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Outbreaks of Rift Valley fever (RVF) occurred in Namibia in 2010 and 2011. Complete genome characterization was obtained from virus isolates collected during disease outbreaks in southern Namibia in 2010 and from wildlife in Etosha National Park in 2011, close to the area where RVF outbreaks occurred in domestic livestock. The virus strains were sequenced using Sanger sequencing (Namibia_2010) or next generation sequencing (Namibia_2011). A sequence-independent, single-primer amplification (SISPA) protocol was used in combination with the Illumina Next 500 sequencer. Phylogenetic analysis of the sequences of the small (S), medium (M), and large (L) genome segments of RVF virus (RVFV) provided evidence that two distinct RVFV strains circulated in the country. The strain collected in Namibia in 2010 is genetically similar to RVFV strains circulating in South Africa in 2009 and 2010, confirming that the outbreaks reported in the southern part of Namibia in 2010 were caused by possible dissemination of the infection from South Africa. Isolates collected in 2011 were close to RVFV isolates from 2010 collected in humans in Sudan and which belong to the large lineage containing RVFV strains that caused an outbreak in 2006–2008 in eastern Africa. This investigation showed that the RVFV strains circulating in Namibia in 2010 and 2011 were from two different introductions and that RVFV has the ability to move across regions. This supports the need for risk-based surveillance and monitoring.

This study reports the monitoring of several emerging viral pathogens in Mauritania, which was carried out by the analysis of bovine and camel samples taken at the slaughterhouse of Nouakchott. Blood and serum were collected by random sampling from 159 camels and 118 cattle in March 2013 at the large animals abattoir in Nouakchott. Serological tests for Rift Valley Fever (RVF), Peste des Petits Ruminants (PPR), West Nile disease (WND), epizootic haemorrhagic disease (EHD) and African horse sickness (AHS) were carried out using commercial ELISA kits. The samples, which resulted positives for PPR, WND and AHS, were tested with the confirmatory virus neutralization test (VNT). According to ELISA results, serological prevalence of RVF was 45% (95% CI 52.3-37.7) in camels and 16% (95% CI 22.6-9.4) in cattle. The difference between the observed prevalences in camels and in cattle was significant (p value ≤ 0.01). PPR was absent in camels and had 12% prevalence (95% CI, 17.86-6.14) in cattle. Furthermore, camels showed 92% (95% CI, 96.1-87.9) prevalence of WNV, 73% (95% CI, 82.3-63.64) of EHD and 3% (95% CI, 5.6-0.4) of AHS. This data are of relevance since provided useful feedbacks on the circulation of the pathogens in field. Moreover, this survey provided new information on the susceptibility of camels to several emerging pathogens and on the possible use of this species as sentinel animal.

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.

Mycoplasma mycoides subsp. mycoides (Mmm) is the etiological agent of contagious bovine pleuropneumonia (CBPP), one of the major diseases affecting cattle in sub-Saharan Africa. Some evidences suggest that the immune system of the host (cattle) plays an important role in the pathogenic mechanism of CBPP, but the factors involved in the process remain largely unknown. The present study aimed to investigate the cell response of bovine polymorphonuclear neutrophils (PMNs) after Mmm in vitro exposure using one step RT-qPCR and Western blotting. Data obtained indicate that gene and protein expression levels of some pro-inflammatory factors already change upon 30 min of PMNs exposure to Mmm. Of note, mRNA expression level in Mmm exposed PMNs increased in a time-dependent manner and for all time points investigated; targets expression was also detected by Western blotting in Mmm exposed PMNs only. These data demonstrate that when bovine PMN cells are triggered by Mmm, they undergo molecular changes, upregulating mRNA and protein expression of specific pro-inflammatory factors. These results provide additional information on host-pathogen interaction during CBPP infection.

