Enhancement by Candida albicans of Staphylococcus aureus, Serratia marcescens, and Streptococcus faecalis in the establishment of infection in mice.

Candida albicans has been shown to stimulate infection in mice by a number of bacteria. Mice inoculated intraperitoneally with as little as 500 colony-forming units of Staphylococcus aureus along with a nonlethal dose of 10(8) colony-forming units of C. albicans (one-third the dose causing 50% mortality in 7 days) developed widespread staphylococcal infection. S. aureus injected alone at that or considerably higher doses did not establish infection. Phage typing methods demonstrated that the staphylococcal infection was due to the bacterial organisms originally injected. A minimum dose of C. albicans between 1.1 X 10(5) and 1.1 X 10(7) colony-forming units was necessary for the amplification of virulence of S. aureus to take place. Serratia marcescens and Streptococcus faecalis inoculated intraperitoneally at small nonlethal doses could also be recovered from blood and tissues 5 days after animals were dually injected with C. albicans but not from animals which were inoculated with the same amounts of these bacteria alone.     

Collagen binding to Staphylococcus aureus.

Staphylococcus aureus can bind soluble collagen in a specific, saturable manner. We have previously shown that some variability exists in the degree of collagen binding between different strains of heat-killed, formaldehyde-fixed S. aureus which are commercially available as immunologic reagents. The present study demonstrates that live S. aureus of the Cowan 1 strain binds amounts of collagen per organism equivalent to those demonstrated previously in heat-killed, formaldehyde-fixed bacteria but has an affinity over 100 times greater, with Kd values of 9.7 X 10(-11) M and 4.3 X 10(-8) M for live and heat-killed organisms, respectively. Studies were also carried out with S. aureus killed by ionizing radiation, since this method of killing the organism seemed less likely to alter the binding moieties on the surface than did heat killing. Bacteria killed by exposure to gamma radiation bound collagen in a manner essentially indistinguishable from that of live organisms. Binding of collagen to irradiated cells of the Cowan 1 strain was rapid, with equilibrium reached by 30 min at 22 degrees C, and was fully reversible. The binding was not inhibited by fibronectin, fibrinogen, C1q, or immunoglobulin G, suggesting a binding site for collagen distinct from those for these proteins. Collagen binding was virtually eliminated in trypsin-treated organisms, indicating that the binding site has a protein component. Of four strains examined, Cowan 1 and S. aureus ATCC 25923 showed saturable, specific binding, while strains Woods and S4 showed a complete lack of binding. These results suggest that some strains of S. aureus contain high-affinity binding sites for collagen. While the number of binding sites per bacterium varied sixfold in the two collagen-binding strains, the apparent affinity was similar. The ability of S. aureus to bind collagen with high affinity may provide a mechanism for bacterial adhesion to host tissue and thereby play a role in the invasive characteristics of this organism.     

Experimental Candida albicans, Staphylococcus aureus, and Streptococcus faecalis pyelonephritis in diabetic rats.

Pyelonephritis was studied after an intravenous injection of Candida albicans, Staphylococcus aureus, or enterococcus in alloxan-diabetic rats and in water-diuresing or non-diuresing nondiabetic rats. The renal microbial populations of C. albicans or S. aureus were found to be greater than 10(5) colony-forming units per g for up to 42 days in diabetic rats, whereas the kidneys tended to become sterile in nondiabetic rats. No significant difference was found in the course of enterococcal pyelonephritis in diabetic versus control rats. The difference in the 50% infective dose for each microorganism between diabetic and control rats was less than or equal to log10. Neither duration of diabetes nor weight loss contributed to the greater and more sustained renal populations of C. albicans and S. aureus in diabetic rats. The inflammatory reaction in kidneys infected with S. aureus or C. albicans was greater in diabetic rats. Fungus balls associated with ureteral obstruction and gross multiple renal abscesses occurred in diabetic, but not in nondiabetic, rats infected with Candida. Growth of C. albicans and S. aureus in vitro in urine from diabetic rats was significantly greater than it was in urine from control rats. Addition of water or glucose to the urine of non-diuresing, nondiabetic rats significantly increased in vitro growth of S. aureus and C. albicans. These studies demonstrate greater severity of infection in the diabetic kidney due to S. aureus and C. albicans, which can be partially explained by decreased inhibitory activity of urine for these organisms in diabetic rats.     

