Search

Abstract Background Control strategies against contagious agalactia (CA), considered one of the most important diseases affecting small ruminants in countries surrounding the Mediterranean, are mainly based on traditional strategies considered suboptimal such as the use of inactivated vaccines and antibiotics. This manuscript analysed the efficacy of an alternative non-mandatory official control programme based on a multi-platform diagnostic panel and biosecurity developed and started in 185 herds placed in a contagious agalactia endemic area of Spain, using the data of 74,080 samples collected and analysed during a 4 years-period (2018–2021). Results Globally, the combined analyses of bulk tank milk (BTM), ear or nasal swabs (in goats or sheep, respectively) and the serology to detect Mycoplasma agalactiae (Ma), allowed the detection of 40.54% of positive farms (n = 75), with Ma the species always detected in sheep (100%) and Mycoplasma mycoides subsp. capri (Mmc, 68,8%), Ma (29,3%) or both of them (1,9%) in goats. Taking into account productive aptitude and vaccination against CA, the use of BTM, ear or nasal swabs, and serology for herd classification demonstrated either a lack of concordance or only weak concordance. In herds that did not vaccinate, the classification of positives by male serology or swab detection showed moderate concordance. Vaccination against AC proved to be a protective factor against the occurrence of herds with bucks or rams testing positive. Conclusions Since the different diagnostic techniques are not interchangeable, it is necessary to apply a multi-platform diagnostic panel for the accurate classification of herds. Based on official classification, strict biosecurity standards, including the prohibition of the entry of animals with unknown health status, allowed the completion of the CA control program.

Mycoplasma bovis (M. bovis) is one of the worldwide most important infectious agents involved in respiratory complex diseases (RCD). In Spain, the endemic presence of subtypes ST-2 and ST-3 with phenotypic differences linked to their susceptibility to fluoroquinolones opened the way to develop control strategies focused on previous diagnosis of the subtype and the use of directed therapies when M. bovis were involved in RCD. Surprisingly, microbiological studies conducted during 2023 evidenced for the first time the presence of Spanish isolates of a new polC-subtype, previously classified as ST-1, recovered from calves with respiratory symptoms and pneumonia in different areas of the country (n = 16). Curiously, the minimum inhibitory concentration (MIC) to a panel of antimicrobials revealed phenotypic differences between these ST-1 isolates when using fluoroquinolones (FLQ). There is no geographical correlation between MIC profiles even for a set of 8 isolates recovered from different animals in the same flock. Sequencing of 4 genes (gyrA, gyrB, parC and parE) encoding quinolone resistance-determining regions (QRDR) evidenced the presence of accumulate mutations in 2 ST-1 isolates with high FLQ MICs, but not in all them (n = 3), thus suggesting that, as previously recorded for ST-2 isolates, other mechanisms should be involved in the acquisition of resistence to these antimicrobials. Additionally, as previously detected in the Spanish ST-2 and ST-3, subtype ST-1 isolates are also resistant to macrolides or lincosamides.

IntroductionThe 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.MethodsA 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.ResultsThe commercial probiotic L2 significantly reduced the concentration of Ma to 0.000 log CFU/mL (p