Mycoplasma infections are commonly associated with respiratory diseases in farm animals. However, few data are currently available about the presence and the etiology of mycoplasma pneumonia in small ruminants in central Italy, a region particularly devoted to pastoralism. The present study aims to investigate retrospectively the causative agents of pneumonia in sheep and goats, a special emphasis being placed upon mycoplasmas. In total, respiratory pathogens were identified in 129 of 380 carcasses (32.36%). Mycoplasmas were detected in a high percentage of cases, alone (59 animals) or in combination with other pathogens (26 animals), Mycoplasma arginini being the most frequent species. No evidence of mycoplasma infection was demonstrated in 44 carcasses; in such animals, Mannheimia haemolytica and Pasteurella multocida were most commonly isolated. Overall, our results suggest that mycoplasma infections can contribute to a relevant portion of respiratory diseases in small ruminants and support the role of Mycoplasma arginini as a frequent cause of pneumonia in sheep. We consider that such data are of value to efficiently manage the health status of animal populations.

Mycoplasma mycoides subsp. mycoides is the etiological agent of contagious bovine pleuropneumonia (CBPP). While several findings on CBPP prevalence in Nigeria were documented, no data were reported about the genomic characterization of Nigerian M. mycoides subsp. mycoides strains. Here, we present the draft genome sequences of two novel M. mycoides subsp. mycoides strains isolated in Nigeria.

Contagious bovine pleuropneumonia (CBPP) is a highly contagious respiratory disease affecting cattle and is widely distributed in the sub-Saharan Africa. The objective of this study was to detect Mycoplasma mycoides subspecies mycoides (Mmm) the causative agent of CBPP from 90 cattle at slaughter using polymerase chain reaction-Restriction fragment length polymorphism. In this study, 450 samples suggestive of CBPP in Maiduguri, Yola and Gombe township abattoirs were processed according to standard protocols. The isolation rate was found to be 3.33% and percentage of identification with PCR-RFLP yielded 1.56%. Subsequently, QIAxcel revealed molecular size of 574 bp for Mycoplasma mycoides subcluster. Further analysis of PCR amplicons with restriction digestion, confirmed the presence of Mmm 16 S rRNA of CAP 21 genomic region with molecular sizes of 180 bp and 380 bp. Thus, the 380 bp fragments delineated Mmm from Mycoplasma mycoides subsp. capri. Three isolates (BL5, BL6 and AL1) were from lungs and four from pleural fluids (APF2, APF8A, APF8B and APF9) were isolated and identified, while a vaccine strain T1/44 was re-detected along with the field isolates. No sample from Gombe had Mmm. In conclusion, the findings of this study have detected the presence of Mmm as causative agent of CBPP. Measures such as surveillance, quarantine and vaccination are hereby recommended for the control of CBPP in Nigeria.

Reactive oxygen species (ROS) are suggested to play a role in the pathogenesis of contagious bovine pleuropneumonia, a severe respiratory disorder caused by Mycoplasma mycoides subsp. mycoides (Mmm). The present study investigated the generation of ROS by different strains of Mmm, as well as their effect on the oxidative response of bovine neutrophils. The production of ROS was indirectly measured using a luminol-based chemiluminescence assay. Our results confirm that Mmm can produce ROS via the metabolism of glycerol, significant differences existing between African and European strains. Mmm was capable of adhering to the external surface of neutrophils. Interestingly, Mmm enhanced the respiratory burst of bovine neutrophils. This activity was particularly pronounced with the African field strain and in presence of glycerol. Taken together, our data argue in favour of a major role for neutrophils as the main source of ROS in contagious bovine pleuropneumonia.

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.

After a May 2011 outbreak of Rift Valley fever among livestock northeast of Etosha National Park, Namibia, wild ruminants in the park were tested for the virus. Antibodies were detected in springbok, wildebeest, and black-faced impala, and viral RNA was detected in springbok. Seroprevalence was high, and immune response was long lasting.

