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Introduction Contagious agalactia (CA) is a disease affecting small ruminants with worldwide distribution and caused by several mycoplasmas, especially M. agalactiae. The main option for systematic diagnosis under monitoring control programmes is the enzyme-linked immunosorbent assay (ELISA) test. Material and Methods This study was designed to appraise the performance of two commercial indirect ELISA tests using M. agalactiae p48 protein and one using total protein, for antibody detection in small ruminants after natural infection with different M. agalactiae strains. We carried out the test evaluation using sera of confirmed M. agalactiae-positive goats with clinical signs. In addition, test agreement was assessed by kappa between the three commercial ELISA tests. Results All three ELISA tests showed high validity scores (Youden’s J: 72.9–84%). The sensitivity values for the P48 protein-based tests were 76.9% and 84.6%, and was 79% for the total protein-based test. The specificity of all tests was 100%. In addition, between the total protein-based ELISA test and the other two ELISA tests based on the P48 protein, the agreement was substantial (kappa: 0.762–0.763) and the agreement between the latter two tests was almost perfect (kappa: 0.93). Conclusion The validity parameters for all tests allowed their application for diagnostic purposes in lactating goats excreting M. agalactiae in milk and presenting clinical signs. The agreements show that any of these ELISA tests could be equally well used for diagnosis in programmes against CA.

Mastitis is a significant disease in dairy ruminants, causing economic losses to the livestock industry and severe risks to public health. Antibiotic therapy is one of the most crucial practices to treat mastitis, although the susceptibility of caprine mastitis pathogens to current antibiotics has not been tested under standard or modified incubation conditions. This work evaluated the in vitro activity of tildipirosin, gamithromycin, oxytetracycline, and danofloxacin against caprine mastitis pathogens incubated following standard conditions of Clinical and Laboratory Standards Institute (CLSI) and deviation method by 25% supplementation with goat serum. Mycoplasma agalactiae, Escherichia coli, Staphylococcus aureus, Streptococcus spp., and coagulase-negative Staphylococci (CNS) were isolated from dairy goats with mastitis in Spain. Minimum inhibitory concentrations (MICs) were determined using the broth microdilution technique. The lowest MIC90 under standard conditions was obtained with danofloxacin for mastitis-causing pathogens. An exception was M. agalactiae, where danofloxacin and oxytetracycline obtained low values. However, after adding serum, gamithromycin showed the lowest MIC50 for S. aureus, Streptococcus spp., and CNS. The lowest MIC50 was obtained with all the antibiotics tested (M. agalactiae. Supplementing with serum resulted in a significant variation in tildipirosin and gamithromycin MIC values for CNS, S. aureus, M. agalagtiae, and E. coli. In brief, the MIC for antibiotics used against mastitis should be determined under conditions closely resembling intramammary infections to obtain representative susceptibility patterns against mastitis pathogens. Caprine mastitis pathogens were broadly susceptible to danofloxacin under standard conditions. The potency of macrolides against caprine mastitis pathogens increases when serum is present in culture media.

Horizontal gene transfer was long thought to be marginal in Mollicutes, but the capacity of some of these wall-less bacteria to exchange large chromosomal regions has been recently documented. Mycoplasma chromosomal transfer (MCT) is an unconventional mechanism that relies on the presence of a functional integrative conjugative element (ICE) in at least one partner and involves the horizontal acquisition of small and large chromosomal fragments from any part of the donor genome, which results in progenies composed of an infinite variety of mosaic genomes. The present study focuses on Mycoplasma bovis, an important pathogen of cattle responsible for major economic losses worldwide. By combining phylogenetic tree reconstructions and detailed comparative genome analyses of 36 isolates collected in Spain (2016 to 2018), we confirmed the mosaic nature of 16 field isolates and mapped chromosomal transfers exchanged between their hypothetical ancestors. This study provides evidence that MCT can take place in the field, most likely during coinfections by multiple strains. Because mobile genetic elements (MGEs) are classical contributors of genome plasticity, the presence of phages, insertion sequences (ISs), and ICEs was also investigated. Data revealed that these elements are widespread within the M. bovis species and evidenced classical horizontal transfer of phages and ICEs in addition to MCT. These events contribute to wide-genome diversity and reorganization within this species and may have a tremendous impact on diagnostic and disease control. IMPORTANCE Mycoplasma bovis is a major pathogen of cattle that has significant detrimental effects on economics and animal welfare in cattle rearing worldwide. Understanding the evolution and the adaptative potential of pathogenic mycoplasma species in the natural host is essential to combating them. In this study, we documented the occurrence of mycoplasma chromosomal transfer, an atypical mechanism of horizontal gene transfer, in field isolates of M. bovis that provide new insights into the evolution of this pathogenic species in their natural host. Although these events are expected to occur at low frequency, their impact is accountable for genome-wide variety and reorganization within M. bovis species, which may compromise both diagnostic and disease control.