Resumen: La pandemia de COVID-19 ha hecho evidente la importancia de la interfaz animal-humano-medio ambiente en la emergencia de zoonosis. A pesar de que el salto de especie se considera un evento raro, el número de enfermedades infecciosas emergentes aumentó de manera significativa en la segunda mitad del siglo xx, siendo estas principalmente de carácter zoonótico y originadas en la fauna silvestre. Entre los determinantes asociados a la emergencia de zoonosis destacan la interacción humana con los ecosistemas, la pérdida de biodiversidad, los cambios en el uso del suelo, el cambio climático, el comercio y el consumo de fauna silvestre, etc. En el proceso del salto de especie existen diferentes fases de adaptación evolutiva entre el patógeno y la especie humana, variando desde su presencia en el reservorio animal sin infección humana hasta enfermedades exclusivamente humanas sin otros reservorios. El conocimiento de la evolución natural de las zoonosis permite identificar los puntos críticos para su control, al tiempo que posibilita identificar posibles candidatos para futuras pandemias. De forma específica, los avances en el conocimiento de los posibles reservorios del SARS-CoV-2 han contribuido a la toma de decisiones durante la pandemia. Por todo ello, y ante la variedad de escenarios que posibilitan el salto de especie y la evolución de los diferentes patógenos en un nuevo huésped, la vigilancia frente a la emergencia de zoonosis debe plantearse bajo la estrategia One Health. Abstract: The new pandemic, COVID-19, highlighted the importance of the animal–human–environment interface in the emergence of zoonoses. The jump of the species barrier is considered a rare event although the number of emerging infectious diseases increased significantly in the second half of the 20th century. Most of them transmitted from wildlife to humans. Several elements can contribute to the emergence of zoonoses: human interaction on ecosystems, biodiversity loss, changes in land use, climate change, trade in or consumption of wildlife. There are different stages in the adaptation of an animal pathogen into a specialized human pathogen, varying from its presence in animals without human infection to a pathogen exclusive to humans. The history of zoonoses allows the identification of critical points for their control, as well as the identification of probable virus or bacteria involved in future pandemics. Concerning COVID-19, the knowledge of the possible reservoirs of SARS-CoV-2 contributed to decision-making during the pandemic. Therefore, and given the variety of scenarios that allow the jump of species barrier or the evolution of pathogens in a new host, One Health strategies for zoonosis emergence surveillance should be implemented.

Contagious agalactia is associated with mastitis, keratoconjunctivitis, arthritis, pneumonia, and septicemia in small ruminants in countries with large dairy industries worldwide. The causative agents belong to four (sub)species of the Mycoplasma genus that have remained essentially susceptible to antimicrobials, including to the widely-used tetracycline family. However, some clinical isolates have been detected that show increased minimum inhibitory concentrations of tetracyclines, although they do not harbor the mutation in the 16SrRNA gene usually associated with resistance. The present work aimed to assess whether efflux pumps, infrequently described in mycoplasmas, could participate in the observed moderate loss of susceptibility. General efflux mechanisms were measured (i) using the fluorescence property of ethidium bromide when accumulated intracellularly and intercalated in the mycoplasma genomes, its active extrusion resulting in a temperature-dependent decrease in fluorescence and (ii) monitoring the growth inhibition of mycoplasmas by subinhibitory concentrations of tetracycline with or without reserpine, a known inhibitor of efflux in other bacteria. Both methods revealed non-specific efflux phenomena in most of the isolates tested, although their efficacy was difficult to quantify. This property could contribute to the acquisition of mutations conferring resistance by maintaining intracellular concentrations of tetracyclines at subinhibitory levels.

Bovine respiratory disease (BRD) is an important viral and/or bacterial disease that mainly affects feedlot calves. The involvement of Mycoplasma bovis in BRD can lead to chronic pneumonia poorly responsive to antimicrobial treatment. Caseonecrotic bronchopneumonia is a pulmonary lesion typically associated with M. bovis. In Spain, M. bovis is widely distributed in the feedlots and circulating isolates are resistant to most antimicrobials in vitro. However, the role of this species in clinical respiratory disease of feedlot calves remains unknown. Furthermore, available data are relative to a fixed panel of antimicrobials commonly used to treat BRD, but not to the specific set of antimicrobials that have been used for treating each animal. This study examined 23 feedlot calves raised in southeast Spain (2016–2019) with clinical signs of respiratory disease unresponsive to treatment. The presence of M. bovis was investigated through bacteriology (culture and subsequent PCR), histopathology and immunohistochemistry. The pathogen was found in 86.9% (20/23) of the calves, mainly in the lungs (78.26%; 18/23). Immunohistochemistry revealed M. bovis antigens in 73.9% (17/23) of the calves in which caseonecrotic bronchopneumonia was the most frequent lesion (16/17). Minimum inhibitory concentration assays confirmed the resistance of a selection of 12 isolates to most of the antimicrobials specifically used for treating the animals in vivo. These results stress the importance of M. bovis in the BRD affecting feedlot calves in Spain.

