Prostatic abscess by Staphylococcus aureus in a diabetic patient

Prostatic abscess is uncommon and difficult to diagnose because the clinical presentation may mimic symptoms of lower urinary tract infection. We report here a case of prostatic abscess in a 50-year-old known diabetic male patient, who presented with urinary retention. Clinical diagnosis was done by clinical presentation and ultrasonography. The causative agents i.e., Staphylococcus aureus was isolated from the aspirate and the patient responded to intravenous Ciprofloxacin therapy. No other surgical intervention was required to treat the patient.     

Protective immunization against Staphylococcus aureus infection in a novel experimental wound model in mice

A novel murine experimental wound infection model was used to assess the efficacy of multi‐component immunization against Staphylococcus aureus infection. Necrotic lesions were induced in mice with venom from Bothrops asper and infected with a low inoculum, 1 × 10² CFU. The wound infection model therefore more resembles a clinical case of S. aureus infection compared with conventional infection models where far more bacteria are required. Before infection, mice were immunized with four recombinant S.aureus proteins expressed from Escherichia coli: (i) domains 1–3 of Extracellular adherence protein (Eap), (ii) Efb – D (fusion protein combining Extracellular fibrinogen binding protein (Efb) and a fibronectin binding domain (D) of the fibronectin binding protein (FnBP) and (iii) clumping factor A (ClfA). In the immunized group, lower bacterial colonization, undisturbed crust formation and significantly faster wound healing were found compared with the unimmunized control group. Efb and Eap have previously been found to impair wound healing and neutralization of these proteins by antibodies restores a more natural wound healing process. This effect is further also enhanced by the proposed opsonic activity of antibodies against ClfA and FnBP.     

耐甲氧西林金黄葡萄球菌小鼠菌血症感染模型的建立与评估

目的 本研究采用临床分离鉴定的ST-239型耐甲氧西林金黄葡萄球菌(MRSA)感染BALB/c小鼠,建立菌血症感染模型.方法 从小鼠的临床症状、生存曲线和主要组织器官的病理学变化进行了时相性监测,并用该模型验证万古霉素对小鼠菌血症的治疗效果.结果 ST-239型MRSA感染的小鼠血液中细菌含量较高,小鼠炎症反应严重并伴有心、肺等多组织器官的损伤;对小鼠使用万古霉素后可显著降低血液中的细菌量,动物存活率明显提高,主要组织器官的病理学症状减轻.结论 该模型的建立将为进一步研究临床分离的MRSA的病原特性、发病机制、治疗方法等提供可靠的实验动物模型.     

Capsule-negative Staphylococcus aureus induces chronic experimental mastitis in mice

Staphylococcus aureus capsular polysaccharides (CP) have been shown to enhance staphylococcal virulence in numerous animal models of infection. Although serotype 5 CP (CP5) and CP8 predominate among S. aureus isolates from humans, most staphylococcal isolates from bovines with mastitis in Argentina are capsule negative. This study was designed to evaluate the effects of CP5 and CP8 expression on the pathogenesis of experimental murine mastitis. Lactating mice were challenged by the intramammary route with one of three isogenic S. aureus strains producing CP5, CP8, or no capsule. Significantly greater numbers of acapsular mutant cells were recovered from the infected glands 12 days after bacterial challenge compared with the encapsulated strains. Histopathological analyses revealed greater polymorphonuclear and mononuclear leukocyte infiltration and congestion in the mammary glands of mice infected with the encapsulated strains compared with the acapsular mutant, and the serotype 5 strain elicited more inflammation than the serotype 8 strain. In vitro experiments revealed that the acapsular S. aureus strain was internalized by MAC-T bovine epithelial cells in significantly greater numbers than the CP5- or CP8-producing strain. Taken together, the results suggest that S. aureus lacking a capsule was able to persist in the murine mammary gland, whereas encapsulated strains elicited more inflammation and were eliminated faster. Loss of CP5 or CP8 expression may enhance the persistence of staphylococci in the mammary glands of chronically infected hosts.     

