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Prevalence of coagulase-positive staphylococci, other than Staphylococcus aureus, in bovine mastitis.

OBJECTIVE: To determine prevalence and relevance of coagulase-positive Staphylococcus hyicus and S intermedius intramammary infections (IMI) in dairy cows and determine the ability of the 4-hour tube coagulase (TC) test to differentiate the coagulase-positive staphylococci (CPS). DESIGN: Prevalence of CPS was determined for primiparous cows (point prevalence and prevalence at first parturition) and multiparous cows (point prevalence) of 2 herd groups: < 6% CPS IMI prevalence = low prevalence (LP); > 10% CPS IMI prevalence = high prevalence (HP). SAMPLE POPULATION: For prevalence, cows of 22 dairy herds. For TC, 1,038 CPS strains isolated from cow milk. PROCEDURE: Speciation of CPS from aseptically collected composite milk samples was performed. Coagulase-positive isolates from 4 cow groups were tested for their ability to coagulate rabbit plasma by 4 hours: LP and HP primiparous cows at parturition, and LP and HP cows any time after first parturition. RESULTS: Of 487 CPS in the prevalence study, 82.1% were S aureus, 17.7% were coagulase-positive S hyicus, and 0.2% were S intermedius. Of all CPS IMI in LP herds, 34% were coagulase-positive S hyicus; of all CPS IMI in HP herds, 9% were coagulase-positive S hyicus. Coagulase-positive S hyicus appeared to persist to the end of lactation in 4 cows (mean linear somatic cell count = 3.7). The TC test was > or = 97% sensitive, < or = 33% specific, and had a predictive value positive range of 60 to 97% for S aureus isolates. CONCLUSION: Coagulase-positive S hyicus appears capable of inducing chronic, low-grade IMI. Staphylococcus intermedius does not appear to be an important mastitis pathogen. The TC test is not valid to use as the sole method to differentiate CPS species.

Animals↗

The effect of sour milk as a postmilking teat dip for mastitis prevention in a dairy herd.

In a preliminary in vitro study, the growth of Staphylococcus aureus was totally inhibited during incubation for 24 h at 35 degrees C-37 degrees C in a solution of cooked commercial milk with 1% of uncooked commercial sour milk ("A piimä"). In a subsequent clinical trial, "A piimä" sour milk with 5% glycerol was used as a postmilking teat dip from February to June. Quarterly milk samples were drawn once a month aseptically from 133 cows. Percentages of pathogen positive samples and somatic cell count (SCC) from teats dipped with the sour milk were compared with those dipped with a commercial iodine teat dip and those of undipped controls. During March-June there were fewer isolations of S. aureus (2.09%) and coagulase-negative staphylococci (2.52%) in the sour-milk group than in the control group (3.09% and 4.07%, respectively). In iodine group, there were fewer isolations of S. aureus (0.83%) but more isolations of coagulase-negative staphylococci (5.26%) than in the control group. During the study period, the percentages of bacterial isolates did not differ statistically significantly between treatments, p = 0.291. The percentage of quarters with a SCC over 125,000 at the end of the study was one third lower in the sour-milk group than in the control group (16.67% and 26.23% respectively) but the difference was not statistically significant (p = 0.074). The results indicate that a sour-milk teat-dip preparation can inhibit new intra mammary infections (IMI).

Animals↗

Occlusal hidden caries: a bacteriological profile.

Eleven teeth from nine patients with occlusal caries lesions beneath an apparently intact enamel surface were treated. Two dentine samples, one from the dentinoenamel junction (Sample A) and one just before all the caries appeared to be removed clinically (Sample B), were taken under aseptic conditions and examined for bacteria. In all cases, the A samples and in ten cases the B samples contained mutans streptococci, lactobacilli or both, while in ten cases the dentine after opening was soft with a light color. This suggests that we are dealing with active caries lesions, which should be treated as such. The salivary data of the patients group were compared with the salivary data of the control group. Although the buffer capacity of the groups were not significantly different (chi-square), further analysis revealed that a significant relationship could be found between the group and the buffer capacity (Kendall's Tau C). While the other salivary components (numbers of mutants streptococci per ml saliva and the secretion rate) showed no significant differences. Unfortunately these occlusal caries lesions are not detected with the present criteria used in epidemiological surveys.

Chi-Square Distribution↗

Antibiotic susceptibility of the bacteria causing odontogenic infections.

