The genetic organization of the capsular polysaccharide biosynthesis region of Actinobacillus pleuropneumoniae serotype 14

The genetic organization of the gene involved in the capsular polysaccharide (CPS) biosynthesis of Actinobacillus pleuropneumoniae serotype 14 has been determined. The DNA region for the CPS biosynthesis of serotype 14 (cps14) comprised 9 open reading frames, designated as cps14AB₁B₂B₃CDEFG genes, encoding Cps14A to Cps14G protein, respectively. Cps14A was similar to CpsA of A. pleuropneumoniae serotypes 1, 4 and 12; the Cps14B₁ and Cps14B₂ were similar to CpsB of A. pleuropneumoniae serotypes 1, 4 and 12, suggesting that CPS structure of A. pleuropneumoniae serotype 14 would belong to Group I including A. pleuropneumoniae serotypes 1, 4, 12 and 15. Surprisingly, the overall nucleotide sequence, deduced amino acid sequence, and the genetic organization of the cps14 were nearly identical to those of Actinobacillus suis. This study will provide the molecular basic knowledge for development of diagnostics and vaccine of A. pleuropneumoniae serotype 14.     

致病性猪链球菌2型的病原分离鉴定及毒力因子的PCR检测

通过对广西某猪场急性死亡的猪进行病原分离,分离到革兰氏阳性球菌,用链球菌快速鉴定试剂条Rapid ID 32 Strep鉴定为猪链球菌2型;并对分离菌进行猪链球菌2型荚膜多糖抗原（cps2J）及其重要毒力因子溶菌酶释放蛋白（MRP）、细胞外蛋白因子（EF）和溶血素（SLY）的多重PCR检测,同时对cps2J基因进行序列测定,与GenBank发表的猪链球菌2型相比,同源性为98.8%,毒力因子MRP、EF和SLY检测均为阳性,证实广西某猪场急性死亡的猪为高毒力猪链球菌2型感染所致。动物试验中,该菌可引起小白鼠部分死亡,家兔体温最高升至40.3℃,最后败血而死。     

9型猪链球菌广东株的分离鉴定和基因序列分析

为了解9型猪链球菌广东分离株毒力因子基因型、基因变异和进化情况，本试验采集发病猪脑脊液、关节液、脾脏、肝脏、淋巴结进行细菌分离培养和生化鉴定，采用PCR方法鉴定其血清型及部分毒力基因，对cps9D、gdh、gapdh与orf2基因进行序列测定和分析，并构建系统发育树。结果显示，分离菌镜检为革兰氏阳性链状球菌，在鲜血琼脂平板中呈β溶血，可发酵大多数糖类，能成功扩增cps9D基因，含重要的保守基因gdh及毒力基因gapdh和orf2，表明分离菌株为9型猪链球菌。分离菌株cps9D、gdh、gapdh、orf2基因核苷酸序列与GenBank上国内外参考菌株的同源性分别为95.9%~100.0%、98.6%~99.8%、98.7%~99.6%和95.5%~99.9%，说明分离菌株与国内外其他9型猪链球菌毒株核苷酸同源性都较高，且与国内外的流行毒株基本一致，未发生太大的变异。系统发育树表明，分离株与国内外来源不同的猪链球菌分离株同源性高，亲缘关系密切。     

A novel fibronectin-binding protein of Streptococcus suis serotype 2 contributes to epithelial cell invasion and in vivo dissemination

Streptococcus suis serotype 2 (SS2) is an important pathogen with zoonotic potential. In this study, a novel in vivo induced protein Ssa, encoded by the functionally unknown gene SSU05_1311, was identified as a surface anchored fibronectin-binding protein. The recombinant Ssa as well as its truncated proteins harboring the N-terminal domain (residues 33–153) could bind to human fibronectin with high-affinity. Isogenic knockout of ssa in SS2 led to decrease of bacterial binding to immobilized fibronectin. SS2 Δssa mutant showed reduced adherence and invasion of human pharyngeal epithelial (HEp-2) cells compared to the wild type strain. Heterologous surface display of Ssa fibronectin-binding domain in Escherichia coli enhanced the bacterial attachment and entry to HEp-2 cells. Less SS2 Δssa mutants than the wild type strain were recovered from the blood and brains of the Balb/c mice infected intranasally. But there was no significant difference between the wild type and the mutant on phagocytosis by macrophages (RAW264.7) and bacterial killing by murine PMNs under opsonizing condition. Our data suggest that Ssa is an important virulence factor for SS2 crossing the mucosal epithelia to disseminate in vivo.     

