猪放线杆菌SHLI株的分离鉴定

从某猪场病死猪肉样变肺脏和严重出血的心脏、肝脏、肾脏及肠系膜淋巴结中分离到一株革兰氏阴性球杆菌。此菌大小０３５ ～０５ ×０５ ～１５μｍ , 单在、成双或短链状。菌落与琼脂有粘连性, 绵羊血琼脂平板上呈β溶血, 能生长于麦康凯琼脂上。发酵碳水化合物产酸不产气, 不发酵甘露醇、卫茅醇和肌醇, 能水解马粟苷和马尿酸钠, 尿酶、硝酸盐还原试验阳性, 不产生靛基质。鉴定为猪放线杆菌（ Ａｃｔｉｎｏｂａｃｉｌｌｕｓｓｕｉｓ） , 并将其定名为猪放线杆菌 Ｓ Ｈ Ｌ Ｉ株。其对小鼠和仔猪有强的致病性, 对豚鼠的致病性较弱。     

Genetic diversity and toxin gene distribution among serovars of Actinobacillus pleuropneumoniae from Australian pigs

Objective

To explore the diversity among isolates of the Actinobacillus pleuropneumoniae serovars most common in Australia (serovars 1, 5, 7 and 15) and to examine the Apx toxin profiles in selected representative isolates.

Design

A total of 250 isolates selected from different farms were examined for their genotypic profiles and a subset of 122 isolates for their toxin profiles.

Methods

The isolates of serovars 1, 5, 7 and 15 selected for this study came from different farms and different Australian states and were submitted for serotyping to the reference laboratory. The overall diversity of the strains was explored with the enterobacterial repetitive intergenic consensus (ERIC) PCR and the presence of the toxin genes was investigated with a toxin PCR assay.

Results

Some degree of variation was observed in the ERIC‐PCR pattern within all four serovars, ranging from 38% to 61% genetic diversity. When looking at the toxin gene profile and, therefore, the predicted ability to produce the expected toxin pattern, one isolate each of serovars 1 (n = 20) and 7 (n = 47) and 17 isolates of serovar 15 (n = 40) showed variation to the expected gene profile.

Conclusion

The variations in toxin gene patterns, as detected by PCR, found in this study could be related to significant changes in the gene sequence or total absence of the gene. Variation in toxin gene sequences has been observed in other countries. This variation in the toxin profile could also explain possible variation in pathogenicity observed in the field.     

Phenotypic and genotypic characterization of Actinobacillus suis sensu stricto isolated from a dairy calf

The species of the genus Actinobacillus have so far been associated with specific animal hosts, and A. suis sensu stricto, an opportunistic pathogen of swine, is rarely isolated from ruminants. We describe here the isolation of A. suis sensu stricto from a newborn calf that died on a dairy farm in Japan. Identification of the isolate was performed by phenotypic and genotypic characterization, with the latter consisting of nucleotide sequence analyses of the 16S rRNA gene plus three housekeeping genes, rpoB, infB and recN.     

Actinobacillus suis infection in pigs in southwestern Ontario

Actinobacillus suis was isolated from tissues of 39 pigs, 2 porcine lungs, and 1 uterine swab submitted for diagnostic evaluation from 24 farms in southwestern Ontario between 1985 and 1988. These isolates represented a gradually increasing incidence of herd outbreaks caused by A. suis in southwestern Ontario. The outbreaks were typified by sudden death in suckling or recently weaned pigs; 87% of the affected pigs examined at the laboratory were between two and 28 days old. Petechial to ecchymotic hemorrhages in the thoracic and abdominal organs accompanied by serofibrinous exudates in both cavities were the most common gross lesions. The lesions were characterized histologically by bacterial thromboembolism and necrosis randomly scattered in thoracic and abdominal organs. Occasionally, bacterial thromboemboli were surrounded by centrifugally radiating, eosinophilic, club-like colonies. Diffuse necrohemorrhagic myocarditis that was more severe in the atria, and diffuse subacute meningoencephalitis, were less frequent but distinctive lesions. Multiple litters were affected in most herd outbreaks, and mortality often approached 50% in affected litters. Although the A. suis organism was susceptible to nearly every antibiotic against which it was tested, the suddenness of herd outbreaks precluded attempts at treatment.     

