Mycoplasma pneumonia of swine (MPS) resistance and immune characteristics of pig lines generated by crossing an MPS pulmonary lesion selected Landrace line and a highly immune capacity selected Large White line

To understand the influence of crossbreeding on Mycoplasma pneumonia of swine (MPS) resistance and immune characteristics, two crossbred lines were characterized. One crossbred line, LaWa, was generated by crossing the MPS pulmonary lesion selected Landrace line (La) and the highly immune‐selected Large White line (Wa). The second crossbred line, LaWb, was generated by crossing the La line and the nonselected Large White line (Wb). The crossbred LbWb line (nonselected Landrace line × nonselected Large White line) and the La line were used as controls. The LaWa and LaWb lines had an intermediate level of MPS lung lesions between La and LbWb lines, although the difference was not statistically significant. After stimulation with sheep red blood cells (SRBCs), the LaWb and LaWa lines showed immune characteristics similar to that of the La line; the number of monocytes in peripheral blood increased, while B cells, T cells, secretion of SRBC‐specific immunoglobulin G, and interleukin (IL)‐13 decreased. Additionally, the number of natural killer (NK) cells and the expression of IL‐4 and IL‐17 were significantly higher in the LaWb and LaWa lines, respectively. These data suggested that crossbreeding of La and Wa lines resulted in the inheritance of some of the selected immune responses.     

Effects of mycoplasmal pneumonia of swine (MPS) lung lesion‐selected Landrace pigs on MPS resistance and immune competence in three‐way crossbred pigs

To clarify the genetic influence of mycoplasmal pneumonia of swine (MPS) lesion‐selected Landrace (La) on MPS resistance and immune characteristics in three‐way crossbred pigs (LaWaDa), the LaWaDa pigs were compared with the non‐selected crossbred (LbWbDb) and purebred (La) pigs. The MPS lesion score in the three lines was as follows: La line < LaWaDa line < LbWbDb line, with significant differences among the lines. The proportions of myeloid cells and T cells were lower and higher, respectively, in the LaWaDa pigs compared with those in the other two lines. Messenger RNA (mRNA) expression of interleukin (IL)‐6, IL‐10, transforming growth factor‐β, and interferon‐γ in peripheral blood was significantly increased after vaccination in the La and LaWaDa lines. IL‐4 mRNA expression in the LaWaDa line was intermediate to the La and LbWbDb lines. Furthermore, principal component analysis for immune traits and MPS lesions was executed to clarify the characteristics of each pig line. These findings suggest that the immune responses in the three pig lines are genetically distinct and that MPS resistance and some immunity characteristics from the La line were transmitted to the three‐way crossbred pigs.     

Immunogenic properties and mycoplasmal pneumonia of swine (MPS) lung lesions in Large White pigs selected for higher peripheral blood immune capacity

Immunogenic properties and mycoplasmal pneumonia of swine (MPS) lung lesions were compared between the immunity‐selected Large White line and the non‐selected Large White line. The selected Large White line showed a higher level of pulmonary MPS lesions compared with the non‐selected Large White line. Subsequent to vaccination, the percentage of natural killer cells and T cells (CD3⁺CD4⁺CD8^‐ and CD3⁺CD4^?CD8⁺ T cells) were significantly increased in the non‐selected line but remained unchanged in the immunity‐selected Large White line. Secretion of Mycoplasma hyopneumoniae vaccine‐specific immunoblogulin G and phagocyte activity in peripheral blood were significantly higher in the immunity‐selected Large White line than in the non‐selected line. Expression of interleukin (IL)‐4 and IL‐6 messenger RNA in hilar lymph nodes was significantly lower in the immunity‐selected Large White line than in the non‐selected line. However, expression of IL‐10 in all immune tissues was significantly higher in the immunity‐selected Large White line. These results suggest that the selection for high immunity was not effective in increasing resistance to MPS lung lesions.     

