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Contagious bovine pleuropneumonia (CBPP) is a respiratory disease of cattle that is listed as notifiable by the World Organization for Animal Health. It is endemic in sub-Saharan Africa and causes important productivity losses due to the high mortality and morbidity rates. CBPP is caused by Mycoplasma mycoides subsp. mycoides ( Mmm) and is characterized by severe fibrinous bronchopneumonia and pleural effusion during the acute to subacute stages and by pulmonary sequestra in chronic cases. Additional lesions can be detected in the kidneys and in the carpal and tarsal joints of calves. Mmm infection occurs through the inhalation of infected aerosol droplets. After the colonization of bronchioles and alveoli, Mmm invades blood and lymphatic vessels and causes vasculitis. Moreover, Mmm can be occasionally demonstrated in blood and in a variety of other tissues. In the lung, Mmm antigen is commonly detected on bronchiolar and alveolar epithelial cells, in lung phagocytic cells, within the wall of blood and lymphatic vessels, inside necrotic areas, and within tertiary lymphoid follicles. Mmm antigen can also be present in the cytoplasm of macrophages within lymph node sinuses, in the germinal center of lymphoid follicles, in glomerular endothelial cells, and in renal tubules. A complete pathological examination is of great value for a rapid presumptive diagnosis, but laboratory investigations are mandatory for definitive diagnosis. The purpose of this review is to describe the main features of CBPP including the causative agent, history, geographic distribution, epidemiology, clinical course, diagnosis, and control. A special focus is placed on gross and microscopic lesions in order to familiarize veterinarians with the pathology and pathogenesis of CBPP.

Trypanosoma equiperdum is the causative agent of dourine, a parasitic venereal disease of equids. In this work, rabbits were infected with T. equiperdum strain OVI; serological tests (complement fixation test, ELISA and immunoblotting), used for the diagnosis of dourine in horses, were applied to study rabbit humoral immune response and to characterise T. equiperdum antigen pattern recognised by antibodies from infected rabbits. Moreover a protein extract of T. equiperdum strain OVI was produced and tested in skin tests on infected rabbits to detect the cell-mediated response induced by T. equiperdum, in order to evaluate its use in the field diagnosis of dourine. Sera of infected rabbits recognized in immunoblotting Trypanosoma protein bands with molecular weight below 37 kDa, providing a serological response comparable with that already observed in dourine infected horses. Moreover the trypanosome protein extract was capable to produce in vivo delayed-type hypersensitivity (DHT Type IV) in rabbits and proved itself to be non-toxic and non-sensitizing.

In Italy, veterinary autogenous vaccines manufacturing is regulated by the legislative decree of the Ministry of Health, March 17th, 1994, n. 287. The production is performed by the network of the ‘Istituti Zooprofilattici Sperimentali’ (IZSs), public health institutes scattered all over the Italian territory. The aim of this research was to evaluate the feasibility of an in vitro method to test the abnormal toxicity of autogenous bacterial vaccines as an alternative to animal models routinely employed. For this purpose, the Istituto Zooprofilattico Sperimentale dell’Abruzzo e del Molise (IZSAM) in partnership with the Istituto Zooprofilattico Sperimentale della Lombardia e dell’Emilia Romagna (IZSLER), evaluated the toxicity of 49 batches of autogenous bacterial vaccines, previously shown to be safe in guinea pigs and mice, on animal model, by means of the methyl tetrazolium (MTT) assay. All vaccines showed cytotoxic effects when tested 1:2 diluted and undiluted; overall, all vaccines lost toxicity at 1:128 dilution. As expected, these findings suggest a different susceptibility of this assay compared to the laboratory animal model. On the other hand, these results do not clarify which components of the vaccines are responsible for the cytotoxic effect. Overall, more experiments are warranted in order to standardize the MTT assay which could be coupled with the trials in laboratory animals.

Contagious bovine pleuropneumonia (CBPP) is a severe disease caused by Mycoplasma mycoides subsp. mycoides (Mmm). Knowledge on CBPP pathogenesis is fragmented and hampered by the limited availability of laboratory animal and in vitro models of investigation. The purpose of the present study is to assess respiratory explants as useful tools to study the early stages of CBPP. Explants were obtained from trachea, bronchi and lungs of slaughtered cattle, tested negative for Mycoplasma spp. and for the major bacterial and viral respiratory pathogens. The interaction of Mmm with explant cells was studied by immunohistochemistry (IHC), double-labelling indirect immunofluorescence (DLIIF) and laser scanning confocal microscopy (LSCM). Mmm capability to survive and proliferate within the explants was evaluated by standard microbiological procedures. Finally, the putative cellular internalization of Mmm was further investigated by the gentamicin invasion assay. IHC and DLIIF indicated that Mmm can colonize explants, showing a marked tropism for lower airways. Specifically, Mmm was detected on/inside the bronchiolar and alveolar epithelial cells, the alveolar macrophages and the endothelial cells. The interaction between Mmm and explant cells was abolished by the pre-incubation of the pathogen with bovine anti-Mmm immune sera. Mmm was able to survive and proliferate in all tracheal, bronchial and lung explants, during the entire time course of the experiments. LSCM and gentamicin invasion assay both confirmed that Mmm can enter non-phagocytic host cells. Taken together, our data supports bovine respiratory explants as a promising tool to investigate CBPP, alternative to cattle experimental infection. Electronic supplementary material The online version of this article (10.1186/s13567-017-0500-z) contains supplementary material, which is available to authorized users.

