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.

NUPR1, a new target in liver cancer: implication in controlling cell growth, migration, invasion and sorafenib resistance.

Sorafenib, an oral multikinase inhibitor, is the only approved agent for the treatment of advanced hepatocellular carcinoma (HCC). However, its benefits are modest, and as its mechanisms of action remain elusive, a better understanding of its anticancer effects is needed. Based on our previous study results, we investigated here the implication of the nuclear protein 1 (NUPR1) in HCC and its role in sorafenib treatment. NUPR1 is a stress-inducible protein that is overexpressed in various malignancies, but its role in HCC is not yet fully understood. We found that NUPR1 expression was significantly higher in primary human HCC samples than in the normal liver. Knockdown of NUPR1 significantly increased cell sensitivity to sorafenib and inhibited the cell growth, migration and invasion of HCC cells, both in vitro and in vivo. Moreover, NUPR1 silencing influenced the expression of RELB and IER3 genes. Unsurprisingly, RELB and IER3 knockdown also inhibited HCC cell viability, growth and migration. Using gene expression profiling of HCC cells following stable NUPR1 knockdown, we found that genes functionally involved in cell death and survival, cellular response to therapies, lipid metabolism, cell growth and proliferation, molecular transport and cellular movement were mostly suppressed. Network analysis of dynamic gene expression identified NF-κB and ERK as downregulated gene nodes, and several HCC-related oncogenes were also suppressed. We identified Runt-related transcription factor 2 (RUNX2) gene as a NUPR1-regulated gene and demonstrated that RUNX2 gene silencing inhibits HCC cell viability, growth, migration and increased cell sensitivity to sorafenib. We propose that the NUPR1/RELB/IER3/RUNX2 pathway has a pivotal role in hepatocarcinogenesis. The identification of the NUPR1/RELB/IER3/RUNX2 pathway as a potential therapeutic target may contribute to the development of new treatment strategies for HCC management.

Expansion of intracellular IFN-γ positive lymphocytes during Mycoplasma agalactiae infection in sheep.

A method to assess the expansion of antigen-specific intracellular IFN-γ positive T cell subsets during the infection will be helpful for a better understanding of mycoplasmal infections physiopathology in the sheep. We analysed the percentage of antigen-specific lymphocytes positive for intracellular IFN-γ during the infection of sheep with Mycoplasma agalactiae by culturing peripheral blood mononuclear cells of infected or uninfected animals with irradiated M. agalactiae. The expansion of antigen-specific IFN-γ positive lymphocytes in infected sheep was initially sustained by CD4(+) T cells at day 15 after infection, when antigen specific IgG start to be detectable, followed by CD8/IFN-γ double positive cells. γδ T-cells were not expanded at any time point analysed. IFNγ(+) T cells disappear 60 days after infection, suggesting that antigen specific IFNγ(+) T cells, mainly detected in the early phase of the disease, could be useful to understand the role of cell-mediated immunity during M. agalactiae infection.

Expression of class II major histocompatibility complex molecules in chronic pulmonary Mycoplasma bovis infection in cattle.

Pulmonary inflammation often results in expression of the class II major histocompatibility complex (MHCII) by both professional antigen-presenting cells (APCs; histiocytes and lymphocytes) and non-professional APCs (respiratory epithelium and endothelium). In this study lesions from 17 cases of bovine chronic pneumonia, associated with Mycoplasma bovis infection, were examined immunohistochemically for M. bovis antigen and MHCII expression. Ten cases of chronic necrosuppurative bronchopneumonia (NBP) were shown to be characterized by abundant perinecrotic M. bovis antigen associated with scant MHCII expression by degenerate leucocytes. Seven cases of chronic catarrhal bronchointerstitial pneumonia (CBP) showed prominent MHCII expression by both professional APCs and respiratory epithelium, in the absence of intralesional M. bovis immunolabelling. The results suggest that prominent MHCII expression by both professional and non-professional APCs plays a role in the pathogenesis of M. bovis-induced CBP. Conversely, the role of MHCII expression in necrosuppurative foci typical of M. bovis-associated NBP can be considered negligible.

Neuropathology of italian cats in feline spongiform encephalopathy surveillance.

Feline spongiform encephalopathy (FSE) is a transmissible spongiform encephalopathy associated with the consumption of feedstuffs contaminated with tissue from bovine spongiform encephalopathy-affected cattle and characterized by the accumulation in the central nervous system of an abnormal isoform of the prion protein (PrP(sc)). Clinically, it presents as a progressive fatal neurologic syndrome that is not easily distinguished from other feline neurologic conditions. Most cases of FSE have been reported in England, where it was first detected in 1990, but a few cases have been reported from other European countries. To identify possible cases of FSE in Italy, the Italian Ministry of Health funded a 2-year surveillance project during which the brains from 110 domestic cats with neurologic signs were evaluated histologically for spongiform encephalopathy and immunohistochemically to detect PrP(sc). Although no cases of FSE were found, the study proved useful in monitoring the Italian cat population for other neurologic diseases: neoplasia (21.8%), toxic-metabolic encephalopathy (18.2%), granulomatous encephalitis (15.5%), suppurative encephalitis (4.6%), trauma (3.6%), circulatory disorders (3.6%), degeneration (2.7%), nonsuppurative encephalitis (2.7%), and neuromuscular diseases (1.8%). No histologic lesions were found in 20% of the brains, and samples from 5.5% of the cats were rejected as unsuitable.

