Enrichment Broth Cultures Research Articles

Natural history of Clostridioides difficile colonization and infection following new acquisition of carriage in healthcare settings: A prospective cohort study.

Limited information is available on the natural history of Clostridioides difficile colonization and infection in patients with new acquisition of C. difficile in healthcare settings. In 3 hospitals and affiliated long-term care facilities, we collected serial perirectal cultures from patients with no diarrhea on enrollment to identify new acquisition of toxigenic C. difficile carriage and determined the duration and burden of carriage. Asymptomatic carriage was defined as transient if only 1 culture was positive with negative cultures before and after or persistent if 2 or more cultures were positive. Clearance of carriage was defined as 2 consecutive negative perirectal cultures. Of 1,432 patients with negative initial cultures and at least 1 follow-up culture, 39 (2.7%) developed CDI without prior detection of carriage and 142 (9.9%) acquired asymptomatic carriage with 19 (13.4%) subsequently diagnosed with CDI. Of 82 patients analyzed for persistence of carriage, 50 (61.0%) had transient carriage and 32 (39.0%) had persistent carriage, with an estimated median of 77 days to clearance of colonization (range, 14 to 133 days). Most persistent carriers had a relatively high burden of carriage and maintained the same ribotype over time, whereas most transient carriers had a low burden of carriage detected only using broth enrichment cultures. In 3 healthcare facilities, 9.9% of patients acquired asymptomatic carriage of toxigenic C. difficile, and 13.4% were subsequently diagnosed with CDI. Most carriers had transient rather than persistent carriage and most patients developing CDI did not have prior detection of carriage.

1416. Assessing the potential for interspecies transmission of Clostridioides difficile on dairy farms

Background Clostridioides difficile (CD) can cause severe colitis in humans and many species of animals. It is thought that farm animals could be a reservoir for CD and that farm workers could therefore be at increased risk of colonization and infection with CD. While pigs and swine farm workers have been shown to be colonized with identical clones of CD, the zoonotic transmission of CD from animals to people has not been definitively demonstrated, and no studies have examined whether dairy farm workers, who are generally in closer contact with animals than swine farmers, are at increased risk of being colonized or infected with CD. The aim of this study was to assess whether dairy calves and farm workers harbored genetically similar isolates of CD.MethodsFirst, we validated a glove-juice protocol to detect CD spores on the hands of farm workers. Volunteers’ hands were inoculated with serially diluted suspensions of non-toxigenic CD organisms, and hand rinsates underwent broth enrichment and anaerobic culture. Second, we collected fecal samples from 5 randomly selected dairy calves (< 7 d of age) from each of 23 farms in southeastern Pennsylvania, northern Maryland, and Delaware. We focused specifically on dairy calves, as the prevalence of CD is highest in this age group. Third, using the glove-juice protocol, we collected hand rinsates from 38 dairy farm workers who work closely with calves. Only 4 of these workers were willing to submit fecal samples along with their hand rinsates. All fecal samples and hand rinsates underwent broth enrichment and anaerobic culture for CD.ResultsValidation of the glove juice protocol showed that CD could be recovered successfully from all hand rinsate dilutions (up to 10-6). When applied to farm workers, this method yielded CD in none of the hand rinsates (0%, 95% CI 0.0-92.2%). CD was also not detected in any of the human fecal samples. However, CD was detected from calf fecal samples on 10 farms (43.5%, 95% CI 20.8%-80.0%).ConclusionWhile the zoonotic transmission of CD cannot be ruled out, our results suggest that contact with dairy animals is not likely to be associated with an increased risk of acquiring CD via the fecal-oral route. The glove-juice protocol appears to be a useful tool for studying the epidemiology of CD in populations where obtaining fecal samples is difficult.Disclosures All Authors: No reported disclosures

High levels of toxigenic Clostridioides difficile contamination of hospital environments: a hidden threat in hospital-acquired infections in Kenya.