Peste des petits ruminants (PPR) virus belongs to the family Paramyxoviridae and represents a major threat to small livestock industry. In recent years, outbreaks of PPR have occurred in Turkey and North Africa. In endemic areas, disease prevention is accomplished using live‑attenuated vaccines. However, the use of live vaccines in non‑endemic regions, such as Europe, is not approved by Veterinary Authorities. In these regions inactivated vaccines are then the only viable alternative. In this study an inactivated vaccine (iPPRVac) was formulated with either a water‑in‑oil emulsion (ISA 71 VG) or with delta inulin adjuvant, alone (AFSA1) or combined with a TLR9 agonist oligonucleotide (AFSA2). These formulations were then tested for immunogenicity on rats. The iPPRV formulation with AFSA2 adjuvant induced 100% seroconversion in rats after 2 injections and was subsequently evaluated in goats. Five goats were immunised twice subcutaneously, 36 days apart with iPPRVac + AFSA2. The immunised goats all seroconverted to PPR by day 9 and remained seropositive until the end of the experimental period (133 days). These data indicate that the rat model is useful in predicting vaccine responses in goats and that inactivated vaccine, when formulated with a delta inulin adjuvant, represents a promising alternative to live attenuated vaccines for PPR vaccination campaigns in non‑endemic areas.

SummaryFour goats were inoculated with an inactivated peste des petits ruminants virus(PPRV) vaccine. Three unvaccinated goats were kept as controls. After 36 days,the four goats were revaccinated. The immune response was monitored by virusneutralization test showing that two doses of the vaccine were able to stimulatestrong immune response in all the vaccinated animals. The vaccinated goat andthe controls were challenged with virulent PPRV intranasally. After PPRV chal-lenge, the three control goats showed fever, viremia and virus excretion throughmucosal surfaces, whereas the vaccinated goats were fully protected against PPRVinfection and replication.IntroductionPeste des petits ruminants (PPR) is a highly contagious andinfectious viral disease which can cause high mortality indomestic and wild small ruminants. Peste des petits rumi-nants virus (PPRV) is a negative-sense single-strandedRNA virus of the Morbillivirus genus. A unique serotype ofPPRV circulates, classified in 4 genetically distinct lineages.The clinical signs are the sudden onset of depression,fever, discharges from the eyes and nose, sores in themouth, breathing disorders and cough and foul-smellingdiarrhoea (Food and Agriculture Organization of the Uni-ted Nations [FAO], 1999). The disease has severe economicimpact in developing countries (FAO, 1999). PPR is ende-mic in many African countries between the Sahara and theEquator. Since 1993, the virus has spread to the Arabianpeninsula, the Middle East and the greater part of theIndian subcontinent (Banyard et al., 2010). PPR infectionwas officially declared in Turkey in 1999 (Ozkul et al.,2002). In 2008, disease appeared for the first time in Mor-occo (Hammouchi et al., 2012). After that, it was recordedin Tunisia in 2011 (Office International des Epizooties[OIE], WAHID Interface, 2011) and 2012 (OIE, WAHIDInterface, 2012a), in Algeria in 2012 (OIE, WAHID Inter-face, 2012b) and 2013 (OIE, 2013) and in Egypt in 2012(OIE, WAHID Interface, 2012c). PPRV isolated in Mor-occo was confirmed as Lineage 4, which was usuallyreported in the Middle East and Asia, but now has alsobeen reported in East Africa (Cosseddu et al., 2013) and inTurkey (Ozkul et al., 2002).The ongoing circulation of PPRV in the European part ofTurkeyaswellasinNorthAfricaraisesconcerninEuropeancountries about the risk for the introduction of this devas-tating disease into the EU. In the event of PPR outbreaks,vaccination may be key measure to prevent more animalsbecoming infected, thereby slowing or stopping the geo-graphic spread of the disease. An efficient and economicallyaffordable DIVA vaccine for PPR is still not available. Fourlive-attenuated vaccines are currently available on the mar-ket: Nigeria/75, Sungri 96, Arasur 87 and Coimbatore 97(Diallo et al., 1989; Saravanan et al., 2010) and used in thevaccination campaigns of many endemic countries (Muniret al.,2013).However, severaldrawbacksmaylimit thepos-sibility to employ these vaccines in case of emergency vacci

The shelf-life of a culture medium is the maximum period of validity for optimum preparation and preservation. Apart from the composition of the medium, the factors that influence shelf-life are sterilisation method, preservation and packaging procedures, storage temperature and exposure to light. The shelf-life of a culture medium is defined by evaluating its basic chemico-physical characteristics so as to obtain the correct growth and characterisation of a specific microorganism. This research was conducted from March to September 2003 on 12 'critical' culture media, i.e. media that had a coded shelf-life of not more than thirty days. Each medium was produced in three separate batches, with a total of 5 940 samples. The purpose of the study was to define a longer period of validity than that coded for each medium by evaluating weight reduction, pH, fertility and sterility. The shelf-life observed for each medium was longer than those coded. The new shelf-life takes into account both the operational needs of complex organisational structures and the efficiency of the medium, depending on its chemico-physical characteristics and storage and preservation methods.