Isolation and characterization of unusual Mycoplasma spp. from captive Eurasian Griffon (Gyps fulvus) in Sicily.

Mycoplasmas have been isolated from birds of prey during clinical examinations, but their significance to the health of raptors is unclear. We report the isolation and characterization of four mycoplasmas found in the upper respiratory tract of four sick Eurasian Griffon (Gyps fulvus) that were housed in a Sicilian rehabilitation center at Ficuzza, near Palermo in Sicily, before reintroduction into the wild. These included Mycoplasma gallinarum, an unidentified mycoplasma highly similar to Mycoplasma glycophilum, and two unidentified mycoplasmas with similarities to Mycoplasma falconis and Mycoplasma gateae.

Bacteriological, serological, pathological and immunohistochemical studies of Mycoplasma bovis respiratory infection in veal calves and adult cattle at slaughter.

Mycoplasma bovis is an important cause of calf pneumonia worldwide. In this study, we examined 140 cattle at slaughter comprising 70 veal calves and 70 beef cattle; 115 animals with pneumonic lesions and 25 without. Lung samples were submitted for bacteriological, histological, and M. bovis-immunohistochemical analyses. Serology for M. bovis was positive in 76% of beef cattle and 100% of veal calves. M. bovis was isolated only from veal calves in 16 out of 64 pneumonic cases. M. bovis was detected by immunohistochemistry in seven bacteriologically positive cases. M. bovis antigen was associated with bronchogenic necrosuppurative or fibrinonecrotizing lesions. Bacteriologically positive and immunohistochemical negative cases were associated with catarrhal bronchointerstitial pneumonia. Results suggest that M. bovis infection may develop into a severe necrosuppurative bronchopneumonia or fibrinonecrotizing pneumonia when associated with a high number of intralesional organisms or, conversely, into a mild catarrhal bronchointerstitial pneumonia when associated with a low number of organisms.

Effect of danofloxacin (Advocin A180) on goats affected with contagious caprine pleuropneumonia.

The efficacy of danofloxacin (Advocin A180) was evaluated for the treatment of contagious caprine pleuropneumonia (CCPP) caused by Mycoplasma capricolum subsp. capripneumoniae. Ten healthy Angora goats, confirmed free of CCPP, were exposed to clinically affected animals from a natural outbreak in Thrace, Turkey. After 14 days exposure, 8 goats showed pyrexia ( > or = 41 degrees C). Shortly after, the Angora goats were divided randomly into two groups. Five of these were injected with danofloxacin (6 mg/kg subcutaneously), which was repeated after 48 h; the five remaining animals received saline. Goats were monitored clinically and blood samples were collected for serology. Animals with severe disease were withdrawn from the trial. Goats completing the study were euthanized at day 42. Lung tissue and bronchial fluid were collected for mycoplasma isolation. All danofloxacin-treated goats showed resolution of clinical disease by the end of the trial. Two saline-treated goats failed to complete the study owing to CCPP. Danofloxacin-treated goats showed fewer lung lesions and had significantly lower combined clinical scores than saline controls (p < 0.001). Danofloxacin was found to be highly effective in the treatment of CCPP in goats.

In vitro susceptibilities of field isolates of Mycoplasma agalactiae to oxytetracycline, tylosin, enrofloxacin, spiramycin and lincomycin-spectinomycin.

The minimum inhibitory concentrations (MICs) of tetracycline, enrofloxacin, tylosin, spiramycin and a lincomycin:spectinomycin 1:2 combination, against 24 Sicilian isolates of Mycoplasma agalactiae, the causative organism of contagious agalactia were determined in vitro by a broth dilution method. Enrofloxacin was the most effective antimicrobial in vitro with a range of MIC values from 0.125 to 0.500 microg/ml and an MIC(50) of 0.203 and MIC(90) of 0.365 microg/ml. Using the MIC(50) and MIC(90) values the remaining four antimicrobials are ranked in order of in vitro effectiveness as follows: tylosin (MIC(50)0.292; MIC(90)0.525 microg/ml) was slightly more effective than tetracycline (MIC(50)0.296; MIC(90)0.533 microg/ml), followed by lincomycin:spectinomycin (MIC(50)0.521; MIC(90)0.938 microg/ml) and spiramycin (MIC(50)1.583; MIC(90)2.850 microg/ml). MIC values above 1.000 microg/ml were obtained using tetracycline, tylosin and spiramycin for some M. agalactiae isolates.