胸膜肺炎放线杆菌对仔猪的感染试验

用作者分离鉴定的胸膜肺炎放线杆菌感染仔猪,临床特征为:呼吸困难,食欲不振,体温升高;病理解剖学变化为:胸腔有大量的积液,心包增厚,并且在肺脏和胸膜粘连胸膜增厚,肺脏萎缩,病灶区呈暗红色,轮廓清晰;病理组织学变化为:肺脏、肝脏和肾脏脉血管均有扩张出血,组织发生坏死.     

猪链球菌2型引致脑炎及败血症的豚鼠模型

选用不同日龄的健康豚鼠 132只 ,分组经不同途径接种对猪具有致病性的猪链球菌 2型江苏分离株 HA980 1,引致脑炎及败血症 ,建立了动物模型。试验表明 ,小于 2 4日龄提前断奶的豚鼠对该菌株较易感 ,最佳感染途径是皮下注射 ,腹腔、鼻腔次之。感染豚鼠表现出典型脑炎的临床症状及病理变化。经重复试验测得该菌株对豚鼠的 L D50 为 5 .5× 10 6cfu,从感染死亡豚鼠脑脊液、大脑、血液及各实质器官均可分离出该病原菌 ,与仔猪自然感染情况相似。该动物模型的建立为进一步探究猪链球菌 2型感染的体内动态分布、感染机制、毒力因子及免疫应答奠定了基础     

Actinobacillus suis infection of horses

Nineteen isolates of Actinobacillus suis were recovered from horses during the period October 1978-December 1980. Animals varied in age from a full term foetus to 12 years. One isolate was obtained from the nose of an apparently healthy horse, the remainder were obtained from still-born foetuses (2), foals dying within a week of birth (5), older animals with respiratory (6) or genital infections (3) or abscesses in the jaw (1). One isolate was obtained from the lung of a 2-week-old foal which had shown diarrhoea. The bacteriological characteristics of the isolates and the pathological lesions present in eight cases are described. The organism has a wide geographical distribution in New Zealand, and in the northern part of the North Island appears to be more common than A. equuli.     

Actinobacillus suis infection of horses

Nineteen isolates of Actinobacilfus suis were recovered from horses during the period October 197%December 1980. Animals varied in age from a full term foetus to 12 years. One isolate was obtained from the nose of an apparently healthy horse, the remainder were obtained from still-born foetuses (2), foals dying within a week of birth (5), older animals with respiratory (6) or genital infections (3) or abscesses in the jaw (1). One isolate was obtained from the lung of a 2-week-old foal which had shown diarrhoea.

The bacteriological characteristics of the isolates and the pathological lesions present in eight cases are described. The organism has a wide geographical distribution in New Zealand, and in the northern part of the North Island appears to be more common than A. equuli.     

Streptococcus suis isolated from pigs in Finland

A total of 58 Streptococcus suis strains were isolated from deceased pigs submitted to the National Veterinary Institute, Regional Laboratory in Kuopio, Finland, over a 3 1/2 year period, most frequently from cases of pneumonia. The bacteria were isolated from cases of meningitis, sepsis, rhinitis, endocarditis and abortion. S. suis was also isolated from nasal cavity, lung and brain of some sick piglets without signs of inflammation. Further S. suis was detected in 12 out of 107 tonsils of healthy fatteners tested. S. suis strains were identified by biochemical methods followed by typing. The most common capsular types were 7, 3 and 2, respectively. Only one type 1 strain and no types 6 and 9 strains were found. All S. suis strains tested were sensitive to penicillin and ampicillin.S. suis is not uncommon in Finnish pig herds. S. suis may be regarded as a potentially pathogenic organism which under certain predisposing conditions may cause serious disease.     