A study was conducted to investigate seroprevalence and risk factors for Brucella species infection in cattle and some wildlife species in communities living at the periphery of the Great Limpopo Transfrontier Conservation Area in south eastern Zimbabwe. Three study sites were selected based on the type of livestock–wildlife interface: porous livestock–wildlife interface (unrestricted); non-porous livestock–wildlife interface (restricted by fencing); and livestock–wildlife non-interface (totally absent or control). Sera were collected from cattle aged ≥ 2 years representing both female and intact male animals. Sera were also collected from selected wild ungulates from Mabalauta (porous interface) and Chipinda (non-interface) areas of the Gonarezhou National Park. Samples were screened for Brucellaantibodies using the Rose Bengal plate test and confirmed by the complement fixation test. Data were analysed by descriptive statistics and multivariate logistic regression modelling. In cattle, brucellosis seroprevalence from all areas was 16.7% (169/1011; 95% CI: 14.5–19.2%). The porous interface recorded a significantly (p = 0.03) higher seroprevalence (19.5%; 95% CI: 16.1–23.4%) compared to the non-interface area (13.0%; 95% CI: 9.2–19.9%).The odds of Brucellaseropositivity increased progressively with parity of animals and were also three times higher (OR = 3.0, 2.0 < OR < 4.6, p < 0.0001) in cows with history of abortion compared to those without.Brucella antibodies were detected in buffaloes; 20.7% (95% CI: 13.9–29.7%) form both study sites, but no antibodies were detected from impalas and kudus. These results highlight the importance of porous interface in the interspecies transmission of Brucella species and that independent infections may be maintained in buffalo populations. Thus, brucellosis control aimed at limiting animal inter-species mixing may help reduce the risk of human brucellosis in interface areas. Further studies should aim at establishing subspecies identity and direction of possible transmission of brucellosis between wildlife and livestock.

Contagious bovine pleuropneumonia (CBPP), caused by Mycoplasma mycoides subsp. mycoides (Mmm) is a severe cattle disease, present in many countries in sub-Saharan Africa. The development of improved diagnostic tests and vaccines for CBPP control remains a research priority. Polyacrylamide gel electrophoresis and mass spectrometry were used to characterize the Triton X-114 soluble proteome of nine Mmm strains isolated from Europe or Africa. Of a total of 250 proteins detected, 67 were present in all strains investigated. Of these, 44 were predicted to be lipoproteins or cytoplasmic membrane-associated proteins and are thus likely to be members of the core in vitro surface membrane-associated proteome of Mmm. Moreover, the presence of all identified proteins in other ruminant Mycoplasma pathogens were investigated. Two proteins of the core proteome were identified only in other cattle pathogens of the genus Mycoplasma pointing towards a role in host–pathogen interactions. The data generated will facilitate the identification and prioritization of candidate Mycoplasma antigens for improved control measures, as it is likely that surface-exposed membrane proteins will include those that are involved in host–pathogen interactions.

A capture operation to ascertain health status in free-ranging buffaloes from six different areas in the Caprivi Strip in the northeast corner of Namibia was conducted in October 2009. Basic information on the ticks and tick-borne pathogens normally found in wildlife from this area are scarce. The objective of this study was to assess the host status of African buffaloes, Syncerus caffer, for ixodid ticks and two selected tick-borne pathogens in the Caprivi Strip, a key area bordering Angola, Zambia, Botswana, and Zimbabwe. Four different tick species have been identified among the 233 collected specimens, and, of 95 tested buffaloes, 54 (57%) were positive for Theileria parva, whereas only 3 (3%) showed evidence of being infected with Ehrlichia ruminantium.