Contagious agalactia (CA) is a disease affecting small ruminants with worldwide distribution and caused by several mycoplasmas, especiallyThis study was designed to appraise the performance of two commercial indirect ELISA tests usingAll three ELISA tests showed high validity scores (Youden's J: 72.9-84%). The sensitivity values for the P48 protein-based tests were 76.9% and 84.6%, and was 79% for the total protein-based test. The specificity of all tests was 100%. In addition, between the total protein-based ELISA test and the other two ELISA tests based on the P48 protein, the agreement was substantial (kappa: 0.762-0.763) and the agreement between the latter two tests was almost perfect (kappa: 0.93).The validity parameters for all tests allowed their application for diagnostic purposes in lactating goats excreting

Simple Summary Contagious agalactia is a multi-faceted disease affecting small ruminants worldwide. It is caused by four different Mycoplasma (sub)-species. In the absence of highly efficient vaccines, its control relies mostly on antibiotic treatment. Tetracyclines are one of the main families used, as they are cheap and often ensure clinical recovery, if not microbial clearance. However, some isolates have shown lowered susceptibilities even without the mutations in the target gene known to result in resistance. We suspected that an active efflux mechanism could be responsible for such lower-susceptibility phenotypes. Using various techniques, we demonstrated that most of the strains we studied did exhibit a capacity to actively extrude various substances including tetracyclines. This might contribute to low resistance profiles. Abstract 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.

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.

Horizontal gene transfer was long thought to be marginal in Mollicutes, but the capacity of some of these wall-less bacteria to exchange large chromosomal regions has been recently documented. Mycoplasma chromosomal transfer (MCT) is an unconventional mechanism that relies on the presence of a functional integrative conjugative element (ICE) in at least one partner and involves the horizontal acquisition of small and large chromosomal fragments from any part of the donor genome, which results in progenies composed of an infinitive variety of mosaic genomes. The present study focuses on Mycoplasma bovis, an important pathogen of cattle responsible for major economic losses worldwide. By combining phylogenetic tree reconstructions and detailed comparative genome analyses of 36 isolates collected in Spain (2016-2018) we confirmed the mosaic nature of 16 field isolates and mapped chromosomal transfers exchanged between their hypothetical ancestors. This study provides evidence that MCT can take place in the field, most likely during co-infections by multiple strains. Because mobile genetic elements (MGEs) are classical contributors of genome plasticity, the presence of phages, insertion sequences (ISs) and ICEs was also investigated. Data revealed that these elements are widespread within the M. bovis species and evidenced classical horizontal transfer of phages and ICEs in addition to MCT. These events contribute to wide-genome diversity and reorganization within this species and may have a tremendous impact on diagnostic and disease control.IMPORTANCEMycoplasma bovis is a major pathogen of cattle with significant detrimental economic and animal welfare on cattle rearing worldwide. Understanding the evolution and the adaptative potential of pathogenic mycoplasma species in the natural host is essential to combating them. In this study, we documented the occurrence of mycoplasma chromosomal transfer, an atypical mechanism of horizontal gene transfer, in field isolates of M. bovis that provide new insights into the evolution of this pathogenic species in their natural host. Despite these events are expected to occur at low frequency, their impact is accountable for genome-wide variety and reorganization within M. bovis species, which may compromise both diagnostic and disease control.

This project has been funded by Ministerio de Economia y Competividad (AGL201676568-R) of Spain. Ana Garcia-Galan Perez is beneficiary of a research fellowship (State Subprogram Training of the State Program for the Promotion of Talent and its Employability, BES-2017-080186)., Thanks are due to J. Carrizosa (IMIDA) and the Caprine Veterinary Farm of the University of Murcia for their assistance with the experiments., In some countries like Spain and France, contagious agalactia (CA) is a highly relevant issue. CA is a mycoplasmosis affecting small ruminants and it is associated with a relevant economic impact on dairy. The poor efficacy of vaccines and their inability to prevent disease transmission is conducive to the use of antibiotics to control CA. However, only a few groups of antimicrobial agents are effective against these species, and selecting an adequate antimicrobial agent following the categorization of antibiotics made by the different international organisms (European Medicine Agency, World Health Organization) in veterinary medicine becomes a difficult task. The PK/PD approach is a useful tool to guide veterinarians on the appropriate targets through a rational selection of the best dose regimen of antimicrobial agents. In this study, marbofloxacin pharmacokinetics was studied after three routes of administration with two long-acting formulations. The minimum inhibitory concentrations (MIC) values of Mycoplasma agalactia isolated from goats affected by CA in Spain were calculated. The results show that systemic exposure achieved in lactating goats following these formulations provides rate of drug release that could be adequate to maintain effective plasma concentrations against M. agalactiae. The PK/PD analysis by Monte Carlo simulation showed that a dosage regimen from 8.47 to 11.57 mg/kg every 24 h could effectively treat goats affected by CA. 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 (AUC(milk)/AUC(plasma)) 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 (fAUC(24)/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., Ministerio de Economia y Competividad of Spain AGL201676568-R, Research fellowship (State Subprogram Training of the State Program for the Promotion of Talent and its Employability) BES-2017-080186

