Trends of cervical tumours amongst women from perspectives of demographic, socioeconomic and geographic indicators: retrospective ecological study in Czechia.

Introduction: For many infectious diseases, women are at higher risk and have a more severe disease course than men for many reasons, including biological differences, social inequalities, and restrictive cultural norms. The study focuses on infections with human papillomaviruses (HPV) in the form of cervical cancer as a gender-specific disease. The main goal is to evaluate cervical tumour incidence trends in the Czech female population in the HPV vaccination period 2012-2020 in relation to selected demographic, socioeconomic, and geographic indicators. Methods: This is a retrospective ecological study. Data from publicly available databases about the incidence and mortality of cervical tumours (C53 Malignant neoplasm of cervix uteri, D06 Carcinoma in situ of cervix uteri according to ICD 10) and HPV vaccination rate were analysed and compared with demographic, socioeconomic and territorial data. Associations were searched using correlation analysis. Results: There was a decreasing trend in the incidence of cervical cancer in the observed period. Regarding cervical tumours (C53, D06) and malignant neoplasm of cervix uteri incidence (C53), the decrease was approximately 11 and 20%, respectively. Differences between regions were observed in incidences and vaccination rates. Based on correlation analysis, indicators connected with urban/rural aspects, such as a share of urban population and population density, were statistically significant. The indicators related to higher cervical cancer incidence are the high unemployment rate of women, the high number of divorces, the high number of abortions, the high share of the urban population, the high number of students, and the high number of women with only primary education. On the other hand, the indicators related to lower cervical cancer incidence are the high gross domestic product (GDP), the high average gross monthly wage per employee, the high employment rate of women, the higher average age of mothers at birth, and the high number of women with tertiary education. Conclusion: Results underline the problem of economically disadvantaged regions and families. Increasing vaccination rates, promoting regular screening for cervical cancer, and supporting awareness in the population, especially in regions with higher incidence rates, should be priorities for public health efforts.

Potential determinants of vaccine hesitancy among celiac disease patients: a single cohort analysis.

Introduction: Though researchers and scholars have greatly emphasized addressing the influencing factors of vaccination hesitancy, little attention has been paid to patients with celiac disease. Addressing the variables hampering attitudes might help direct appropriate patient advocacy and doctor-patient communication endeavors to encourage vaccination among celiac disease patients. The present investigation seeks to explore the coverage against vaccine-preventable diseases, vaccination attitudes, and related possible factors among celiac disease patients in the Pakistani setting. Methods: A self-reported online survey was conducted in Islamabad, Pakistan, for celiac disease patients aged 18 and above. The questionnaire was completed by 226 participants, with a response rate of 43.8%. The influencing variables for vaccination hesitancy were examined, and 95% confidence intervals for the crude and adjusted odds ratios were computed. Results: Among the study population, the majority were females, with a ratio of 75.66%. A prominent proportion of 69.03% was observed for influenza vaccination, while 39.82% were unable to recall all of the vaccinations they had previously received. Only 7% of the patients were considered to have a negative attitude toward vaccination, compared to an estimated 76% who were in favor of it. The significantly positive influencing factors observed toward vaccination were being well-educated (graduate, master, or above), possible recurrence of vaccine-preventable diseases with declining vaccination coverage (adjusted OR: 13.36), and increased confidence in vaccines from health care experts compared to electronic media (adjusted OR: 8.41). Contrarily, practicing complementary and alternative medicines (adjusted OR: 5.59), willingness to get vaccinated again in the future (adjusted OR: 15.59), and prior negative perspectives (adjusted OR: 1.01) were the determinants with a significant negative association. Discussion: In conclusion, the outcomes of the current work raise the possibility that health practitioners may be accountable for inappropriately prescribing vaccines to this demographic since 77% of the participants had a favorable attitude toward vaccination. These findings could serve as a springboard for creating targeted immunization efforts to raise vaccination coverage against vaccine-preventive diseases among celiac disease patients.

Screening of Antibiotic and Virulence Genes from Whole Genome Sequenced Cronobacter sakazakii Isolated from Food and Milk-Producing Environments.