Introduction The contribution of Clostridioides difficile (formerly Clostridium difficile ) to the burden of hospital-associated infections (HAIs) remains undetermined in many African countries.Aim This study aimed to identify a sensitive and readily adaptable C. difficile detection assay and to evaluate the C. difficile HAI risk in Kenya.Methodology Sterile swabs in neutralizing buffer were used to sample equipment or surfaces that patients and clinical staff touched frequently. These swabs were either plated directly on chromogenic agar or cultured in an enrichment broth before plating. The swab suspensions, enrichment broth and plate cultures were screened by quantitative PCR (qPCR) to determine the most efficient detection method. The HAI risk was evaluated by testing the C. difficile -positive samples by qPCR for the A, B and binary toxins.Results C. difficile was detected on 4/57 (7.0 %) equipment and surfaces by direct culture. The additional enrichment step increased the detection rate 10-fold to 43/57 (75.4 %). In total, 51/57 (89.5 %) environmental samples were positive for C. difficile detected through either culture or qPCR. The genes encoding the primary toxins, tcdA and tcdB, were detected on six surfaces, while the genes encoding the binary toxins, cdtA and cdtB, were detected on 2/57 (3.5 %) and 3/57 (5.3 %) surfaces, respectively. Different C. difficile toxin gene profiles were detected: the tcdA+/tcdB− gene profile on 4/10 (40 %) high-touch surfaces, tcdA−/tcdB+ on 3/10 (30 %) surfaces, tcdA+/tcdB+/cdtA+/cdtB+ on 2/10 (20 %) surfaces and tcdA−/tcdB+/cdtB+ on one high-touch surface.Conclusion The widespread contamination of hospital environments by toxigenic C. difficile gives a strong indication of the high risk of C. difficile infections (CDIs). The two-step culture process described can easily be adapted for monitoring hospital environment contamination by C. difficile .

Development of a qPCR for the detection and quantification of Salmonella spp. in sheep feces and tissues.

Salmonella spp. are common causes of disease in intensive livestock production systems, and contamination of foodstuffs is of significant concern for public health. Therefore, the identification and quantification of Salmonella spp. is important for monitoring the level of fecal shedding or tissue colonization in infected animals and animal products. We developed and evaluated a quantitative PCR (qPCR) method on spiked sheep tissue and fecal samples for the detection and quantification of Salmonella spp. Without the use of a pre-enrichment step, the qPCR limit of detection (LOD) results for sheep fecal (4 × 104-6 × 103 cfu/g) and tissue (4 × 105-4 × 103 cfu/g) samples were not adequate for detection purposes. With the inclusion of a 6-h pre-enrichment step in buffered peptone water (BPW), the LOD was 9 cfu/g (2.57 × 101 copies/g) in sheep feces, and 5.4 cfu/g (3.22 copies/g) sheep tissue. Comparison of the 6-h BPW qPCR method with a 24-h mannitol-selenite-cystine broth enrichment culture method using spiked samples revealed a sensitivity of 91% and 92%, respectively, and a specificity of 100% for both methods. The correlation was significant between the quantity (copies/mL) of Salmonella spp. in BPW at 6 h and at 0 h, allowing semiquantitative analysis. Our results demonstrate that, following inclusion of a 6-h pre-enrichment step in BPW, qPCR is semiquantitative with improved LODs of Salmonella spp. in sheep fecal and tissue samples.

Diagnostic Accuracy of Presurgical Staphylococcus aureus PCR Assay Compared with Culture and Post-PCR Implementation Surgical Site Infection Rates