Antibodies to capsular polysaccharides are not protective against experimental Staphylococcus aureus endocarditis.

The protective efficacy of antibodies to the Staphylococcus aureus capsular polysaccharide was examined in a rat model of catheter-induced endocarditis. Capsular antibodies were induced either by active immunization with killed S. aureus or by passive immunization with hyperimmune rabbit antiserum to S. aureus. Control rats were injected with phosphate-buffered saline or passively immunized with normal rabbit serum or rabbit antiserum to a nonencapsulated strain. Animals with indwelling catheters were challenged intravenously with 5 x 10(4) to 4 x 10(6) CFU of the homologous S. aureus strain (capsular serotype 5 strain Reynolds or serotype 1 strain SA1 mucoid). Both immunized and control rats developed S. aureus endocarditis. The numbers of S. aureus cells recovered from the blood and aortic valve vegetations of immunized rats were similar to those of control rats, indicating that capsule-specific antibodies were not protective. To determine whether the presence of an indwelling catheter interfered with antibody-mediated protection against S. aureus endocarditis, catheters were removed 2 h after insertion in additional groups of rats. An inoculum of 10(8) CFU of strain Reynolds was needed to provoke endocarditis in rats catheterized for 2 h, compared with 5 x 10(4) CFU for rats with indwelling catheters. Passively transferred capsular antibodies were not protective since both immunized and nonimmunized animals developed endocarditis, and quantitative cultures of blood and valvular vegetations revealed no differences between immunized and control animals. The findings of this study indicate that antibodies to the capsular polysaccharide are not protective in the rat model of experimental S. aureus endocarditis.     

Vitamin A deficiency predisposes to Staphylococcus aureus infection.

We have investigated the consequences of vitamin A deficiency in a rat model of T-cell-dependent and superantigen-mediated Staphylococcus aureus arthritis. After intravenous inoculation of enterotoxin A-producing staphylococci, the vitamin-A-deficient rats showed a decreased weight gain compared with the paired fed controls despite equal food consumption. The control rats developed arthritis in the first few days after bacterial inoculation, with a peak frequency at day 5, and then gradually recovered; however, the frequency of arthritis 18 days after bacterial inoculation was 86% among the vitamin A-deficient rats and 44% among the control rats. During this period, 3 of 10 deficient rats and 1 of 10 control rats died. Further in vitro analysis revealed that T-cell responses to S. aureus were significantly higher in the vitamin A-deficient rats than in the control animals. In contrast, B-cell reactivity, measured as immunoglobulin levels, autoantibody levels, and specific antibacterial antibody levels in serum, did not differ between the groups. Interestingly, the innate host defense mechanisms against S. aureus were also profoundly affected by vitamin A deficiency. Thus, despite a larger number of circulating phagocytic cells in the vitamin-A-deficient group, the capacity to phagocytize and exert intracellular killing of S. aureus was significantly decreased in comparison with the control rats. Furthermore, serum from the vitamin A-deficient rats inoculated with Staphylococcus aureus displayed decreased complement lysis activity. Our results suggest that the increased susceptibility to S. aureus infection observed in the vitamin-A-deficient rats is due to a concerted action of antigen-specific T-cell hyperactivity, impaired function of the phagocytes, and decreased complement activity.     

Phagocytic killing of encapsulated and microencapsulated Staphylococcus aureus by human polymorphonuclear leukocytes.