La pandemia Covid-19 ha motivado la adopción de medidas excepcionales en todo el mundo, a efectos de limitar los contagios y el colapso de los sistemas sanitarios. El cierre de comercios y otras actividades consideradas no esenciales, o las limitaciones al movimiento, ha generado un impacto económico en el sector de los pequeños rumiantes, poniendo de manifiesto la necesidad del profesional veterinario de disponer de las competencias necesarias para valorar económicamente el impacto de las enfermedades en los colectivos. Este trabajo analiza, en primer lugar, la formación en economía que reciben los estudiantes del Grado de Veterinaria en las diferentes facultades españolas, comparándola con la que reciben los estudiantes en el Grado de Ingeniería Agrícola. En segundo lugar, se ha diseñado una encuesta para egresados y estudiantes de último curso de veterinaria, en referencia a este tipo de competencias, su aplicación en la sanidad animal y su repercusión en el desempeño profesional. Los datos evidencian que el tiempo empleado para adquirir las competencias en economía de los veterinarios es menor (3-6 créditos en 5 años) que el que disponen los ingenieros agrícolas (12 créditos en 4 años). Los resultados de la encuesta revelan que, si bien se registran diferencias significativas cuantitativamente entre egresados y estudiantes, ambos grupos coinciden en la escasa formación recibida para valorar el impacto de las enfermedades en los colectivos, así como en la necesidad de formarse en aspectos de gestión económica una vez finalizados sus estudios de grado para el desempeño de su labor profesional, competencias que deberían ser reforzadas en el grado. Entre los egresados, la opinión es similar, independientemente de los años de desempeño profesional. The Covid-19 pandemic led to extreme control measures around the world aiming to halting the number of new infections. Non-essential activities closures and population confinement had an economic impact on the small ruminant sector, highlighting the need for veterinarians to have some skills to assess the economic impact of diseases on flocks. Firstly, this study analyzed the economic training received by the veterinary students at the Spanish faculties, also comparing it with the ones received by students of the agricultural engineering degree. Secondly, a survey in reference to the acquisition of this type of competences and its application for animal health was designed and applied for graduates and final-year veterinary students. The data showed that the ECTS taken to acquire the economic skills of veterinarians is less (3-6 ECTS in 5 years) than that of agricultural engineers (12 ECTS in 4 years). The results of the survey also showed that, although there are significant differences between graduates and students, both them are largely in agreement on the little training received to assess the impact of diseases, and on the need for an additional training after completing their studies. Therefore, these skills should be reinforced in the degree. Among the graduates, the opinion is similar, regardless of the years of professional experience.

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.

Simple Summary Mycoplasma bovis is an important infectious agent in cattle. The pathogen may cause a wide range of clinical signs, including mastitis, arthritis, pneumonia and reproductive disorders. Artificial insemination with contaminated semen may be a source of infection in infection-free areas or herds. Hence, the antimicrobials used in the preparation of seminal doses should be re-evaluated, or alternative measures to antimicrobials should be tested. This in vitro study aims to evaluate novel strategies to reduce the risk of the transmission of the pathogen through contaminated semen during artificial insemination. Hence, we assess the effect of the addition of (1) the antimicrobial enrofloxacin, (2) the antimicrobial doxycycline, or (3) a probiotic of human origin that contains acid lactic bacteria of the genus Lactobacillus, on the viability of Mycoplasma bovis in diluted bull semen in a Tris-citrate-fructose medium. The data show that the pathogen is negatively affected by the addition of 0.125 μg/mL of enrofloxacin, 0.0625 μg/mL of doxycycline, or the probiotic at a concentration of 3.24 × 106 or 3.24 × 108 colony-forming units/mL in diluted semen. Our results are promising in the field, as they may support new strategies to reduce the risk of the transmission of Mycoplasma bovis through artificial insemination. Abstract 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.

Official milk prices in the Spanish small ruminant sector were used for 5 years (2015&ndash, 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&rsquo, 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&ndash, 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 (&minus, 19% vs. &minus, 2%) during the first 7 months. Finally, there was little or no readjustment of the workforce in the small ruminant flocks.