Nasal colonization with Staphylococcus aureus is a well-referenced risk factor for postoperative surgical site infections (SSIs). Our health care system that performs >40,000 surgeries per year assessed both the diagnostic accuracy of the BD MAX StaphSR assay (MAX StaphSR), a PCR-based test that detects and differentiates S. aureus and methicillin-resistant S. aureus (MRSA), compared with our standard of care culture and the subsequent clinical impact on SSIs 1 year after implementation. In addition, residual specimens were tested by broth-enriched culture. Performance parameters for all methods were determined using latent class analysis. Direct culture was the least sensitive for S. aureus (85.1%) and MRSA (76.7%), whereas the MAX StaphSR assay and broth-enriched culture had similar sensitivities (96.7%) for MRSA. Prospective assessment using MAX StaphSR during a 1-year, postimplementation period revealed a lower rate of SSIs per 100 targeted surgeries (0.3) compared with MRSA-only screening (1.10) and no screening (2.28) (P<0.05 for StaphSR versus MRSA-only screening and StaphSR versus no testing). MRSA and methicillin-sensitive S. aureus SSIs occurred equally (n = 14 each). The MAX StaphSR assay provided accurate detection of both S. aureus and MRSA nasal colonization in presurgical patients, allowing infection prevention measures, including presurgical prophylaxis, to be implemented in a timely and consistent manner to avoid SSIs.

Optimal detection of extended-spectrum β-lactamase producers, carbapenemase producers, polymyxin-resistant Enterobacterales, and vancomycin-resistant enterococci from stools

This study compared the impact of different approaches, namely, nonenrichment, nonselective enrichment, and selective (antibiotic-containing) enrichment steps, for detecting extended-spectrum β-lactamase producing Enterobacterales (ESBL-E), carbapenemase-producing Enterobacterales (CPE), polymyxin-resistant Enterobacterales (PMR-E), and vancomycin-resistant enterococci (VRE) from spiked stools. The use of a nonselective 18-h enrichment broth culture significantly improved the recovery rate of all types of resistant bacteria after their plating onto selective media. In addition, the detection of ESBL-E, CPE, PMR-E, and VRE was further improved when using an enrichment step using antibiotic-supplemented broths respectively supplemented with cefotaxime (0.1 μg/mL), ertapenem (0.1 μg/mL), colistin (0.5 μg/mL), and vancomycin (1 μg/mL). Therefore, we showed here that a screening strategy based on a selective broth enrichment step significantly contributes to an increased rate of detection of multidrug-resistant bacteria, which may be crucial in term of improvement of infection control.

Group B Streptococcus DNA Copy Numbers Measured by Digital PCR Correlates with Perinatal Outcomes.

Group B Streptococcus (GBS) is a one of the main causes of perinatal disease, yet the method for GBS detection, broth-enriched culture, is time-consuming and has low sensitivity and accuracy. We aimed to develop a GBS digital PCR (GBS-dPCR) assay for detecting GBS colonization. More rapid and accurate detection of GBS colonization could increase GBS diagnosis and treatment closer to delivery. A single-center, retrospective, case-controlled study was performed. A total of 182 rectovaginal swabs from pregnant women, who were undergoing prenatal screening by broth-enriched culture, were evaluated using GBS-dPCR targeting the cfb gene of GBS. Pregnant women with GBS colonization were followed up for correlation analysis between GBS DNA copy numbers and perinatal outcomes. The results of the GBS-dPCR assay were compared to those from the broth-enriched culture, which is the gold standard for GBS detection. The sensitivity and specificity of GBS-dPCR were 98% and 92.5%, respectively. By discrepant result analysis, the specificity of GBS-dPCR was raised to 97.4%. The incidence of premature rupture of membrane (PROM) and neonatal infection were statistically significantly positively correlated with GBS DNA copy numbers. GBS-dPCR has the advantage of directly detecting GBS colonization from swabs with high specificity and sensitivity, while reducing turnaround time (<4 h). Analysis of clinical samples with GBS-dPCR shows that GBS DNA copy numbers are positively correlated with the incidence of PROM and neonatal infection, suggesting that dPCR is a promising method for detection of GBS colonization during pregnancy.

Multi-Year Persistence of Verotoxigenic Escherichia coli (VTEC) in a Closed Canadian Beef Herd: A Cohort Study.

