Contagious agalactia: costs and control revisited.

The economic costs of contagious agalactia (CA) to the small ruminant dairy industry are not well known but include losses due to mortality, lowered milk production, spoiled products, abortions and animal welfare problems, as well as diagnosis and treatment. This paper reports financial estimates made in southern Europe, including a study on small- and large-scale farming systems in Italy, indicating that the financial losses are high and underestimated. Furthermore, the current control strategies, including chemotherapy and vaccination, in selected countries in Europe are described. In some countries, disease control is hampered by excessively strict veterinary legislation which discourages farmers and private veterinarians from notifying outbreaks because it leads to the prohibition of milk sales and can result in delays in lifting restrictions. In addition, new European Union legislation may downgrade the importance of CA, which will have implications for international research efforts. Finally, a series of recommendations are provided that cover the proper notification and handling of CA outbreaks, including movement control, current diagnostics, treatment, vaccination and disinfection.

Si le coût économique exact de l'agalaxie contagieuse pour le secteur ovin et caprin de production laitière n'est pas connu, on sait néanmoins qu'il recouvre les pertes dues à la mortalité dans les cheptels, à une chute de la production de lait, aux produits altérés, aux avortements et aux problèmes de bien-être animal, en plus des coûts du diagnostic et des traitements. Les auteurs font état d'estimations financières réalisées en Europe méridionale, dont une étude sur les exploitations familiales et les élevages de grande taille en Italie, qui coïncident dans le constat de pertes financières à la fois importantes et sous-estimées. Les auteurs décrivent également les stratégies de lutte mises en place actuellement par plusieurs pays d'Europe, en particulier l'antibiothérapie et la vaccination. Dans certains pays, les efforts de lutte sont entravés par une législation vétérinaire excessivement rigoureuse qui dissuade les éleveurs et les vétérinaires privés de notifier les foyers car cela entraîne l'interdiction de vendre le lait issu des troupeaux infectés et retarde la levée des mesures de restriction. En outre, la nouvelle réglementation de l'Union européenne risque d'abaisser l'importance de l'agalaxie contagieuse, ce qui aura des conséquences sur les efforts mobilisés par la recherche au niveau international. Pour conclure, les auteurs formulent plusieurs recommandations en vue d'une notification et gestion appropriées des foyers d'agalaxie contagieuse, notamment pour ce qui concerne le contrôle des mouvements d'animaux, les méthodes actuelles de diagnostic, le traitement, la vaccination et la désinfection.

Aunque no se conocen bien los costos económicos que la agalaxia contagiosa inflige a la industria lechera de pequeños rumiantes, se sabe que las pérdidas por mortalidad, mengua de la producción lechera, productos echados a perder, abortos y problemas de bienestar animal son un factor importante, sin olvidar los gastos de diagnóstico y tratamiento. Los autores dan cuenta de cálculos económicos realizados en Europa meridional, en particular a raíz de un estudio de pequeñas y grandes explotaciones ganaderas de Italia, que llevaron a la conclusión de que las pérdidas económicas son cuantiosas y están subestimadas. Además, los autores describen los métodos de lucha aplicados actualmente en determinados países de Europa, que incluyen tratamiento medicamentoso y vacunaciones. En algunos países la lucha contra la enfermedad se ve lastrada por una legislación veterinaria demasiado estricta, que no alienta a productores y veterinarios privados a notificar brotes porque ello conduce a la prohibición de las ventas de leche y puede demorar el levantamiento de las restricciones. Por otra parte, hay nuevos textos legislativos de la Unión Europea que quizá vengan a restar importancia a la agalaxia contagiosa, lo que repercutiría en las actividades internacionales de investigación. Por último, los autores formulan una serie de recomendaciones referidas a cuestiones que van desde la correcta notificación y gestión de los brotes de agalaxia contagiosa hasta el control de los desplazamientos, pasando por los procedimientos vigentes de diagnóstico o los métodos de tratamiento, vacunación y desinfección.

