Carriage of Corynebacterium pyogenes by the cattle nuisance flies Hydrotaea irritans (Fallén) and Musca autumnalis (De Geer)

Two species of cattle-visiting Muscidae were experimentally contaminated with C. pyogenes, a pathogen involved in the aetiology of summer mastitis. Surface contamination persisted for at least 4 days. Since M. autumnalis would not feed on media containing C. pyogenes the bacterium did not persist internally. All C. pyogenes were eliminated from the gut of H. irritans in 4 days. H. irritans is thus more likely to transmit C. pyogenes than is M. autumnalis but only by mechanical transfer, and is not a true vector.     

Endometrial biopsy in Holstein-Friesian dairy cows. III. Bacteriological analysis and correlations with histological findings.

This study examines the results of bacterial culture from 159 endometrial biopsy samples from 97 commercial dairy cows and correlations between bacteriological and histological findings. Bacteria were isolated from approximately 80% of biopsies taken at day 26 and day 40 postpartum. Eleven percent of biopsies were positive for both aerobic and anaerobic culture. Streptococci, Escherichia coli and Actinomyces pyogenes were the most common isolates. Isolation of A. pyogenes from a biopsy at day 26 was positively correlated with isolation of anaerobic bacteria and segmented cell inflammation in the same biopsy, and with subsequent isolation of A. pyogenes at day 40. There was a strong association between isolation of A. pyogenes and anaerobes at day 26 with increased uterine lesions at day 40. Isolation of alpha hemolytic streptococci (AHS) was negatively correlated with isolation of A. pyogenes and with inflammation. Actinomyces pyogenes and AHS showed opposite associations with mononuclear cell inflammation and lymphocytic foci.     

化脓隐秘杆菌肝素结合蛋白的提取及铁结合蛋白的黏附特性分析

肝素可剂量依赖性地抑制化脓隐秘杆菌(Trueperella pyogenes)黏附宿主细胞.本研究旨在认识化脓隐秘杆菌肝素结合蛋白及其黏附特性,采用肝素琼脂糖凝胶从化脓隐秘杆菌裂解物中提取蛋白,运用蛋白质质谱和免疫印迹对所提取的蛋白质进行鉴定.采用免疫印迹检测所提取蛋白质与自然感染化脓隐秘杆菌山羊血清的反应原性.制备重...     

Molecular characterization of the pore-forming toxin, pyolysin, a major virulence determinant of Arcanobacterium pyogenes

Arcanobacterium pyogenes is a common inhabitant and opportunistic pathogen of domestic animals. The pathogenesis of this organism in a range of suppurative diseases is not well understood. However, the development of genetic techniques to study this organism has allowed advances in the analysis of A. pyogenes virulence factors. A major step in this analysis was the identification and cloning of the A. pyogenes hemolytic exotoxin, pyolysin (PLO). PLO is the most divergent member of the cholesterol-binding pore-forming family of toxins. PLO is also divergent in a C-terminal undecapeptide motif which is almost invariant among other members of the family. This divergent undecapeptide motif is required for the full cytolytic activity of PLO and is also responsible for its oxygen-resistant nature. Insertional inactivation of the plo gene results in a significant reduction in virulence in an intraperitoneal mouse model of infection. The virulence of the plo mutant can be restored by providing PLO in trans, suggesting that PLO is a major virulence factor in A. pyogenes pathogenesis in mice. Results of previous vaccination trials with crude antigens against A. pyogenes infection in domestic animals and mice have been equivocal at best. However, a recombinant PLO-based subunit vaccine protected mice from experimental A. pyogenes infection, indicating that PLO is also an important host protective antigen. These results provide promise that the dogma that domestic animals are recalcitrant to vaccination against A. pyogenes infection may prove false.     

Isolation of Arcanobacterium (Actinomyces) pyogenes from cases of feline otitis externa and canine cystitis.

Arcanobacterium pyogenes is a normal inhabitant of the mucous membranes of domestic animals, such as cattle, sheep, swine, and goats. It is also an opportunistic pathogen in these animals, where it causes a variety of purulent infections involving the skin, joints, and visceral organs. Two recent cases of isolation of A. pyogenes from companion animals are reported. In the first case, a cat presented with a chronic otitis externa, from which A. pyogenes was isolated in pure culture. The second case involved a dog with a urinary tract infection, where A. pyogenes was isolated from urine as the predominant bacterial species. In both cases, the A. pyogenes isolates were presumptively identified by macrobiochemical tests, and then their identities were confirmed by polymerase chain reaction analysis and 16S rRNA gene sequencing.     