In this study, fecal samples were collected from a closed beef herd in Alberta, Canada from 2012 to 2015. To limit serotype bias, which was observed in enrichment broth cultures, Verotoxigenic Escherichia coli (VTEC) were isolated directly from samples using a hydrophobic grid-membrane filter verotoxin immunoblot assay. Overall VTEC isolation rates were similar for three different cohorts of yearling heifers on both an annual (68.5 to 71.8%) and seasonal basis (67.3 to 76.0%). Across all three cohorts, O139:H19 (37.1% of VTEC-positive samples), O22:H8 (15.8%) and O?(O108):H8 (15.4%) were among the most prevalent serotypes. However, isolation rates for serotypes O139:H19, O130:H38, O6:H34, O91:H21, and O113:H21 differed significantly between cohort-years, as did isolation rates for some serotypes within a single heifer cohort. There was a high level of VTEC serotype diversity with an average of 4.3 serotypes isolated per heifer and 65.8% of the heifers classified as “persistent shedders” of VTEC based on the criteria of >50% of samples positive and ≥4 consecutive samples positive. Only 26.8% (90/336) of the VTEC isolates from yearling heifers belonged to the human disease-associated seropathotypes A (O157:H7), B (O26:H11, O111:NM), and C (O22:H8, O91:H21, O113:H21, O137:H41, O2:H6). Conversely, seropathotypes B (O26:NM, O111:NM) and C (O91:H21, O2:H29) strains were dominant (76.0%, 19/25) among VTEC isolates from month-old calves from this herd. Among VTEC from heifers, carriage rates of vt1, vt2, vt1+vt2, eae, and hlyA were 10.7, 20.8, 68.5, 3.9, and 88.7%, respectively. The adhesin gene saa was present in 82.7% of heifer strains but absent from all of 13 eae+ve strains (from serotypes/intimin types O157:H7/γ1, O26:H11/β1, O111:NM/θ, O84:H2/ζ, and O182:H25/ζ). Phylogenetic relationships inferred from wgMLST and pan genome-derived core SNP analysis showed that strains clustered by phylotype and serotype. Further, VTEC strains of the same serotype usually shared the same suite of antibiotic resistance and virulence genes, suggesting the circulation of dominant clones within this distinct herd. This study provides insight into the diverse and dynamic nature of VTEC populations within groups of cattle and points to a broad spectrum of human health risks associated with these E. coli strains.

Development of an optimized broth enrichment culture medium for the isolation of Clostridium difficile

Clostridium difficile is a spore forming bacterium and the leading cause of colitis and antibiotic associated diarrhoea in the developed world. Effective recovery of spores, particularly in low numbers, is imperative to obtain accurate prevalence data, due to the low number of spores found within non-clinical samples (<20/ml). Through comparison of C. difficile enrichment media, this study showed the importance of selecting an effective enrichment media. Commonly used broths, such as Cooked Meat broth, promote significantly less growth than other available broths such as Brain Heart Infusion broth, BHI. The optimization of BHI using selective antibiotics, moxalactam and norfloxacin, and sodium taurocholate at a concentration of 0.4%, allowed for high growth rate (0.465 h-1), short lag times (<14 h) and recovery of spores at low concentrations. The optimized broth, designated BHIMN-T, out-performed other commonly used broths so can be recommended for future studies.

Comparing the Yield of Staphylococcus aureus Recovery with Static versus Agitated Broth Incubation.

Given the lack of standardization of methodologies for microbial recovery from built environments, we sought to compare the yield of Staphylococcus aureus with a broth enrichment method when incubated in agitated versus static conditions. Five unique strains of S. aureus at five different concentrations were cultured to compare direct plating, agitated broth enrichment, and static broth enrichment culture methods. All samples were incubated at 35° in ambient air. The lowest concentration recovered across three replicates and five strains did not differ between culture methods (Fisher's exact test, p=0.50); notably, recovery of S. aureus was equivalent between static and agitated broth incubation. When broth enrichment was used (both static and agitated), the burden of S. aureus growth was higher (by semiquantitative assessment of 4-quadrant streaking) compared to the direct plating culture method. Optimizing strategies for microbial recovery is essential, particularly in areas of lower biomass, given the paucity of research concerning microbial communities of built environments. The results of this study, in conjunction with other experiments investigating microbiomes of built environments, can help inform protocols for standardizing culturing methods within built environments.