猪链球菌2型Ⅲ型溶血素的研究

为了探讨猪链球菌2型(Streptococcus suis serotype 2,SS2)的Ⅲ型溶血素是否具有溶血活性以及Ⅲ型溶血素在SS2致病过程中的作用,本研究利用同源重组基因敲除法成功构建了SS205ZY的Ⅲ型溶血素(slyrp)基因缺失突变菌株△slyrp及双基因缺失突变菌株△sly/△slyrp,并比较了野生菌株和基因缺失突变菌株的溶血能力以及对小鼠的致病力.结果表明,slyrp基因敲除后可导致SS2裂解红细胞的能力有所下降,而双基因缺失突变菌株△sly/△slyrp的溶血能力完全丧失;slyrp基因敲除后对小鼠的致病力没有影响.结果提示猪链球菌2型Ⅲ型溶血素具有一定的溶血能力,该Ⅲ型溶血素在SS2感染过程中,对溶血素(sly)起协同作用,不是SS2主要的毒力相关基因.     

The genetic organization of the capsular polysaccharide biosynthesis region of Actinobacillus pleuropneumoniae serotype 15

Nucleotide sequence determination and analysis of the cps gene involved in the capsular polysaccharide biosynthesis of Actinobacillus pleuropneumoniae serotype 15 revealed the presence of three open reading frames, designated as cps15ABC genes. At the protein level, Cps15A and Cps15B showed considerably high homology to CpsA (67.0 to 68.7%) and CpsB (31.7 to 36.8%), respectively, of A. pleuropneumoniae serotypes 1, 4 and 12, revealing the common genetic organization of the cps among serotypes 1, 4, 12 and 15. However, Cps15C showed no homology to any proteins of A. pleuropneumoniae serotypes, indicating that cps15C may be specific to serotype 15. This study will provide the basic molecular knowledge necessary for the development of diagnostics and a vaccine for A. pleuropneumoniae serotype 15.     

猪链球菌PCR检测技术研究进展

猪链球菌是猪的一种重要病原菌,并且也会引起人的链球菌病。有35个荚膜血清型(1/21、~34),通常自发病或死亡猪体分离获得1,2,7,9型和14型菌株,其中2型是毒力最强的血清型。根据已知猪链球菌16 SrRNA及溶血素(sly)、谷氨酸脱氢酶(gdh)、荚膜多糖(cps)、胞壁蛋白或溶菌酶释放相关蛋白(mrp)、胞外因子(epf)编码基因序列设计特异性引物,建立猪链球菌群和1(14),2(1/2),7型和9型特异性PCR或多重PCR,建立2型致病性菌株和1型高致病性菌株毒力鉴定PCR或多重PCR,用于检测和鉴别临床病料和细菌分离物中的猪链球菌,具有高敏感性和高特异性,与其他致病菌及其他血清的猪链球菌型无交叉反应,为疫病诊断及流行病学的研究提供了快速、简便和有用的工具。     

血清2型猪链球菌河南分离株毒力基因的鉴定及其cps2j全基因序列分析

为了解血清2型猪链球菌(S.suis2)河南分离株的毒力基因型及cps2j全基因突变情况,本研究扩增S.suis2毒力基因gdh、cps2j、mrp、ef 、sly、orf2和fbps,并对s.suis2的cps2j进行全基因序列测定和同源性分析.结果表明,gdh、cps2j、mrp、ef、sly、orf2和fbps基因在8个S.suis2分离株中的检出率分别为100％(8/8)、100％(8/8)、62.5％(5/8)、75％(6/8)、87.5％(7/8)、100％(8/8)和100％(8/8);其中cps2j全基因核苷酸及推导的氨基酸序列与来源不同的S.suis2分离株同源性分别达98％和97％以上,以该基因构建的系统发育树表明8株S.suis2河南分离株与国内外不同参考株同源性高,亲缘关系密切.     

Atypical Actinobacillus pleuropneumoniae isolates that share antigenic determinants with both serotypes 1 and 7.