Correlated response of peripheral blood cytokines with selection for reduced mycoplasma pneumonia of swine lesions in Landrace pigs

Mycoplasma pneumonia of swine (MPS) is responsible for significant economic losses in the swine industry. We selected Landrace pigs for reduced MPS pulmonary lesions over five generations, and measured concentrations of the following cytokines: interleukin (IL)‐10, IL‐13, IL‐17, tumor necrosis factor (TNF)‐α and interferon (IFN)‐γ to estimate their correlation with MPS lesions. Sheep red blood cells (SRBC) were injected twice intramuscularly at 70 and 95 kg body weight. Blood serum samples were collected after 1 week of secondary SRBC inoculation and cytokine concentrations were analyzed by ELISA. Genetic parameters and breeding values were estimated. The heritability estimates of IL‐10, IL‐13, IL‐17, TNF‐α and IFN‐γ were 0.20 ± 0.06, 0.12 ± 0.06, 0.27 ± 0.07, 0.20 ± 0.10 and 0.05 ± 0.03, respectively. Genetic correlations of IL‐17 and TNF‐α with pulmonary MPS lesions were high (‐0.86 ± 0.13 and 0.69 ± 0.29, respectively) and those of IFN‐γ and IL‐13 with MPS lesions were moderately negative (‐0.45). Through selection, the breeding values of IL‐17 and IFN‐γ increased substantially and those of TNF‐α decreased. These results suggest that innate and cellular immunity are more important for the suppression of pulmonary lesions in MPS than humoral‐mediated immunity, such as antibody response.     

Immunological characterization of peripheral blood leukocytes using vaccine for mycoplasmal pneumonia of swine (MPS) in swine line selected for resistance to MPS

This study was conducted to evaluate immunological changes in peripheral blood leukocytes in pigs that were genetically selected for their improved resistance to mycoplasmal pneumonia of swine (MPS), using MPS vaccine as an antigen. Twelve castrated MPS‐selected Landrace pigs were compared with the same number of pigs from a nonselected line by using a time‐course analysis at the hematological level. After the second sensitization with MPS vaccine, the percentages of B cells, CD4⁺ T cells, and natural killer (NK) cells in total leukocytes were lower in the selected line than in the nonselected line, whereas the percentage of granulocytes in total leukocytes increased in the MPS‐selected line. We also assessed the proliferative ability of peripheral blood mononuclear cells (PBMCs) stimulated with Mycoplasma hyopneumoniae, lipopolysaccharide or concanavalin A, and found that although the proliferative ability of the PBMC was not different between the two lines at a steady state, the nonselected line showed a significantly higher proliferative ability after sensitization with MPS vaccine than the selected line regardless of antigens used. These results thus indicate that the selection of pigs on the basis of MPS resistance changes their immunophenotype, and would give us beneficial information for the prevention of MPS infection.     

Correlated responses of respiratory disease and immune capacity traits of Landrace pigs selected for Mycoplasmal pneumonia of swine (MPS) lesion

Five generations of Landrace pigs selected for average daily gain, backfat thickness, Mycoplasmal pneumonia of swine (MPS) lesion score, and plasma cortisol levels, was executed to decrease the MPS lesion score. Genetic parameters and correlated genetic responses for respiratory disease and peripheral blood immune traits were estimated in 1395 Landrace pigs. We estimated the negative genetic correlation of MPS lesion score with phagocytic activity (PA) at 7 weeks of age (‐0.67). The breeding values of PA at 7 weeks of age and 105 kg body weight and the correlated selection response of the ratio of granular leukocytes to lymphocytes at 105 kg body weight were significantly increased, and sheep red blood cell‐specific antibody production (AP) was significantly decreased in a selection‐dependent manner. Increasing of natural immunological indicators (e.g. PA) and decreasing of humoral immunological indicator (e.g. AP) were observed due to genetically decreasing MPS lesion score.     

