Accuracy of methods using somatic cell count and N-acetyl-beta-D-glucosaminidase activity in milk to assess the bacteriological cure of bovine clinical mastitis

This study examined the capability of milk somatic cell count (SCC) and NAGase activity to discriminate between quarters that had been cured versus those that had not been cured at 4 wk after antimicrobial therapy for clinical mastitis. The distribution of microorganisms that were isolated before therapy from 630 quarters with mastitis was as follows: 225 strains of Staphylococcus aureus, 96 strains of coagulase-negative staphylococci, 152 strains of streptococci (Streptococcus dysgalactiae and Streptococcus uberis), and 157 strains of coliform bacteria. Bacteriological cure rates were 35% for mastitis caused by Staph. aureus, 75% for mastitis caused by coagulase-negative staphylococci, 66% for mastitis caused by streptococci, and 72% for mastitis caused by coliforms. Diagnostic accuracy of milk SCC and NAGase and their interquarter ratios for predicting bacteriological status of the control samples was assessed by calculating sensitivity, specificity, and accuracy and by means of receiver operating characteristic analysis. The efficiency of milk SCC and NAGase for predicting bacteriological cure was greatest for cows that had been infected with Staph. aureus. The main problem in detecting coagulase-negative staphylococci was low sensitivity, and the main problem in detecting streptococci and coliforms was low specificity. Receiver operating characteristic analysis is not completely suitable for the detection of mastitis because reference method bacteriology and indirect tests can never fully agree. To assess the recovery of cows from mastitis caused by Staph. aureus, bacteriology should be supplemented with an examination of milk SCC or NAGase activity at threshold values such as those presented here.     

Staphylococcal strains involved in bovine mastitis are inhibited by Staphylococcus aureus antimicrobial peptides

The inhibitory activity of five bacteriocin (Bac)-producer strains of Staphylococcus aureus was tested against bacteria pathogenic for cattle. Sixty-five epidemiologically unrelated strains of Staph. aureus involved in bovine mastitis were used as indicators in an agar diffusion test. Bacteriocins produced by four strains could inhibit only a limited number of test organisms. However, all 65 indicator strains proved to be susceptible to the combined action of both bacteriocins encoded by pRJ9, a Bac plasmid found in strain A53. Therefore, the bacteriocins produced by this strain may represent new antimicrobial peptides with potential applications in the prevention and treatment of bovine mastitis.     

Characteristics of the staphylococci isolated from the udder of cows with mastitis

A total of 175 strains of Staphylococcus aureus and 67 strains of Staphylococcus epidermidis were studied, isolated from 486 samples of milk secretion taken aceptically from the individual quarters of the udder of cows affected with subclinical and purulent (clinical) mastitis. The staphylococci were referred to as the causative agent of mastitis in case they were the only microflora in the seedings of the investigated material. Tests were applied as given in Fig. 1 to characterize the strains. It was found that mastitis in cows could be due to both plasma coagulating staphylococci (Staphylococcus aureus) and coagulase-negative Staphylococcus epidermidis organisms. The two Staphylococcus species were isolated from cows with clinical and subclinical mastitis. The division between pathogenic and nonpathogenic Staphylococcus strains by the plasma-coagulating symptom proved impossible, and this made it necessary to use other tests for pathogenicity. It became evident that the thing Staph. aureus and Staph. epidermidis had in common when isolated from cows with mastitis was the production of a gold-like pigment and delta hemolysin. Similarly to Staph. aureus isolated animals, the bovine Staph. epidermidis organisms did not possess fibrinolysin and rarely produced hemolysin. The isolated organisms belonging to the coagulase-positive staphylococci corresponded by their basic properties to Staphylococcus aureus var. bovis as described in the literature. The cultures of Staphylococcus epidermidis isolated under similar conditions showed in a considerable per cent of the cases somewhat different behaviour.     