In the present study, the characterization of 3 atypical isolates of Actinobacillus pleuropneumoniae is presented. Two isolates (1B and 27E) showed positive reactions in coagglutination, immunodiffusion, and indirect hemagglutination tests for serotypes 1 and 7, whereas the third isolate (26B) reacted with antisera to serotypes 1, 4, and 7. These atypical isolates of A. pleuropneumoniae possessed a capsular polysaccharide (CPS) antigenically related to serotype 1 as well as an O-chain lipopolysaccharide antigenically related to serotype 7 or to serotypes 4 and 7, as shown by the use of monoclonal antibodies. Results of toxin profile and virulence assays for mice and pigs showed them to be more related to A. pleuropneumoniae serotype 7 field isolates. All 3 isolates induced antibodies mainly against serotype 7/4 O-long-chain lipopolysaccharide (LC-LPS) and, to a lesser extent, to the CPS of serotype 1, in experimentally infected pigs. Diagnostic laboratories that use a LC-LPS-based enzyme-linked immunosorbent assay (ELISA) for serodiagnosis of A. pleuropneumoniae infection in swine would probably diagnose herds infected with these atypical isolates as being infected by A. pleuropneumoniae serotypes 7 or 4, whereas those that use a CPS-based ELISA would probably consider them as infected by A. pleuropneumoniae serotype 1.     

广东省1株猪2型链球菌的分离鉴定及部分生物学特性研究

为确诊引起广东省某规模化猪场保育仔猪发生死亡的病因,本试验采集病猪肝脏、脾脏、心脏、脑、关节液和淋巴结等病料进行细菌分离,并对分离菌株进行纯化培养、菌落形态观察、革兰氏染色、形态学观察、生化试验,用血清型特异性引物进行PCR扩增,并进行毒力基因检测、药敏试验、生长曲线测定及动物试验。结果显示,分离菌株在成牛血清琼脂平板上呈圆形、灰白色、表面光滑、边缘整齐的菌落,镜检为革兰氏阳性球菌,呈链状排列。该菌株的生化试验结果符合链球菌的特征,PCR扩增结果显示猪链球菌保守基因片段（gdh和gapdh）和猪2型链球菌特异的cps2J基因片段均为阳性,表明该分离菌株为猪2型链球菌。毒力基因检测结果表明,该菌株毒力因子基因型为gdh⁺/gapdh⁺/epf⁺/mrp⁺/sly⁺/orf2⁺/fbps⁺。分离菌株用营养肉汤培养6 h达到对数生长期,培养10 h的菌液D_{600 nm}值为0.310。药敏试验结果表明,该菌株对青霉素、阿莫西林、阿莫西林-克拉维酸钾等药物敏感,但对强力霉素耐药。动物试验结果表明,感染分离株的BALB/c小鼠未出现死亡且无明显临床症状,毒力较低。本研究为该猪场链球菌病的防控提供了重要参考信息。     

Mutants in the lipopolysaccharide of Brucella ovis are attenuated and protect against B. ovis infection in mice

Brucella spp. are Gram-negative bacteria that behave as facultative intracellular parasites of a variety of mammals. This genus includes smooth (S) and rough (R) species that carry S and R lipopolysaccharides (LPS), respectively. S-LPS is a virulence factor, and mutants affected in the S-LPS O-polysaccharide (R mutants), core oligosaccharide or both show attenuation. However, B. ovis is naturally R and is virulent in sheep. We studied the role of B. ovis LPS in virulence by mutating the orthologues of wadA, wadB and wadC, three genes known to encode LPS core glycosyltransferases in S brucellae. When mapped with antibodies to outer membrane proteins (Omps) and R-LPS, wadB and wadC mutants displayed defects in LPS structure and outer membrane topology but inactivation of wadA had little or no effect. Consistent with these observations, the wadB and wadC but not the wadA mutants were attenuated in mice. When tested as vaccines, the wadB and wadC mutants protected mice against B. ovis challenge. The results demonstrate that the LPS core is a structure essential for survival in vivo not only of S brucellae but also of a naturally R Brucella pathogenic species, and they confirm our previous hypothesis that the Brucella LPS core is a target for vaccine development. Since vaccine B. melitensis Rev 1 is S and thus interferes in serological testing for S brucellae, wadB mutant represents a candidate vaccine to be evaluated against B. ovis infection of sheep suitable for areas free of B. melitensis.     