Genetic selection for resistance to mycoplasmal pneumonia of swine (MPS) in the Landrace line influences the expression of soluble factors in blood after MPS vaccine sensitization

We recently developed a Landrace line that is resistant to mycoplasmal pneumonia of swine (MPS) infection by genetic selection for five generations, and we reported that the immunophenotype of this line is different from that of the non‐selected line in terms of changes in peripheral blood leukocyte population after MPS vaccination. This study followed up previous findings demonstrating changes in soluble factors in blood, namely, hormones, Mycoplasma hyopneumoniae‐specific immunoglobulin G (IgG), and cytokines. These two lines were injected with MPS vaccine on days ?7 and 0 after blood sampling on those days, and blood samples were collected on days ?14, ?7, 0, 2, 7 and 14. We found changes in the levels of many hormones and cytokines in both lines. However, we found that only growth hormone (GH) and interferon (IFN)‐γ levels were statistically different between these two lines. GH concentration was reduced (day 0) and IFN‐γ concentration was increased (day 14) in the MPS‐selected line compared with the non‐selected line, despite unchanged IFN‐γ messenger RNA expression in blood cells. Although detailed mechanisms underlying these phenotypes remain unsolved, these traits would be useful to improve MPS resistance in pig production and provide an insight into MPS infection.     

Halothane anesthesia of several hours' duration in swine of the German Landrace breed, selected for resistance to malignant hyperthermia

The malignant hyperthermia syndrome is widely spread in pigs of the German Landrace. This has precluded the use of halothane anesthesia. The advent of homozygous halothane-negative strains stimulated us to test whether such animals were suitable for longduration halothane anesthesia. Nine sows of the German Landrace (n = 4, homozygous halothane-positive (H+); n = 5, homozygous halothane-negative (-)) were used. Anesthesia was induced by Azaperon/Metomidat-HCL followed by intubation and exposure to halothane. All 4 halothane-positive animals died within 45-90 min following the onset of halothane administration, while all 5 halothane-negative animals survived a 4-5 h deep anesthesia without problems. Thus, homozygous hylothane-positive strains of German Landrace pigs may be surgically anesthetized for long durations with halothane anesthesia.     

Drug resistance in strains of Escherichia coli isolated from the intestinal tract of pigs in Norway.

Faecal samples from 95 healthy pigs and samples of jejunal content from 85 piglets suffering from colienterotoxaemia were tested for the presence of drug resistant E. coli strains. Practically all pigs in both groups harboured E. coli strains resistant to one or more of the 6 antibiotics/chemotherapeutic agents tested (Oxytetracycline, streptomycin, sulphaisodimidin, neomycin, ampicillin, chloramphenicol). Almost 100% of healthy and approx. 90% of diseased pigs harboured strains resistant to Oxytetracycline, streptomycin and sulphaisodimidin. Pigs with strains resistant to neomycin, ampicillin and chloramphenicol were less frequently found. The predominant coliform flora consisted of E. coli strains” resistant to Oxytetracycline, streptomycin and sulphaisodimidin in 71% to 81% of diseased pigs and in 47% to 69% of the healthy pigs. In diseased pigs ¾ of the animals had a coliform flora dominated by neomycinresistant E. coli strains.Of the 721 resistant E. coli strains isolated from healthy pigs, 11% were single resistant while the corresponding figure for the 518 resistant strains isolated from diseased pigs was 6%. Thus 89% and 94% of strains showed simultaneous resistance to 2 or more antibiotics. E. coli strains resistant to 3 or more drugs were found in approx. 60% and 70% of the isolates from healthy and diseased animals, respectively. Oxytetracycline/streptomycin/sulphaisodimidin resistance was most commonly found, approx. 22% and 38% of the strains from healthy and diseased pigs, respectively, showing this resistance pattern.Transmission of drug resistance which was examined in E. coli strains originating from the diseased pigs was demonstrated in approx. 76% of the isolates. The incidence of drug resistance transfer in single, double, triple and quadruple resistant strains was 11%, 68%, 97% and 98%, respectively.     