DS-Nh as an experimental model of atopic dermatitis induced by Staphylococcus aureus producing staphylococcal enterotoxin C

DS-Nh mice raised under conventional conditions spontaneously develop dermatitis similar to human atopic dermatitis (AD), which is associated with staphylococcal infection. In the present study, we show that Staphylococcus aureus producing staphylococcus exotoxin C (SEC) was recovered from the culture of the skin lesions of DS-Nh mice with AD-like dermatitis and that the serum levels of anti-SEC antibodies from these mice were elevated. We describe here how to promote experimental AD by epicutaneous injection with SEC-producing S. aureus to DS-Nh mice. In order to assess the role of SEC in the pathogenesis of AD, the mitogenic activity, TCRBV repertoire analysis and the production of IL-4 and IFN-gamma from spleen mononuclear cells (MNC) from DS-Nh stimulated by SEC were compared with those due to SEA, SEB and TSST. The weakest was the mitogenic activity of SEC, and higher IL-4 responses and lower IFN-gamma responses to SEC showed correlation with TCRBV8S2-positive T cells, which were selectively stimulated by SEC. We also demonstrate that SEC-producing S. aureus was able to survive in DS-Nh after intradermal injection. These results suggest a possible role for SEC in the pathogenesis of AD through host-S. aureus relationships.     

Regulation of Staphylococcus aureus type 5 capsular polysaccharides by agr and sarA in vitro and in an experimental endocarditis model

The expression of antiphagocytic polysaccharide capsules is an important pathogenetic step in establishing Staphylococcus aureus infections. Using a green fluorescent protein reporter gene (gfp) system, we examined the expression and genetic regulation of the cap5 promoter (capsular polysaccharide 5 genes) by two major global regulators of S. aureus (agr and sarA) in vitro and in a rabbit endocarditis model. In vitro, cap5 expression substantially increased during the post-exponential phase in parental, as well assarA mutant constructs. However, cap5 expression was greatly reduced in agr and agr/sarA double mutants. In the endocarditis model, the extent of cap5 expression in vegetations infected with the parental strain was substantially higher than that observed with the agr/sarA double mutants (P<0.05). Similar trends were noted in renal, but not splenic abscesses. Collectively, these data suggest that agr positively regulates cap5 expression both in vitro and in vivo, while the contribution of sarA to cap5 regulation, although modest, is readily discerned in vivo in agr minus background. In addition, the regulation ofcap5 expression by these global regulators may vary in distinct anatomic niches in vivo.     

Gastrointestinal colonization by methicillin-resistant Staphylococcus aureus in immunosuppressed mice.

ICR mice were inoculated intranasally with methicillin-resistant Staphylococcus aureus (MRSA) N133, and the inoculated MRSA was quantitatively recovered from the ceca and feces. The viable counts of the MRSA recovered from ceca correlated well with those from feces. Some mice eliminated MRSA from the cecum by 14 days after inoculation. Intraperitoneal administration of cyclophosphamide at a dose of 200 mg/kg 3 days before inoculation inhibited the elimination of the MRSA from both ceca and feces. All mice treated with cyclophosphamide excreted more than 10(4) CFU of the MRSA per g of feces for at least 70 days, indicating persistent colonization of the MRSA in the gastrointestinal tract. Some beta-lactam antibiotics decreased the colonization level, but others did not. The colonization level was suggested to depend on the antibacterial activity of the antibiotic against the MRSA and the degree of disturbance of intestinal flora by the antibiotic.     