Elucidating the role of ApxI in hemolysis and cellular damage by using a novel apxIA mutant of Actinobacillus pleuropneumoniae serotype 10

Exotoxins produced by Actinobacillus (A.) pleuropneumoniae (Apx) play major roles in the pathogenesis of pleuropneumonia in swine. This study investigated the role of ApxI in hemolysis and cellular damage using a novel apxIA mutant, ApxIA336, which was developed from the parental strain A. pleuropneumoniae serotype 10 that produces only ApxI in vitro. The genotype of ApxIA336 was confirmed by PCR, Southern blotting, and gene sequencing. Exotoxin preparation derived from ApxIA336 was analyzed for its bioactivity towards porcine erythrocytes and alveolar macrophages. Analysis results indicated that ApxIA336 contained a kanamycin-resistant cassette inserted immediately after 1005 bp of the apxIA gene. Phenotype analysis of ApxIA336 revealed no difference in the growth rate as compared to the parental strain. Meanwhile, ApxI production was abolished in the bacterial culture supernatant, i.e. exotoxin preparation. The inability of ApxIA336 to produce ApxI corresponded to the loss of hemolytic and cytotoxic bioactivity in exotoxin preparation, as demonstrated by hemolysis, lactate dehydrogenase release, mitochondrial activity, and apoptosis assays. Additionally, the virulence of ApxIA336 appeared to be attenuated by 15-fold in BALB/c mice. Collectively, ApxI, but not other components in the exotoxin preparation of A. pleuropneumoniae serotype 10, was responsible for the hemolytic and cytotoxic effects on porcine erythrocytes and alveolar macrophages.     

The role of rpoS,hmp, and ssrAB in Salmonella enterica Gallinarum and evaluation of a triple-deletion mutant as a live vaccine candidate in Lohmann layer chickens

Salmonella enterica Gallinarum (SG) causes fowl typhoid (FT), a septicemic disease in avian species. We constructed deletion mutants lacking the stress sigma factor RpoS, the nitric oxide (NO)-detoxifying flavohemoglobin Hmp, and the SsrA/SsrB regulator to confirm the functions of these factors in SG. All gene products were fully functional in wild-type (WT) SG whereas mutants harboring single mutations or a combination of rpoS, hmp, and ssrAB mutations showed hypersusceptibility to H₂O₂, loss of NO metabolism, and absence of Salmonella pathogenicity island (SPI)-2 expression, respectively. A triple-deletion mutant, SGΔ3 (SGΔrpoSΔhmpΔssrAB), was evaluated for attenuated virulence and protection efficacy in two-week-old Lohmann layer chickens. The SGΔ3 mutant did not cause any mortality after inoculation with either 1 × 10⁶ or 1 × 10⁸ colony-forming units (CFUs) of bacteria. Significantly lower numbers of salmonellae were recovered from the liver and spleen of chickens inoculated with the SGΔ3 mutant compared to chickens inoculated with WT SG. Vaccination with the SGΔ3 mutant conferred complete protection against challenge with virulent SG on the chickens comparable to the group vaccinated with a conventional vaccine strain, SG9R. Overall, these results indicate that SGΔ3 could be a promising candidate for a live Salmonella vaccine against FT.     

OmpA is a virulence factor of Riemerella anatipestifer

Riemerella anatipestifer infection is probably the most economically important disease of farm ducks worldwide. The pathogen R. anatipestifer causes septicemia anserum exsudativa in ducks, but little is known about the molecular basis of its pathogenesis and the virulence factors involved. In this study, by deleting ompA gene from R. anatipestifer serotype 2 strain Th4, we constructed a mutant strain Th4ΔompA to investigate whether R. anatipestifer OmpA is an important virulence factor. Results showed that although the growth curve, bacterial and colony morphology of Th4ΔompA in tryptic soybean broth (TSB) or on TSB agar were similar to its parent strain Th4, the adhesion and invasion capacities of mutant strain to Vero cells were decreased significantly. Furthermore, the median lethal dose (LD(50)) of both strains was determined to measure the virulence with 10-day-old Cherry Valley ducklings. The results showed that LD(50) of Th4ΔompA mutant was >10(10) colony forming units (CFU), it was attenuated significantly in comparison with that of Th4 which LD(50) was 4.41 × 10(8) CFU. Additional analysis indicated that blood bacterial loading of ducklings infected with the Th4ΔompA mutant were much lower than those of Th4-infected ducklings. The results demonstrate that OmpA is a virulence factor of R. anatipestifer, and that it may act as an adhesin.     

