Regional Eradication of Mycoplasma hyopneumoniae From Pig Herds and Documentation of Freedom of the Disease

The objectives of this study were to 1) screen all sow herds in a region for M. hyopneumoniae, 2) to effectuate an eradication programme in all those herds which were shown to be infected with M. hyopneumoniae, and 3) to follow the success of the screening and the eradication programmes. The ultimate goal was to eradicate M. hyopneumoniae from all member herds of a cooperative slaughterhouse (153 farrowing herds + 85 farrowing-to-finishing herds + 150 specialised finishing herds) before year 2000. During 1998 and 1999, a total of 5067 colostral whey and 755 serum samples (mean, 25 samples/herd) were collected from sow herds and analysed for antibodies to M. hyopneumoniae by ELISA. Antibodies were detected in 208 (3.6%) samples. Two farrowing herds (1.3%) and 20 farrowing-to-finishing herds (23.5%) were shown to be infected with M. hyopneumoniae. A programme to eradicate the infection from these herds was undertaken. During March 2000, a survey was made to prove the success of the screening and the eradication programmes. In total, 509 serum samples were collected randomly from slaughtered finishing pigs. Antibodies to M. hyopneumoniae were not detected in 506 of the samples, whereas 3 samples were considered suspicious or positive. Accordingly, 3 herds were shown to be infected. One of the herds was previously falsely classified as non-infected. Two of the herds were finishing herds practising continuous flow system (CF). Unlike finishing herds which practice all-in/all-out management routines on herd level, CF herds do not get rid of transmissible diseases spontaneously between batches, for which reason a screening was made in the rest of the CF herds (total n = 7). Consequently, 2 more infected herds were detected. In addition to the results of the survey, a decreasing prevalence of lung lesions at slaughter (from 5.2% to 0.1%) and lack of clinical breakdowns indicated that all member herds were finally free from M. hyopneumoniae in the end of year 2000.     

Post-translational processing targets functionally diverse proteins in Mycoplasma hyopneumoniae

Mycoplasma hyopneumoniae is a genome-reduced, cell wall-less, bacterial pathogen with a predicted coding capacity of less than 700 proteins and is one of the smallest self-replicating pathogens. The cell surface of M. hyopneumoniae is extensively modified by processing events that target the P97 and P102 adhesin families. Here, we present analyses of the proteome of M. hyopneumoniae-type strain J using protein-centric approaches (one- and two-dimensional GeLC–MS/MS) that enabled us to focus on global processing events in this species. While these approaches only identified 52% of the predicted proteome (347 proteins), our analyses identified 35 surface-associated proteins with widely divergent functions that were targets of unusual endoproteolytic processing events, including cell adhesins, lipoproteins and proteins with canonical functions in the cytosol that moonlight on the cell surface. Affinity chromatography assays that separately used heparin, fibronectin, actin and host epithelial cell surface proteins as bait recovered cleavage products derived from these processed proteins, suggesting these fragments interact directly with the bait proteins and display previously unrecognized adhesive functions. We hypothesize that protein processing is underestimated as a post-translational modification in genome-reduced bacteria and prokaryotes more broadly, and represents an important mechanism for creating cell surface protein diversity.     

Mycoplasma hyopneumoniae and Mycoplasma hyosynoviae Infection in Cases of Fibrinous Pericarditis in Slaughter Pigs

Pericarditis, acute or subacute, is found at post mortem meat inspection of baconers in about 0.02 to 0.04% of slaughtered pigs in Denmark. The pathological findings are usually restricted to the pericardial sac. The pericardial sac is filled with a fibrinous exudate, which may be blood stained. In some cases massive granulation tissue formation is seen underlying the fibrinous exudate. Other constantly occurring, but less aggravating lesions, are chronic catarrhal bronchopneumonia and increased volume of serosanguinous synovial fluid in the large joints of the limbs. Lesions usually seen as sequelae to septicaemia have not been observed. The lesions seem to be part of a pathological entity which may be involved in the pathogenesis of fibrinous pericarditis in baconers.     