Home Environmental Contamination Is Associated with Community-associated Methicillin-resistant Staphylococcus aureus Re-colonization in Treated Patients

BackgroundStrategies to interrupt household transmission of community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) that target human colonization show mixed results. Our aim was to determine whether home environmental contamination and pet carriage with MRSA were associated with re-colonization or persistent colonization of index patients diagnosed with CA-MRSA skin or soft-tissue infection (SSTI).MethodsIndex patients from a randomized controlled trial (NCT00966446) that tested household-wide decolonization of people were eligible to participate in this substudy. Before randomization, eight environmental sites and all pets were sampled in the home. Patients were treated by their physician for the initial SSTI between diagnosis (visit 0) and the home visit (visit 1). They provided swabs every 2 weeks for 3 months (7 visits). After broth-enrichment culture, MRSA isolates were PCR-confirmed and spa-typed.ResultsOf 88 index patients recruited from the main trial, 64 (73%) provided swabs for ≥3 visits and were included in this analysis. At visit 1, 41 (64%) households were MRSA contaminated and 6 (9%) had MRSA-positive pet(s). All MRSA-positive pets lived in homes with MRSA environmental contamination. After visit 1, 42 (66%) index patients and their household members were block-randomized to nasal mupirocin and chlorhexidine body wash decolonization. Thirty-seven (58%) index patients had two consecutive negative swabs (de-colonized); 13 (35%) of these later were MRSA-positive (re-colonized). Patients with home contamination had higher rates of re-colonization than those without (Cox proportional hazard ratio 6.0 [95% CI: 1.2, 30.6], P < 0.03). Persistent colonization (all or all but one swab positive) was identified in 6 (9%) of index patients and was associated with identification of matching spa-types in environmental and subsequent human MRSA isolates (P < 0.05).ConclusionIn patients with MRSA SSTI, MRSA-contaminated homes, and potentially MRSA-positive pets, are associated with re-colonization and persistent colonization. Future studies are needed to determine whether environmental decontamination can improve the success of household decolonization interventions.Disclosures All authors: No reported disclosures.

Modified use of real-time PCR detection of group B Streptococcus in pregnancy.

The CDC recommends antenatal screening of vaginal/rectal samples for Streptococcus agalactiae at 35-37 weeks' gestation, with intra-partum antibiotic prophylaxis for positive cases. We tested a modified use of the Cepheid Xpert GBS real-time PCR kit on enrichment cultures from 554 vaginal/rectal swabs compared to the current subculturing gold standard method. Swabs were inoculated on polymyxin nalidixic acid agar plates, and Todd-Hewitt enrichment broth cultures were examined daily for growth. Todd-Hewitt broth culture was also used for Xpert GBS. There was 92.06 % agreement between the subculture and PCR methods. Sensitivity of Xpert GBS was 100 %, specificity was 89.40 %, positive predictive value was 75.96 % and negative predictive value was 100 %. Colonization incidence was higher with younger (≤24 years) or older (≥35 years) maternal age. Modified use of the Cepheid Xpert GBS would assist rapid diagnosis of S. agalactiae colonization and facilitate timely and appropriate assignment to intra-partum antibiotic prophylaxis.

Screening for group B Streptococcus (GBS) at labour onset using PCR: accuracy and potential impact – a pilot study