Morbidity and mortality associated with Arcanobacterium pyogenes in a group of captive blackbuck (Antilope cervicapra).

Arcanobacterium pyogenes was associated with necrotizing pneumonia; mandibular osteomyelitis; peritonitis; and hepatic, pulmonary, renal, and subcutaneous abscessation in a group of captive blackbuck (Antilope cervicapra). Males were more frequently (73.3%) affected than females. Infection with A. pyogenes was fatal or necessitated euthanasia in 15 of 16 (93.7%) cases. Deaths associated with A. pyogenes occurred most frequently (60%) during winter.     

Effect of damage to the teat end on the experimental induction of mastitis in dry cows with Corynebacterium pyogenes

The teat ends of 12 dry cows were contaminated with Corynebacterium pyogenes. To determine whether a pre-existing (an)aerobic bacterial infection of the udder was a predisposing factor for a C pyogenes mastitis they included infected and uninfected quarters. Anaerobic bacteria could not be found and mastitis was not induced. When the teats were contaminated with C pyogenes after the teat ends had been injured 30 of the quarters became infected, and anaerobic bacteria were demonstrated in many quarters.     

Pyometra in camels: case report.

Pyometra was recorded in five camels and Actinomyces pyogenes (Corynebacterium pyogenes) was isolated from pus of the affected camels.     

Influence of Escherichia coli and Arcanobacterium pyogenes isolated from bovine puerperal uteri on phenotypic and functional properties of neutrophils

When cows develop endometritis after birth, Escherichia coli and Arcanobacterium pyogenes are usually the most prominent bacteria present in bovine uterine lochial secretions. A. pyogenes alone is rarely found in the course of a disturbed puerperium. This was confirmed in this study, since average and high-grade uterine contaminations were always associated with the presence of both bacteria. The contamination grade was positively correlated with uterine polymorphonuclear granulocyte (PMN) numbers and negatively correlated with blood PMN numbers. Whether E. coli and A. pyogenes affect the phenotype and function of bovine PMN in a similar or differential way was subject to in vitro studies. PMN were tested in the presence of washed bacterial fragments or culture supernatants taken as a source for soluble and/or secreted bacterial products. Fragments and soluble products differed only quantitatively in their effects on PMN. Usually, long-time exposure (24h) of PMN to fragments induced the strongest effects. Accelerated death of granulocytes was only moderately induced by both E. coli and A. pyogenes products. Both E. coli and A. pyogenes products induced the enhanced expression of a membrane molecule detected by mAb IL-A110 and of CD11b. Expression of other surface structures remained largely unchanged (MHC class I, CD11c). Functional parameters of PMN (phagocytosis; generation of reactive oxygen species, ROS; antibody-independent cellular cytotoxicity, AICC) generally declined after pre-incubation for 24h with products of E. coli or A. pyogenes. Interestingly, soluble products of A. pyogenes stimulated the phagocytosis of PMN. However, co-incubation with E. coli products abrogated this stimulatory effect. The results supply evidence for similar modes of action of the gram-negative E. coli and the gram-positive A. pyogenes on bovine PMN. Alterations in PMN function and phenotype are mainly triggered by direct contact between bacterial fragments and PMN. Inhibition experiments with polymyxin B demonstrated that E. coli-mediated effects were not solely due to the action of lipopolysaccharide. The dominant functional depression of neutrophils by E. coli products strengthens the suggestion that the earlier appearance of E. coli in the uterus may support the co-infection of this organ by A. pyogenes at later times.     

Pyothorax associated with a Mycoplasma sp and Arcanobacterium pyogenes in a kitten

Pyothorax associated with a Mycoplasma sp and Arcanobacterium pyogenes was diagnosed at necropsy in a 1-month-old female Van kitten. The pleural cavity contained approximately 50 mL of blood-tinged, reddish-brown, nonodourous fluid bilaterally. Gram positive coccobacilli were seen in the exudate from necrotic plaques on the pleurae. Mycoplasma sp and A pyogenes were isolated from a sample of the fluid in the pleural cavity. The concomitant presence of Mycoplasma sp and A pyogenes could be considered another variation on the polymicrobial nature of pyothorax and associated pleural lesions in cats.     