Phagocytosis by human polymorphonuclear leukocytes (PMNs) is an important host defense against infections caused by Staphylococcus aureus. Using an in vitro assay, we compared the opsonic requirements for phagocytic killing of prototype strains of encapsulated (type 1) and microencapsulated (type 5 and type 8) S. aureus by human PMNs. More than 85% of broth-grown, logarithmic-phase type 5 and 8 S. aureus organisms were killed by PMNs incubated with fresh normal human, rabbit, or guinea pig serum with complement activity. Under similar conditions, the highly encapsulated type 1 strain was not killed. Both encapsulated and microencapsulated strains were opsonized for phagocytosis by heat-inactivated serum raised in rabbits to killed bacteria. Opsonization by homologous serum was required for phagocytosis of the type 1 strain. In contrast, microencapsulated type 5 and 8 S. aureus organisms were killed by heat-inactivated rabbit serum raised to type 5, type 8, or nonencapsulated isolates; this result suggested that antibodies to the capsule or to cell wall components other than the capsule could opsonize these organisms for phagocytosis. The specificity of the assay was confirmed with capsule type 5-specific monoclonal antibodies, which were opsonic only for the type 5 S. aureus isolate. These studies indicate that, unlike the highly encapsulated type 1 strain, broth-grown microencapsulated S. aureus strains do not resist opsonophagocytic killing in vitro by normal serum.     

Reassessing the role of Staphylococcus aureus clumping factor and fibronectin-binding protein by expression in Lactococcus lactis.

Since Staphylococcus aureus expresses multiple pathogenic factors, studying their individual roles in single-gene-knockout mutants is difficult. To circumvent this problem, S. aureus clumping factor A (clfA) and fibronectin-binding protein A (fnbA) genes were constitutively expressed in poorly pathogenic Lactococcus lactis using the recently described pOri23 vector. The recombinant organisms were tested in vitro for their adherence to immobilized fibrinogen and fibronectin and in vivo for their ability to infect rats with catheter-induced aortic vegetations. In vitro, both clfA and fnbA increased the adherence of lactococci to their specific ligands to a similar extent as the S. aureus gene donor. In vivo, the minimum inoculum size producing endocarditis in > or =80% of the rats (80% infective dose [ID80]) with the parent lactococcus was > or =10(7) CFU. In contrast, clfA-expressing and fnbA-expressing lactococci required only 10(5) CFU to infect the majority of the animals (P < 0.00005). This was comparable to the infectivities of classical endocarditis pathogens such as S. aureus and streptococci (ID80 = 10(4) to 10(5) CFU) in this model. The results confirmed the role of clfA in endovascular infection, but with a much higher degree of confidence than with single-gene-inactivated staphylococci. Moreover, they identified fnbA as a critical virulence factor of equivalent importance. This was in contrast to previous studies that produced controversial results regarding this very determinant. Taken together, the present observations suggest that if antiadhesin therapy were to be developed, at least both of the clfA and fnbA products should be blocked for the therapy to be effective.     

Impaired lung defenses against Staphylococcus aureus in mice with hereditary deficiency of the fifth component of complement.

Mice of the C5-deficient DBA/2J and B10.D2/oSnJ inbred strains were aerosolized with Staphylococcus aureus, and the pulmonary clearance of bacteria was determined 4 h later. Both C5-deficient strains had a significantly decreased lung clearance of S. aureus compared with their genetically closest C5-sufficient relatives, DBA/1J and B10.D2/nSnJ strains, respectively. Serum hemolytic activity and pulmonary clearance of S. aureus were also investigated in F1 and F2 progenies (DBA/1J X DBA/2J and DBA/2J X B10.D2/oSnJ). Serum hemolytic activity was present in all F1 (DBA/1J X DBA/2J) mice, and their pulmonary clearance of S. aureus was no different from that of the C5-sufficient parents (DBA/1J). The absence of serum hemolytic activity (absence of C5) in all mice from F1 and F2 (DBA/2J X B10.D2/oSnJ) and 20% of the F2 (DBA/1J X DBA/2J) was related to a decreased lung clearance of S. aureus. These results are consistent with an autosomal recessive pattern of heredity for the murine abnormality in pulmonary clearance of S. aureus.     