Specific humoral immune responses in a clinical trial on cattle for vaccines against contagious bovine pleuropneumonia (CBPP) were investigated. The trial included a subunit vaccine consisting of five recombinant putative variable surface proteins of the infectious agent Mycoplasma mycoides subsp. mycoides small colony type ( M. mycoides SC) compared to the currently approved attenuated vaccine strain T1/44 and untreated controls. Humoral immune responses to 65 individual recombinant surface proteins of M. mycoides SC were monitored by a recently developed bead-based array assay. Responses to the subunit vaccine components were found to be weak. Animals vaccinated with this vaccine were not protected and had CBPP lesions similar to those of the untreated controls. In correlating protein-specific humoral responses to T1/44-induced immunity, five proteins associated with a protective immune response were identified by statistical evaluation, namely, MSC_1046 (LppQ), MSC_0271, MSC_0136, MSC_0079, and MSC_0431. These five proteins may be important candidates in the development of a novel subunit vaccine against CBPP.

Mycoplasma mycoides subsp. mycoides is generally considered one of most pathogenic Mycoplasma species, and it is the etiological agent of contagious bovine pleuropneumonia (CBPP). Here, we present the annotated genome sequence of M. mycoides subsp. mycoides Italian strain 57/13, isolated in 1992 during CBPP outbreaks in Italy.

The authors studied the persistence of infection in 46 ewes experimentally infected with Brucella melitensis biovar 3 and monitored through three subsequent reproductive cycles. The entire experimental period lasted for 151 weeks. Infection of ewes and elimination of Brucella in milk, or its presence in vaginal discharges, persisted throughout the duration of the trial, as demonstrated by recurrent elimination of Brucella in milk and vaginal discharges. Brucella melitensis was recovered from the tissues of one ewe killed at the end of the trial. The strain was recovered from vaginal swabs and milk following parturition in the third reproductive cycle from an ewe that had aborted in the first cycle but was not pregnant in the second cycle. From a public health point of view, the periodical recovery of Brucella from the milk during the entire trial period illustrated that brucellosis in sheep remains a continuous occupational risk and a significant public health problem for consumers of fresh milk and milk products. That risk may persist for at least 3 years following the initial infection of the flock. Lamb antibody titres became negative in all lambs within 5 months after birth. This suggested that serological tests on lambs may have no practical diagnostic significance if performed during the first 5 months of life. Nevertheless, the birth of three infected lambs suggested that the phenomenon of latent carrier state may represent another way for B. melitensis to persist in a flock.

Contagious bovine pleuropneumonia (CBPP), a highly infectious and fatal disease of cattle present in many countries in sub-Saharan Africa, is usually controlled by mass vaccinations. However, vaccination against CBPP is known to cause site reactions in a percentage of cattle especially in primary vaccinations. In Zambia, a record of site reactions was kept for seven consecutive years from 2005 to 2011 to establish the level of the problem. In some areas, after 3 years of consecutive vaccination campaigns, immunization could not be implemented for a period of 2 years because of logistical difficulties or owner resistance. Whereas in the three preceding years when animals were vaccinated annually, site reactions were in the range of 6.2 %; on resumption of vaccination in the herds that had not been immunized for 2 years, site reactions averaged 21.3 %. This data shows that the T1/44 vaccine may cause severe local reactions in cattle if there is any break in annual vaccinations. It is therefore important for authorities to ensure that the cattle at risk of contracting CBPP are regularly vaccinated to avoid discouraging farmers from presenting their animals.

During May–July 2010 in Namibia, outbreaks of Rift Valley fever were reported to the National Veterinary Service. Analysis of animal specimens confirmed virus circulation on 7 farms. Molecular characterization showed that all outbreaks were caused by a strain of Rift Valley fever virus closely related to virus strains responsible for outbreaks in South Africa during 2009–2010.

Contagious bovine pleuropneumonia (CBPP), a severe respiratory disease of cattle caused by Mycoplasma mycoides subsp. mycoides (Mmm) is endemic in many African countries due to fragmented veterinary services and the lack of an efficient vaccine and sensitive diagnostics. More efficient tools to control the disease are needed, but to develop the tools, a better understanding of host–pathogen interactions is necessary. The aim of this study was to characterize the kinetics of the humoral immune response against 65 Mmm surface antigens for an extended period in cattle that survived a primary infection with Mmm. We describe clinical and haematological outcomes, and dissect the humoral immune response over time, to specific antigens and compared the antibody responses between different pathomorphological outcomes.No antigen-specific antibodies correlating with protection were identified. Interestingly we found that animals that developed Mycoplasma-containing sequestra had significantly higher antibody levels against proteins comprising the surface proteome than the animals that cleared Mycoplasma from their lungs. Based on these data we suggest that high antibody titres might play a role in the establishment of pathomorphological changes, such as vasculitis, which should be investigated in future studies. Beneficial antibody specificities and cellular immune responses need to be identified in order to foster the development of an improved vaccine in the future.

