Immunology of experimental staphylococcal mastitis in mice

Anti-staphylococcal serum, inoculated intramammarily, significantly reduced the number of bacteria recovered from mammary glands in which staphylococcal mastitis had been induced. In addition, there was an alleviation of some of the pathological clinical signs following the immune-serum treatment. Intramammary inoculated normal serum and systemically inoculated immune serum failed to elicit any protection.

The role of cell-mediated immunity in relation to staphylococcal mastitis was not clearly established.     

Bacterial adherence in mastitis caused by Escherichia coli.

The possible role of bacterial adherence in the pathogenesis of experimental mastitis in the mouse was examined with four strains of Escherichia coli. Two of these strains had a known adhesion antigen (K88) and two did not. The K88 antigen did not play a significant role in the virulence or infectivity of E. coli either in the murine or bovine mammary gland. Two E. coli strains, W1 (K88+) and J2 (K88-) were virulent in the mouse but did not adhere to epithelial cells. Both these strains produced clinical mastitis in the cow. A third strain, D282 (K88-), produced mild disease in the mouse but was avirulent in the cow. The fourth strain, 233/ID (K88+), was avirulent in both the mouse and the cow. Strains D282 and 233/1D were killed rapidly by bovine serum whilst J2 and W1 were more resistant. All strains were more sensitive than the control resistant strain E. coli P4, which is known to be highly virulent for the lactating udder.     

Recombinant bovine soluble CD14 reduces severity of experimental Escherichia coli mastitis in mice

Endotoxin, or lipopolysaccharide (LPS), is responsible for pathogenesis of infections induced by Gram-negative bacteria, such as E. coli. The cellular response to LPS is modulated by interactions among LPS, LPS-binding protein (LBP) and CD14. Accumulated evidence shows that the soluble form of CD14 (sCD14) competes with membrane-bound CD14 (mCD14) for LPS and plays a pivotal role in regulating bacterial infection and septic shock caused by Gram-negative bacteria. Recombinant bovine sCD14 (rbosCD14) was produced by transfected insect sf/9 cells and its biological function was evaluated in mice. Eighty-one 8-week old BALB/cj female mice were randomly assigned to two groups, and injected intraperitoneally with either LPS (8 microg/g of body weight, n = 41) or LPS plus rbosCD14 (6.8 microg/g of body weight, n = 40). Survival rate at 24 h after injection for mice injected with either LPS or LPS plus rbosCD14 was 30 and 72%, respectively (P < 0.01). At 48 h survival rate was 7 and 37%, respectively (P < 0.01). To investigate the protective effect of rbosCD14 on experimentally induced mastitis in mice, two abdominal contralateral mammary glands of 7 lactating BALB/cj mice were injected through the teat canal with 10-20 colony-forming units (CFU) of Escherichia coli. One gland simultaneously received rbosCD14 (6 microg) and the other saline. At 24 h after challenge, glands that received rbosCD14 had less swelling and hemorrhaging, significantly lower bacterial counts (P < 0.05) and lower concentrations of TNF-alpha (P < 0.05). Results indicate that rbosCD14 is biologically functional and reduces mortality in mice from endotoxin shock and severity of intramammary infection by E. coli.     

Mycoplasmosis: experimental mastitis. Demonstration of antibody in milk

According to several authors specific antibodies can be demonstrated in serum of cows with mycoplasma mastitis. For practical reasons, and for pathogenetic studies, it would be of interest if presence of antibody in milk could be established. To elucidate this and to examine the pathogenicity of the type strain, PG 11, of M. bovigenitalium, the experiment described below was conducted.     

细菌黏附研究进展

细菌黏附(bacterial adherence)是一种普遍存在和十分重要的生物现象,也是细菌寄生活动的基础和前提条件.细菌黏附是一个非常复杂的细菌与宿主细胞相互作用的过程.细菌表面的黏附素与宿主细胞表面的相应受体结合之后,细菌便能牢固地黏附于宿主细胞上,并在该局部组织定居,大量繁殖,产生毒素或侵入深部,损伤或破坏组织,引起疾病或造成隐性感染.细菌的黏附作用需有许多物质参与,但最基本的物质是黏附素和受体.     

A freeze-etching study on experimental murine mastitis.

Streptococcal mastitis was produced experimentally in mice inoculated by the intramammary route; freeze-etched preparations from the affected mammary glands were studied by electron microscopy. The inoculated cocci were seen free in the acinar lumen, within luminal phagocytes and within cells of the epithelium. No significant pathological changes were noted in the junctional complexes between secretory epithelial cells. The results were comparable to those obtained by ultrathin sectioning and indicated that, while cocci can transfer from the acinar lumen into the substance of the epithelium and towards a subepithelial location, the junctional complexes between epithelial cells present a potential barrier to movement through the intercellular spaces.     

Predisposing factors for bacterial mastitis in ewes

Objective of this review paper is the appraisal of predisposing factors for bacterial mastitis in ewes. Factors that predispose ewes to mastitis can be classified into non‐animal‐related factors (environmental and climatological factors, housing, nutrition, milking practices) and animal‐related factors (anatomic, genetic, litter size—ethological factors, number and stage of lactation period, health problems). There are clear management—environment—animal interactions in mastitis development in ewes, which underline its multifactorial nature. Research and studies regarding risk factors are important, in order to develop strategies for their elimination, control or correction. Control measures for bacterial mastitis, which attempt to eliminate predisposing factors for infection, can thus be developed and implemented.     