Diabetes mellitus in sub-saharan Africa

The value of total coliforms, faecal coliforms and faecal streptococci in predicting the presence of Salmonella spp. and the numbers of Staphylococcus aureus and Candida albicans in sewage polluted coastal water were assessed. All indicators had strong positive association with Salmonella and moderate positive correlations with Staph. aureus and C. albicans. Total coliforms correlated better with salmonellas and Staph. aureus than did the two faecal groups. Regression analysis revealed that total coliforms have a better value as predictors of the presence of Salmonella and Staph. aureus, while faecal coliforms are better predictors of C. albicans, in moderately polluted areas. The conclusion reached is that enumeration of total coliforms is sufficient to predict the presence of Salmonella spp. or Staph. aureus in sea water moderately affected by sewage pollution, without the additional measurement of faecal coliforms and faecal streptococci.     

The students'' page. Are we making good use of this space?

Parallel quantitative determination executed in 92 herds at 107 sampling dates was focused on the counts of major pathogens (Staphylococcus aureus, Streptococcus agalactiae) in bulk milk samples and on the prevalence of the above pathogens in a herd by examination of individual milk samples. The counts of main pathogens were also determined in terms of quantity in rinsing water before milking and in bulk samples in 5 herds with pipeline milking. Tab. I shows the qualitative analysis of the relation. Sensitivity of the method is satisfactory for the pathogens observed (95% and 91%, resp.), but method is less specific for Staph. aureus (67% against 92% Str. agalactiae). Figs. 1 and 2 show coordinate graphs of the results obtained while Figs. 3 and 4 document the distribution of frequency of the particular values in data sets. The values do not exhibit normal distribution (P < 0.01). Spearman's coefficient of rank correlation of bulk milk and individual examinations amounted to 0.823 and 0.900 for Staph. aureus and Str. agalactiae, respectively. Four mathematical models were tested in the course of quantitative analysis, describing the relation between bulk milk examination and individual examination: (1) linear regression, (2) linear regression with fixed starting point, (3) logarithmic regression and (4) irrational function. A model based on the equation of irrational function (4) was found to be best: y = a + bx + c square root of x + k +/- a1 + i(t)c1 square root of x + k1. Tab. II shows the parameters of the equation for examined microorganisms. Correlation coefficients for the above equation are r = 0.733 and r = 0.842 for Staph. aureus and Str. agalactiae, respectively. Prediction curves (Figs. 5 to 8) and confidence regions of prediction curves were also determined for the best model, and a prediction table was constructed (Tab. III). It was confirmed that the milking machines were not a significant source of direct contamination of bulk milk samples with the examined pathogens (Tab. V).     

Pyogenic liver abscesses and acute schistosomiasis mansoni: report on 3 cases and experimental study

Three children with acute schistosomiasis mansoni developed pyogenic liver abscesses. The abscesses were diagnosed by ultrasonography and confirmed during laparotomy. Staphylococcus aureus were the sole bacteria isolated from the abscesses. An experimental study was carried out in mice to establish whether schistosomiasis is a predisposing cause for pyogenic liver abscesses. Seventeen mice (group 1) were infected with 40 Schistosoma mansoni cercariae (LE strain) and 60 d later inoculated intravenously with a strain of Staph. aureus, isolated from a patient with bacteraemia; 17 mice infected with Sch. mansoni (group 2), 19 infected with bacteria alone (group 3), and 18 uninfected mice (group 4), served as controls. Thirteen group 1 mice (77%) developed multiple liver abscesses while none was observed in the controls. These results indicate that acute schistosomiasis mansoni concurrent with Staph. aureus bacteraemia favours the colonization of the liver by bacteria and the development of pyogenic hepatic abscesses.     

Conversion of chronic staphylococcal mastitis to acute gangrenous mastitis after neutropenia in blood and bone marrow produced by an equine anti-bovine leukocyte serum