Este artículo se encuentra disponible en la siguiente URL: https://www.mscbs.gob.es/biblioPublic/publicaciones/recursos_propios/resp/revista_cdrom/VOL94/C_ESPECIALES/RS94C_202003022.pdf Las aves acuáticas silvestres representan el principal reservorio natural de los virus influenza y han participado en el reordenamiento tanto de virus pandémicos como de los virus responsables de los brotes de gripe aviar en las especies domésticas y silvestres. Con el objetivo de conocer los determinantes implicados, en el presente trabajo se revisaron los casos humanos de influenza aviar asociados al manejo de aves silvestres y la utilización de los equipos de protección personal, así como las bases de la vigilancia de la influencia aviar altamente patógena en aves silvestres en España. Las evidencias existentes permiten concluir que la transmisión directa de virus influenza desde las aves silvestres al ser humano es un evento raro. No obstante, nuestro contexto epidemiológico se encuentra influido por el cambio climático y queda marcado por la presencia de rutas migratorias desde territorios donde la infección puede estar presente. Por ello, y ante las implicaciones clínicas, económicas y para la salud pública que dichas infecciones pueden tener, los diferentes colectivos expuestos a las aves silvestres (veterinarios, biólogos, ornitólogos, conservacionistas, técnicos de campo, agentes medioambientales, cetreros, cazadores, etc.) deberían conocer las posibles fuentes de contagio y manejar correctamente los equipos de protección personal. Al mismo tiempo, es importante que dichos grupos conozcan la situación sanitaria actualizada respecto a la influenza aviar, para adaptar sus actividades en consecuencia y poder aplicar las medidas de protección de forma proporcionada a la misma, amén de aportar una valiosa información para los programas de vigilancia. / Wild waterfowl are considered the main natural reservoir of influenza viruses and they have contributed to the reassortment of both pandemic viruses and viruses responsible for outbreaks of avian influenza in wild and domestic species. In order to determinate the factors involved, we reviewed the human cases of avian influenza related to the management of wild birds, the use of personal protective equipment, as well as the basis of surveillance programs of highly pathogenic avian influenza in wild birds in Spain. The direct transmission of influenza virus from wild birds to humans is a rare event. However, our epidemiological context is influenced by climate change and marked by the presence of migratory routes from territories where infection may be present. Thus, and due to the clinical, economical and public health implications that such infections may have, the different groups exposed to wild birds (veterinarians, biologists, ornithologists, conservationists, field technicians, environmental officers, falconers, hunters, etc.) should know which are the possible sources of infection and how to handle the personal protective equipment. Besides, it is important that those groups know the current sanitary situation regarding avian influenza so they can consequently adapt their activities and employ proper protective measures, in addition to providing valuable information for surveillance programs.

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&rsquo, 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 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 &mu, g/mL of enrofloxacin or 0.0625 &mu, g/mL of doxycycline and to the addition of the probiotic at a concentration of 3.24 &times, 106 colony forming units (CFU)/mL or 3.24 &times, 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 &lt, 0.05). Our results may support new strategies for reducing the risk of M. bovis transmission through AI.

Mycoplasma capricolum subsp. capricolum (Mcc) is one of the causative agents of contagious agalactia, and antimicrobial therapy is the most commonly applied measure to treat outbreaks of this disease. Macrolides and lincosamides bind specifically to nucleotides at domains II and V of the 23S rRNA. Furthermore, rplD and rplV genes encode ribosomal proteins L4 and L22, which are also implicated in the macrolide binding site. The aim of this work was to study the relationship between mutations in these genes and the acquisition of macrolide and lincosamide resistance in Mcc. For this purpose, in vitro selected resistant mutants and field isolates were studied. This study demonstrates the appearance of DNA point mutations at the 23S rRNA encoding genes (A2058G, A2059G and A2062C) and rplV gene (Ala89Asp) in association to high minimum inhibitory concentration values. Hence, it proves the importance of alterations in 23S rRNA domain V and ribosomal protein L22 as molecular mechanisms responsible for the acquisition of macrolide and lincosamide resistance in both field isolates and in vitro selected mutants. Moreover, these mutations enable us to provide an interpretative breakpoint of antimicrobial resistance for Mcc at MIC 0.8 μg/ml.

The extensive use of antimicrobials for disease control has caused a remarkable decrease in antimicrobial susceptibility of different animal mycoplasma species, including Mycoplasma agalactiae (M. agalactiae), the main causative agent of contagious agalactia. However, the molecular mechanisms behind M. agalactiae resistance to macrolides and lincomycin have not yet been elucidated. The aim of the present study was to investigate the association between minimum inhibitory concentration (MIC) values of different antimicrobials and mutations in the 23S rRNA gene and ribosomal proteins L4 and L22, analysing both field isolates (n=50) and in vitro selected resistant mutants of M. agalactiae. The obtained MIC results of the studied field isolates demonstrate an increasing development of tylosin resistance in this bacterium, in comparison to previous studies. Interestingly, predicted amino acid changes in L22 (Ser89Leu and Gln90Lys/His) were the first variations observed when MICs of M. agalactiae started to increase (tylosin MIC ≥0.8μg/ml), whereas mutations at positions 2058 or 2059 of domain V of the 23S rRNA gene appeared from MIC values of 1.6μg/ml. These results were consistent in both field isolates and in vitro selected mutants of M. agalactiae. Thus, although in other mycoplasma species resistance to macrolides and lincosamides had been mainly related to mutations in the 23S rRNA gene, this work demonstrates the role of alterations in ribosomal protein L22 in decreased susceptibility of M. agalactiae. Moreover, these mutations can be used as molecular markers to set an interpretative breakpoint of antimicrobial resistance for M. agalactiae.