Carriage of Staphylococcus aureus by Free-Living Wild Animals in Spain

The presence of methicillin-susceptible Staphylococcus aureus (MSSA) was analyzed in different free-living wild animals to assess the genetic diversity and predominant genotypes on each animal species. Samples were taken from the skin and/or nares, and isolates were characterized by spa typing, multilocus sequence typing (MLST) and antimicrobial susceptibility testing. The proportion of MSSA carriers were 5.00, 22.93, 19.78, and 17.67% in Eurasian griffon vulture, Iberian ibex, red deer, and wild boar, respectively (P = 0.057). A higher proportion of isolates (P = 0.000) were recovered from nasal samples (78.51%) than skin samples (21.49%), but the 9.26% of red deer and 18.25% of wild boar would have been undetected if only nasal samples had been tested. Sixty-three different spa types were identified, including 25 new spa types. The most common were t528 (43.59%) in Iberian ibex, t548 and t11212 (15.79% and 14.04%) in red deer, and t3750 (36.11%) in wild boar. By MLST, 27 STs were detected, of which 12 had not been described previously. The most frequent were ST581 for Iberian ibex (48.72%), ST425 for red deer (29.82%), and ST2328 for wild boar (42.36%). Isolates from Eurasian griffon vulture belong to ST133. Host specificity has been observed for the most frequent spa types and STs (P = 0.000). The highest resistance percentage was found against benzylpenicillin (average, 22.2%), although most of the S. aureus isolates were susceptible to all antimicrobial tested. Basically, MSSA isolates were different from those MRSA isolates previously detected in the same animal species.     

Investigating the Role of Free-Ranging Wild Boar (Sus scrofa) in the Re-Emergence of Enzootic Pneumonia in Domestic Pig Herds: A Pathological,Prevalence and Risk-Factor Study

Enzootic pneumonia (EP) caused by Mycoplasma hyopneumoniae has a significant economic impact on domestic pig production. A control program carried out from 1999 to 2003 successfully reduced disease occurrence in domestic pigs in Switzerland, but recurrent outbreaks suggested a potential role of free-ranging wild boar (Sus scrofa) as a source of re-infection. Since little is known on the epidemiology of EP in wild boar populations, our aims were: (1) to estimate the prevalence of M. hyopneumoniae infections in wild boar in Switzerland; (2) to identify risk factors for infection in wild boar; and (3) to assess whether infection in wild boar is associated with the same gross and microscopic lesions typical of EP in domestic pigs. Nasal swabs, bronchial swabs and lung samples were collected from 978 wild boar from five study areas in Switzerland between October 2011 and May 2013. Swabs were analyzed by qualitative real time PCR and a histopathological study was conducted on lung tissues. Risk factor analysis was performed using multivariable logistic regression modeling. Overall prevalence in nasal swabs was 26.2% (95% CI 23.3–29.3%) but significant geographical differences were observed. Wild boar density, occurrence of EP outbreaks in domestic pigs and young age were identified as risk factors for infection. There was a significant association between infection and lesions consistent with EP in domestic pigs. We have concluded that M. hyopneumoniae is widespread in the Swiss wild boar population, that the same risk factors for infection of domestic pigs also act as risk factors for infection of wild boar, and that infected wild boar develop lesions similar to those found in domestic pigs. However, based on our data and the outbreak pattern in domestic pigs, we propose that spillover from domestic pigs to wild boar is more likely than transmission from wild boar to pigs.     

小兴安岭南坡春季野猪活动规律

2005年4~5月,在小兴安岭南坡利用无线电遥测技术对笼式活捕的12头野猪(Sus scrofa ussuricua)的活动规律进行144h的监测。结果表明,小兴安岭南坡野猪春季的活动高峰主要集中于8:00~18:00时,5个活动强度峰值分别为76.52%、89.23%、91.56%、78.79%和84.46%;活动规律表现为白天活动而夜间休息;活动时间略少于休息时间;雄性野猪日活动量大于雌性(P=0.021,t=2.474),差异显著,成体野猪的日活动量和亚成体差异不显著(P=0.287,t=-1.089),家族群野猪日活动量显著小于孤野猪(P=0.039,t=-2.194)。     