Disbalance of calcium regulation-related genes in broiler hearts induced by selenium deficiency

Dietary selenium (Se) deficiency may influence the calcium (Ca) homeostasis in broilers. Our objective was to investigate the effects of Se deficiency on Ca regulation-related genes in broiler hearts. In the present study, 1-day-old broilers were fed either a commercial diet (as control group) with 0.15?mg/kg Se or a Se-deficient diet (as L group) with 0.033?mg/kg Se for 35 days. We examined the mRNA expression levels of 15 Ca regulation-related genes (ITPR 1, ITPR 2, ITPR3, RyR2, RyR3, SERCA1s, SLC8A1, PMCA1, CACNA1S, TRPC1, TRPC3, stromal interacting molecule 1, ORAI1, calmodulin (CaLM) and calreticulin (CRT) in broiler hearts. Then, Kyoto Encyclopedia of Genes and Genomes analysis, protein–protein interactions (PPI) analysis and correlation analysis were performed to analyse the relationships between these genes. The results showed that the mRNA expression levels of ITPR 1, ITPR 2, RyR2, RyR3, SERCA1s, SLC8A1, PMCA1, CACNA1S, CaLM and CRT were generally decreased by Se deficiency, while mRNA expression levels of TRPC1, TRPC3, stromal interacting molecule 1, ORAI1 and ITPR3 were increased by Se deficiency. Kyoto Encyclopedia of Genes and Genomes and PPI analysis showed that these Ca regulation-related genes are involved in the Ca signalling pathway and a total of 15 PPIs with a combined score of >0.4 were obtained. In conclusion, the results demonstrated that Se deficiency might cause heart injury via modulating the Ca-related pathway genes, and then induce Ca²⁺ overload in the heart of broilers.     

Involvement of staphylococcal protein A and cytoskeletal actin in Staphylococcus aureus invasion of cultured human oral epithelial cells

Following the coincidental discovery that beta-actin isolated from renal epithelial cells was precipitated by staphylococcal protein A (SPA), the possibility that SPA and cytoskeletal actin filaments may be involved in Staphylococcus aureus infection of epithelial cells was considered. Therefore, to clarify the potential role of SPA and actin filaments in S. aureus infection, the invasion efficiency of S. aureus was determined quantitatively by measuring the number of cfu of viable organisms recovered from cultured KB cells. S. aureus invasion was found to be time dependent (0-60 min) and increased linearly when increasing numbers of bacteria were added (10(4)-10(6) cfu/ml). However, significant variation in the level of invasion was noted in protein A-deficient S. aureus Wood 46. Cytochalasin B inhibited the invasion efficiency of S. aureus in a dose-dependent manner. The present study suggests that interaction of staphylococcal protein A and cytoskeletal actin filaments is involved in the S. aureus invasion of cultured KB cells, and this process may contribute, in part, to the intracellular movement, cell-to-cell spread and dissemination of S. aureus within human oral epithelial cells in vivo.     

Complement activation influences Staphylococcus aureus adherence to endothelial cells

The ability of Staphylococcus aureus to adhere to endothelial cells (EC) is a critical step in the development of metastatic infection. The role of complement in S. aureus binding to EC remains uninvestigated. Log-phase S. aureus, expressing minimal capsule, was incubated with serum under various conditions, washed, and then incubated at 37 degrees C for 30 min with cultured human umbilical vein EC (ATCC CRL-1730). Adherence was scored visually after staining with acridine orange. Incubation in 10% heat-inactivated human serum increased adherence to endothelial cells by 488% compared to organisms incubated in buffer. Incubating S. aureus in complement-active normal human serum (NHS) decreased binding to EC by 58% compared to organisms incubated in heat-inactivated serum. The importance of active complement was confirmed by experiments using serum with added EDTA or cobra venom factor, a protein that depletes C3. The expression of capsule by S. aureus strongly interfered with adherence. It has been shown that an important protein for S. aureus adhesion to EC is fibronectin. S. aureus adherence to purified fibronectin increased by 511% after incubation in heat-inactivated serum, compared to that of organisms incubated in buffer. This decreased by 56% in complement-active serum, suggesting that inhibition of S. aureus adherence to EC is due, in part, to complement-mediated diminished binding to fibronectin. Interestingly, when EC were exposed to S. aureus-activated serum and then washed, binding by S. aureus was 234% higher than that of EC exposed to NHS. Thus, complement-activated EC have increased S. aureus binding, while complement on the bacterial surface markedly reduces adherence.     