Gap analysis of Mycoplasma bovis disease, diagnosis and control: An aid to identify future development requirements.

There is a worldwide problem of disease caused by Mycoplasma (M.) bovis in cattle; it has a significant detrimental economic and animal welfare impact on cattle rearing. Infection can manifest as a plethora of clinical signs including mastitis, pneumonia, arthritis, keratoconjunctivitis, otitis media and genital disorders that may result in infertility and abortion. Current diagnosis and control information are reviewed and analysed to identify gaps in knowledge of the causative organism in respect of the disease pathology, diagnosis and control methods. The main considerations are as follows: no vaccines are commercially available; antimicrobial resistance is increasing; diagnostic and antimicrobial sensitivity testing needs to be improved; and a pen-side test would facilitate more rapid diagnosis and implementation of treatment with antimicrobials. More data on host susceptibility, stress factors, immune response and infectious dose levels are required. The impact of asymptomatic carriers, M. bovis survival in the environment and the role of wildlife in transmitting the disease also needs investigation. To facilitate development of vaccines, further analysis of more M. bovis genomes, its pathogenic mechanisms, including variable surface proteins, is required, along with reproducible disease models.

Survey of contagious caprine pleuropneumonia in goat herds in the Thrace region of Turkey.

Contagious caprine pleuropneumonia (CCPP) was detected for the first time on the European continent in the Thrace region of Turkey in 2002 following outbreaks of an unusually severe respiratory disease in goats. Mycoplasma capricolums ubspecies capripneumoniae (Mccp), the causative agent, was identified in many infected herds throughout the region by serological, bacteriological and molecular biological techniques. However, as no quantitative data on the prevalence and distribution of the disease have been gathered in the intervening years, the extent of infection is unknown. Consequently, in 2014, a random sample of 2,400 goats was drawn from a population of over 167,000 goats in the region by multistage sampling. Serum samples were collected and tested by a monoclonal antibodybased competitive enzyme-linked immunosorbent assay (cELISA). The overall prevalence of CCPP in Thrace was found to be just over 8%. The prevalence in the individual provinces was approximately 18%, 7%, 8% and 6% for Çanakkale, Edirne, K?rklareli and Tekirda?, respectively, while Istanbul province appeared to be CCPP-free. The results showed that CCPP has spread throughout the region and has become endemic, and may pose a potential risk to the neighbouring countries of Greece and Bulgaria. Mccp was regularly detected from lung samples of suspect goats until 2015 but since then there has been no further detection from clinical samples.

La pleuropneumonie contagieuse caprine (PPCC) était détectée pour la première fois sur le continent européen en Thrace orientale (Turquie) en 2002, à la suite de foyers d'une maladie respiratoire particulièrement sévère affectant le cheptel caprin. L'agent causal identifié par diverses méthodes sérologiques, bactériologiques et de biologie moléculaire dans plusieurs troupeaux infectés de la région était Mycoplasma capricolum sous-espèce capripneumoniae (Mccp). Cependant, en l'absence de données quantitatives sur la prévalence et la distribution de la maladie depuis ces foyers, l'extension de l'infection est restée inconnue. Pour y remédier, un échantillonnage stratifié et aléatoire de2 400 chèvres parmi une population de plus de 167 000 chèvres de la région a été réalisé en 2014. L'analyse par la méthode immunoenzymatique de compétition (cELISA) faisant appel à des anticorps monoclonaux des sérums collectés a montré que la prévalence globale pour la Thrace orientale était un peu au-dessus de 8 %. Dans les provinces de Çanakkale, d'Edirne, de K?rklareli et de Tekirda?, la prévalence était respectivement de 18 %, 7 %, 8 % et 6 % tandis qu'Istanbul était indemne de PPCC. Ces résultats montrent que la PPCC est devenue endémique dans l'ensemble de la région et représente un risque potentiel pour les pays avoisinants, c'est-à-dire la Grèce et la Bulgarie. Jusqu'en 2015, Mccp a été régulièrement détectée dans les échantillons de poumon prélevés chez des chèvres suspectées infectées mais depuis cette date aucun échantillon clinique n'a été testé positif.