M. agalactiae is the main causative agent of contagious agalactia, against which antimicrobial treatment is the main applied control measure. Quinolones are an effective group of antimicrobials inhibiting the growth of M. agalactiae, but in the last years, various reports have demonstrated an increase of resistance in field isolates due to its massive use. Nevertheless, the molecular mechanisms involved in the acquisition of fluoroquinolones resistance in M. agalactiae have not been elucidated yet. Therefore, the aim of this work was to analyze the presence of DNA variations that could be related to changes in fluoroquinolone susceptibility. For this purpose, three M. agalactiae strains were selected to obtain in vitro resistant mutants against enrofloxacin, marbofloxacin and moxifloxacin and afterwards, partial sequences of their gyrA, gyrB, parC and parE genes were analyzed. In addition, a set of field isolates with different MIC values were also studied. Changes related to variations in fluoroquinolones susceptibility were found in gyrB, parC and parE. Specifically, gyrB genes were affected at the predicted amino acid position 424, four amino acid changes were detected in parC (positions 78, 79, 80 and 84) and two substitutions were reported in parE (amino acid positions 429 and 459). Mutations at predicted positions 424 of gyrB and 429 of parE are novel DNA changes which had not been previously described and, on the whole, parC was the first gene showing alterations when changes in susceptibility to fluoroquinolones occurred. Thus, this gene is the most suitable target for a rapid study of fluoroquinolone resistance in field isolates of M. agalactiae.

Quinolones interact with bacterial DNA gyrase and topoisomerase IV, the subunits of which are encoded by gyrA/gyrB and parC/parE, respectively. The aim of this study was to evaluate the relationship between changes in these genes and quinolone susceptibility of Mycoplasma capricolum subsp. capricolum (Mcc). Using in vitro selected resistant mutants and field isolates from goats, predicted amino acid changes in gyrA, gyrB and parC were associated with higher minimum inhibitory concentration values for quinolones. Alterations in parC predicted amino acid sequences were most frequently associated with quinolone resistance in Mcc.

En las sociedades occidentales, la tenencia de mascotas en el hogar se incrementa anualmente debido a los beneficios emocionales, físicos y psíquicos que aportan. Junto a estos beneficios, debe considerarse también la existencia de diferentes enfermedades que pueden transmitirse desde los animales al resto de convivientes en el hogar, con especial importancia para aquellas personas pertenecientes a grupos de riesgo. El aparente desafío riesgo-beneficio que provoca la tenencia de mascotas deberá solventarse considerando el estado de salud de los miembros del hogar para la selección de la especie y la recomendación de medidas higiénicas y de manejo específicas. De forma general, el conocimiento y la preocupación de los propietarios por las zoonosis asociadas a sus mascotas son escasos, incluso en los hogares donde hay personas de alto riesgo. Además, las limitaciones en los procedimientos de comunicación con los diferentes profesionales sanitarios pueden ocasionar que las situaciones de riesgo pasen desapercibidas. Por todo ello, se impone la necesidad de establecer protocolos, tanto intra como interprofesionales, para proporcionar un adecuado flujo de información que asegure la comunicación con el propietario de acuerdo con las particularidades sanitarias de los convivientes de cada hogar.

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&ndash, 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 in a 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.

The unexpected presence of Mycoplasma auris was evidenced in mastitis samples collected from a dairy goat herd under surveillance programmes of contagious agalactia (CA), a serious disease affecting Mediterranean small ruminant herds. Thus, control programs based on permanent analysis of milk from bulk tank and clinical mastitis, as well as ear swabs, are currently conducted in affected areas. In order to analyze the presence of CA- causing species, three, 11 and 18 of these samples were respectively collected from the same goat herd. Mycoplasmas were isolated in four milk samples from clinical mastitis and two ear swabs samples and no other pathogen was detected. The presence of CA-causing mycoplasmas was discarded by PCR. However, M. auris was identified in all the positive samples after the amplification and posterior PCR product sequencing of a partial fragment of their 16S-ribosomal ARN gene. As far as we know, these are the first isolations of M. auris from milk of goats with clinical mastitis.