野猪MC4R基因的克隆及变异初步研究

黑素皮质素受体4是在人类肥胖研究中发现的重要调节因子,参与调节动物的体重、采食量和能量稳态,缺失MC4R基因的突变纯合体小鼠出现遗传性肥胖。为了进一步揭示其群体遗传变异,寻找新的遗传标记,本研究对野猪(Sus scrofa ussuricus)MC4R基因进行了克隆(GenBank accession NoDQ388767)和序列分析,并对所发现的错义突变进行了基于限制性内切酶HindⅢ的PCR-RFLP分析。序列分析表明野猪与民猪MC4R基因的编码区序列完全相同,与大白猪相比存在4个SNPs;对14头野猪的酶切多态性分析表明该突变位点是多态位点,并且3种基因型的分布符合Hardy-Weinberg定律。结果表明,野猪具有独特的遗传信息。     

Vaccination Against Porcine Circovirus-2 Reduces Severity of Tuberculosis in Wild Boar

Tuberculosis (TB) in wild boar (Sus scrofa) may be affected by coinfections with other pathogens, such as porcine circovirus type 2 (PCV2). Therefore, sanitary measures focused on controlling PCV2 could be useful in reducing the impact of TB in this wild suid. The aim of this study was to explore whether vaccination against PCV2 targeting young animals affects TB prevalence and TB severity in wild boar. The study was conducted on a game estate in mid-western Spain. Seventy animals of ages ranging from 4 to 8 months were captured, individually identified, vaccinated against PCV2 and released, forming a vaccinated group. Not-captured animals cohabiting with the vaccinated wild boar constituted the control group. Animals from both groups were hunted between 2013 and 2016 and a TB diagnosis based on pathological assessment and microbiological culture was made in all of them. The effect of PCV2 vaccination on TB prevalence and severity was explored using generalized lineal models. Whereas TB prevalence was similar in vaccinated and control groups (54.55 vs. 57.78%), vaccinated animals showed less probabilities to develop generalized TB lesions. Furthermore, mean TB severity score was significantly lower in vaccinated animals (1.55 vs. 2.42) suggesting a positive effect of PCV2 vaccination.     

Characterization of Newcastle Disease Viruses in Wild and Domestic Birds in Luxembourg from 2006 to 2008

Newcastle disease virus (NDV) is one of the most important viral diseases of birds. Wild birds constitute a natural reservoir of low-virulence viruses, while poultry are the main reservoir of virulent strains. Exchange of virus between these reservoirs represents a risk for both bird populations. Samples from wild and domestic birds collected between 2006 and 2010 in Luxembourg were analyzed for NDV. Three similar avirulent genotype I strains were found in ducks during consecutive years, suggesting that the virus may have survived and spread locally. However, separate introductions cannot be excluded, because no recent complete F gene sequences of genotype I from other European countries are available. Detection of vaccine-like strains in wild waterbirds suggested the spread of vaccine strains, despite the nonvaccination policy in Luxembourg. Among domestic birds, only one chicken was positive for a genotype II strain differing from the LaSota vaccine and exhibiting a so-far-unrecognized fusion protein cleavage site of predicted low virulence. Three genotype VI strains from pigeons were the only virulent strains found. The circulation of NDV in wild and free-ranging domestic birds warrants continuous surveillance because of increased concern that low-virulence wild-bird viruses could become more virulent in domestic populations.     

Cloning and purification of recombinant proteins of Mycoplasma hyopneumoniae expressed in Escherichia coli

Mycoplasma hyopneumoniae, the etiological agent of swine enzootic pneumonia, is an important pathogen in the swine industry worldwide. Vaccination is the most cost-effective strategy for controlling and prevention of this disease. However, investigations on pathogenicity mechanisms as well as current serological detection methods and the development of new recombinant subunit vaccines are hampered by the lack of known and well characterized species-specific M. hyopneumoniae antigens. In this work, 54 predicted genes encoding proteins with potential to be used as subunit vaccine or antigens in diagnostic tests were selected, amplified by PCR and cloned into Escherichia coli expression vectors. Recombinant protein expression, solubility and yields were analyzed. The majority of the recombinant proteins were expressed in inclusion bodies. After solubilization with urea or N-lauroyl sarcosine, recombinant proteins were purified by Ni²⁺ affinity chromatography. This approach allowed purification of thirty recombinant M. hyopneumoniae proteins which will be evaluated as vaccine candidates and/or as antigens to be used in diagnostic tests.     