Variation in resistance to the Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) in Pietrain and Miniature pigs

Porcine reproductive and respiratory syndrome (PRRS) is one of the economically most important diseases of swine. Viraemia and the prolonged persistence of the virus are among the most critical factors. Virus replication and severity of disease vary with virus isolates, and there is rising evidence for a genetic component of the host susceptibility. Dissecting the genetic basis of resistance/susceptibility to PRRS virus (PRRSV) might lead to improved knowledge on the molecular mechanisms of PRRS and the establishment of genetic markers for future disease control. The aim of this study was to establish a porcine model with emphasized genetic differences in PRRSV susceptibility. Seven ‘Wiesenauer Miniature’ pigs (MI), a local German breed and eight commercial Pietrain (PI) pigs were challenged with 10⁵ TCID₅₀ of an attenuated PRRSV strain (Ingelvac^® PRRSV MLV). Clinical status, viraemia and seroconversion of the pigs were compared. No clinical signs were observed during the experiment. Viraemia peaked on day 6 p.i., with 100% of viraemic pigs in PI and on day 12 p.i with 87% of viraemic MI. Viraemia lasted for up to 35 days in MI and for at least 72 days in PI. This surprising result was confirmed by a second study with another four MI. MI and PI showed maximum virus titres of 10^2.5 TCID₅₀/ml of serum and 10^4.5 TCID₅₀/ml, respectively, indicating a virus replication in MI of approximately 3.3% that of PI over the complete period. MI were more efficient in antibody production. With such pronounced breed differences, the model is of high relevance for the genetic dissection of PRRS pathogenesis and susceptibility.     

“非瘟”与“新冠肺炎”双重防控压力下我国养猪业所面临的困境及现代生物技术和育种技术应用的迫切性

目前我国养猪业正经受防控非洲猪瘟和新冠肺炎双重疫情的挑战。本文分析了我国种猪资源保护利用以及育种工作的现状和所面临的问题,并建议我国尽快利用现代生物技术保护我国优良猪种种质资源,尽快推广和应用现代育种技术,提高种猪生产性能,加快我国生猪产能的恢复。     

国内现有猪口蹄疫O型灭活疫苗对2010年流行毒株的抵抗力

在2010年上半年,猪O型口蹄疫不断发生,作为国内几个生产厂家的猪口蹄疫现用疫苗种毒的提供者,我组进行了猪源流行毒对该种毒灭活疫苗免疫猪的攻毒保护试验及观察。结果如下:1)、猪源流行毒致猪发病速度慢。猪源流行毒接种猪后,致猪发病的时间在3～5天,而疫苗株致猪发病时间是接种后2～3天;2)、该疫苗毒株能有效抵抗猪源流行毒的攻击,PD50大于6;3)、免疫保护和ELISA抗体水平没有线性相关性;4)、免疫保护与140S抗原量有相关性。     

An abattoir survey of pneumonia and pleuritis in slaughter weight swine from 9 selected herds. II. Enzootic pneumonia of pigs: microbiological findings and their relationship to pathomorphology

Lungs from 191 slaughter pigs with gross lesions indicative of enzootic pneumonia of pigs (EPP) and 80 grossly normal lungs, all originating from 9 different herds, were subjected to microbiological and pathological examinations. The microbiological studies included both bacterial and mycoplasmal culture and also testing for Mycoplasma hyopneumoniae antigen in tissue by indirect immunofluorescent technique. M. hyopneumoniae, Pasteurella multocida and Mycoplasma hyorhinis were detected in 83%, 43% and 37% of the pneumonic lungs, respectively. Mycoplasma flocculare was the most frequently isolated organism in the non-pneumonic lungs. The greatest amounts of macroscopic pneumonia (25.2%) were recorded in lungs with all the three agents M. hyopneumoniae, P. multocida and M. hyorhinis present. The amounts of pneumonia in lungs with M. hyopneumoniae alone and in concurrence with P. multocida, were 9.3% and 15.6%, respectively. M. hyorhinis was also, in this study, associated with higher frequency of diffuse pleuritis. These findings indicate that M. hyorhinis might be involved in the pathogenesis of pneumonia in slaughter pigs. Ninety-six per cent of the isolates of P. multocida from pneumonic lungs could be characterized as type A. In the herds which had the most severe pneumonia problems, toxin production was detected in 83% of the P. multocida strains while only 28% were toxigenic in herds with subclinical to moderate pneumonia problems.     