AIMS: An evaluation is made of bacterial species and susceptibility to various antibiotics used in application to odontogenic infections of periapical location and in pericoronitis of the lower third molar, with the aim of optimizing the antibiotherapy of such infections and thus preventing unnecessary side effects and over-treatment. MATERIAL AND METHODS: Sixty-four patients with odontogenic infection were selected on the basis of a series of inclusion and exclusion criteria. Samples were collected from lesions under maximally aseptic conditions, avoiding oral saprophytic contamination. The samples were cultured and incubated under aerobic and anaerobic conditions, followed by bacteriological identification and antibiotic susceptibility testing. RESULTS: A total of 184 bacterial strains were isolated and identified, comprising grampositive facultative anaerobes (68%), gramnegative strict anaerobes (30%) and grampositive facultative anaerobes (2%). Regardless of the origin of the odontogenic infection, the causal bacteria yielded the best results in terms of increased sensitivity and lesser resistance with amoxicillin / clavulanate and amoxicillin, respectively (p<0.05). DISCUSSION: There are increasingly numerous reports in the literature of growing bacterial resistance to antibiotics in infectious processes affecting non-buccodental territories. This same tendency has not been observed in relation to oral infections, though important resistance has been documented for certain concrete antibiotics. According to our results, the common-use antibiotics with the greatest sensitivity and lowest resistance were shown to be amoxicillin/clavulanate followed by amoxicillin alone.

Acetamides↗

Aseptic meningitis after neurosurgery: a demonstration of bacterial involvement.

OBJECTIVE: To evaluate the presence of bacteria in samples from patients suffering from 'aseptic' meningitis following craniotomy. METHODS: Prospective study in which cerebrospinal fluid (CSF) from patients suffering from post-craniotomy meningitis and negative control patients were submitted to conventional culture and to polymerase chain reaction (PCR) using bacterial 16S rRNA universal primers, followed in some cases by DNA sequencing of the PCR product and phylogenetic analysis. RESULTS: CSF from patients with either culture-positive or culture-negative meningitis yielded positive amplifications, whereas no amplification was obtained with CSF from control patients. All positive signals were confirmed by Southern hybridization with a prokaryote 16S RNA-specific probe. Six PCR products, of which three were collected from later cases of culture-negative meningitis, were cloned and sequenced. Sequence analysis suggested affinities with Pseudomonas in three cases, with Escherichia in two cases and with Rhodococcus in one case. CONCLUSIONS: Many cases of culture-negative (aseptic) meningitis are probably bacterial meningitis and justify antibiotic treatment. The bacteria responsible for these cases of culture-negative meningitis might have peculiar growth requirements in vitro.

Journal Article↗

Recurring synovitis as a possible reason for aseptic loosening of knee endoprostheses in patients with rheumatoid arthritis.

We evaluated histologically samples of synovial tissue from the knees of 50 patients with rheumatoid arthritis (RA). The samples were taken during revision for aseptic loosening. The findings were compared with those in 64 knees with osteoarthritis (OA) and aseptic loosening and in 18 knees with RA without loosening. The last group had been revised because of failure of the inlay or the coupling system of a constrained prosthesis. All the patients had had a total ventral synovectomy before implantation of the primary prosthesis. In all three groups a foreign-body reaction and lymphocellular infiltration were seen in more than 80% of the tissue samples. Deposits of fibrin were observed in about one-third to one-half of the knees in all groups. Typical signs of the reactivation of RA such as rheumatoid necrosis and/or proliferation of synovial stromal cells were found in 26% of knees with RA and loosening, but not in those with OA and loosening and in those with RA without loosening. Our findings show that reactivation of rheumatoid synovitis occurs after total knee replacement and may be a cofactor in aseptic loosening in patients with RA.

Aged↗

Bacterial isolates from blood cultures of children with suspected septicaemia in Calabar, Nigeria.

BACKGROUND: Septicaemia is a common cause of morbidity and mortality among children in the developing world. This pattern has changed little in the past decade. Physical signs and symptoms, though useful in identifying possible cases have limited specificity. Definitive diagnosis is by bacteriologic culture of blood samples to identify organisms and establish antibiotic susceptibility. These results are usually not available promptly. Therefore a knowledge of epidemiologic and antimicribial susceptibility pattern of common pathogens is useful for prompt treatment of patients. This report highlights the pattern of bacterial isolates in our environment from a retrospective study of our patients' records. METHODS: One thousand, two hundred and one blood samples were analysed from children aged 0-15 years, admitted into the children's wards of the University of Calabar Teaching Hospital, Calabar, Nigeria with features suggesting septicaemia. Samples were collected under aseptic conditions and cultured for aerobic and anaerobic organisms. Isolates were identified using bacteriologic and biochemical methods and antibiotic sensitivity determined by agar diffusion method using standard antibiotic discs. RESULTS: Bacteria was isolated in 552 (48.9%) of samples with highest rates among newborns (271 : 50.8). The most frequent isolates were Staphylococcal aureus (48.7%) and Coliforms (23.4%). Results showed high susceptibilities to the Cephalosporins (Ceftriazone- 100%:83.2%, Cefuroxime-100%:76.5%) and Macrolides (Azithromycin-100%:92.9%) for S. aureus and coliforms respectively. This study underscores the importance of septicaemia as a common cause of febrile illness in children and provides information on common prevalent aetiologic agents and drug susceptibilities of the commonest pathogens. CONCLUSION: Staphylococcus aureus and coliforms were the leading causes of septicaemia in children in this locality, and the third generation cephalosporins and azithromycin were shown to be effective against these pathogens.