Pyruvate kinase is necessary for Brucella abortus full virulence in BALB/c mouse

Brucellosis, caused by a facultative intracellular pathogen Brucella, is one of the most prevalent zoonosis worldwide. Host infection relies on several uncanonical virulence factors. A recent research hotpot is the links between carbon metabolism and bacterial virulence. In this study, we found that a carbon metabolism-related pyruvate kinase (Pyk) encoded by pyk gene (locus tag BAB_RS24320) was associated with Brucella virulence. Determination of bacterial growth curves and resistance to environmental stress factors showed that Pyk plays an important role in B. abortus growth, especially under the conditions of nutrition deprivation, and resistance to oxidative stress. Additionally, cell infection assay showed that Pyk is necessary for B. abortus survival and evading fusion with lysosomes within RAW264.7 cells. Moreover, animal experiments exhibited that the Pyk deletion significantly reduced B. abortus virulence in a mouse infection model. Our results elucidated the role of the Pyk in B. abortus virulence and provided information for further investigation of Brucella virulence associated carbon metabolism.     

Sample pooling for real-time PCR detection and virulence determination of the footrot pathogen <Emphasis Type="Italic">Dichelobacter nodosus</Emphasis>

Dichelobacter nodosus is the principal cause of ovine footrot and strain virulence is an important factor in disease severity. Therefore, detection and virulence determination of D. nodosus is important for proper diagnosis of the disease. Today this is possible by real-time PCR analysis. Analysis of large numbers of samples is costly and laborious; therefore, pooling of individual samples is common in surveillance programs. However, pooling can reduce the sensitivity of the method. The aim of this study was to develop a pooling method for real-time PCR analysis that would allow sensitive detection and simultaneous virulence determination of D. nodosus. A total of 225 sheep from 17 flocks were sampled using ESwabs within the Swedish Footrot Control Program in 2014. Samples were first analysed individually and then in pools of five by real-time PCR assays targeting the 16S rRNA and aprV2/B2 genes of D. nodosus. Each pool consisted of four negative and one positive D. nodosus samples with varying amounts of the bacterium. In the individual analysis, 61 (27.1%) samples were positive in the 16S rRNA and the aprV2/B2 PCR assays and 164 (72.9%) samples were negative. All samples positive in the aprV2/B2 PCR-assay were of aprB2 variant. The pooled analysis showed that all 41 pools were also positive for D. nodosus 16S rRNA and the aprB2 variant. The diagnostic sensitivity for pooled and individual samples was therefore similar. Our method includes concentration of the bacteria before DNA-extraction. This may account for the maintenance of diagnostic sensitivity. Diagnostic sensitivity in the real-time PCR assays of the pooled samples were comparable to the sensitivity obtained for individually analysed samples. Even sub-clinical infections were able to be detected in the pooled PCR samples which is important for control of the disease. This method may therefore be implemented in footrot control programs where it can replace analysis of individual samples.     

Virulence determinants of Salmonella Gallinarum biovar Pullorum identified by PCR signature-tagged mutagenesis and the spiC mutant as a candidate live attenuated vaccine

Salmonella Gallinarum biovar Pullorum (S. Gallinarum biovar Pullorum) is the causative agent of pullorum disease (PD) in chickens which results in considerable economic losses to the poultry industries in developing countries. PCR-Signature Tagged Mutagenesis was used to identify virulence determinants of S. Gallinarum biovar Pullorum and novel attenuated live vaccine candidates for use against this disease. A library of 1800 signature-tagged S. Gallinarum biovar Pullorum mutants was constructed and screened for virulence-associated genes in chickens. The attenuation of 10 mutants was confirmed by in vivo and in vitro competitive index (CI) studies. The transposons were found to be located in SPI-1 (2/10 mutants), SPI-2 (3/10), the virulence plasmid (1/10) and non-SPI genes (4/10). One highly attenuated spiC mutant persisted in spleen and liver for less than 10 days and induced high levels of circulating antibody and protective immunity against oral challenge in young broiler chickens. The spiC mutant is a potential new vaccine candidate for use with chickens against this disease.     