Corynebacterium pyrogenes; a biochemical and serological study

The study comprises 136 strains of Corynebacterium pyogenes originating from cattle (105), swine (20), sheep (1), and insects (10). For comparison 2 strains of human origin and 1 strain of Gorynebacterium hemolyticum were examined.One of the bovine strains was atypical, being gelatinase-negative, otherwise the strains of Cb. pyogenes were found to be biochemically identical apart from minor deviations in fermentation patterns (Table 1). Neither were antigenic differences demonstrated (gel diffusion analyses, Figs. 1 and 2).Both of the human strains agreed biochemically with Cb. pyogenes (Table 1). By gel diffusion cross analyses one of them was found to be identical with Cb. pyogenes, the other not, though anti-genically related to it (Fig. 2).Gb. hemolyticum deviated biochemically as well as serologically from Gb. pyogenes, but the 2 organisms shared antigenic determinants (Fig. 3).     

Susceptibility of Arcanobacterium pyogenes from different sources to tetracycline, macrolide and lincosamide antimicrobial agents

Chlortetracycline, oxytetracycline, and the macrolide, tylosin, are extensively used for growth promotion and disease prophylaxis in the cattle and swine industries in the US. Arcanobacterium pyogenes, a common inhabitant of the mucosal surfaces of cattle and swine, is also a pathogen associated with a variety of infections in these animals. A broth microdilution technique was used to determine the antimicrobial susceptibility of 48 A. pyogenes isolates to macrolides, lincosamides and tetracyclines. The MIC50 and MIC90 for chlortetracycline were 0.12 and 8 mg/l, respectively. Similarly, the MIC50 and MIC90 for oxytetracycline were 0.25 and 8 mg/l, while the MIC50 and MIC90 for tetracycline were 0.25 and 16 mg/l, respectively. The MIC50 and the MIC90 were < or = 0.06 and >64 mg/l, respectively, for erythromycin, tylosin and clindamycin. This resistance pattern indicated that some of these A. pyogenes isolates may carry an MLS(B) resistance determinant. A. pyogenes isolates (12.5%) were resistant to erythromycin, and this percentage doubled when MICs were performed following induction with erythromycin. Of the 48 A. pyogenes isolates, 25 and 41.7% were resistant to MLS(B) antimicrobial agents and the tetracycline derivatives, respectively. MLS(B) resistance was present in 22.2 and 35.3% of A. pyogenes isolates of bovine (n=27) or porcine (n=17) origin. In contrast, 70.6% of porcine isolates were resistant to the tetracyclines, compared with 25.9% of bovine isolates. These data suggest that a large proportion of A. pyogenes field isolates may be resistant to these commonly used antimicrobial agents.     

利血平对化脓隐秘杆菌大环内酯类外排基因mefA表达的影响

化脓隐秘杆菌(Trueperella pyogenes,T.pyogenes)是一种能够引起动物和人化脓性感染的重要病原菌。随着抗生素的广泛使用,该菌已对临床常用的大环内酯类、四环素类等药物产生不同程度的耐药性。本试验拟探讨外排泵抑制剂利血平对T.pyogenes大环内酯类外排基因mefA mRNA及蛋白表达的影响,将为细菌外排泵抑制剂的研究奠定理论基础。以携带mefA基因的T.pyogenes分离株(17,20号)为试验菌株,采用荧光定量PCR及Western blot技术,检测利血平作用前后各菌株mefA基因mRNA及蛋白表达量。结果表明,1/2 MIC利血平作用2株耐药菌株36 h后,大环内酯类抗生素红霉素、罗红霉素、泰乐菌素、阿奇霉素和替米考星对携带mefA基因的T.pyogenes的MIC值均有不同程度的降低。17和20号分离株外排基因mefA mRNA表达水平较药物作用前均极显著降低(利血平作用前是作用后的5.18,17.54倍),外排蛋白MefA表达量较药物作用前也显著降低(利血平作用前是作用后的1.08,1.70倍),且基因水平和蛋白水平的变化与其耐药表型的变化趋势一致;提示利血平可能是通过抑制外排基因mefA的转录和翻译而消减T.pyogenes对大环内酯类抗生素的耐药性。     

Evaluation of the API Coryne test system for identification of Actinomyces pyogenes.