Adolescent↗

Six years' experience of using the BacT/ALERT system to screen all platelet concentrates, and additional testing of outdated platelet concentrates to estimate the frequency of false-negative results.

BACKGROUND AND OBJECTIVES: Approximately 1 in every 2000 units of platelets is contaminated with bacteria. The BacT/ALERT automated blood culture system can be used to screen platelet concentrates (PCs) for bacterial contamination. MATERIALS AND METHODS: Data were collected from May 1998 until May 2004. The number of PCs tested during this period was 36 896, most of which were produced from pools of four buffy-coats. On the day following blood collection or platelet apheresis, a 5-10 ml sample of the PC was aseptically transferred to a BacT/ALERT culture bottle for detection of aerobic bacteria. The sample was monitored for bacterial growth during the entire storage period of the PC (6.5 days). When a positive signal was generated, the culture bottle, the PC and the erythrocyte concentrates were tested for bacterial growth. In order to determine the frequency of false-negative BacT/ALERT signals, 1061 outdated PCs were tested during the period from May 2002 to May 2004. RESULTS: Eighty-eight positive signals were detected by the BacT/ALERT system, of which 12 were interpreted as truly positive. Fourteen signals were interpreted as truly false positive. Thirty-three signals were interpreted to be probably false positive. Two of 1061 outdated units tested positive, and Bacillus spp. and Staphylococcus epidermidis, respectively, were isolated from these PCs. CONCLUSIONS: Between 0.03% and 0.12% of the PCs were contaminated with bacteria. BacT/ALERT is an efficient tool for monitoring PCs for bacterial contamination; however, it is important to realize that false-negative results may occur.

Bacillus↗

Microbial contamination potential of solutions in prefilled disposable syringes used with a syringe pump.

The sterility of trypticase soy broth (TSB) that was frozen and thawed in disposable plastic syringes and infused via syringe pump was studied to determine whether ambient air or personnel-transferred contaminants compromised the sterility of the solution. Samples of TSB (10, 20, 30 mL) were prepared aseptically in syringes of three different brands--150 samples for each volume (50 for each manufacturer). The syringes were placed in zip-lock bags, stored for 24 hours at -15 to -20 degrees C, and thawed for three hours. Both positive and negative controls were used. For the test samples, infusion sets were connected to the syringes under aseptic conditions, and the solution was infused via syringe pump in ambient air into polyvinyl chloride minibags before incubation. The remaining samples were prepared in the same manner as the test solutions except that they were intentionally challenged with Bacillus subtilis introduced distal to the plunger. All samples were inspected visually for turbidity after a 14-day incubation period. There was no growth in any of the test infusion samples or in samples that were intentionally contaminated. The negative controls showed no growth; all of the positive controls showed growth. The sterility of solutions frozen in disposable plastic syringes does not appear to be compromised by touch contamination of the plunger shaft or by airborne microorganisms settling on the infusion system.

Bacteria↗

Determination of Brucella spp. in raw milk and Turkish white cheese in Kirikkale.

In this study, a total of 105 samples including 35 raw milk, 35 cows' milk cheese, and 35 ewes' milk cheese samples were obtained from Kirikkale city and investigated for the presence and contamination level of Brucella species. For this reason, milk and cheese samples were collected in aseptic conditions and brought to the laboratory under cold conditions. The method suggested from FARREL was used for the isolation and identification of Brucella spp. In addition, Most Probable Number Technique was used to determine the contamination level in Brucella spp. positive samples. According to analysis findings, B. melitensis was isolated from 5 (14.2%) of 35 ewes' milk cheese samples at the level of 3.6 x 10(1)-9.3 x 10(3) MPN/g. Brucella spp. were not detected in any of raw milk and cows' milk cheese samples. In conclusion this study reinforced the previous reports that ewes' milk cheese is an important source of Brucella spp. and they have been risk for public health.