Development and design of a novel in vivo chamber implant for the analysis of microbial virulence and assessment of antimicrobial therapy.

An accurate reflection of the pathogenicity of microorganisms and the therapeutic effects of antimicrobial agents on their growth necessitates testing within an in vivo environment. We have developed a novel diffusion chamber, incorporating two 0.22 microns membrane filters, for the growth of in vivo organisms. The chamber, which is implanted intraperitoneally into the rat, has an external sampling portal. This portal allows multiple and sequential sampling of the microbial inoculum without killing the rat, thus significantly reducing the total number of animals used in such studies. In addition, the chamber is superior to other reported implants since it is well tolerated, reusable, easily constructed and can be used within two days of implantation. Staphylococcus epidermidis and a toxic shock syndrome toxin-1 (TSST-1) producing strain of S. aureus have been successfully grown within in vivo chambers, with 10(8)-10(9) organisms per millilitre being recovered within 48 h. Scanning electron microscopy revealed clusters of staphylococci and fibrous material adhering to the inner surface of the filters, with numerous phagocytic cells attached to the outer side. Western immunoblotting indicated that higher levels of TSST-1 were produced by S. aureus grown in vivo as opposed to cells grown in vitro.     

Disulfide-bonded outer membrane proteins in the genus Legionella.

Staphylococcus aureus has been classified into at least eight different capsular types by using polyclonal rabbit antisera specific for their associated capsular polysaccharides. We produced and characterized monoclonal antibodies reactive with two serologically distinct capsular types, types 5 and 8, which account for more than 70% of all S. aureus bacteremias. These type-specific, monoclonal antibodies reacted with S. aureus clinical isolates possessing the homologous capsular type and exhibited no cross-reactivity against S. aureus clinical isolates possessing the heterologous capsular type, nontypeable S. aureus clinical isolates, Staphylococcus epidermidis clinical isolates, or a variety of gram-negative organisms. The anti-type 8 monoclonal antibodies also reacted with purified capsular polysaccharide derived from the prototype type 8 S. aureus strain.     

Specific immune response in the respiratory tract after administration of an oral polyvalent bacterial vaccine.

An oral killed polyvalent bacterial vaccine was assessed in a double-blind trial involving healthy volunteers. Three courses of oral vaccine were given over a 2-month period; each course contained 10(10) Haemophilus influenzae and 7 X 10(9) Staphylococcus aureus organisms. Immunity was assessed by monitoring antibody in saliva and serum over a 3-month period. No evidence of a nonspecific effect on immune parameters (immunoglobulin levels and Escherichia coli antibody) was detected in saliva or serum. An increase in H. influenzae antibody in saliva was detected in 55% of subjects receiving the vaccine compared with 6.7% of the placebo group. Antibody was associated with immunoglobulin A (IgA), IgG, and IgM, but the greatest increases over preimmunization levels were detected in the IgA class. No increase in serum antibody levels was detected. Subjects with higher preimmunization levels of salivary antibody to H. influenzae were less likely to respond to the oral bacterial vaccine. No increase in S. aureus antibody was detected in saliva or serum.     

Capsular antibodies induce type-specific phagocytosis of capsulated Staphylococcus aureus by human polymorphonuclear leukocytes.