Synergistic effect of linezolid with fosfomycin against Staphylococcus aureus in vitro and in an experimental Galleria mellonella model

Background/purposesTreatment of Staphylococcus aureus infections is challenging owing to widespread multidrug resistance. There is now considerable interest in the potential of combination therapies. Although linezolid/fosfomycin combination appears to be a promising treatment option based on in vitro data, further preclinical work is needed. In this study, the Galleria mellonella system was employed to study the in vivo efficacy of this combination in order to determine whether it should be explored further for the treatment of S. aureus infections.MethodsThe antimicrobial activity of linezolid and fosfomycin alone and in combination was assessed versus four S. aureus. Synergy studies were performed using the microtitre plate chequerboard assay and time-kill methodology. The in vivo activity of linezolid/fosfomycin combination was assessed using a G. mellonella larvae model.ResultsThe combination of linezolid and fosfomycin was synergistic and bacteriostatic against four tested strains. Treatment of G. mellonella larvae infected with lethal doses of S. aureus resulted in significantly enhanced survival rates when low-dose of combination has no significant differences with high-dose combination (P > 0.05), G. mellonella hemolymph burden of S. aureus suggest that combination therapy with rapid and sustained bacteriostatic activity compared monotherapy.ConclusionThis work indicated that linezolid combination with fosfomycin has synergistic effect against S. aureus in vitro and in an experimental G. mellonella model, and it suggests that high-dose of linezolid and fosfomycin may not necessary.     

Rapid differentiation of methicillin-resistant Staphylococcus aureus and methicillin-susceptible Staphylococcus aureus from blood cultures by use of a direct cefoxitin disk diffusion test

A total of 276 blood culture bottles with Staphylococcus aureus were tested by direct cefoxitin disk diffusion testing; 105 (38.1%) had zone sizes of ≤17 mm (all 105 had methicillin-resistant S. aureus [MRSA]), 18 (6.5%) had zone sizes that measured 18 mm (17 had MRSA and 1 had methicillin-susceptible S. aureus [MSSA]), 8 (2.9%) had zone sizes that measured 19 mm (6 had MRSA and 2 had MSSA), 8 (2.9%) had zone sizes that measured 20 mm (6 had MRSA and 2 had MSSA), and 137 (49.6%) had zone sizes of ≥21 mm (all 137 had MSSA). Detection of MRSA/MSSA in blood cultures could be reported 10 to 24 h earlier for 88% of cultures with total accuracy.     

Role of monocytes in experimental Staphylococcus aureus endocarditis

In the pathogenesis of bacterial endocarditis (BE), the clotting system plays a cardinal role in the formation and maintenance of the endocardial vegetations. The extrinsic pathway is involved in the activation of the coagulation pathway with tissue factor (TF) as the key protein. Staphylococcus aureus is a frequently isolated bacterium from patients with BE. We therefore investigated whether S. aureus can induce TF activity (TFA) on fibrin-adherent monocytes, used as an in vitro model of BE. We also assessed in vivo in rabbits with catheter induced vegetations, the effect of S. aureus infection on vegetational TFA. In vitro experiments showed that adherent S. aureus induced TFA on fibrin-adherent monocytes which was optimal at a bacterium/monocyte ratio of 1 to 1. Monocyte damage occurred when this ratio exceeded 4 to 1 (visually) or 6 to 1 (propidium iodide influx) Consequently, TFA decreased. In vivo S. aureus led to very high bacterial numbers in the vegetations and a significant increase of their weight. However, TFA of infected vegetations was the same as of sterile ones. This may be due to the high bacteria to monocyte ratio as well as bacterium-induced monocyte damage. Teicoplanin treatment of infected rabbits reduced bacterial numbers in the blood and in the vegetations. Two-day treatment resulted in an increase of vegetational TFA, but after four-day treatment vegetational TFA dropped, most probably due to a suboptimal bacterium/monocyte ratio. S. aureus endocarditis in etoposide (Vepesid)-treated rabbits, leading to a selective monocytopenia, caused a rapid death of the animals. In these rabbits no vegetations were found at all. We conclude that, like Streptococcus sanguis and Staphylococcus epidermidis, S. aureus is able to induce TFA in fibrin-adherent blood monocytes. In addition, monocytes have a protective effect during the course of S. aureus endocarditis.     