Contagious bovine pleuropneumonia (CBPP) is a disease of economic importance that is widely distributed in sub-Saharan African and contributes significantly to cattle morbidity and mortality. Lack of resources to implement eradication measures has led to the disease becoming endemic in most areas in sub-Saharan Africa where governments have little resources and the majority of the people are poor. Usually, control and eradication of such diseases as CBPP is treated as a public good by governments and to achieve this, governments are usually assisted by nongovernment organisations, bilateral government programmes and international donors. The private sector, which usually is companies that run businesses to make profit, although not very well established in sub-Saharan Africa could play a big role in the eradication of CBPP in the region. This could play a dual role of promoting investment and also eradicate livestock diseases which have proved a menace in the livestock sector. This paper highlights the role played by the private sector in the control of CBPP in Zambia.

Clinical Cases of Contagious Bovine Pleuropneumonia (CBPP) among Calves in Vaccinated Cattle Herds in Zambia: A Case Study An observational study was conducted to ascertain the clinical manifestation of Contagious Bovine Pleuropneumonia (CBPP) in the Western Province of Zambia following an outbreak in an endemic region. It was observed that both calves and adult cattle in 9 herds that were not vaccinated with CBPP T1/44 vaccine reported mortality while 24 herds that had consistently been vaccinated against CBPP only reported mortalities in calves between 2 months and 8 months of age. This observation indicates that in CBPP endemic areas, calves may acquire the disease from the carriers in the infected herds.

In order to get a reliable estimate of brucellosis prevalence in Eritrean dairy cattle, a cross-sectional study was carried out in 2009. The survey considered the sub-population of dairy cattle reared in modern small- and medium-sized farms. Samples were screened with the Rose Bengal test (RBT) and positive cases were confirmed with the complement fixation test (CFT). A total of 2.77%(417/15 049; Credibility Interval CI: 2.52% – 3.05%) of the animals tested in this study were positive for antibodies to Brucellaspecies, with a variable and generally low distribution of positive animals at regional level. The highest seroprevalence was found in the Maekel region (5.15%; CI: 4.58% – 5.80%), followed by the Debub (1.99%; CI: 1.59% – 2.50%) and Gash-Barka (1.71%; CI: 1.34% – 2.20%) regions. Seroprevalence at sub-regional levels was also generally low, except for two sub-regions of Debub and the sub-region Haicota from the Gash-Barka region. Seroprevalence was high and more uniformly distributed in the Maekel region, namely in the Asmara, Berik and Serejeka sub-regions. Considering the overall low brucellosis prevalence in the country, as identified by the present study, a brucellosis eradication programme for dairy farms using a test-and-slaughter policy would be possible. However, to encourage the voluntary participation of farmers to the programme and to raise their awareness of the risks related to the disease for animals and humans, an extensive public awareness campaign should be carefully considered, as well as strict and mandatory dairy movement control.