The veterinary profession implies a greater risk of infection by zoonotic pathogens than the overall population. The aim of the present work was to evaluate the impact of zoonoses on the occupational health of veterinarians reviewing the published surveys addressing this subject. Following these inquiries, between 4% and 64.3% of the surveyed professionals acknowledge to have suffered at least one zoonotic disease, and dermatophytosis is the most frequently described zoonosis. In addition, from a qualitative point of view, it is necessary to highlight the occurrence of different diseases of important clinical seriousness and the existence of infections by antimicrobial-resistant pathogens. Due to the absence of updated studies in Spain, the present work reflects on the need to recognize the health determinants associated to the veterinary profession within the characteristics of our labour market. The available data suggest that the occurrence of zoonoses as occupational diseases is underestimated in Spain. Therefore, performing surveys on occupational health, documenting and publishing cases and reviewing the risks and the impact of zoonotic diseases on the veterinary profession would contribute to the description and notification of such diseases on behalf of the healthcare administration and would also become an essential tool in occupational risk prevention.La profesión veterinaria presenta mayor riesgo de infección frente a agentes patógenos de carácter zoonósico que la población general. Con el objetivo de valorar el impacto de las zoonosis, el presente trabajo revisa las encuestas publicadas sobre salud laboral en la profesión veterinaria. En dichas encuestas, el rango de profesionales que refieren haber padecido al menos una zoonosis oscila entre el 4% y el 64,3%, siendo la dermatofitosis la zoonosis comunicada con mayor frecuencia. Además, desde el punto de vista cualitativo, cabe destacar la comunicación de diferentes procesos de especial gravedad clínica, así como la existencia documentada de infecciones por agentes patógenos resistentes a los antimicrobianos. Dada la ausencia de estudios actualizados en España, se reflexiona sobre la necesidad de conocer los determinantes de salud asociados a las características del mercado laboral de la profesión en nuestro entorno. Los datos existentes sugieren que la consideración legal de enfermedad profesional para las zoonosis se encuentra infraestimada en nuestro país. Por todo ello, la realización de encuestas de salud laboral, la documentación y publicación de los casos, así como la revisión de los riesgos y el impacto de las zoonosis en la profesión veterinaria, pueden contribuir a la calificación y notificación de dichas enfermedades por parte de la administración sanitaria, al tiempo que suponen una herramienta fundamental en el ámbito de la prevención de riesgos laborales.

One of the most economically impacting diseases that affects small domestic ruminants in arid lands of Africa and Asia is contagious caprine pleuropneumonia (CCPP), whose causative agent is Mycoplasma capricolum subspecies (subsp.) capripneumoniae (Mccp). The exclusively respiratory clinical-lesional picture caused by Mccp includes the presence of pleuropneumonia with unilateral hepatization, adhesions, pleuritis and pleural fluid accumulation. Its improved diagnosis has confirmed the spread of the disease across these continents (e.g. Kenya, Ethiopia, Tanzania or Pakistan), also confirming its presence on other typical species of these arid zones (e.g. gazelles or mouflons). This report reviews the most important features of CCPP and examines in detail the most novel aspects of its epidemiology, diagnosis and control. In brief, the repercussions of two other mycoplasmal emergent diseases also affecting small ruminants of arid zones are also addressed: atypical pneumonia caused by Mycoplasma ovipneumoniae, which participates with other microorganisms in the domestic small ruminant respiratory syndrome occurred in semi-arid areas of Europe (e.g. Portugal or Spain), and infectious keratoconjunctivitis, whose main responsible agent is Mycoplasma conjunctivae. This last disease mostly affects wild ruminants worldwide with recent reports of clinical outbreaks in countries as Saudi Arabia, Iran, Australia, India or European zones as the Pyrenees or the Alps, where it is considered endemic.

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.

Background 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. Objectives The present study utilizes a systematic review of the literature to identify zoonoses described in veterinary students. Data sources Web of Science and PubMed. Results 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. Limitations 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. Conclusions/implications 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.

Contagious agalactia (CA) is a serious disease of small dairy ruminants that has a substantial economic impact on the goat and sheep milk industries. The main aetiological agent of the disease is Mycoplasma agalactiae, although other species, such as Mycoplasma mycoides subsp. capri, Mycoplasma capricolum subsp. capricolum and Mycoplasma putrefaciens, are pathogenic in goats. There are two clinical-epidemiological states of CA in sheep and goats; herds and flocks may exhibit outbreaks of CA or may be chronically infected, the latter with a high incidence of subclinical mastitis and only occasional clinical cases. The complex epidemiology of CA is related to the genetic characteristics and mechanisms of molecular variation of the Mycoplasma spp. involved, along with presence of CA-mycoplasmas in wild ruminant species. In goats, the situation is particularly complex and asymptomatic carriers have been detected in chronically infected herds. The coexistence of other non-pathogenic mycoplasmas in the herd further complicates the diagnosis of CA and the design of efficient strategies to control the disease. Routes of infection, such as the venereal route, may be involved in the establishment of chronic infection in herds. Current challenges include the need for improved diagnostic methods for detection of chronic and subclinical infections and for the design of more efficient vaccines.