The effect of incorporation of cloxacillin in liposomes on treatment of experimental staphylococcal mastitis in mice

The effect of incorporation of cloxacillin in liposomes on the treatment of staphylococcal mastitis was assessed bacteriologically 18 h after treatment of experimental infections in mice caused by two strains of Staphylococcus aureus. Intramammary treatments were cloxacillin incorporated in liposomes, cloxacillin in combination with liposomes, empty liposomes, cloxacillin in saline and saline alone. In none of the experiments did entrapment of cloxacillin within liposomes enhance its antibacterial effects. Electron microscopic studies demonstrated the presence of liposomes in neutrophils which also contained staphylococci. The results support the hypothesis that intracellular staphylococci are metabolically dormant and therefore not susceptible to the action of inhibitors of cell wall synthesis such as cloxacillin.     

金黄色葡萄球菌诱发小鼠乳腺炎模型的建立

近年来,国内外开始选用小鼠建立乳腺炎病理模型,成功解决了使用奶牛或奶山羊等动物成本太高或操作不便以及无法进行标准化管理的问题[1-2].但其在诱发小鼠乳腺炎的操作中对小鼠乳腺组织易造成人为的损伤,不能真实模拟奶牛乳腺炎的自然发病机制[3].本试验用乳腺炎病牛奶样中分离到的金黄色葡萄球菌人工诱发小鼠乳腺炎,成功建立了小鼠乳腺炎模型,为研究乳腺的免疫学、病理学、药动学、药效学和临床治疗等提供了试验平台,报道如下.     

Vaccination against experimental staphylococcal mastitis in ewes

Experiments were carried out in ewes using a new vaccine developed for the prevention of mastitis caused by Staphylococcus aureus. The vaccine comprised three major components: (i) killed S aureus cells which had been cultured to induce synthesis of pseudocapsule; (ii) toxoided staphylococcal beta haemolysin and (iii) the adjuvant dextran sulphate. Ewes systemically vaccinated twice during pregnancy developed significantly elevated circulating levels of IgG1 and IgG2 anti-pseudocapsule antibody, as well as increased serum titres of anti-beta haemolysin. Five different strains of S aureus were used to challenge both vaccinated and control ewes by the intramammary route during the ensuing lactation. The incidence of acute gangrenous mastitis and nonacute, clinical mastitis was significantly lower in vaccinated than in control groups after challenge with each strain. Vaccinated ewes produced significantly more milk than control ewes after challenge with four of the five strains of S aureus.     

Absence of encapsulation in strains of Staphylococcus aureus isolated from bovine mastitis

Thirty of 104 strains of Staphylococcus aureus isolated from clinical cases of bovine mastitis in England grew as diffuse colonies in serum soft agar (SSA), 45 grew as mixed diffuse and compact colonies and 29 yielded compact colonies only. The compact strains grew as diffuse colonies in SSA after one passage in the mammary gland of mice. However, none of the strains had an unstained halo when examined by the India ink technique and there was a 99.99 per cent reduction in the viable numbers of the bacteria in 30 representative strains 24 hours after inoculation into the peritoneal cavity of mice. By contrast the truly encapsulated strain M had an unstained halo by the India ink technique and resisted phagocytic killing in the peritoneal cavity. It is concluded that these strains from cases of mastitis are not encapsulated and that growth as diffuse colonies in SSA is not a reliable test of encapsulation.     

Accuracy of clinicians in predicting the bacterial cause of clinical bovine mastitis

We examined the ability of clinicians to predict the causative organism of bovine mastitis in our practice. We obtained 118 milk culture results from 112 mastitic cows and compared the culture results to the predictions of clinicians at the time of milk sample collection. Sixty of 118 culture results were accurately predicted. The positive predictive value for coliform mastitis was 42% and the negative predictive value was 79% in a study population with a 31% prevalence of coliform mastitis. The proportion of coliform positive cultures accurately predicted was significantly greater than that expected by chance (p < 0.02).     

Vaccination against experimental staphylococcal mastitis in dairy heifers

Vaccination-challenge experiments were carried out with dairy heifers using new, killed cell-toxoid-adjuvant Staphylococcus aureus vaccines. The organisms in the vaccines were cultured under conditions which simulated in vivo growth and induced expression of a pseudocapsule. Dextran sulphate which promotes synthesis of IgG2 antibody was included in the vaccines as the primary adjuvant. Vaccinated heifers developed very high levels of both IgG1 and IgG2 anti-pseudocapsule antibody in serum, however, titres of neutralising antibody against toxoided haemolysins were generally low. Vaccinated and unvaccinated control heifers were challenged by intramammary infusion of three virulent strains of S aureus in four experiments. Vaccinated heifers were more resistant to clinical mastitis following challenge than were controls, and the vaccinates had significantly greater milk production than controls following challenge. The most promising vaccine had dextran sulphate combined with mineral oil as the adjuvant injected intramuscularly.