Background Contagious Bovine Pleuropneumonia (CBPP), caused by Mycoplasma mycoides subsp. mycoides, is widespread in sub-Saharan Africa. The current live vaccine T1/44 has limited efficacy and occasionally leads to severe side effects in the animals. A better understanding of the immune responses triggered by Mycoplasma mycoides subsp. mycoides and their role in disease progression will help to facilitate the design of a rational vaccine. Currently, knowledge of cytokines involved in immunity and immunopathology in CBPP is rather limited. The aim of this study was to characterize the in vivo plasma concentrations of the cytokines TNF-α, IFN-γ, IL-4, IL-10 and the overall role of CD4+ T cells in the development of cytokine levels during a primary infection. Plasma cytokine concentrations in two groups of cattle (CD4+ T cell-depleted and non-depleted cattle) experimentally infected with Mycoplasma mycoides subsp. mycoides were measured and their relationship to the clinical outcomes was investigated. Results Plasma cytokine concentrations varied between animals in each group. Depletion of CD4+ T cells did not induce significant changes in plasma levels of TNF-α, IL-4, and IL-10, suggesting a minor role of CD4+ T cells in regulation or production of the three cytokines during the time window of depletion (1-2 weeks post depletion). Unexpectedly, the IFN-γ concentrations were slightly, but statistically significantly higher in the depleted group (p < 0.05) between week three and four post infection. Three CD4+ T cell-depleted animals that experienced severe disease, had high levels of TNF-α and IFN-γ. Only one severely diseased non-depleted animal showed a high serum concentration of IL-4 post infection. Conclusions Comparison of most severely diseased animals, which had to be euthanized prior to the expected date, versus less severe diseased animals, irrespective of the depletion status, suggested that high TNF-α levels are correlated with more severe pathology in concomitance with high IFN-γ levels.

The present study describes the use of microarray technology for rapid identification and differentiation of Mycoplasma mycoides subsp. mycoides from other mycoplasmas that may be pathogenic to ruminants, including those of the Mycoplasma mycoides cluster, genetically and antigenically strictly correlated with Mycoplasma mycoides subsp. mycoides. A microarray containing genetic sequences of 55 different bacterial species from Acholeplasma, Mycoplasma, Spiroplasma and Ureaplasma genera was constructed. Sequences to genes of interest were collected in FASTA format from NCBI. The collected sequences were processed with OligoPicker software. Oligonucleotides were then checked for their selectivity with BLAST searches in GenBank. The microarray was tested with ATCC/NCTC strains of Mycoplasma spp. of veterinary importance in ruminants including Mycoplasma belonging to the mycoides cluster as well as Mycoplasma mycoides subsp. mycoides and Mycoplasma mycoides subsp. capri field strains. The results showed that but one ATCC/NCTC reference strains hybridized with their species-specific sequences showed a profile/signature different and distinct from each other. The heat-map of the hybridization results for the nine genes interrogated for Mycoplasma mycoides subsp. mycoides demonstrated that the reference strain Mycoplasma mycoides subsp mycoides PG1 was positive for all of the gene sequences spotted on the microarray. CBPP field, vaccine and reference strains were all typed to be M. mycoides subsp. mycoides, and seven of the nine strains gave positive hybridization results for all of the nine genes. Two Italian strains were negative for some of the genes. Comparison with non-Mycoplasma mycoides subsp. mycoides reference strains showed some positive signals or considerable homology to Mycoplasma mycoides subsp. mycoides genes. As expected, some correlations were observed between the strictly genetically and antigenically correlated Mycoplasma mycoides subsp. mycoides and Mycoplasma mycoides subsp. capri strains. Specifically, we observed that some Italian Mycoplasma mycoides subsp. mycoides strains were positive for two out of the three Mycoplasma mycoides subsp. capri genes, differently from what has been observed for other European or African Mycoplasma mycoides subsp. mycoides strains. This study highlighted the use of microarray technology as a simple and effective method for a single-step identification and differentiation of Mycoplasma mycoides subsp. mycoides from other mycoplasmas that may be pathogenic to ruminants, including those of the Mycoplasma mycoides cluster, genetically and antigenically strictly correlated with Mycoplasma mycoides subsp. mycoides. The opportunity to discriminate several mycoplasmas in a single analysis enhances diagnostic rapidity and may represent a useful tool to screen occasionally mycoplasmas affecting animal farming in territories where diagnostic laboratory support is limited. The heat-map of the hybridization results of the comparative genomic hybridizations DNA-designed chip clearly indicates that the microarray performs well for the identification of the tested Mycoplasma mycoides subsp. mycoides reference and field strains, discriminating them from other mycoplasmas.