The present study was designed to evaluate the accuracy of the API Coryne test system for identification of Actinomyces pyogenes. The test system correctly identified 36 of 42 A. pyogenes and 4 of 5 comparatively studied Arcanobacterium haemolyticum-cultures. The biochemical profiles of the remaining 6 A. pyogenes- and 1 A. haemolyticum-cultures were not included in the analytical profile index. None of the cultures were misidentified. According to the API database (ATB Plus V 1.5.4.) the unidentified cultures could be correctly identified as A. pyogenes and A. haemolyticum respectively. A greater repertoire of A. pyogenes specific biochemical profiles incorporated into the analytical profile index would improve the applicability of this test system for veterinary diagnostics.     

Isolation and partial characterisation of bacteria recovered from abscesses of normally slaughtered pigs

One hundred and one pig abscesses of different localisation were examined. Bacteria were re-covered from all abscesses. In 6’8 abscesses a mixed aerobic and anaerobic flora was found. In 22 abscesses solely aerobes and in 11 solely anaerobes were found. Gorynebaoterium pyogenes and Fuso-bacterium necrophorum were found in 61 and 45 abscesses, respectively. Hemagglutination litres of G. pyogenes strains were higher with pig erythrocytes than with sheep erythrocytes. Three of 4 isolates of Staphylococcus aureus produced enterotoxin type A.Key words: pig abscess, Corynebacterium pyogenes, Fuso-bacterium necrophorum, hemagglutination     

Genomic characterization of Arcanobacterium pyogenes isolates recovered from the uterus of dairy cows with normal puerperium or clinical metritis

Arcanobacterium pyogenes is considered to be the most relevant bacterium involved in the establishment of puerperal uterine infection in cattle due to its persistence in utero, resistance to treatment and synergic action with Gram negative anaerobes. Once the infection is established, A. pyogenes is responsible for the persistence of the infection. The objective of this study was to characterize A. pyogenes field isolates recovered from the uterus of cows with either normal puerperium or clinical metritis, in an attempt to identify factors that might be associated with the establishment and persistence of the disease. This characterization was based on BOX-PCR typing and on screening of eight virulence factor genes (plo, nanP, nanH, cbpA, fimA, fimC, fimE, fimG) by conventional PCR. Finally, a relationship between clonal types, virulence factors and presence of disease was investigated. A. pyogenes clonal types identified from isolates recovered from the uterus of postpartum dairy cows differed among herds. Although some clonal types were strictly associated with the development of clinical metritis, others were identified from isolates recovered from normal puerperium and clinical metritis cows. Moreover, the presence of the eight virulence factor genes was not related with the ability to induce clinical metritis, suggesting that the type of A. pyogenes may not be a determinant factor in the development of the disease. We suggest that host intrinsic factors, the synergism between A. pyogenes and other bacteria and the differential gene expression of virulence factor genes may play a more relevant role in the establishment of puerperal uterine infections.     

Infection following challenge of the lactating and dry udder of dairy cows with Actinomyces pyogenes and Peptostreptococcus indolicus

Challenge of 12 mammary glands of cows in mid-lactation with 10(7) colony forming units (cfu) of Peptostreptococcus indolicus on two occasions led to clinical mastitis in only four quarters. The bacteria were rarely recovered and disappeared from the secretion within 14 days. In challenges 7 days prior to drying off eight of 12 quarters became infected and at drying off all quarters challenged became infected. The infections established at drying off persisted well into the dry period. P. indolicus infection was also established in all of 12 dry glands challenged, but usually eliminated at calving or early in the next lactation. Isolation of P. indolicus was accompanied in about one-third of cases by changes in the appearance of the secretion. Intramammary challenge with Actinomyces (formally Corynebacterium) pyogenes led to clinical and subclinical infections in nine of 12 lactating glands and in all of six dry glands. Dry period infections with A. pyogenes were more severe and rarely eliminated even by antibiotic therapy. Infections during lactation were often eliminated either naturally or by antibiotic therapy. Intermittent recovery of A. pyogenes from the lactating mammary gland, without clinical signs of infection, was possible for up to 90 days after challenge. Combined infections with A. pyogenes and P. indolicus were clinically more severe with a higher frequency of systemic involvement. It was shown that in the non-lactating gland an acute mastitis, similar to 'summer mastitis' could be established either by simultaneous inoculation with A. pyogenes and P. indolicus or by subsequent inoculation of quarters excreting P. indolicus with A. pyogenes.     