En 2002 se detectó por primera vez la pleuroneumonía contagiosa caprina en el continente europeo, a raíz de una serie de brotes de enfermedad respiratoria de inusitada gravedad que afectaron a cabras de la región turca de Tracia. Empleando técnicas serológicas, bacteriológicas y de biología molecular, se detectó en muchos rebaños infectados de toda la región la presencia del agente causal, Mycoplasma capricolum subespecie capripneumoniae (Mccp). Sin embargo, se desconoce el alcance de la infección, porque en los años transcurridos desde entonces no se han reunido datos cuantitativos sobre la prevalencia y distribución de la enfermedad. Por consiguiente en 2014, a partir de una población de más de 167.000 cabras de la región, se obtuvo por muestreo multietápico una muestra aleatoria de 2.400 ejemplares, a los que se extrajeron muestras de suero que fueron sometidas a un ensayo inmunoenzimático de competición (ELISAc) con anticuerpos monoclonales. Se calculó que la prevalencia global de pleuroneumonía contagiosa caprina en Tracia era algo superior al 8%. Por provincias, la prevalencia resultó de alrededor de un 18%, un 7%, un 8% y un 6% en las provincias de Çanakkale, Edirne, K?rklareli y Tekirda?, respectivamente, mientras que la provincia de Estambul resultó estar libre de la infección. Estos resultados demostraron que la enfermedad se ha extendido por toda la región, ha cobrado carácter endémico y puede amenazar a los países vecinos, como Grecia y Bulgaria. Hasta 2015 se detectó regularmente la presencia de Mccp en muestras pulmonares de cabras sospechosas, pero desde entonces no se ha vuelto a observar tal cosa en ninguna muestra clínica.

Search for OIE-listed ruminant mycoplasma diseases in Afghanistan.

BACKGROUND: Little is known about the occurrence of important diseases of ruminants in Afghanistan because of the conflict affecting the country over the last 40 years. To address this discrepancy, ruminant herds in Afghanistan were screened for OIE-listed mycoplasma diseases, contagious bovine (CBPP) and caprine pleuropneumonias (CCPP). RESULTS: Of the 825 samples from 24 provinces tested for serological evidence of CBPP caused by Mycoplasma mycoides subsp.mycoides, 20 (3.4%) had ELISA values greater than the positive threshold of 50% though all were less than 55%. Repeat testing of these suspect sera gave values below 50. A smaller number of sera (330) from cattle in nine provinces were also tested by the rapid latex agglutination test (LAT) for CBPP, 10 of which were considered suspect. However, no positive bands were seen when immunoblotting was carried out on all sera that gave suspect results. Serological evidence of Mycoplasma bovis was detected in half of 28 herds in eight provinces. The cause of CCPP, M. capricolum subsp. capripneumoniae was not detected in any of the 107 nasal swabs and lung tissue collected from goats in seven provinces though sample handling and storage were not optimal. However, strong serological evidence was detected in goat herds in several villages near Kabul some of which were over 50% seropositive by LAT and ELISAs for CCPP; immunoblotting confirmed positive results on a selection of these sera. CONCLUSIONS: The data presented here provide a first assessment of the occurrence of the two OIE listed mycoplasma diseases in Afghanistan. From the results of the testing bovine sera from the majority of provinces there is no evidence of the presence of CBPP in Afghanistan. However the samples tested represented only 0.03% of the cattle population so a larger survey is required to confirm these findings. Serological, but not bacterial, evidence was produced during this investigation to show that CCPP is highly likely to be present in parts of Afghanistan.

16S rRNA gene mutations associated with decreased susceptibility to tetracycline in Mycoplasma bovis.