Contagious agalactia is a mycoplasmosis affecting small ruminants that have become an important issue in many countries. However, PK/PD studies of antibiotics to treat this problem in lactating goats affected by Mycoplasma (M.) agalactiae, the main CA-causing mycoplasma are almost non-existent. The aims of this study were to evaluate the plasma and milk disposition of marbofloxacin in lactating goats after intravenous (IV), subcutaneous (SC) and subcutaneous poloxamer P407 formulations with and without carboxy-methylcellulose (SC-P407-CMC and SC-P407) administration. Marbofloxacin concentrations were analysed by the High Performance Liquid Chromatography (HPLC) method. Minimum inhibitory concentrations (MIC) of M. agalactiae field isolates from mastitic goat’s milk were used to calculate surrogate markers of efficacy. Terminal half-lives of marbofloxacin after IV, SC, SC-P407 and SC-P407-CMC administration were 7.12, 6.57, 13.92 and 12.19 h in plasma, and the half-lives of elimination of marbofloxacin in milk were 7.22, 7.16, 9.30 and 7.74 h after IV, SC, SC-P407 and SC-P407-CMC administration, respectively. Marbofloxacin penetration from the blood into the milk was extensive, with Area Under the Curve (AUCmilk/AUCplasma) ratios ranged 1.04–1.23, and maximum concentrations (Cmax-milk/Cmax-plasma) ratios ranged 0.72–1.20. The PK/PD surrogate markers of efficacy fAUC24/MIC and the Monte Carlo simulation show that marbofloxacin ratio (fAUC24/MIC > 125) using a 90% of target attainment rate (TAR) need a dose regimen between 8.4 mg/kg (SC) and 11.57 mg/kg (P407CMC) and should be adequate to treat contagious agalactia in lactating goats.

Official milk prices in the Spanish small ruminant sector were used for 5 years (2015–2019) to analyze the effect caused by the coronavirus disease (COVID-19) crisis in 2020. Meat price fluctuations were also studied using the weekly prices officially provided by some of the main agrarian markets of the country (n = 6) in 2019 and 2020. Moreover, the sales and prices of three protected geographical indications (PGI) of lamb meat served to study the marketability when the products are sold or not under these quality labels in a crisis context. According to Spanish Government’s official communications, 2020 was divided in three periods of study (pre-COVID-19, total confinement and post-confinement). The evolution of employment in this subsector in 2020, as a direct consequence of this crisis, was also analyzed considering data provided by producers. Results showed an intra-annual seasonal effect for milk prices in 2020 for both livestock species, as observed in previous years. However, a negative economic impact on goat milk prices due to the pandemic was checked during the confinement and post-confinement months. Sheep milk prices remained stable. Lamb and goat kid meat prices showed a similar trend in comparison with 2019 during the pre-COVID-19 period. The total confinement period recorded a short interval of 1–2 weeks in which the prices declined, before the suspension of quotations in many markets. In contrast, once confinement was completed, meat prices for both ruminant species rapidly reached levels that existed before the coronavirus crisis. Overall data suggested the protective effect of the PGI marks on lamb meat. Lambs with a PGI had better 2020 prices than non-PGI lambs (+8%), regardless of the period analyzed. Moreover, with fewer lambs sold in 2020, there was a relevant drop in sales of non-PGI lambs vs. PGI (−19% vs. −2%) during the first 7 months. Finally, there was little or no readjustment of the workforce in the small ruminant flocks.