A method for creating an experimental model of abscess

A technique of 7-day formation of an intramuscular abscess which is clinically and morphologically similar to that frequently encountered in clinical practice has been tried on 39 rabbits. The standard model proposed involves the following stages: trauma of the spinal erector 2.5-3 cm to the right or to the left from the midline between the second and fourth lumbar vertebra which is produced by a lumbar puncture under either anesthesia with a pulp extractor brought into the muscle up to 1.5 cm and rotating at an angle of 45 degrees; 24 h after the puncture the skin at its site is to be dissected involving subcutaneous fat, superficial and lumbar fasciae, the incision being 1.4-1.6 cm long and parallel to the body axis. The operation is terminated with an introduction of ribbon gauze (10-12 mm in diameter) previously saturated with 24 h suspension culture of staphylococcus (10 ml, concentration of the microorganisms 1.5.10(6] into a pocket made by the clamp tip in the muscle's depth (1.3-1.6 cm) with the help of the handles which produced the opening of 1.2 cm wide. The endpoint is the wound tight layer-by-layer suture.     

Detection of encapsulation in Staphylococcus aureus by use of antiserum agar

We examined an antiserum agar method to study its reliability in screening Staphylococcus aureus strains for capsule production. The encapsulated S. aureus Smith diffuse strain was compared with its nonencapsulated variant, Smith compact, in CCY medium containing 0.5% NaCl and 5.0% Smith diffuse rabbit antiserum. A halo was visible surrounding colonies of the Smith diffuse strain but not the Smith compact strain. On this same medium, the protein A-producing Cowan I strain possessed a halo that was visible on photographs. Single high-salt medium is known to inhibit protein A production, halo formation by the strains was also compared in 7.5% NaCl medium. The halo surrounding the Cowan I strain was not present when the salt content of the medium was increased. In contrast, the halo surrounding the Smith diffuse strain persisted in the 7.5% NaCl medium. By use of this medium, the antiserum agar technique may be valuable for the identification of encapsulated staphylococci without appreciable interference from protein A.     

Diminished virulence of an alpha-toxin mutant of Staphylococcus aureus in experimental brain abscesses

Staphylococcus aureus is one of the major etiologic agents of brain abscesses in humans, occasionally leading to focal neurological deficits and even death. The objective of the present study was to identify key virulence determinants contributing to the pathogenesis of S. aureus in the brain using a murine brain abscess model. The importance of virulence factor production in disease development was demonstrated by the inability of heat-inactivated S. aureus to induce proinflammatory cytokine or chemokine expression or brain abscess formation in vivo. To directly address the contribution of virulence determinants in brain abscess development, the abilities of S. aureus strains with mutations in the global regulatory loci sarA and agr were examined. An S. aureus sarA agr double mutant exhibited reduced virulence in vivo, as demonstrated by attenuated proinflammatory cytokine and chemokine expression and bacterial replication. Subsequent studies focused on the expression of factors that are altered in the sarA agr double mutant. Evaluation of an alpha-toxin mutant revealed a phenotype similar to that of the sarA agr mutant in vivo, as evidenced by lower bacterial burdens and attenuation of cytokine and chemokine expression in the brain. This suggested that alpha-toxin is a central virulence determinant in brain abscess development. Another virulence mechanism utilized by staphylococci is intracellular survival. Cells recovered from brain abscesses were shown to harbor S. aureus intracellularly, providing a means by which the organism may establish chronic infections in the brain. Together, these data identify alpha-toxin as a key virulence determinant for the survival of S. aureus in the brain.     