Mycoplasma bovis isolates with decreased susceptibilities to tetracyclines are increasingly reported worldwide. The acquired molecular mechanisms associated with this phenomenon were investigated in 70 clinical isolates of M. bovis. Sequence analysis of the two 16S rRNA-encoding genes (rrs3 and rrs4 alleles) containing the primary binding pocket for tetracycline (Tet-1 site) was performed on isolates with tetracycline hydrochloride MICs of 0.125 to 16 µg/ml. Mutations at positions A965T, A967T/C (Escherichia coli numbering) of helix 31, U1199C of helix 34, and G1058A/C were identified. Decreased susceptibilities to tetracycline (MICs, ≥2 µg/ml) were associated with mutations present at two (A965 and A967) or three positions (A965, A967, and G1058) of the two rrs alleles. No tet(M), tet(O), or tet(L) determinants were found in the genome of any of the 70 M. bovis isolates. The data presented correlate (P<0.0001) the mutations identified in the Tet-1 site of clinical isolates of M. bovis with decreased susceptibility to tetracycline.

Global multilocus sequence typing analysis of Mycoplasma bovis isolates reveals two main population clusters.

Mycoplasma bovis is a major bovine pathogen associated with bovine respiratory disease complex and is responsible for substantial economic losses worldwide. M. bovis is also associated with other clinical presentations in cattle, including mastitis, otitis, arthritis, and reproductive disorders. To gain a better understanding of the genetic diversity of this pathogen, a multilocus sequence typing (MLST) scheme was developed and applied to the characterization of 137 M. bovis isolates from diverse geographical origins, obtained from healthy or clinically infected cattle. After in silico analysis, a final set of 7 housekeeping genes was selected (dnaA, metS, recA, tufA, atpA, rpoD, and tkt). MLST analysis demonstrated the presence of 35 different sequence types (STs) distributed in two main clonal complexes (CCs), defined at the double-locus variant level, namely, CC1, which included most of the British and German isolates, and CC2, which was a more heterogeneous and geographically distant group of isolates, including European, Asian, and Australian samples. Index of association analysis confirmed the clonal nature of the investigated M. bovis population, based on MLST data. This scheme has demonstrated high discriminatory power, with the analysis showing the presence of genetically distant and divergent clusters of isolates predominantly associated with geographical origins.

First isolation of Mycoplasma iowae in grey partridge flocks.

Mycoplasma iowae, an occasional pathogen of turkeys, was isolated for the first time from captive grey partridges (Perdix perdix). Clinical signs including respiratory and intestinal disorder were seen in birds of all ages but mainly in those kept housed during rearing. Mortality rates averaged over 20% during the year. Treatment with antibiotics and antiparasitic drugs produced only a transient improvement in condition. The gross pathology findings included poor body growth, lack of development of the breast muscles, abnormalities in the keel development, and bone fragility. Some birds showed infraorbital sinusitis with serous or fibrinous exudates and catarrhal tracheitis, while others presented serofibrinous airsacculitis and splenomegaly. Laboratory investigations revealed pure cultures of M. iowae in the gut as well as sinus and air sacs. While other organisms such as coccidia, Trichomonas, Escherichia coli, Clostridium perfringens, and Aspergillus spp. were detected, the similarity of the disease with that seen in turkeys infected with M. iowae strongly suggests that this mycoplasma may be the primary pathogen here. The presence of M. iowae in game birds commonly released into the wild could have serious implications particularly in areas where industrial poultry farms are concentrated.

Isolation, molecular characterization and antimicrobial susceptibilities of isolates of Mycoplasma agalactiae from bulk tank milk in an endemic area of Spain.