The human pandemic COVID-19 caused by the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) started in China in 2019 and has rapidly spread around the world, leading to extreme control measures such as population confinement and industry activity closure. Although small ruminants are not sanitary affected by this virus, the short-term economic impact derived by COVID-19 on Spanish flocks is estimated in this study, using data provided by producers and two major slaughterhouses. Milk prices of dairy goat flocks suffered a substantial drop in April 2020, close to 4.5 cts EUR/liter compared to the previous month. In contrast, the monthly milk prices in sheep remained almost stable during this period, and even increases of more than EUR 6 cts were reported in comparison with the previous year. Nevertheless, economical differences are reported by areas where producers could receive a higher income, close to EUR 0.3 per liter of milk. Global data provided by feedlots affecting 2750 Spanish flocks evidenced a lamb price drop ranging from 16.8% to 26.9% after the pandemic arrival; in line with the data directly reported by a limited sample of producers (ranging from 11.0% to 23.7%). The goat kid meat market also suffered a reduction in prices per kg, near 12.5%; although, for some flocks, losses reached up to 40%. In the same line, 2 slaughterhouses reported a sudden sacrifice drop around 27% for lambs and goat kids sacrifices in April, in contrast with the usual sacrifice figures from the beginning of 2020. Moreover, our study showed a temporary and unexpected retention of lambs and goat kids at farms due to a reduction in animals slaughtered during this period. In conclusion, data evidenced a considerable negative economic impact on Spanish small ruminant flocks, throughout the first 60 days after COVID-19’s pandemic declaration. Further studies are needed to evaluate the long-term economic consequences, in order to establish contingency plans and avoid the collapse of small ruminant industries when a crisis of these characteristics occurs.

Mycoplasma bovis is an important bovine pathogen causing pneumonia, mastitis, and arthritis and is responsible for major economic losses worldwide. In the absence of an efficient vaccine, control of M. bovis infections mainly relies on antimicrobial treatments, but resistance is reported in an increasing number of countries. To address the situation in Spain, M. bovis was searched in 436 samples collected from beef and dairy cattle (2016–2019) and 28% were positive. Single-locus typing using polC sequences further revealed that two subtypes ST2 and ST3, circulate in Spain both in beef and dairy cattle, regardless of the regions or the clinical signs. Monitoring of ST2 and ST3 isolates minimum inhibitory concentration (MIC) to a panel of antimicrobials revealed one major difference when using fluoroquinolones (FQL): ST2 is more susceptible than ST3. Accordingly, whole-genome sequencing (WGS) further identified mutations in the gyrA and parC regions, encoding quinolone resistance-determining regions (QRDR) only in ST3 isolates. This situation shows the capacity of ST3 to accumulate mutations in QRDR and might reflect the selective pressure imposed by the extensive use of these antimicrobials. MIC values and detection of mutations by WGS also showed that most Spanish isolates are resistant to macrolides, lincosamides, and tetracyclines. Valnemulin was the only one effective, at least in vitro, against both STs.

Mycoplasma 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.

En el contexto actual de la enseñanza, el profesor ha dejado fundamentalmente de transmitir conocimientos para pasar a involucrarse de una forma mucho más activa en el proceso de aprendizaje, cuyo papel fundamental debe recaer en el alumno. La aplicación del método de estudio de casos facilita que éstos asuman el papel de protagonistas activos de su propio aprendizaje, potenciando la adquisición de competencias disciplinares, genéricas y académicas. En este trabajo, presentamos las experiencias realizadas en dos contextos educativos muy diferentes: la docencia práctica de la asignatura de Enfermedades Infecciosas de la materia de Sanidad Animal en la Licenciatura de Veterinaria y en segundo lugar, exponemos su utilización como herramienta en las clases teóricas de la asignatura de Economía de 1º y 2º de Bachiller. Este trabajo presenta las pautas generales de la metodología utilizada y las principales conclusiones obtenidas de su aplicación en el proceso de aprendizaje.