A stock strain of Staphylococcus aureus of mastitis origin, characterized by alpha-, beta-, and delta-toxins, was used to produce chronic mastitis of 20 to 300 days' duration in 6 lactating mammary quarters of 4 cows. Early acute Streptococcus agalactiae mastitis was produced in 1 additional mammary quarter of 1 cow. Equine anti-bovine leukocyte serum (EABLS) was administered to all cows by continuous intravascular drip for 12 to 32 hours. Neutropenia in blood and partial depletion of neutrophil reserve in bone marrow were produced. Chronic subclinical staphylococcal mastitis in 2 quarters of 1 cow changed to gangrenous mastitis by the 40th hour after EABLS administration and led to death of the cow. The disappearance of neutrophil leukocytes from the milk was followed by uninhibited multiplication of S aureus. Probably, staphylococcal leukocidins accelerated the destruction of neutrophils in the milk as S aureus multiplication became intensified. In another quarter of the same cow that was infected with Str agalactiae, neutrophil leukocytes were present in milk as long as 3 days after their disappearance from blood and bone marrow. This may give some indication of the extravascular life-span of the neutrophil in the udder in mastitis. The 2nd cow died at the 16th hour from the start of EABLS administration and at a time when gangrenous mastitis was in the initial stages of development. The S aureus-infected quarters of the 2 remaining cows did not become gangrenous. Administration of EABLS to these 2 cows did not significantly reduce the numbers of neutrophil leukocytes entering the milk of the 3 S aureus-infected quarters. It is concluded that continuous diapedesis of neutrophil leukocytes into the milk in chronic staphylococcal mastitis protects the gland against the development of gangrenous mastitis in the presence of a strain of S aureus capable of alpha-toxin production.     

Staphylococci outside the hospital. Staphylococcus aureus in sheep

Biochemical properties were studied in Staph. aureus strains obtained from the anterior nares of healthy sheep and from the udders of ewes suffering from purulent mastitis. Of the total number of 84 isolated staphylococcal strains 75 (89.3%) were classified as the C biotype. These undoubtedly sheep-adapted staphylococci produced pigment and beta hemolysin, they were growing on crystal violet agar as the negative type in violet colonies lacking both fibrinolysin and alpha hemolysin. All of them coagulated human plasma within one hour after inoculation. In bovine plasma 27 strains (36%) formed the coagulum within 3 hours, 16 (21.3%) within 24 hours, and the remaining 32 strains (42.7%) only within 72 hours. Mannitol was fermented after five days only by 33 cultures (44%). The staphylococci were sensitive to the applied antibiotics without exception. All these sheep-adapted staphylococci had analogous biochemical features to the earlier discussed staphylococcal strains obtained by the authors from the nasal cavities of cattle. Next two strains were denoted as deficit variants of the C biotype because of their lack of pigment. Of quite a different character were 3 strains (3.6%) of the A biotype and one strain identified as the E biotype. The former were presumably transferred to sheep from man while the latter from a dog. The remaining 3 strains could not be subdivided according to the classificatory criteria used here.     

Sensitivity of Streptococcus agalactiae and Staphylococcus aureus to antibiotics in the Slovak Socialist Republic in the year 1974

In 1974, the State veterinary institutes in the Slovak Socialist Republic studied the sensitivity of 4420 strains of Streptococcus agalactiae and 1056 strains of Staphylococcus aureus to eight antibiotics. The strains were isolated from milk samples obtained from dairy cows suffering from mastitis. 100 per cent of the strains of Streptococcus agalactiae was sensitive to ampicillin, 99.23% to erythromycin, 98.70% to oxytetracycline, 93.02% to bacitracin, 90.77% to chloramphenicol, 41.97% to penicillin, 12.39% to neomycin, 9.73% to streptomycin. As to the strains of Staphylococcus aureus, 98.68 were sensitive to chloramphenicol, 98.50% to ampicillin, 97.92% to erythromycin, 94.98% to oxytetracycline, 93.85% to neomycin, 92.67% to bacitracin 87.50% to streptomycin, and 46.24% to penicillin. The results are discussed in relation to the use of antibiotics in the treatment of mastitis in dairy cows.     

Endogenous opioid tone in patients with rheumatoid arthritis

A monoclonal antibody giving a dominant reaction with the group-specific polysaccharide of streptococcus group B in an ELISA test has been developed. The purified polysaccharide exhibited a high positivity with reference anti B streptococcal antiserum in the ELISA test. Cross-tests of antibodies with other groups of streptococci provided a minimum cross-reaction only in the case of G streptococci. Monoclonal antibodies were prepared using Streptococcus agalactiae S 589 MT strain isolated from a case of bovine mastitis which does not express Ia, Ib, II, III, IV and V type antigens, nor C, R and X protein antigens.     