The presence of goat males carriers of the main causal agents of contagious agalactia, Mycoplasma agalactiae (Ma) and Mycoplasma mycoides subsp. capri (Mmc), is a high risk factor for the spread of the infection in artificial insemination centres. The present study was designed to assess the efficacy of a 5 day course of marbofloxacin given intramuscularly (2 mg kg−1 day−1) in terms of its capacity to reverse the carrier state of goat buck populations naturally infected with one or both mycoplasmas (Mmc, n = 25; Mmc + Ma, n = 2). Before and after treatment, ear swabs and semen samples were culture- and PCR-checked for both mycoplasmas. We also evaluated the impact of this treatment on semen quality (sperm count, motility and daily sperm production) by monitoring these variables of 594 semen samples collected during pre- and post-treatment periods of 48 days each. All bucks examined tested positive for the presence of Ma and Mmc before and after treatment though Mycoplasma spp. never was identified in semen. Sperm motility was significantly reduced in response to treatment (from 72.9% to 56.9%), although some recovery was observed at the end of the study (40 days after treatment). Ejaculate volume and concentration remained unchanged throughout the study. Our findings indicate that systemic marbofloxacin therapy is unable to eliminate mycoplasmas from the external auditory canal of male goats, and that it also causes a transient detrimental effect on sperm motility. Other strategies to control the presence of mycoplasma carriers in goat artificial insemination centres need to be assessed.

Mycoplasma mycoides subsp. capri (Mmc) is one of the main causative agents of caprine contagious agalactia. Besides, the absence of accurate control methods eases its dispersion between different herds within endemic areas of this disease. In this context, there is a need to implement molecular typing schemes which offer valuable information useful to establish control measures and enables the surveillance of this pathogen. The aim of this study was to assess the genetic variability of different strains of Mmc from a contagious agalactia endemic area through multilocus sequence typing (MLST). For this purpose, five house-keeping genes (fusA, glpQ, gyrB, lepA, rpoB) from 39 field isolates were analysed. These isolates were obtained from different geographic areas of Spain, between the years 2004 and 2015. The results obtained in this study suggest that the selected MLST scheme could be a useful technique to monitor the genetic variability of Mmc in endemic areas. Despite the significant differences found between the assessed field isolates, they could be classified according to their geographical origin. Moreover, it was also possible to detect genetic differences between Mmc strains coming from the same herd at the same sampling time, which may need to be taken into consideration when designing or arranging prophylactic strategies.

Coxiella (C.) burnetii is the etiological agent of the zoonotic disease known a Q fever. This agent can infect multiple hosts although its pathogenic potential in wild ruminants has been poorly studied. The polymerase chain reaction and the serological test detected the presence of C. burnetii in a population of North African gazelles (n = 355), comprising dorcas gazelle (Gazella dorcas neglecta), dama gazelle (Nanger dama mhorr) and Cuvier's gazelle (Gazella cuvieri) which, some of them, they recently aborted. Serological tests for Brucella spp., C. burnetii, Chlamydophila abortus, border disease pestivirus, and Toxoplasma spp. were performed together with specific cultures to detect Salmonella spp., Listeria spp., and Campylobacter spp. and a polymerase chain reaction for C. burnetii on serum and vaginal swabs samples collected from a representative number of animals (n = 65). These tests only detected the presence of C. burnetii in 18 specimens (27.3%). C. burnetii was the only pathogen detected, with eight animals testing positive on the polymerase chain reaction, 15 on the serological test, and five on both the tests. This article reveals the presence of C. burnetii during a medium and late-stage abortions occurred in a population of North African gazelles. The presence of C. burnetii as causal agent of abortions in Cuvier's gazelles has never been reported. The consequences of the findings are discussed here, showing the need to adopt urgent control measures to prevent the spread of C. burnetii in captive populations that are essential for the conservation of these endangered species.

This study was designed to monitor the presence of Mycoplasma agalactiae and Mycoplasma mycoides subsp. capri (Mmc) in 66 dairy goat herds of a genetic improvement programme in a region of Spain where contagious agalactia is endemic. Over a whole lactation period, 300 bulk tank milk and 381 milk samples from goats with clinical mastitis were subjected to polymerase chain reaction (PCR) to detect the two mycoplasma species. The presence of mycoplasmas (either species or both) was detected in 66.7% of the herds and M. agalactiae was identified in 95.45% of these positives herds. In a given infected herd, mycoplasmas were not continuously detected over the whole study period. Our findings indicate that in an endemic area, M. agalactiae and Mmc can be monitored through PCR analysis of mastitic milk and bulk tank milk (BTM) samples. Over a lactation period we recommend testing multiple BTM samples on a herd. No relationship was observed between the use of inactivated mycoplasma vaccines and the PCR detection of both mycoplasmas.