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.

During final phases of eradication programmes, strains of Mycobacterium sp. not belonging to the tuberculosis complex increase their relative frequency and are responsible for positive skin test reactions. Moreover, the specificity of any indirect diagnostic test, such as the skin test, is never completely accurate, therefore even when tuberculosis infection is completely eradicated, a number of false positive reactions are to be expected. The aim of this paper is to evaluate the performances of traditional isolation/typing techniques, automatic isolation/typing techniques based on fluorimetric detection of bacterial growth (Bactec), skin tests and the -interferon test. Samples examined for the evaluation of test sensitivities originated from 154 infected animals belonging to 32 infected herds. Samples used as negative controls in the evaluation of test specificities originated from 86 animals of nine officially infection-free herds. The automatic isolation/typing technique based on fluorimetric detection of bacterial growth showed higher sensitivity than the traditional isolation typing technique. Moreover, it allowed a safer processing of bacterial cultures, decreasing the risk for laboratory workers. The observed performance of the gamma-interferon test was considered beneficial in that it increased the sensitivity of individual diagnosis within an infected herd, especially in 'problem herds', but its poor specificity did not improve detection of infected herds compared to the skin test.

[EN] In a rabbit production facility, health monitoring for enteropathies was performed in 15 production cycles for 20 mo. For each cycle, up to a hundred 35 d old rabbits weaned the same day were randomly selected, reared in the same fattening unit, but separately from the source batch and fed with the same feed except for antimicrobial supplementation. Clinical symptoms and enteric lesions of the selected group were recorded, using two checklists with binomial response (yes/no answer to a list of 54 clinical and enteric variables). The day after weaning, one week later, at the beginning of the enteric symptoms and 4-5 d after the start of the symptoms, inocula from the small intestine and caecum of selected animals were subjected to microbiological, C. spiroforme, Eimeria oocyst and rotavirus antigen detection tests. Representative samples of E. coli and C. perfringens isolates were tested, respectively, for serotype, biotype, eae, afr/2 genes and for a, b1, b2, e, i and enterotoxin toxin genes. The answers to the clinical-pathological variables were subjected to statistical analysis with a cluster analysis programme in order to obtain homogeneous, statistically significant groups of diseased animals (clusters). Then, the clusters were statistically associated with the laboratory outcomes. The cluster to which the enterotyphlitis lesions significantly contributed was associated with E. coli detection, E. coli O103 serotype detection and C. spiroforme ("several elements" variable). C. spiroforme ("rare elements" variable) was significantly associated with a cluster, characterised by a pathological profile consisting of bloating/rumbling noise and liquid content in stomach and caecum, without enteric inflammation. C. perfringens was significantly associated with a cluster, characterised by a pathological profile consisting of dilation/liquid content of small intestine, caecal impaction and mucoid content in the colon. Eighteen out of twenty-fi ve C. perfringens strains, examined for their toxin genotypes, proved to be toxin type A, while 7 out of 25 strains showed the a and b2 toxin genes in combination. The rotavirus antigen and Eimeria oocysts were detected from healthy rabbits (specimens of the day after weaning and one week later) in about 15% of specimens examined, but their presence in the sick animals was not significantly associated with any cluster., This study was supported by a financial contribution from Avitalia, Unione Nazionale Associazioni di Produttori Avicunicoli, Forlì, Italy, as part of the programme entitled “Miglioramento della qualità, della gestione dell’offerta delle produzioni cunicole e di rafforzamento dei rapporti di filiera. Azione 4.3”. Our thanks go to breeder Leta Covelli and Dasco srl for supplying the rabbits, to our colleague Romolo Salini and to Fabrizio Agnoletti of the Istituto Zooprofilattico Sperimentale del Veneto, Trevise, Italy