Animals↗

Epidemiological study of non-contagious intramammary infections in nine commercial dairy herds following a Staphylococcus aureus control programme.

Changes in prevalence in intramammary infection, by pathogen type, in herds applying a stringent contagious mastitis control programme was studied. Enrollment of 1651 lactating cows and collection of milk samples was made in this ancillary study to a cohort study of the dynamics of mastitis prevalence after adoption of a strict contagious mastitis control programme that targeted the elimination of mastitis caused by Staphylococcus aureus. Nine commercial dairies in Italy were used. Aseptic collection of milk samples from all lactating cows was performed at the time of enrollment, from all cows within 7-14 days of entering the lactating herd after the date of enrollment, and from all lactating cows at 2, 4, 7, 10 and 14 months after the date of enrollment. Prevalence of intramammary infection by pathogen type was determined from culture of milk samples. Application of the strict contagious mastitis programme did not lead to an increased risk of non-contagious mastitis. The risk of coliform, environmental streptococcal and coagulase-negative staphylococcal intramammary infections decreased after adoption of the programme. The data reported herein indicate that the overall risk for any intramammary infections decreases with adoption of a strict contagious mastitis programme, and that such a programme therefore does not necessarily lead to an increase in environmental mastitis.

Animals↗

Application of polymerase chain reaction and subsequent phylogenetic analysis to the diagnosis of enteroviral infection in the central nervous system.

BACKGROUND: Enteroviral infections of the central nervous system (CNS) are often difficult to diagnose, even if consistent conventional laboratory methodologies are used. OBJECTIVES: To clarify the efficiency of two polymerase chain reaction (PCR) methods for the sensitive detection of enteroviruses and for the identification of enteroviral genotypes based on phylogenetic analysis of the amplified genome sequences, and to facilitate the diagnosis of enteroviral infection in CNS. STUDY DESIGN: Cerebrospinal fluid (CSF), throat swab, rectal swab, and/or serum samples were collected from 171 patients with aseptic meningitis and 67 patients with febrile seizures. The samples were tested for the presence of enteroviruses by cell culture and PCR methods for the detection and identification of enteroviruses. RESULTS: In 111 (64.9%) of 171 patients with aseptic meningitis, enteroviruses were isolated by cell cultures from any site. In 143 (83.6%) patients, including 110 of 111 patients with aseptic meningitis, the enteroviral genome was detected in CSF by PCR. No enterovirus was isolated from any site for the 67 patients with febrile seizures. PCR detected the enteroviral genome in CSF samples from 13 (61.9%) of 21 patients who developed febrile seizures in the summer (June-August). Phylogenetic analysis of amplified genome sequences showed that the major pathogens of febrile seizures in summer were group A coxsackieviruses, which are usually difficult to isolate by cell culture. CONCLUSION: PCR methods for the detection and identification of enteroviruses were useful for the diagnosis of enteroviral infection in CNS.

Central Nervous System Viral Diseases↗

Microbial growth in small-volume pharmaceuticals.

The ability of aseptically filled pharmaceuticals to support microbial growth was tested on 43 small-volume products (mainly parenterals). These were inoculated with a variety of microorganisms which were known to be associated with contamination of pharmaceutical products. In general, Gram-negative bacteria were found to be much more likely to grow than Gram-positive. It was possible for an inoculum of a few cells to multiply to levels up to 10(7)/mL. The presence of preservatives also influenced the likelihood of growth, 12 out of 19 (63%) of the pharmaceuticals without preservatives supporting growth of one or more microorganisms; only 3 out of 24 (12%) of those with preservatives supported growth. The importance of these observations is discussed with reference to formulation of aseptically filled products, environmental sampling and the quality of cleanroom conditions necessary for production. It is suggested that those pharmaceuticals which are designed to be sterile but not to be terminally sterilized, should be tested before production begins, for their ability to support microbial growth. In this way, the risks involved in aseptically filling can be ascertained. A test is proposed in which "indicator" microorganisms would predict the likelihood of pharmaceutical formulations supporting growth.

Drug Contamination↗

Application of PCR for various neurotropic viruses on the diagnosis of viral meningitis.