Time-frequency analysis of heart murmurs. Part II: Optimisation of time-frequency representations and performance evaluation

OBJECTIVE: We hypothesized that carriage of Staphylococcus aureus among continuous ambulatory peritoneal dialysis (CAPD) patients was influenced by their spouses. Furthermore, this carrier status was compared to previous Staph. aureus peritonitis episodes in order to identify the influence of Staph. aureus carriage on peritonitis rate. DESIGN: A combined prospective surveillance study (Staph. aureus carriage) and retrospective chart review (Staph. aureus peritonitis). SETTING: A single peritoneal dialysis unit in a county hospital. PATIENTS AND METHODS: Cultures from patients (n = 32) and spouses (n = 16) were obtained twice, with a 1-month interval, from the anterior nares, the umbilical, and one groin area. All positive cultures were phage typed. Retrospective chart review of all episodes of Staph. aureus peritonitis among the patients was carried out. RESULTS: Twelve of 32 patients (37.5%) and 5 of 16 spouses (31%) evaluated were carriers. Half of the spouses of patients who were Staph. aureus carriers, were also carriers, as opposed to 20% of spouses of noncarrier patients (p = 0.30). Patients and spouses always shared the same phage type. Among patients, Staph. aureus was found in the nose only (n = 9), in all three regions (n = 2), and extranasally only (n = 1). If only one nasal culture was used to establish carriage, the sensitivity and negative predictive value would be 92% and 95%, respectively. A trend toward a higher incidence (p = 0.062) of Staph. aureus peritonitis was found among carriers (patients), 0.37 versus 0.28 peritonitis episode/dialysis-year. CONCLUSIONS: Only one positive nasal culture was necessary when carriage of Staph. aureus was to be established. Staph. aureus carriage was found more often in patients who had previously suffered from Staph. aureus peritonitis. The phage types isolated remained fairly constant, and the patients and spouses often had the same carrier state and shared the same phage types, although transmission does not always take place.     

Contagious mastitis

Contagious mastitis is defined. The major mastitis pathogens are Streptococcus agalactiae, Staphylococcus aureus, Corynebacterium bovis, Mycoplasma sp, and Streptococcus dysgalactiae. These pathogens are discussed relative to prevalence, virulence factors, pathology, and control. These control measures include milking time hygiene, segregation, culling, vaccination, and treatment.     

Diagnosis and therapy of splenic abscess exemplified by 3 unusual cases

The total enumeration and biotyping of staphylococci isolated from the rumen content of deer is presented in this study. The total counts of staphylococcal isolates from deer reached 1.5 x 10(3) cfu/ml. Of 960 colonies, 904 were taxonomically identified. Coagulase-negative staphylococci mainly Staphylococcus simulans and Staph. xylosus were the most frequently isolated species, followed by Staph. warneri, Staph. lentus, Staph. saprophyticus, Staph. epidermidis and Staph. hominis. Moreover, the atypical species were isolated, such as Staph. chromogenes, Staph. capitis, Staph. caseolyticus and Staph. auricularis. Except Staph. aureus, no coagulase-positive staphylococci were found. The strain identified showed high resistance to vancomycin (61.2%) and to chloramphenicol (41.9%). The resistance to erythromycin and bacitracin was demonstrated in 16.1% and 35.5% of isolates. On the other hand, 99% of isolates were sensitive to gentamycin, 96% to linkomycin, 93% to methicilin and 87% of isolates were sensitive to bacitracin and virginiamycin. The mean value of lactic acid production was 0.446 +/- 0.026 mol/l. The majority of strains belonged to ureolytic bacteria with low urease production. However, eight different species showed urease activity higher than 30 nkat/ml. No bacteriocin active strain was detected by indicator organisms used. Although the percentage of nonidentifiable strains was high (64.7%), the detection of different species improved and confirmed species variability in the rumen microbial ecosystem of deer as well as in total. Moreover, additional tests are under way, especially those regarding bacteriocin production.     

Results of bacitracin sensitivity tests on 250 beta hemolytic streptococcal strains

On blood agar plates prepared from tryptose blood agar base (Difco) containing 5% defibrinated sheep blood cultures were done. After overnight incubation plates were examined and beta hemolytic streptococci tested with Taxo A (Difco) disks. 234 of them came out sensitive.     