奶牛源化脓隐秘杆菌的分离鉴定及其毒力基因的检测

本研究旨在分离鉴定来自北京某奶牛场死亡奶牛肺脏的1株疑似病原菌CVCC 3982,并测定其致病性。通过分离培养获得疑似病原菌,采用Biolog鉴定系统和16S rDNA序列分析对其进行了鉴定,人工接种CD-1小鼠测定了其致病性,合成引物对其主要毒力基因进行了检测。结果显示,该疑似病原菌为革兰氏阳性杆菌,β溶血,Biolog鉴定结果显示其为化脓隐秘杆菌,其16S rDNA序列与化脓隐秘杆菌模式菌株NCTC 5224的同源性达100%,系统发育分析显示其与化脓隐秘杆菌处于同一分支。腹腔注射该菌可致小鼠死亡。分离菌株基因组中含有溶血素(PLO)基因,神经氨酸酶H(NanH)基因,神经氨酸酶P(NanP)基因,菌毛基因(fimA、fimC和fimE),但缺失胶原结合蛋白(CbpA)基因和菌毛fimG基因。结果表明该分离菌株为化脓隐秘杆菌且具有致病性。     

Cause, occurrence, and clinical signs of mastitis and anorexia in cows in a Wisconsin study

Microbiologic culture revealed the following cause of mastitis and anorexia in 145 cows in Wisconsin to be Escherichia coli, 66 cows; Klebsiella spp, 3; Corynebacterium pyogenes, 27; streptococci, 21; staphylococci, 20; yeasts, 1; and no bacterial growth, 7. Mastitis was detected with approximately equal frequency throughout the year. Escherichia coli was isolated throughout the year, but was more common and was the predominant organism during the summer. Corynebacterium pyogenes was isolated most often in winter and spring; streptococci in fall, winter, and spring; and staphylococci throughout the year. Corynebacterium pyogenes caused most of the mastitis in nonlactating cows. Escherichia coli, C pyogenes, streptococci, and staphylococci were isolated with about equal frequency at parturition, whereas E coli was the predominant cause of mastitis in early and late lactation. Of cases of mastitis, 27% were seen 10 days before and after parturition. Local and systemic clinical signs of infection were similar for all causes, except that C pyogenes caused more (P less than 0.01) malodorous and purulent milk than did other organisms and was isolated more commonly from quarters with injured teats. Recovery was significantly (P less than 0.01) higher in cows with E coli infections, compared with recovery in cows with gram-positive organism infections. Cows with C pyogenes infections frequently had quarters that ultimately ceased lactation. A few cows were recumbent at initiation of antimicrobial therapy and a few were not eating 24 hours later; however, 50% of these cows recovered. Criteria such as season of year, stage of lactation, appearance of milk and udder, and appetite permitted the cause (gram-negative or gram-positive organisms) of the mastitis to be predicted with 77% accuracy.     

Adherence of streptococcal isolates from cattle and horses to their respective host epithelial cells

Adherence of Streptococcus dysgalactiae isolates from cattle and S equi isolates from horses to their respective host epithelial cells was compared with the adherence of S pyogenes to human epithelial cells. The adherence was quantitatively determined by use of fluorescein-labeled streptococci. All 3 streptococcal species adhered selectively to their respective host cells. The mechanism of adherence was evaluated by binding studies with adhesive plasma protein, fibronectin. Although all 3 streptococcal species bound fibronectin, S dysgalactiae and S equi interacted preferentially with a 210-kilodalton (kD) C-terminal fragment of fibronectin, whereas S pyogenes bound only a 29-kD N-terminal fragment. A synthetic peptide Gly-Arg-Gly-Asp-Ser, representing the host cell attachment site of fibronectin, partially inhibited the binding of fibronectin and of its 210 kD fragment to S dysgalactiae, but not to S equi. The binding of fibronectin and its 29-kD fragment to S pyogenes was not inhibited by Gly-Arg-Gly-Asp-Ser. These differences in binding activities corresponded to the ability of fibronectin to mediate the adherence of the streptococci to the epithelial cells: fibronectin strongly inhibited the adherence of S pyogenes and S equi to the epithelial cells, but only weakly inhibited that of S dysgalactiae.