Mycoplasma hyopneumoniae temporal trends of infection and pathological effects in wild boar populations

Mycoplasma hyopneumoniae (Mhyo) is the principal etiological agent of enzootic pneumonia (EP), one of the most economically important diseases in the pig production industry worldwide. Although swine and wild boars (Sus scrofa) share susceptibility to Mhyo infection, information regarding the dynamics of Mhyo infections and pathology in wild boars is currently limited. Therefore, the aim of this study was to determine the spatiotemporal dynamics of Mhyo infections in free-living wild boars through serological ELISA testing. Additionally, the presence of EP-like gross lesions and their association with the presence of Mhyo, based on a PCR assay, were assessed. Over a period of 5 years, antibodies against Mhyo were detected in 655 (30.0 %) of the 2,177 analyzed wild boars. A generalized temporal increase of seroprevalence coupled with an increase of mean antibody titers of seropositive individuals was found. Moreover, a similar seroprevalence between age classes associated with higher antibody titers of younger individuals indicated the wild boars were infected during the early stages of their life. Out of 99 lungs tested, 43 showed EP-like lesions and 45 were PCR positive for the presence of Mhyo DNA. The lung lesion scores were related to the sex and age of the wild boars, with young individuals having higher lung scores than others, and there was a positive association with the Mhyo PCR-positive status. The temporal increase of the Mhyo seroprevalence and infection association with EP-like lesions, when coupled with the spreading of wild boar populations, raises concerns on the epidemiological role of this species.     

Disentangling Natural From Hunting Mortality in an Intensively Hunted Wild Boar Population

Abstract We assessed age-specific natural mortality (i.e., excluding hunting mortality) and hunting mortality of 1,175 male and 1,076 female wild boar (Sus scrofa) from Chǎteauvillain-Arc en Barrois (eastern France), using a 22-year dataset (1982–2004) and mark-recapture-recovery methods. Overall yearly mortality was >50% for all sex and age-classes. Low survival was mostly due to high hunting mortality; a wild boar had a >40% of chance of being harvested annually, and this risk was as high as 70% for adult males. Natural mortality rates of wild boar were similar for males and females (approx. 0.15). These rates were comparable to rates typical of male ungulates but high for female ungulates. Wild boar survival did not vary across sex and age-classes. Despite high hunting mortality, we did not detect evidence of compensatory mortality. Whereas natural mortality for males was constant over time, female mortality varied annually, independent of fluctuations in mast availability. Female wild boar survival patterns differed from those reported in other ungulates, with high and variable natural mortality. In other ungulates, natural mortality is typically low and stable across a wide range of environmental conditions. These differences may partly reflect high litter sizes for wild boar, which carries high energetic costs. High hunting mortality may induce a high investment of females in reproduction early in life, at the detriment to survival. Despite high hunting mortality, the study population increased. Effective population control of wild boar should target a high harvest rate of piglets and reproductive females.     

实时荧光定量PCR在猪肺炎支原体检测中的应用

实时荧光定量PCR技术是一种利用荧光检测方法来定量核酸的技术,具有高度的灵敏性、特异性和精确性.综述了荧光定量PCR技术的基本原理及其在猪肺炎支原体检测中的应用.     

海南大田自然保护区野猪的生境选择

对海南大田自然保护区内野猪(Sus scrofa)的生境选择进行了研究。结果表明,野猪偏好选择低平地热带草原和灌丛草地,通常选择草本植物高、草本生物量大、草本覆盖度高、灌木矮小、1m以下当年枝比例高、隐蔽程度相对较低、接近水源的地点作为取食地,而选择灌木高大且密度高、灌木覆盖度高、草本植物高、隐蔽程度好、接近水源和远离人为干扰的地点作为卧息地。在雨季,野猪卧息地的灌木覆盖度和隐蔽度均比旱季高,而取食地的草本生物量也比旱季高。主成分分析表明,取食地第Ⅰ主成分的贡献率达35.51%,其中绝对值较大的有乔木密度、灌木高度、灌木覆盖度、1m以下当年枝比例、草本覆盖度和隐蔽级;卧息地第Ⅰ主成分的贡献率为39.57%,其中绝对值较大的出现在灌木高度、灌木覆盖度、1m以下当年枝比例、草本高度、草本覆盖度、草本生物量、距水源距离和隐蔽级。