This pilot study assessed the diagnostic accuracy and potential impact of a rapid PCR-based screening test for the detection of group B Streptococcus (GBS) at the onset of labour for the purpose of optimising intrapartum antibiotic prophylaxis (IAP). Vaginal and rectal swabs from a convenience sample of 158 women were analysed by conventional broth-enriched culture and a rapid PCR test. Overall, GBS carriage was 18.98% by culture and 19.62% by PCR. PCR for the detection of GBS had a sensitivity of 93.1%, specificity of 96.67% and area under the curve (AUC) of 0.95. Only 19.3% GBS-positive women received IAP. Three-fourths of babies born to GBS-positive mothers did not receive surveillance for early-onset GBS disease. Of the women who received IAP, only 32.5% were GBS carriers. Seventy-four percent of the GBS-positive mothers delivered more than 5 h after recruitment, which gives adequate swab to delivery interval for appropriate antibiotic prophylaxis in labour.Impact statementWhat is already known about this subject: Appropriate intra-partum treatment of colonized mothers reduces the risk of GBS transmission to neonates. Universal ante partum screening of pregnant women or IAP based on risk factors in labour for GBS prevention fail to accurately identify and treat the woman who actually harbors GBS in the birth canal in labour. A PCR based rapid test, allows for real-time assessment of GBS carriage in labour.What this study adds: This study highlights the fact that a large number of GBS carriers in labour, who could potentially infect their babies, do not receive IAP, and most of their babies do not receive added surveillance in the neonatal period for EOGBS disease. It also confirms that PCR testing at onset of labour is a highly sensitive and reliable test that identifies the women who are GBS carriers in labour and hence need IAP.What the implications are of these findings for clinical practice and/or further research: Timely provision of IAP for the appropriate woman is possible by adopting universal GBS screening at the onset of labor using GBS-PCR. This would involve additional costs to health care facilities and added work to laboratory personnel.

Occurrence of Plasmid- and Chromosome-Carried mcr-1 in Waterborne Enterobacteriaceae in China.

The aim of this study was to investigate the prevalence of the polymyxin resistance gene mcr-1 in Enterobacteriaceae from environmental water sources in Hangzhou, China. Colistin-resistant bacteria were isolated from environmental water samples using an enrichment broth culture method, were screened for mcr-1, and then were analyzed for the location and transferability of mcr-1 Isolates positive for mcr-1 were further examined to determine their susceptibility profiles and were screened for the presence of additional resistance genes. Twenty-three mcr-1-positive isolates (16 Escherichia coli, two Citrobacter freundii, two Klebsiella oxytoca, two Citrobacter braakii, and one Enterobacter cloacae) were isolated from 7/9 sampling locations; of those, eight mcr-1-positive isolates also contained β-lactamase-resistance genes, eight contained qnrS, and 10 contained oqx No mcr-2-positive isolates were identified. The majority of isolates demonstrated a low to moderate level of colistin resistance. Transconjugation was successfully conducted from 14 of the 23 mcr-1-positive isolates, and mcr-1 was identified on plasmids ranging from 60 to 220 kb in these isolates. Conjugation and hybridization experiments revealed that mcr-1 was chromosome-borne in only three isolates. Pulsed-field gel electrophoresis showed that the majority of E. coli isolates belonged to different clonal lineages. Multilocus sequence typing analysis revealed that sequence type 10 (ST10) was the most prevalent, followed by ST181 and ST206. This study demonstrates the utility of enrichment broth culture for identifying environmental mcr-1-positive isolates. Furthermore, it highlights the importance of responsible agriculture and clinical use of polymyxins to prevent further widespread dissemination of polymyxin-resistant pathogens.

Multiplexed Single Intact Cell Droplet Digital PCR (MuSIC ddPCR) Method for Specific Detection of Enterohemorrhagic E. coli (EHEC) in Food Enrichment Cultures.

Foodborne illness attributed to enterohemorrhagic E. coli (EHEC), a highly pathogenic subset of Shiga toxin-producing E. coli (STEC), is increasingly recognized as a significant public health issue. Current microbiological methods for identification of EHEC in foods often use PCR-based approaches to screen enrichment broth cultures for characteristic gene markers [i.e., Shiga toxin (stx) and intimin (eae)]. However, false positives arise when complex food matrices, such as beef, contain mixtures of eae-negative STEC and eae-positive E. coli, but no EHEC with both markers in a single cell. To reduce false-positive detection of EHEC in food enrichment samples, a Multiplexed, Single Intact Cell droplet digital PCR (MuSIC ddPCR) assay capable of detecting the co-occurrence of the stx and eae genes in a single bacterial cell was developed. This method requires: (1) dispersal of intact bacteria into droplets; (2) release of genomic DNA (gDNA) by heat lysis; and (3) amplification and detection of genetic targets (stx and eae) using standard TaqMan chemistries with ddPCR. Performance of the method was tested with panels of EHEC and non-target E. coli. By determining the linkage (i.e., the proportion of droplets in which stx and eae targets were both amplified), samples containing EHEC (typically greater than 20% linkage) could be distinguished from samples containing mixtures of eae-negative STEC and eae-positive E. coli (0–2% linkage). The use of intact cells was necessary as this linkage was not observed with gDNA extracts. EHEC could be accurately identified in enrichment broth cultures containing excess amounts of background E. coli and in enrichment cultures derived from ground beef/pork and leafy-green produce samples. To our knowledge, this is the first report of dual-target detection in single bacterial cells using ddPCR. The application of MuSIC ddPCR to enrichment-culture screening would reduce false-positives, thereby improving the cost, speed, and accuracy of current methods for EHEC detection in foods.