The use of sequential organ failure assessment parameters in an awake porcine model of severe Staphylococcus aureus sepsis

The human sequential organ failure assessment (SOFA) scoring system is used worldwide in intensive care units for assessing the extent of organ dysfunction/failure in patients with severe sepsis. An increasing number of septic cases are caused by Gram‐positive bacteria as Staphylococcus aureus. The aim of the current study was to apply the human SOFA parameters in an awake, porcine model of severe S. aureus sepsis. Five pigs were inoculated intravenously with S. aureus and two control animals were sham‐inoculated. Extensive clinical monitoring and sequential blood sampling was obtained and analysed for SOFA parameters. Dysfunction/failure was observed in the respiratory, haemostatic and hepatic system of all infected animals, together with initial cardiovascular dysfunction. The pulmonary system was the first to fail clinically, which corresponds with similar human findings, whereas the liver was affected earlier in pigs compared to humans. The use of human SOFA parameters was valuable in identifying dysfunctional/failing organs and showed consistency between this porcine model and human severe sepsis. Applying SOFA parameters in this model increased the relevance for comparison to clinical methods of evaluating human severe sepsis. Changes in SOFA parameters may in future porcine studies serve as a target for monitoring the effect of therapeutic intervention.     

Correlation of histopathologic and bacteriologic changes with cytokine expression in an experimental murine model of bacteremic Staphylococcus aureus infection.

Staphylococcus aureus infections are often life threatening. Relatively little is known about the host response to these infections, in particular, the role played by cytokines. We established a mouse model of bacteremic S. aureus infection to correlate bacteriologic findings and pathologic changes with cytokine gene expression. Bacterial density in blood and tissue was highest at 1 h and minimal by 48 h. Despite the rapid clearance of bacteria, pathologic abnormalities and inflammatory cytokines were detected after clearance of the bacteria. The number of infiltrating inflammatory cells, as well as the size of inflammatory foci, increased with time. Interstitial accumulation of inflammatory cells and tissue damage, such as microabscesses, edema, and necrosis progressed following clearance of bacteria from the tissues. Levels of tumor necrosis factor and interleukin-1 protein in serum were detectable at 1 h and peaked at 4 h. Interleukin-6 protein expression showed different kinetics, with low levels detected at 1 h and increasing levels at 72 h postinfection. Tumor necrosis factor and the interleukins were expressed in inflammatory and noninflammatory cells in lung, liver, and heart tissues. Leukocytes in the infected tissues were highly reactive with antibodies to the three cytokines, suggesting that activated leukocytes are a major source of inflammatory cytokines after staphylococcal infection. Expression of interleukin-1 and interleukin-6 in tissue-specific cells and endothelial cells was also detected in infected tissues, indicating that cells other than leukocytes contribute to the elevated cytokine levels in this model. Once initiated, expression of inflammatory cytokines contributes to the pathogenesis of S. aureus disease.     

Surface proteins of Staphylococcus aureus play an important role in experimental skin infection

Staphylococcus aureus is the most common cause of skin infections that range from mild diseases up to life‐threatening conditions. Mechanisms of S. aureus virulence in those infections remain poorly studied. To investigate the impact of S. aureus surface proteins on skin infection, we used mouse models of skin abscess formation and skin necrosis, induced by a subcutaneous injection of bacteria. In the skin abscess model, a sortase‐deficient S. aureus strain lacking all of its cell‐wall anchored proteins was less virulent than its wild‐type strain. Also, strains specifically lacking protein A, fibronecting binding proteins, clumping factor A or surface protein SasF were impaired in their virulence. When a model of dermonecrosis was studied, the S. aureus surface proteins could not be shown to be involved. In summary, surface proteins play an important role in virulence of S. aureus skin abscess infections, but not in formation of skin necrosis.     

Community-acquired methicillin-resistant Staphylococcus aureus liver abscess in a patient with end-stage renal disease.

Community-acquired methicillin-resistant Staphylococcus aureus (MRSA) infections are an emerging problem. Although most liver abscesses are caused by Escherichia coli and Klebsiella pneumoniae, S. aureus can occasionally be isolated as the pathogenic organism. Liver abscess caused by MRSA is rarely reported. Here, we report a case of liver abscess due to MRSA in a 34-year-old man with end-stage renal disease and a 13-year history of hemodialysis treatment.