Mycoplasma capricolum subsp. capricolum is one of the causative agents of contagious agalactia (CA). Nevertheless, there is still a lack of information about its antimicrobial susceptibility and genetic characteristics. Therefore, the aim of this work was to study the antimicrobial and genetic variability of different Mycoplasma capricolum subsp. capricolum field isolates. For this purpose, the growth inhibition effect of 18 antimicrobials and a multilocus sequence typing (MLST) scheme based on five housekeeping genes (fusA, glpQ, gyrB, lepA and rpoB) were performed on 32 selected field isolates from Italy and Spain.The results showed a wide range of growth inhibitory effects for almost all the antimicrobials studied. Macrolides presented lower efficacy inhibiting Mcc growth than in previous works performed on other CA-causative mycoplasmas. Erythromycin was not able to inhibit the growth of any of the studied strains, contrary to doxycycline, which inhibited the growth of all of them from low concentrations. On the other hand, the study of the concatenated genes revealed a high genetic variability among the different Mcc isolates. Hence, these genetic variations were greater than the ones reported in prior works on other mycoplasma species.

Veterinary students face diverse potential sources of zoonotic pathogens since the first years of their academic degree. Such sources include different animal species and pathologic materials which are used at university facilities as well as commercial clinics, farms and other external facilities.The present study utilizes a systematic review of the literature to identify zoonoses described in veterinary students.Web of Science and PubMed.Of the 1,254 titles produced by the bibliographic search, 62 were included in this review. Whereas 28 of these articles (45.2%) described individual cases or outbreaks, the remaining 34 (54.8%) reported serological results. The zoonotic etiological agents described were bacteria, in 39 studies (62.9%), parasites, in 12 works (19.4%), virus, in 9 studies (14.5%) and fungi, in 2 (3.2%) of the selected articles. The selected literature included references from 24 different countries and covered the time period of the last 55 years.The fact that common cases of disease or cases of little clinical importance without collective repercussions are not usually published in peer-reviewed journals limits the possibility to reach conclusions from a quantitative point of view. Furthermore, most of the selected works (66.1%) refer to European or North American countries, and thus, the number of cases due to pathogens which could appear more frequently in non-occidental countries might be underestimated.The results of the present systematic review highlight the need of including training in zoonotic diseases since the first years of Veterinary Science degrees, especially focusing on biosecurity measures (hygienic measures and the utilization of the personal protective equipment), as a way of protecting students, and on monitoring programs, so as to adequately advise affected students or students suspicious of enduring zoonoses.

Between 2006 and 2008, an outbreak of Infectious Keratoconjunctivitis (IKC) affected Pyrenean chamois Rupicapra p. pyrenaica, an endemic subspecies of mountain ungulate that lives in the Pyrenees. The study focused on 14 mountain massifs (180,000 ha) where the species' population is stable. Cases of IKC were detected in ten of the massifs and, in five of them, mortality was substantial. The outbreak spread quickly from the first location detected, with two peaks in mortality that affected one (2007) and three (2008) massifs. In the latter, the peak was seasonal (spring to autumn) and, in the former, the outbreak persisted through winter. To identify the outbreak's aetiology, we examined 105 Pyrenean chamois clinically affected with IKC. TaqMan rt-PCR identified Mycoplasma conjunctivae in 93 (88.5%) of the chamois. Another rt-PCR detected Chlamydophila spp. in 14 of chamois, and 12 of those had mixed infections with mycoplasmas. In the period 2000-2007, the chamois population increased slightly (λ 1.026) but decreased significantly during the IKC outbreak (λ 0.8, 2007-2008; λ 0.85, 2008-2009) before increasing significantly after the outbreak (λ 1.1, 2009-2010). Sex-biased mortality shifted the adult sex ratio toward males (from 0.6 to 0.7 males per female) and reduced productivity slightly. Hunting was practically banned in the massifs where chamois experienced significant mortality and allowed again after the outbreak ended. Long-term monitoring of wild populations provides a basis for understanding the impacts of disease outbreaks and improves management decisions, particularly when species are subject to extractive exploitation.