Bovine mastitis pathogens in New York and Pennsylvania: prevalence and effects on somatic cell count and milk production

Milk samples were collected from 108,312 dairy cows during 1601 farm visits made between January 1991 and June 1995. The herd visits were made by personnel from the Central Laboratory of the Quality Milk Promotion Services at Cornell University (Ithaca, NY) to farms located in central New York and northern Pennsylvania. Dairy Herd Improvement Association records were available for 32,978 cows in 327 herds. Intramammary infections, as defined by positive milk cultures, were present in 48.5% of all cows and in 36.3% of cows in herds enrolled in the Dairy Herd Improvement Association. Over 75% of the intramammary infections were caused by Streptococcus agalactiae, Streptococcus spp. other than Strep. agalactiae, Staphylococcus aureus, and coagulase-negative staphylococci. Mean days in milk at the time of diagnosis, linear score of the somatic cell count, cost of milk loss per lactation, and milk production effects were calculated for 24 etiologic agents of bovine mastitis.     

Incidence of nasal carriers of Staphylococcus aureus in and outside hospital environment and antibiotic sensitivity of isolated staphylococcus strains

This study was carried out on 100 nasal swabs collected from medical personnel (nurses and doctors) and patients inside hospital environment and also from 50 individuals outside hospital. The swabs were inoculated on different culture media for isolation of /staphylococci which were further identified as S. aureus either by classic bacteriologic methods or by one of rapid screening test of S. aureus. The isolated strains were tested for antibiotic sensitivity to some of B-Lactam antibiotics and to other antibiotics. The results showed that significantly higher percentage of coagulase + ve Staph. were isolated from newborn nursery (90%), operating theatre (71.4%) and hemodialysis unit (60%) than those isolated from intensive care unit, cancer chemotherapy, surgery, chest, internal medicine departments (25%, 26.6%, 31.2%, 33.3%, 50%) respectively. It also showed significant difference in isolation rate between persons at the hospital (patients, doctors and nurses) 44% and controls (normal population) 26%. Most isolates of coagulase + ve Staph. were resistant to penicillin G (93.2%), Streptomycin (77.3%), tetracycline (61.4%) and sensitive to cefamandole (95.4%). All coagulase+ve Staph. isolates were resistant to sulphonamide and methicillin and all sensitive to vancomycin.     

Microbiology of liver and spleen abscesses

To study the aerobic and anaerobic microbiology of liver and spleen abscesses and correlate the results with predisposing factors, potential causes and routes of infection, clinical and laboratory data of 48 patients with liver abscesses and 29 with spleen abscesses treated between 1970 and 1990 were reviewed retrospectively. In liver abscesses, a total of 116 isolates (2.4 isolates/specimen) was obtained; 43 were aerobic and facultative species (0.9 isolates/specimen) and 73 were anaerobic species or microaerophilic streptococci (1.5 isolates/specimen). Aerobic bacteria only were isolated from 12 (25%) abscesses, anaerobic bacteria only from eight (17%), and mixed aerobic and anaerobic bacteria from 28 (58%); polymicrobial infection was present in 38 (79%). The predominant aerobic and facultative isolates were Escherichia coli (11 isolates), Streptococcus group D (8), Klebsiella pneumoniae (5) and Staphylococcus aureus (4). The predominant anaerobes were Peptostreptococcus spp. (18 isolates), Bacteroides spp. (13), Fusobacterium spp. (10), Clostridium spp. (10) and Prevotella spp. (4). There were 12 isolates of micro-aerophilic streptococci. S. aureus and beta-haemolytic streptococci were associated with trauma; Streptococcus group D, K. pneumoniae and Clostridium spp. with biliary disease; and Bacteroides spp. and Clostridium spp. with colonic disease. In splenic abscesses, a total of 56 isolates (1.9 isolates/specimen) was obtained; 23 were aerobic and facultative species (0.8 isolates/specimen), 31 were anaerobic species or micro-aerophilic streptococci (1.1 isolates/specimen) and two were Candida albicans. Aerobic bacteria only were isolated from nine (31%) abscesses, anaerobic bacteria from eight (28%), mixed aerobic and anaerobic bacteria from 10 (34%) and C. albicans in two (7%); polymicrobial infection was present in 16 (55%). The predominant aerobic and facultative isolates were E. coli (5 isolates), Proteus mirabilis (3), Streptococcus group D (3), K. pneumoniae (3) and S. aureus (4). The predominant anaerobes were Peptostreptococcus spp. (11 isolates), Bacteroides spp. (5), Fusobacterium spp. (3) and Clostridium spp. (3). S. aureus, K. pneumoniae and Streptococcus group D were associated with endocarditis, E. coli with urinary tract and abdominal infection, Bacteroides spp. and Clostridium spp. with abdominal infection and Fusobacterium spp. with respiratory infection.     