Capsular types 5 and 8, which account for about 70% of Staphylococcus aureus strains isolated from the blood of patients, resisted in vitro phagocytosis by human polymorphonuclear leukocytes (PMN). Antisera and monoclonal antibody to type 5 and 8 capsular polysaccharides (CPS) induced type-specific in vitro phagocytosis of capsulated organisms by PMN. Antibodies directed against the O-acetyl moiety of the type 8 CPS were more effective in inducing phagocytosis of type 8 organisms by PMN. Either type-specific antiserum or monoclonal antibody reactive with the native O-acetylated type 8 CPS was most effective in inducing in vitro phagocytosis of type 8 organisms by PMN. These results provide further evidence that CPS of S. aureus are associated with host immunity to this organism.     

Transfer of R factors from Escherichia coli to salmonellas in the rumen of sheep.

Adult sheep were given into the rumen c. 10(8) cells each of donor strains of E. coli containing an R factor and prospective salmonella-recipient organisms and were maintained on a diet of lucerne chaff; the animals excreted the organisms, remained healthy, and no transfer of the R factor was detected. When the animals were starved for 48 h before inoculation, the ruminal environment was altered so that, on resumption of feeding, small numbers (c. 10(2)-10(4) cells--less than one cell per ml of rumen fluid) of the introduced organisms were able to multiply and reach sufficient numbers for the transfer of R factors to occur within the rumen. One animal, given 7-8 X 10(3) cells of recipient S. lomita after starvation for 48 h, became a carrier of this organism. A second animal, given 4-4 X 10(2) cells of S. typhimurium after starvation for 48 h, developed acute, fatal salmonellosis 5 days later; at the time of death, large numbers of salmonella organisms (c. 10(9) cells per g) were present in the faeces; these included many (c. 10(6) cells per g) that had received the R factor by transfer in vivo. These results indicate that short periods of starvation may enhance the transfer of R factors and possibly other plasmids between suitable micro-organisms in vivo, and may increase the susceptibility of animals to pathogenic micro-organisms.     

The secondary immune response to Staphylococcus aureus vaccines in efferent popliteal lymph of sheep.

The secondary immune response to live and killed Staphylococcus aureus vaccines was studied in efferent popliteal lymph of sheep. Animals were immunized with either live or killed S. aureus intracutaneously on the lateral hock, in an area draining into the popliteal lymph node. Six weeks later, an efferent popliteal lymphatic vessel in the vaccinated leg was cannulated, and 48 hr after surgery a second inoculation (identical to the primary) was placed in the skin adjacent to the primary vaccination lesion. A dramatic decrease in lymphocyte output ('cell shutdown') was observed in lymph collected from sheep given the secondary inoculation of live S. aureus during the first 8 hr after inoculation. However, only a moderate decrease in lymphocyte output occurred in lymph from animals receiving killed S. aureus or from control animals. The proportion of eosinophils in lymph collected from animals given live S. aureus increased to a peak (14% of total leucocytes in lymph) between 6 hr and 8 hr, and returned to prechallenge levels by 24 hr post-inoculation. The percentage of neutrophils in lymph peaked between 8 hr and 1 day after injection of live bacteria. This granulocyte response was not observed in animals given killed S. aureus or control animals. IgM-, IgG1- and IgG2- containing cells (-cc) in lymph were quantified by indirect immunofluorescence. Animals given live S. aureus produced lymph with greater numbers of Ig-cc of these isotypes than those given killed organisms. The ratio of IgG2-cc:IgG1-cc was significantly greater in lymph from animals given live S. aureus from Day 2 to Day 6 post-challenge. IgM and IgG1 anti-staphylococcal antibody levels increased in lymph collected from all vaccinated animals, but only sheep given live S. aureus showed any increase in levels of IgG2 antibody.     

Reduced adherence to traumatized rat heart valves by a low-fibronectin-binding mutant of Staphylococcus aureus.