AIM: To isolate and characterize strains of Mycoplasma agalactiae from bulk tank and silo ewes' milk. METHODS AND RESULTS: Thirteen mycoplasma isolates were obtained from samples of sheep milk taken from bulk tank and large silos and identified as Myc. agalactiae by PCR-DGGE. The isolates were typed by pulsed field gel electrophoresis (PFGE), SDS-PAGE and immunoblot. The in vitro activity of 13 antimicrobials of veterinary interest was tested against these isolates. Results showed that the most effective compounds against Myc. agalactiae in vitro were clindamycin, an antibiotic not previously described as a suitable contagious agalactia (CA) treatment, with Minimum Inhibitory Concentration (MIC) values of <0·12 µg ml(-1) , and quinolones, with MIC values <0·12-0·5 µg ml(-1) , which are used as standard treatments against CA. CONCLUSIONS: Based on the in vitro assay, clindamycin, quinolones, tylosin and tilmicosin would be appropriate antimicrobials for CA treatment. The isolates were mostly resistant to erythromycin, indicating that it would not be a suitable choice for therapy. The isolates showed common molecular and protein profiles by PFGE and SDS-PAGE, with minor differences observed by immunoblot analysis, suggesting a clonal relationship among them. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrated the importance of the appropriate selection of antimicrobials for treatment of CA.

Serological survey to determine the occurrence of respiratory Mycoplasma infections in the Polish cattle population.

Mycoplasma bovis and Mycoplasma mycoides subspecies mycoides small colony (MmmSC) are causes of bovine mycoplasmosis and contagious bovine pleuropneumonia (CBPP), respectively, and are responsible for serious economic losses in cattle around the world. CBPP was last reported in Poland in 1939 but bovine mycoplasmosis is believed to be endemic. A survey of 3670 serum samples for antibodies to M bovis and MmmSC from 361 herds in 16 Polish provinces Poland between 2007 and 2010 found no evidence of CBPP. The seroprevalence of M bovis, however, appeared high with 76.7 per cent of samples giving a positive reaction in the ELISA test, which did not appear to reflect the clinical disease status of the cattle. Adjusting the sensitivity of the test reduced the prevalence to 28.2 per cent and reflects the levels reported in other European countries.

Mycoplasma neophronis sp. nov., isolated from the upper respiratory tract of Canarian Egyptian vultures (Neophron percnopterus majorensis).

Six strains with the typical characteristics of mycoplasmas were isolated from the tracheae of six Canarian Egyptian vultures (Neophron percnopterus majorensis). The results of biochemical, serological and molecular genetic studies showed that the isolates were nearly identical and that they could be considered as representing a novel species of the genus Mycoplasma. Colonies possessed the typical fried-egg appearance and electron micrographs revealed a pleomorphic cellular morphology with the lack of a cell wall. The isolates hydrolysed arginine and required sterol for growth but did not ferment glucose or hydrolyse urea. We propose that the isolates be assigned to a novel species,Mycoplasma neophronis sp. nov. The type strain is G.A.(T) ( = DSM 24097(T) = ATCC BAA-2157(T)). The antiserum of strain G.A.(T) has been deposited in the Mollicutes collection at Purdue University (Indiana, USA).

Investigations into an outbreak of corvid respiratory disease associated with Pasteurella multocida.

The possible cause of disease and mortality in corvids on an outdoor pig unit in the north of England between August 2007 and March 2008 was investigated. Nine carrion crows (Corvus corone corone) and nine rooks (Corvus frugilegus), comprising five live-caught birds with clinical signs of respiratory disease, one live-caught bird without respiratory disease, and 12 birds submitted dead were examined. Clinical signs, gross and histopathological examination, microbiology and toxicology indicated that Pasteurella multocida infection was the cause of disease. Molecular and serotyping analyses showed that P. multocida isolates (obtained from live-caught birds with clinical respiratory disease) were all capsular type F with a mix of somatic serotypes 3, 4 and 7. Immunohistochemistry increased the diagnostic sensitivity of the analysis and detected P. multocida within the pulmonary lesions of all affected live-caught birds and 10 of 12 birds found dead. These findings suggest that wild corvids in the UK can suffer from lung pathology associated with P. multocida and, as potential vectors of P. multocida, may pose a risk to domestic poultry.

Bovine mycoplasmosis: silent and deadly.