Phenotypic detection of mec A-positive staphylococcal blood stream isolates: high accuracy of simple disk diffusion tests

Detection of oxacillin-resistance in staphylococci by phenotypic methods remains problematic. Although standardized susceptibility test methods are adequate for Staphylococcus aureus, many are less satisfactory for the coagulase-negative staphylococci (CNS). We have studied 108 consecutive blood culture isolates of staphylococci. The mec A gene was detected by PCR in one S. aureus and 55 CNS isolates. Susceptibility testing was performed as follows: oxacillin (1-microgram), ceftizoxime (30-microgram), and cephalothin (30-microgram) by disk diffusion; oxacillin, ceftizoxime, cephalothin, methicillin, ampicillin, ampicillin/ sulbactam, penicillin, cefazolin, imipenem, and meropenem by the broth microdilution method. In addition, isolates were tested by the oxacillin agar screen plate method. The single oxacillin-resistant S. aureus strain was detected by all oxacillin susceptibility test methods and by the ceftizoxime disk and MIC methods. Two oxacillin-susceptible S. aureus were intermediate (minor error) by ceftizoxime broth microdilution (MIC, 16 micrograms/mL). The most sensitive, simple phenotypic methods for detection of oxacillin-resistant CNS (mec A positive) were as follows: oxacillin disk diffusion at 98%, oxacillin screen plate at 91%, oxacillin broth microdilution at 87%, ceftizoxime disk diffusion at 100%, ceftizoxime broth microdilution at 87%, and methicillin broth microdilution at 83%. These results indicate that oxacillin and ceftizoxime disk diffusion tests are the most accurate phenotypic methods in routine clinical use for detection of oxacillin-resistant CNS. Oxacillin broth microdilution MIC testing (2% NaCl supplement) would perform more satisfactorily (100% sensitivity) with an adjusted interpretive breakpoint at < or = 0.5 microgram/mL, in contrast to the lower accuracy of the "so-called" reference agar screen test.     

Prevention of bovine mastitis by a premilking teat disinfectant containing chlorous acid and chlorine dioxide

The objective of this study was to evaluate the efficacy of a premilking teat disinfectant for the prevention of mastitis in dairy cows under natural exposure conditions. Predipping was compared with a negative control using a split udder experimental design. All teats were dipped after milking with the same teat dip. Percentage of quarters newly infected by major mastitis pathogens was 34% lower in quarters with teats predipped and postdipped than in quarters with teats postdipped only. New IMI by Streptococcus uberis and Staphylococcus aureus were significantly lower in quarters with teats predipped and postdipped than in quarters with teats postdipped only. Differences in incidence of clinical mastitis between treatment groups approached significance. Predipping and postdipping were no more effective against Gram-negative bacteria, coagulase-negative Staphylococcus species, and Corynebacterium bovis than postdipping only. No chapping or irritation of teats was observed, and no adverse effects were noted using the test product as a premilking and postmilking teat disinfectant. Results of this study suggest that predipping and then postdipping with the test product was a more effective procedure against major mastitis pathogens than postdipping only.     

An agar medium for direct enumeration of Staphylococcus aureus: pork plasma medium for S. aureus (PPSA)

A selective agar medium (pork plasma medium for S. aureus (PPSA)) enables the direct enumeration of coagulase-positive staphylococci. This medium is based on the Baird-Parker agar without egg yolk and is supplemented with pig plasma. Colonies of Staphylococcus aureus are surrounded by a halo of precipitated fibrin. When foods such as dairy products contain large numbers of egg yolk-negative strains of S. aureus, the PPSA agar has the advantage over egg yolk containing media such as Baird-Parker agar that fewer suspect colonies have to be confirmed.