2019年湖南省部分地区鸡、鸭、猪源大肠杆菌耐药性调查

旨在调查2019年湖南省部分地区不同来源的大肠杆菌对抗生素的耐药水平,为养殖合理用药提供参考。从4个市畜禽养殖场采集猪、鸡和鸭粪便285份,使用麦康凯琼脂培养基和伊红美蓝培养基对大肠杆菌筛选,利用基质辅助激光解析电离飞行时间质谱进行鉴定,采用微量肉汤稀释法检测16种抗生素对大肠杆菌的最小抑菌浓度(MIC)。共分离出203株大肠杆菌(鸡源146株、鸭源20株和猪源37株),对四环素和氨苄西林耐药率最高,分别达到89.2%和84.7%;对头孢他啶、奥格门丁、黏菌素耐药率较低,分别为2.5%、2.0%和0.5%;所有菌株均对美罗培南敏感。抗3种及以上抗生素的菌株数目占比为87.7%,而仅5.42%的菌株对所有检测的抗生素敏感。另外,来源于减抗示范养殖场的大肠杆菌对大部分抗生素的耐药率,与来源于非减抗养殖场大肠杆菌相比未见显著差异。上述结果表明,湖南省动物源大肠杆菌的耐药水平仍处在较高水平,重视兽药使用管理并持续开展对大肠杆菌的耐药性监测十分必要。     

贵州地区紫云花猪腹泻病原菌的分离鉴定及耐药性分析

为了鉴定贵州地区紫云花猪腹泻病原菌,分析其耐药性,试验采用细菌学常规鉴定、16SrRNA基因测序、K-B药敏纸片法对从贵州地区不同养猪场采集的患腹泻紫云花猪肝脏、肛拭子、粪便等样品196份进行细菌分离鉴定,并分析其耐药性。结果显示,从采集的196份样品中分离得到了60株大肠杆菌、35株沙门菌、20株志贺菌,分离的60株大肠杆菌对庆大霉素、阿莫西林、强力霉素等5种药物的耐药率在70.0%以上,对其他药物耐药率在15.0%~40.0%之间,耐11、10种药物的菌株最多,分别占分离菌株31.7%、23.3%;分离的35株沙门菌对氨苄西林、阿莫西林、氟苯尼考等5种药物的耐药率在58.3%以上,对其他药物耐药率在48.6%~17.1%之间,耐10、9种药物的菌株最多,分别占分离菌株25.7%、22.9%;分离的20株志贺菌对氨苄西林、阿莫西林、新霉素等9种药物的耐药率在65.0%以上,对其他药物的药物耐药率在25%~35%之间,耐9、8种药物的菌株最多,分别占分离菌株30.0%、20.0%。本研究为该地区紫云花猪细菌性腹泻的防控提供研究基础。     

Antimicrobial resistance in Enterococcus faecium and Enterococcus faecalis isolates of swine origin from eighteen provinces in China