Group B streptococcus colonisation in pregnant women at Dr. George Mukhari Hospital, South Africa

The aim of the study was to estimate group B streptococcus (GBS) colonisation in pregnant mothers using selective enrichment broth and solid media for culturing GBS. Vaginal and rectal swabs were collected from 413 pregnant women for GBS culture at recruitment stage. Direct plating and enrichment broth culture methods were compared by using the same swab samples. The swabs were cultured on colistin nalidixic agar (CNA) plate and incubated at 37°C and examined after 18-24 h. The samples which were culture negative on a CNA agar plate were then inoculated into a Todd-Hewitt enrichment broth to recover any GBS present that was not recovered on the solid agar. With the CNA agar plate, the samples were cultured separately to enable identification of colonised sites such as vaginal sites or rectal sites. Rectal and vaginal swabs were inoculated into Todd-Hewitt enrichment broth at the same time in the same tube. The GBS colonisation rate in pregnant women was 30.9% (128/413). The CNA agar plate recovered 45.3% (58/128) of the GBS isolates, whereas 54.7% (70/128) isolates were recovered from Todd-Hewitt broth. Pregnant women of various ages were found to be at risk of GBS colonisation. The colonisation rate was however highest among women of 25–29 age groups as compared with other age groups. Detection of group B streptococcus improved when both rectal and vaginal swabs were collected for laboratory analysis. The simultaneous use of Todd-Hewitt broth and CNA plate also improved the yield of group B streptococcus.

A Novel Risk Factor Associated With Colonization by Carbapenemase-Producing Enterobacteriaceae: Use of Proton Pump Inhibitors in Addition to Antimicrobial Treatment

OBJECTIVE To study the association between gastrointestinal colonization of carbapenemase-producing Enterobacteriaceae (CPE) and proton pump inhibitors (PPIs). METHODS We analyzed 31,526 patients with prospective collection of fecal specimens for CPE screening: upon admission (targeted screening) and during hospitalization (opportunistic screening, safety net screening, and extensive contact tracing), in our healthcare network with 3,200 beds from July 1, 2011, through December 31, 2015. Specimens were collected at least once weekly during hospitalization for CPE carriers and subjected to broth enrichment culture and multiplex polymerase chain reaction. RESULTS Of 66,672 fecal specimens collected, 345 specimens (0.5%) from 100 patients (0.3%) had CPE. The number and prevalence (per 100,000 patient-days) of CPE increased from 2 (0.3) in 2012 to 63 (8.0) in 2015 (P<.001). Male sex (odds ratio, 1.91 [95% CI, 1.15-3.18], P=.013), presence of wound or drain (3.12 [1.70-5.71], P<.001), and use of cephalosporins (3.06 [1.42-6.59], P=.004), carbapenems (2.21 [1.10-4.48], P=.027), and PPIs (2.84 [1.72-4.71], P<.001) in the preceding 6 months were significant risk factors by multivariable analysis. Of 79 patients with serial fecal specimens, spontaneous clearance of CPE was noted in 57 (72.2%), with a median (range) of 30 (3-411) days. Comparing patients without use of antibiotics and PPIs, consumption of both antibiotics and PPIs after CPE identification was associated with later clearance of CPE (hazard ratio, 0.35 [95% CI, 0.17-0.73], P=.005). CONCLUSIONS Concomitant use of antibiotics and PPIs prolonged duration of gastrointestinal colonization by CPE. Infect Control Hosp Epidemiol 2016;1418-1425.