红霉素与四环素耐药基因在猪链球菌临床分离株中的检测

为了解临床分离的48株猪链球菌对大环内酯类药物及四环素耐药基因的分布,用微量稀释法测定48株临床分离的猪链球菌对大环内酯类、四环素、β-内酰胺类及头孢类9种抗生素的药物敏感性,建立PCR方法对耐药菌株大环内酯类耐药基因ermA/B/C、mefA/E、msrD、mphB、23S rRNA,L4,L22和四环素耐药基因tetM、tetO、tetL、tetK及与Tn916转座子相关的int和xis基因进行检测。结果表明,31株2型猪链球菌中大环内酯类药物耐药率为3.23%,17株9型猪链球菌红霉素耐药率为88.24%,泰乐菌素、磷酸替米考星、阿奇霉素的耐药率均为70.59%。48株猪链球菌对四环素均耐药,但对青霉素、阿莫西林、头孢曲松钠、氨苄西林均敏感。大环内酯类耐药基因主要以ermB为主,占75%(12/16),mefA/E、msrD占25%(4/16),16株红霉素耐药菌株中,tetM、tetO、int、xis的检出率分别为25%(4/16)、62.5%(10/16)、31.25%(5/16)和31.25%(5/16),没有检测到ermA、ermC、mphB、tetL、tetK。所有红霉素耐药菌株均未检测到23S rRNA、L4和L22突变。     

Serological characterisation of Actinobacillus pleuropneumoniae isolated from pigs in 1993 to 1996

OBJECTIVES: To clarify the serological identity of the prototype strain of a group of Actinobacillus pleuropneumoniae isolates that could not be serotyped in previous studies and to establish the serovar of 378 isolates of A pleuropneumoniae obtained from pigs in Australia over the period 1993 to 1996. DESIGN: After initial validation, QGD and IHA tests were used to characterise the prototype isolate (HS143) selected to represent the cross-reacting isolates that were found in a previous study. Next, 378 recent field isolates of A pleuropneumoniae were characterised using the existing gel diffusion serotyping technique and/or the IHA or QGD tests. RESULTS: The indirect haemagglutination test was shown to be capable of correctly recognising the reference strain for all serovars except serovar 11. While the quantitative gel diffusion test was not as effective as indirect haemagglutination, it could recognise serovar 11. When the two tests were used to examine the prototype strain (HS143) of the cross-reactive isolates, the results indicated that HS143 is serologically distinct from all 12 of the recognised serovars of A pleuropneumoniae. However, as HS143 was subsequently identified as serovar 12 by one of the leading international reference laboratories, the antiserum to isolate HS143 was used as the serovar 12 antiserum. A total of 346 of the 378 A pleuropneumoniae field isolates examined could be confidently serotyped with almost 90% of the isolates being either serovar 1 (104 isolates); serovar 7 (83 isolates) or serovar 12 (142 isolates). A range of other serovars and some cross-reactive isolates made up the remainder of the isolates. CONCLUSION: The serovar 12 antiserum produced against the international reference strain (1096) does not recognise Australian serovar 12 isolates. The antiserum raised against isolate HS143 does recognise the Australian serovar 12 isolates. The dominant serovars of A pleuropneumoniae infecting Australian pigs are (in decreasing order) serovars 12, 1 and 7.     

2型猪链球菌的血清学鉴定

本试验首次证实了２型猪链球菌在我国的存在，采用玻片凝集、琼脂扩散及毛细管沉淀等３项试验，对１９９０年以来广东某猪场断奶仔猪暴发流行的败血症、脑膜炎及关节炎病猪中分离的１５株链球菌进行了血清学鉴定。结果表明：１５株链球菌均为２型猪链球菌；用高压法提取抗原进行沉淀反应，其结果比酸热法提取抗原具有简便、敏感、特异性强的特点。     

猪传染性胸膜肺炎放线杆菌PCR检测试剂盒的研制

根据GenBank中猪传染性胸膜肺炎放线杆菌apxⅣA基因序列,设计了1对引物,通过对PCR反应条件进行优化,研制了检测猪传染性胸膜肺炎放线杆菌的PCR试剂盒。该试剂盒扩增的阳性条带为600 bp,特异性与敏感性结果显示,该PCR检测试剂盒的最低核酸检测量为50 CFU/mL,而对金黄色葡萄球菌、链球菌、多杀性巴氏杆菌、鼠伤寒沙门氏菌、副猪嗜血杆菌、大肠杆菌的扩增结果均为阴性。-20℃至少可保存12个月,且重复性良好。应用该PCR试剂盒对41份临床样本进行了检测,其PCR检测结果与细菌学检测结果相一致。结果表明,猪传染性胸膜肺炎放线杆菌PCR检测试剂盒能够对APP临床样品进行快捷、灵敏、准确的检测。     