Isogenic strains of Staphylococcus aureus, differing in fibronectin binding, were constructed for studies of the contribution of fibronectin binding to the in vivo pathogenesis of staphylococcal disease. Mutagenesis of S. aureus 879R4S was accomplished by mating with Enterococcus faecalis FA378 that carried the transposon Tn918. Four low-fibronectin-binding mutants were identified that bound 24- to 35-fold less fibronectin than the parent strain did. A spectinomycin-resistant strain, R4SSp, was transduced by a bacteriophage (80 alpha) lysate propagated on a low-binding mutant of 879R4S to produce R4SSp/1536, which bound less fibronectin, contained a single copy of the transposon, and grew on spectinomycin-containing medium. Using a rat model of endocarditis, we determined the distribution of S. aureus R4SSp and its transductant in normal and cardiac catheterized rats. Cultures of heart tissue showed that catheterized rats challenged with the fibronectin-binding parent strain had over 250-fold more organisms in the left heart than did rats challenged with the low-binding transductant. The ability to bind fibronectin had no effect on the number of S. aureus cells cultured from other tissues. These data suggest that the ability of S. aureus to bind fibronectin is an important factor in establishing adherence to damaged heart valves in vivo.     

In-vitro interaction of azithromycin and fluoroquinolones against gram-positive and gram-negative bacteria

Objective: To determine the combined in-vitro effects of azithromycin plus the fluoroquinolone ofloxacin or lomefloxacin against gram-positive and gram-negative bacteria.
Methods: Fractional inhibitory (FIC) and fractional bactericidal concentration indices of azithromycin and the fluoroquinolone were determined using a microtiter-checkerboard method. Clinical isolates of Staphylococcus aureus, Streptococcus pneumoniae, Neisseria gonorrhoeae, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Pseudomonas cepacia, Haemophilus influenzae, Xanthomonas maltophilia and Acinetobacter calcoaceticus were studied. Fourteen strains of S. aureus were also studied in time-kill curves with azithromycin (4 mg/L), lomefloxacin (6 mg/L) and the two in combination.
Results: No synergism or antagonism was found in inhibitory assays. However, bactericidal assays revealed antagonism with some strains of S. aureus, S. pneumoniae, X. maltophilia, A. calcoaceticus, P. aeruginosa, P. cepacia, K. pneumoniae and E. coli. Kill-curve results with 14 strains of S. aureus showed no antagonism with four strains of methicillin-resistant S. aureus (MRSA), and antagonism with one strain of MRSA and seven methicillin-susceptible S. aureus (MSSA).
Conclusions: In-vitro exposure to combinations of azithromycin and a fluoroquinolone does not produce a synergistic effect. Antagonism was found in bactericidal assays against some gram-negative bacteria and MSSA; caution is therefore recommended in the use of macrolides and quinolones against these organisms.     

Antibody response to teichoic acid and peptidoglycan in Staphylococcus aureus osteomyelitis.

An enzyme-linked immunosorbent assay was used to evaluate the immunoglobulin G (IgG) response to Staphylococcus aureus crude teichoic acid (TA) and peptidoglycan (PG) in both rabbits and patients with osteomyelitis. In rabbits with experimental S. aureus osteomyelitis, elevated levels of IgG to TA were present in 13/18 (72%) of the serum samples obtained at 4 and 10 weeks postinfection. In contrast, only 5/18 (28%) of these sera were found to be positive for antibodies to PG. Of a total of 39 patients with confirmed S. aureus osteomyelitis (11 acute, 28 chronic), IgG to TA was elevated in 17 (44%), whereas antibodies to PG were found to be increased in only 1 (3%). Cross-reacting antibodies to S. aureus TA were detected in only 1/18 (6%) of the patients with osteomyelitis caused by organisms other than S. aureus. These studies indicate that IgG to TA is more prevalent than IgG to PG in patients with staphylococcal osteomyelitis. Although these results are encouraging, a larger number of patients is required for an adequate evaluation of the TA enzyme-linked immunosorbent assay for the diagnosis and management of suspected S. aureus osteomyelitis.