The role of inapparent carriers of Mycoplasma agalactiae and the strategies used to colonise the external ear canal in goats remain unclear. This study examined the ability of M. agalactiae to colonise the ears of goats infected experimentally by the intramammary route. The right mammary glands of 15 lactating goats were inoculated with 10(10) colony forming units (cfu) of M. agalactiae. The goats were randomly assigned to three groups of five animals each and sampled at slaughter at 5, 15 or 45 days post-infection (dpi). A further four goats served as uninfected controls. Right and left ear swabs were collected for detection of M. agalactiae by culture before and after sacrifice. M. agalactiae was detected in 19/20 (95%) ear swabs from goats sampled at 15 and 45dpi, whereas all ear swabs collected before inoculation, ear swabs collected from the group sampled at 5dpi and ear swabs from control goats at the time of sacrifice were negative for M. agalactiae. Blood samples collected at 6, 12, 24, 48 and 72h post-infection for detection of M. agalactiae by culture were also negative. There were differences in the antigenic profiles of isolates recovered from the ears compared to the 7MAG strain used to inoculate the animals and most isolates from the mammary gland, milk and supramammary lymph nodes.

Goat milk samples routinely obtained by milk distributing companies and quality control laboratories are preserved using azidiol (AZ) or bronopol (BR). This study was designed to determine if these treated goat milk samples are suitable for Mycoplasma ( M. ) agalactiae detection by PCR. The effects of these preservatives on the limits of M. agalactiae direct PCR detection were established using samples inoculated with mycoplasma cultures. Samples inoculated with a determined concentration of M. agalactiae showed no differences among the 3 preservation methods and in all cases the limits of detection of the PCR assays were 10 3 CFU ml. In addition, 300 bulk tank milk (BTM) samples from 45 herds from an infected area were each divided into three aliquots subjected to different preservation treatments (no preservative, NP, AZ or, BR) and then PCR tested. Of the 300 BTM samples, 43 different ones scored a positive result for M. agalactiae in at least one condition distributed in this way, 32 NP aliquots (74.4% of the positive samples), 27 BP aliquots (62.8%) and 27 AZ aliquots (62.8%). Among the three preservation methods, no significant differences were found ( p > 0.05) and the difference in results was attributed to the sensitivity increasement when analyzing more than one time a BTM sample. These results suggest that the PCR technique can be used as a rapid diagnostic method to detect M. agalactiae in AZ or BR preserved goat milk samples. The possible use of such samples for health programmes would have considerable cost benefits.

Mycoplasma bovis is a serious, worldwide-spread but often overlooked pathogen causing respiratory disease, mastitis, and arthritis in cattle. In this study we characterize the protein and antigenic profiles of M. bovis field strains isolated in Bosnia and Herzegovina by sodium dodecyl sulphate polyacrylamide gel electrophoresis and immunoblotting, and analyze possible variations among these strains. Greater differences occurred when comparing field strains with the reference strain PG45. One field strain isolated from lung samples of a heifer was markedly different from strains isolated from nasal swabs taken from cattle raised in another geographic region. A possible correlation may exist between protein and antigen profiles of M. bovis field strains, geographic regions and anatomical sites of isolation. Mycoplasma bovis, cattle, antigen, immunoblotting, protein, SDS-PAGE Mycoplasma bovis is the most pathogenic bovine mycoplasma in the parts of the world considered to be free of contagious bovine pleuropneumonia. This pathogen primarily causes calf pneumonia, mastitis and arthritis. Other symptoms are seen less frequently. Diseases caused by M. bovis are found worldwide, causing significant economic losses. Yet, there are no effective control measures or standard protocols for their routine application available (Nicholas and Ayling 2003). Although M. bovis, like other mycoplasmas, possesses a small genome size, it is still highly invasive and able to colonize organ systems. It is now clear that one of the possible explanations for its ability to avoid bodily defence mechanisms and adapt to different environments in the host is due to phenotypic variations in mycoplasmas (Wise 1993; Razin et al. 1998; Minion 2002). Rosengarten et al. (1994) demonstrated highly variable membrane surface proteins (Vsps) of M. bovis which represent a family of antigenically and structurally related lipoproteins. These Vsps represent the predominant antigens recognized by the host immune response during infection, and they can undergo high frequency changes in size and expression. This system is responsible for the survival of these pathogens in the presence of cytolytic antibodies and for their adaptation to specialized environments within their respective hosts (Behrens et al. 1994; Razin et al.1998). A full understanding of phenotypic variation among M. bovis strains should be of great value when developing diagnostic methods and vaccines for the control of infections caused by this organism. Recently, M. bovis has been isolated, for the first time, in Bosnia and Herzegovina (Rifatbegovic et al. 2007). The aim of this study was to characterize the protein and antigenic profile of these M. bovis field strains by SDS-PAGE (sodium dodecyl sulphatepolyacrylamide gel electrophoresis) and immunoblot and to analyze possible variations among these strains.