BACKGROUND: Epidemiological studies have indicated that the majority of cases of aseptic meningitis result from viral infections. However, specific viral pathogens for aseptic meningitis can be identified in only some cases even if consistent conventional diagnostic methodologies rare used. OBJECTIVES: To clarify the etiological agents of aseptic meningitis by means of polymerase chain reaction (PCR) for various neurotropic viruses. STUDY DESIGN: Cerebrospinal fluid (CSF) samples were collected from 73 children suspected of having meningitis from November 1991 to December 1994. The samples were examined for infectious viruses by cell culture and for viral genomes by PCR. RESULTS AND CONCLUSIONS: Of 45 samples from patients diagnosed with aseptic meningitis, positive PCR results for enterovirus, mumps virus, cytomegalovirus, and varicella-zoster virus were obtained from respectively 25, 14, 1, and 1. Viral pathogens were thus identified in 41 (91.1%) of the 45 CSF samples. By the combination of PCR methods with conventional virological methods, the diagnosis of viral meningitis was established in 97.8% of the 45 cases. Our findings prove that the application of PCR methods is useful for etiological study of aseptic meningitis, and that the vast majority of cases of aseptic meningitis result from viral infection.

Cerebrospinal Fluid↗

Enteroviral polymerase chain reaction in the investigation of aseptic meningitis.

We used a nested polymerase chain reaction (nPCR) to seek evidence for enteroviruses in clinical samples from patients with symptoms of aseptic meningitis. When compared with conventional virus isolation methods on a total of 366 samples collected during 1994-1995, an increase in positivity from 6% to 27% was shown. The results indicate that nPCR would be a valuable aid to the laboratory diagnosis of enteroviral infections as it can detect those enteroviruses that cannot be identified by current isolation methods.

Adolescent↗

Penetration of cefpodoxime into uterine and vaginal secretions from postpartum women after a single oral dose of cefpodoxime proxetil.

We evaluated the usefulness of cefpodoxime proxetil (CPDX-PR) in the treatment of puerperal infection and obtained the following results. (i) The susceptibilities of 124 clinical isolates from 85 uterine lochia samples were determined. The MIC at which the growth of Streptococcus agalactiae, Escherichia coli, and Bacteroides fragilis isolates was inhibited by 90% (MIC90) was 0.39 micrograms/ml or less. The MIC90 for Staphylococcus aureus was 3.13 micrograms/ml. (ii) Seven puerperal women received 200 mg of CPDX-PR orally. The CPDX concentration in the lochias in the uterine cavity was not statistically different from that in the vagina, suggesting that the vaginal samples, which can be obtained more safely and aseptically, can be substituted for the uterine samples. The CPDX concentration in cubital venous blood reached a peak of 1.61 micrograms/ml at 3 hours after CPDX-PR administration. The CPDX concentration in the lochias gradually increased and reached a peak of 1.20 micrograms/ml in the uterine cavity and 1.27 micrograms/ml in the vagina at 5 h after drug administration and gradually declined thereafter. These results suggest that CPDX-PR, with its good transfer to the lochia and its potent antimicrobial activity, is a promising drug for the prophylactic and therapeutic treatment of puerperal infections caused by susceptible organisms.

Administration, Oral↗

Automatic diluter for bacteriological samples.

The described apparatus, carrying 190 tubes, allows automatic and aseptic dilution of liquid or suspended-solid samples. Serial 10-fold dilutions are programmable from 10(-1) to 10(-9) and are carried out in glass tubes with screw caps and split silicone septa. Dilution assays performed with strains of Escherichia coli and Bacillus stearothermophilus permitted efficient conditions for sterilization of the needle to be defined and showed that the automatic dilutions were as accurate and as reproducible as the most rigorous conventional dilutions.

Bacteriological Techniques↗

Nonantibiotic approach in control of bovine mastitis during dry period.

Twenty cows were used to determine effectiveness of a nonantibiotic method for control of intramammary infection during the dry period. Right mammary quarters of cows were infused with colchicine, endotoxin, or both at 1 day prior to or at the end of lactation. Left quarters were uninfused controls within group. Samples (n = 1105) of foremilk were obtained aseptically from quarters for bacteriological analysis. Quarters were sampled during the last week of lactation, early nonlactating period, prepartum, at parturition, and 1 and 2 wk postpartum. Primary pathogens isolated were esculin + streptococci, coagulase + staphylococci, and coliform bacteria. Isolation of primary pathogens was reduced 50% in right infused quarters during the 1st wk of involution as compared to left uninfused quarters. Thirteen of 80 quarters were infected at the end of lactation (6 right and 7 left), and 20 of 80 quarters (9 right and 11 left) were infected at calving. Sixteen of the 20 infections at calving were new during the dry period. Treatment reduced infection during early involution but failed to prevent new infection during the remainder of the dry period.

Animals↗