Enterococcus faecium and E. faecalis are important human pathogens and also served as sentinel organisms for monitoring systems of antimicrobial resistance in both animals and humans. In this study, 106 E. faecium and 56 E. faecalis isolates were collected from 61 pig farms in 18 proveinces of China. Antimicrobial susceptibility was determined for 9 clinically important antibiotics and 3 antimicrobial growth promoters. The Enterococcus isolates showed high prevalence of resistance to medically important antibiotics, such as ampicillin (50.9% for E. faecium and 19.6% for E. faecalis), chloramphenicol (24.5% for E. faecium and 41.1% for E. faecalis), erythromycin (83.0% for E. faecium and 91.1% for E. faecalis), tetracycline (79.2% for E. faecium and 100% for E. faecalis), quinupristin/dalfopristin (26.4% for E. faecium) and ciprofloxacin (73.6% for E. faecium and 66.1% for E. faecalis). Resistance to tigecycline, linezolid and vancomycin was very rare. The resistance status of three representative in-feed antibiotics bacitracin, nosiheptide and enramycin was firstly investigated with Enterococcus as indicator bacteria. The Enterococcus isolates showed extremely high frequency of bacitracin resistance (96.7% for E. faecium and 87.8% for E. faecalis), while no nosiheptide and enramycin resistance was observed. Pulsed-field gel electrophoresis (PFGE) analysis showed that a majority of E. faecium and E. faecalis strains showed unrelated profiles, indicating high heterogeneity among the Enterococcus isolates. Our study provided basic data on the antimicrobial resistance of E. faecium and E. faecalis isolates.     

柱花草对炭疽病的抗病性研究进展

综述了柱花草Stylosanthes spp.与炭疽病Stylo anthracnose互作的抗病性研究进展,着重于柱花草与抗炭疽病互作过程中细胞学及组织学的抗病生理反应、生化反应、诱导抗病性及防卫反应等.     

肉鸡肺动脉高压综合征抗病育种研究

为了研究肉鸡肺动脉高压综合征(PHS)的抗病育种情况,试验采用静脉注射纤维素颗粒的方法诱发PHS,比较发病前后非易感组肉鸡和易感组肉鸡右心室与全心室重量比(RV/TV)值,心电图Ⅱ导联S波、R波和RS综合波波幅,红细胞比容(PCV),血红蛋白(HB)和血清蛋白含量。结果表明:静脉注射纤维素颗粒明显提高了肉鸡PHS的发病率。发病后腹水组肉鸡PCV、HB含量和RV/TV值升高,血清蛋白含量降低,心电图Ⅱ导联S波、R波和RS综合波波幅均升高,符合肉鸡PHS的特点;但发病前易感组肉鸡与非易感组肉鸡相比,PCV、HB含量、血清蛋白含量和心电图Ⅱ导联S波、R波和RS综合波波幅均无显著差异。     

以免疫活性串联片段FB为抗原检测FMDV抗体的间接ELISA方法

将口蹄疫病毒免疫串联片段FB克隆至原核表达载体pBAD／TOPO中，得到重组质粒pBAD-FB，将此重组质粒转化到受体茵TOPIO中，以不同浓度的阿拉伯醛糖进行诱导，并在不同诱导时间进行采样，经处理后做SDS-PAGE、Western-blotting分析。结果发现以终浓度为0．02g／L的阿拉伯醛糖进行诱导，4h后表达量可达高峰，其大小约为26ku，扫描结果显示，FB融合蛋白的表达量占细菌总蛋白的28．9％，能与抗FMDV抗体发生特异性反应，融合蛋白以包涵体和可溶形式存在。将融合蛋白的可溶性组分用500g／L Ni-NTA树脂过柱纯化后作为包被抗原，成功地建立了检测血清中FMD抗体的间接ELISA方法。融合蛋白的最佳包被浓度是40μg／mL；标准阳性血清的最适稀释倍数为1：160；最佳封闭液为50g／L脱脂奶粉和PBS；最佳封闭时间为1h。用建立的ELISA方法对采集的118份样品进行检测，并与全病毒包被抗原ELISA、间接血凝诊断试剂盒和UBI VP1抗体检测ELISA试验盒的检测结果比较，证明所建立的方法具有良好的特异性、敏感性和可重复性。