附红细胞体自然感染猪血液流变学

选择临床健康的2月龄附红细胞体自然感染仔猪15头,每周采血1次,连续4次,检测血样的RBC、PCV、ηb10s-1、ηb150 s-1、ηp、ηre10 s、ηre150 s-1、AI、RI、TK.检测结果如下随着猪附红细胞体感染强度的增强,ηb10 s-1、ηre10s-1、ηre150 s-1、AI、RI、TK明显增高,而RBC、PCV、ηb150 s-1、ηp无明显变化,表明猪感染附红细胞体后,血液粘度增高,红细胞聚集性增高,红细胞变形能力降低,进而影响脏器组织的血液灌注和微循环功能.这些血液流变学改变,构成了血栓、炎症、变性、水肿、坏死等的病理学基础.     

人感染猪链球菌2型的研究进展

猪链球菌2型是一种重要的人畜共患病原菌,可引起人的脑膜炎、败血症等疾病.2005年6月发生的四川疫情是由猪链球菌2型引起的人猪共同感染,流行广,发展快,人猪患病数量多.本文从病原、流行特点、感染途径、临床表现、实验室鉴定(包括培养特性、形态特征、生化特性、血清凝集、毒力基因)等方面做了详细的综述,初步探讨了2型猪链球菌病的防治方法.     

Evaluation of a single dose versus a divided dose regimen of amoxycillin in treatment of Actinobacillus pleuropneumoniae infection in pigs

The theory of a time-dependent effect of amoxycillin was examined in a model of porcine Actinobacillus pleuropneumoniae (Ap)-infection using clinically relevant dosage regimens. Twenty hours after infection of fourteen pigs, when clinical signs of pneumonia were present, one group of pigs received a single dose of amoxycillin (20 mg/kg, i.m.), whereas another group received four doses of 5 mg/kg injected at 8-h intervals. A similar AUC of the plasma amoxycillin concentration versus time curve was obtained in the two groups, whereas the maximum concentration was threefold higher using the single high dose. Plasma amoxycillin was above the MIC for twice as long using the fractionated dosage scheme. The condition of the animals was evaluated by clinical and haematological observations combined with quantification of biochemical infection markers: C-reactive protein, zinc and ascorbic acid. Within 48 h of treatment, the pigs in both treatment groups recovered clinically. No significant differences in the time-course of clinical observations or plasma concentrations of the biomarkers of infection were observed between the two treatments. In conclusion, the efficacy of these two dosage regimens of amoxycillin was not significantly different in treatment of acute Ap-infection in pigs.     

Evaluation of a single dose versus a divided dose regimen of danofloxacin in treatment of Actinobacillus pleuropneumoniae infection in pigs

A single versus a divided dose regimen of danofloxacin was evaluated in treatment of porcine Actinobacillus pleuropneumoniae infection using clinical observations combined with biochemical infection markers: C-reactive protein, zinc and ascorbic acid. Twenty hours after experimental infection, the 18 pigs received danofloxacin intravenously as a single dose of 2.5mg/kg or four doses of 0.6 mg/kg administered at 24h intervals. These dosage regimens resulted in similar AUCs of the plasma danofloxacin vs time curve. The maximum concentration was 3.5-fold higher using the single dose regimen, while the time with concentrations above the MIC was 2.5-fold longer using the fractionated regimen. Using the single dose regimen, temperature was normalised 32 h post-infection. In contrast, normalisation was delayed until 44 h post-infection using four low doses and a relapse with elevated temperatures at 52 and 68 h was observed. No other significant differences between the treatments were found, neither regarding clinical, haematological nor biochemical observations. The use of the more convenient single dose regimen was appropriate, as it was at least equivalent to the fractionated regimen.