Detection of haemoparasites in cattle by reverse line blot hybridisation with a note on the distribution of ticks in Sicily.

A reverse line blot hybridisation (RLB) of 21 oligonucleotides with polymerase chain reaction (PCR) amplified regions of 16S rRNA (Ehrlichia/Anaplasma group) or 18S rRNA (Babesia/Theileria group) genes of haemoparasites detected Theileria annulata, T. buffeli/orientalis, Babesia bovis, B. bigemina, B. divergens, Ehrlichia bovis, Anaplasma marginale, A. centrale and unknown species within the Rickettsia tribe.A very high prevalence of mixed infections was detected, which indicated that animals infected with Babesia spp. were also infected with Theileria spp. and/or Anaplasma spp. The tick distribution appeared to be seasonal with Hyalomma marginatum as the most frequently observed tick and Boophilus annulatus and Ixodes ricinus as the least frequently observed ticks. Other species identified in the 818 ticks collected during the five sampling periods between April 1998 and November 1999 included H. lusitanicum, Rhipicephalus sanguineus group, R. bursa, Dermacentor marginatus, Haemaphysalis punctata, B. annulatus and I. ricinus.

Integrated molecular diagnosis of Theileria and Babesia species of cattle in Italy.

A reverse line blot hybridization (RLB) test was developed to specifically identify six Theileria spp. (T. annulata, T. parva, T. mutans, T. velifera, T. taurotragi, and T. buffeli/orientalis) and three Babesia spp. (B. bovis, B. bigemina, and B. divergens). No cross reaction was observed with other livestock pathogens (such as Anaplasma marginale, A. centrale, A. ovis, Cowdria ruminantium, Trypanosoma brucei, T. congolense, and T. vivax). This method was used to test bovine blood samples collected in Sicily in April and November, 1998. Preliminary results indicated that T. annulata and T. buffeli/orientalis were the main species observed in cattle blood. Babesia species represented 1.8% and 23.5% in April and November, respectively.

Bovine babesiosis in Sicily: preliminary study on pathology.

In this study, the authors report the preliminary pathological features of an outbreak of spontaneous babesiosis detected in a group of thirty-five, five to six months-old Simmenthal calves imported in Sicily in November 1998. Five of them showed fever, anaemia, haemoglobinuria, inappetence, depression, thirst, swollen lymph-nodes and poor body conditions a few days after being introduced in the farm. Several ticks were present particularly at the basis of the horns and near the ears of the sick animals. Blood smears, stained with May-Grunwald-Giemsa, showed several pyriform merozoites in the red blood cells referred to Babesia bigemina parasites; all ticks collected were identified as Dermacentor marginatus. Gross post-mortem examination and histopathological observations revealed severe involvement of kidneys, liver, lungs, spleen and lymph-nodes. The pathogenesis of the referred features in a spontaneous fatal case observed is discussed.

Clinical and laboratory studies on theileriosis outbreaks in Sicily, Italy.

A total of 153 blood samples were collected all over the Sicilian island in 15 farms. 29.4% and 25.5% were PCR positive for Theileria annulata and T. buffeli/orientalis respectively. 15.7% of cattle blood samples showed a mixed infection with both Theileria species. Each farm showed an infection rate ranging from 0% to 100%. Hyalomma marginatum and Boophilus annulatus were the most common ticks collected in April and November 1998 respectively. The aim of the present work was to investigate on haemoparasite species spread in Sicily. Ticks vectors involved in the transmission, distribution, risk areas and seasons, state of healthy carrier, are also discussed in this paper.

Characterization of Theileria species by PCR using specific target sequences.

Theileriosis is an infectious disease in tropical countries and in the Mediterranean area. It is caused by Theileria, a haemoprotozoan, transmitted by vectors belonging to the Ixodidae. In Southern Italy and in Sicily the infection is due mainly to T. annulata, but in some cases other species are involved in the disease. The authors describe a method to identify theileriosis in cattle blood samples, using PCR and hybridization techniques. Different primer sets were used to amplify different DNA target sequences, both genus and species specific. Blood samples from cattle were collected in Sicily. The DNA extracted from blood samples was employed first to detect the presence of the 18S ribosomal subunit gene specific for Theileria genus. Successively the positive samples were analysed to identify the species, T. annulata or T. buffeli/orientalis, using as target sequences for amplification respectively a fragment of the TAMS-1 and p33/34 antigens gene. Here the authors describe for the first time the presence of T. buffeli/orientalis infection in Sicilian herds. In fact 66% of positive blood samples were T. buffeli/orientalis infected.