Performance of the BD MAX™ instrument with Check-Direct CPE real-time PCR for the detection of carbapenemase genes from rectal swabs, in a setting with endemic dissemination of carbapenemase-producing Enterobacteriaceae

Carbapenemase-producing Enterobacteriaceae (CPE) represent an increasing public health issue and the early detection of colonization by CPE can help the implementation of infection control measures among inpatients. In this study, BD MAX Check-Direct CPE screen, with two different Master Mixes (BDMix and CPMix), using the automatic BD MAX™ instrument, was evaluated for the detection of blaKPC, blaOXA-48, blaVIM and blaNDM genes, in comparison to selective broth enrichment and direct culture from rectal swabs. Among a total of 557 rectal swabs samples, 29 (5.2%) tested positive for CPE (23 for blaKPC, 5 for blaVIM and one for blaOXA-48). The sensitivity, specificity, positive and negative likelihood ratios values were 93.1%, 97.3%, 34.5 and 0.07, for BMix, and 100%, 97.1 %, 34.5 and 0 for CPMix, respectively. Five samples were positive with molecular methods only. The turn-around time was reduced from 18-24 hours (direct culture), or 48 h (broth enrichment) to only 3 h.

Group B streptococcus colonisation in pregnant women at Dr. George Mukhari Hospital, South Africa

The aim of the study was to estimate group B streptococcus (GBS) colonisation in pregnant mothers using selective enrichment broth and solid media for culturing GBS. Vaginal and rectal swabs were collected from 413 pregnant women for GBS culture at recruitment stage. Direct plating and enrichment broth culture methods were compared by using the same swab samples. The swabs were cultured on colistin nalidixic agar (CNA) plate and incubated at 37°C and examined after 18-24 h. The samples which were culture negative on a CNA agar plate were then inoculated into a Todd-Hewitt enrichment broth to recover any GBS present that was not recovered on the solid agar. With the CNA agar plate, the samples were cultured separately to enable identification of colonised sites such as vaginal sites or rectal sites. Rectal and vaginal swabs were inoculated into Todd-Hewitt enrichment broth at the same time in the same tube. The GBS colonisation rate in pregnant women was 30.9% (128/413). The CNA agar plate recovered 45.3% ...

Diagnosis and Treatment of Sialectasis in a Domestic Rabbit (Oryctolagus cuniculus)

An approximately 10-year-old female spayed rabbit (Oryctolagus cuniculus) was presented with a subcutaneous mass of the ventral submandibular and cervical region. Initial culture results from fine-needle aspirates revealed Gram-positive cocci; cultures following antibiotic therapy identified Cryptococcus spp. (not Cryptococcus neoformans), Actinomyces spp., unidentified yeast, and alpha-hemolytic Streptococcus spp. The mass did not resolve following antibiotic treatment and the rabbit was referred for further evaluation. A computed tomography scan revealed a large (9 × 5 × 5cm3) fluid-filled rim-enhancing mass in the left submandibular and intermandibular region that extended caudally into the cervical region. The mass and attached skin were surgically excised, and the tissue was submitted for histopathology and culture. The histopathologic diagnosis of the mass was severe, cystic, heterophilic, lymphoplasmacytic dochitis, and cellulitis consistent with sialectasis. Enrichment broth culture grew Candida albicans. The rabbit made a rapid postoperative recovery. No signs of recurrence were noted at multiple examinations until 1 year following the surgical procedure, when a small (0.5 × 0.5 × 0.5cm3) mass was identified at the previous location and contained fluid consistent with saliva. Culture of this fluid was negative. About 1 year following the initial surgery, computed tomography images were consistent with mild fluid dilation of the remaining tract of the salivary duct; surgical exploration and complete excision of the duct and salivary gland were recommended but declined by the owner.