Lack of a type-2 glycosyltransferase in the fish pathogen Flavobacterium psychrophilum determines pleiotropic changes and loss of virulence

Flavobacterium psychrophilum is an important fish pathogen, responsible for Cold Water Disease, with a significant economic impact on salmonid farms worldwide. In spite of this, little is known about the bacterial physiology and pathogenesis mechanisms, maybe because it is difficult to manipulate, being considered a fastidious microorganism. Mutants obtained using a Tn4351 transposon were screened in order to identify those with alteration in colony morphology, colony spreading and extracellular proteolytic activity, amongst other phenotypes. A F. psychrophilum mutant lacking gliding motility showed interruption of the FP1638 locus that encodes a putative type-2 glycosyltransferase (from here on referred to as fpgA gene, Flavobacterium psychrophilum glycosyltransferase). Additionally, the mutant also showed a decrease in the extracellular proteolytic activity as a consequence of down regulation in the fpgA mutant background of the fpp2-fpp1 operon promoter, responsible for the major extracellular proteolytic activity of the bacterium. The protein glycosylation profile of the parental strain showed the presence of a 22 kDa glycosylated protein which is lost in the mutant. Complementation with the fpgA gene led to the recovery of the wild-type phenotype. LD₅₀ experiments in the rainbow trout infection model show that the mutant was highly attenuated. The pleiotropic phenotype of the mutant demonstrated the importance of this glycosyltranferase in the physiology and virulence of the bacterium. Moreover, the fpgA mutant strain could be considered a good candidate for the design of an attenuated vaccine.     

Serotype distribution and production of muramidase-released protein, extracellular factor and suilysin by field strains of Streptococcus suis isolated in the United States

Streptococcus suis is an important swine pathogen and a zoonotic agent. Differences in virulence have been noted among the 33 described serotypes, serotype 2 being considered the most virulent. In this study, we aimed at assessing the serotype distribution and the production of virulence-associated markers by strains recovered from diseased pigs in the United States (U.S.). Results showed that among the 100 strains evaluated, serotype 3 (20% of the isolates) and serotype 2 (17%) were the most prevalent. We then investigated the presence in these isolates of the genes sly, epf and mrp, encoding the virulence-associated markers suilysin (SLY), extracellular factor (EF) and muramidase-released (MRP) protein, respectively. The effective production of the markers by the strains was also verified. Results showed that the presence of the gene did not always correlate with actual expression of the respective protein. In the case of MRP, this was due, in most cases, to frameshift mutations at the 5′ end of the gene resulting in premature stop codons. The most prevalent phenotypes among U.S. strains were MRP⁺EF⁻SLY⁻ (40%) and MRP⁻EF⁻SLY⁺ (35%). Serotype distribution greatly differed from that reported in several European countries, as did the production of virulence markers, particularly for serotype 2. On the other hand, our results for the U.S. S. suis isolates are similar to those reported for Canadian strains, suggesting a common status in North America.     

猪链球菌14型的分离鉴定及生物学特性研究

为了检测确定2019年5月河南某规模化猪场一栋保育仔猪发病猪群的病原,本研究从送检的发病猪关节液中分离获得1株细菌。通过细菌纯化培养、革兰氏染色、形态学观察及猪链球菌gdh基因PCR扩增,确定该分离菌株为猪链球菌。用猪链球菌分型引物对该菌株进行PCR扩增分型鉴定及软件比对分析,结果表明该分离菌株为猪链球菌14型,与猪链球菌JS14株（GenBank登录号：CP002465.1）同源性为100%。毒力基因检测结果表明,该菌株的同时携带有epf、mrp、sly、fbps、orf2毒力基因,属于高致病性菌株。小鼠致病性试验结果也证明该菌株是一株高致病性猪链球菌。药物敏感性试验结果显示,该菌株对β内酰胺类和喹诺酮类药物敏感,对氨基糖苷类、四环素类、大环内酯类和磺胺类高度耐药,表现出多重耐药现象。对该菌株进行5大类24种耐药基因检测,该菌株同时携带有bla_TEM、aadA1、strA、strB、aacC2、aphA1、tet（B）、gyrA、parC、sul2耐药基因。该研究为后续进一步开展猪链球菌14型流行特点和致病机制研究奠定了基础,为猪链球菌14型临床防控提供了理论依据,同时具有重要的公共卫生意义。