The first international conference on bovine mycoplasmosis, held in Saskatoon, Canada, in 2009, marked an official recognition of the importance of mycoplasmas in bovine respiratory disease, mastitis, arthritis and other disorders. Furthermore, bovine mycoplasmosis has been identified as a major emerging infectious disease by the European Community 7th Framework Programme under the EMIDA ERA-NET, which is funding a three-year study into the development of improved diagnosis and control of bovine mycoplasmosis involving the UK, Germany, Italy and Israel. These developments coincide with the publication of the complete sequence of the Mycoplasma bovis genome, which offers fresh insight into the properties of this tiny wall-less bacterium. With this in mind, it is an appropriate time to review our knowledge of cattle diseases caused by mycoplasmas.

Multilocus sequence typing of Mycoplasma agalactiae.

Mycoplasma agalactiae is the main cause of contagious agalactia, a serious disease of sheep and goats, which has major clinical and economic impacts. We have developed a multilocus sequence typing (MLST) scheme using the sequenced genomes of the M. agalactiae strains PG2 and 5632. An MLST scheme based on the genes gltX, metS, gyrB, tufA and dnaA was designed and in total 3468 bp of sequence were analysed for each strain. MLST offers a highly discriminatory typing method for M. agalactiae and was capable of subdividing 53 strains into 17 distinct sequence types, largely according to geographical origin. MLST detected unexpected diversity in recent isolates from Spain, identifying two novel outliers, and enabled typing of novel Mongolian isolates for the first time. Genetic diversity in the sequenced regions was largely due to mutation, with recombination playing a much smaller role. A web-accessible database has been set up for this MLST scheme for M. agalactiae: http://pubmlst.org/magalactiae/. MLST offers a robust, objective molecular epidemiological tool for M. agalactiae that that enables interlaboratory comparison of data.

Outbreak of bovine clinical mastitis caused by Mycoplasma bovis in a North Italian herd.

This report describes an outbreak of Mycoplasma bovis mastitis affecting 45 cows in a herd of 122 dairy cattle in Northern Italy. Clinically, the outbreak was characterized by agalactia, multiple swollen and painless quarters, high milk somatic cell count and unresponsiveness to conventional antibiotic therapy. M. bovis was isolated from the milk samples of all the 32 affected cows tested and from the mammary tissue of three affected cows that underwent necropsy. No other pathogens were isolated from these samples. Lesions in two of the necropsied cows were characterized by mild chronic suppurative mastitis and galactophoritis. The other necropsied cow showed a chronic necrosuppurative and pyogranulamaous galactophoritis, a condition not previously associated with M. bovis. M. bovis was detected immunohistochemically in the lumen of the affected mammary ducts suggesting that ascending infection via the teat canal was the likely route of transmission. No other intralesional pathogens were demonstrated microscopically.

Experimental infection of goats with an unusual strain of Mycoplasma mycoides subsp. capri isolated in Jordan.

Goats were infected experimentally with a mycoplasma (the "Irbid" strain) isolated previously from a goat with contagious agalactia in northern Jordan. The strain was unusual in that, although it had been identified by molecular methods as Mycoplasma mycoides subsp. mycoides LC/Mycoplasma mycoides subsp. capri, it showed no inhibition of growth by any of the hyperimmune rabbit antisera conventionally used to speciate members of the Mycoplasma mycoides cluster. Animals were infected either intratracheally or by aerosol and placed "in-contact" with other goats. After 2 weeks, those infected intratracheally became febrile, showing a nasal discharge and slight conjunctivitis, followed a week later by respiratory distress and polyarthritis; lesions seen at necropsy included coagulative necrotic pneumonia, fibrinous pleurisy with pleural exudate, and inflammatory exudates, necrosis and fibrosis in the joints. Animals infected by aerosol showed much milder clinical signs, including nasal discharge and occasional swollen joints. In the "in-contact" goats, seroconversion was first seen after 7 weeks, accompanied by coughing and laboured respiration; lesions in this group consisted of fibrinous pneumonia with focal areas of necrosis and abundant pleural exudate.