interferon tau (IFNτ) or microRNA (miRNA) and early pregnancy factors (EPF) are examples of these promising new methods still under developmen