The objective of this study was to use whole-genome sequencing (WGS) to screen for genes encoding for antibiotic resistance, fitness and virulence in Cronobacter sakazakii strains that had been isolated from food and powdered-milk-producing environments. Virulence (VGs) and antibiotic-resistance genes (ARGs) were detected with the Comprehensive Antibiotic Resistance Database (CARD) platform, ResFinder and PlasmidFinder tools. Susceptibility testing was performed using disk diffusion. Fifteen presumptive strains of Cronobacter spp. were identified by MALDI-TOF MS and ribosomal-MLST. Nine C. sakazakii strains were found in the meningitic pathovar ST4: two were ST83 and one was ST1. The C. sakazakii ST4 strains were further distinguished using core genome MLST based on 3678 loci. Almost all (93%) strains were resistant to cephalotin and 33% were resistant to ampicillin. In addition, 20 ARGs, mainly involved in regulatory and efflux antibiotics, were detected. Ninety-nine VGs were detected that encoded for OmpA, siderophores and genes involved in metabolism and stress. The IncFIB (pCTU3) plasmid was detected, and the prevalent mobile genetic elements (MGEs) were ISEsa1, ISEc52 and ISEhe3. The C. sakazakii isolates analyzed in this study harbored ARGs and VGs, which could have contributed to their persistence in powdered-milk-producing environments, and increase the risk of infection in susceptible population groups.

A comparison of heart failure patients with reduced ejection fraction in the Moravian Midlands Registry with the LCZ696 patients in the Paradigm-HF trial.

BACKGROUND AND AIMS: There are limited data on real clinical practice in heart failure patients in the Czech Republic. We analysed the clinical parameters from the Moravian Midlands Registry (MMR) and compared them to LCZ696 patients in the Paradigm-HF trial. The Moravian Midlands Registry is a retrospective patient database from two outpatient cardiology centres in the Czech Republic. The Paradigm-HF is a large-scale prospective randomized multicentre trial with more than 8000 individuals with stabilized chronic heart failure. METHODS: A retrospective analysis of heart failure with reduced ejection fraction patients from two outpatient cardiology centres in the Czech Republic from October 2016 to December 2019. RESULTS: Patients in the MMR were younger (60.5 ± 10.7 vs 63.8 ± 11.5 years, P<0.05), had a higher body mass index (30.3 ± 5.0 vs 28.1 ± 5.5, P<0.05) and higher serum creatinine level (101.9 ± 36.0 vs 99.9 ± 26.5 µmol/L, P<0.05). MMR patients had lower left ventricular ejection fraction (27.8 ± 6.9 vs 29.6 ± 6.1%, P<0.05). The serum N-terminal pro-B-type natriuretic peptide, [2563.5 (377-3536) vs 1631 (885-3154), was non significantly higher P=0.07]. Pharmacotherapy use differed for mineralocorticoid antagonist (91.4% in MMR vs 54.2% in Paradigm-HF), and digoxin (13.5% vs 29.2%). Beta-blocker use was similar (96.2% vs 93.1%) as was angiotensin-converting enzyme (ACE) inhibitors - (71.2% vs 78.0%) and angiotensin-receptor blockers - ARB (27.9% vs 22.2%). Dosages of the commonly used ACE inhibitors at the screening visit (Paradigm-HF) / before angiotensin receptor-neprilysin inhibitor administration (MMR) differed significantly only for ramipril (7.0 ± 3.1 mg vs 4.8 ± 2.9 mg, P<0.05), dosages of ARB were - losartan (67.1 ± 30.2 vs 39.6 ± 32.0 mg, P=0.09) and valsartan (181.5 ± 71.1 vs 130.9 ± 82.2 mg, P=0.07). There was a substantial difference in device-based therapy (ICD in 60.6%, CRT 25.9% in MMR vs 14.9% and 7.0% in Paradigm-HF). CONCLUSION: The differences between the groups for the majority of clinical parameters compared were minimal, except for younger age, higher body mass index and serum creatinine level and lower left ventricular ejection fraction and substantially lower dosage of administered ramipril prior to commencing sacubitril/valsartan therapy. There was a higher prevalence of implantable cardioverter-defibrillators (ICD) and cardiac resynchronization therapy (CRT) in the MMR group.

Predominance of Acinetobacter pseudolwoffii among Acinetobacter species in domestic animals in the Czech Republic.

The aim of this study was to map the spectrum of microorganisms belonging to the genus Acinetobacter in domestic animals with a specific focus on the prevalence of Acinetobacter pseudolwoffii. Additionally, the susceptibility of isolates to antimicrobial agents was determined. In the period from January 1, 2014, to August 31, 2015, a total of 9 544 samples originating from gross lesions and pathological processes of animals exhibiting clinical symptoms of the disease were examined across 41 districts in the Czech Republic. The examinations were carried out using culture methods involving meat-peptone blood agar and Endo agar under aerobic conditions at a temperature of 37 ± 1 °C for 18-24 hours. Isolates were confirmed using molecular phenotypic method MALDI-TOF MS with the MBT Compass Library Revision L 2020 covering 3 239 species/entries (9 607 MSP) from Bruker Daltonics company. Out of the 108 isolates (prevalence 1.13%), 14 species of Acinetobacter spp. were identified, with 5 isolates remaining unclassified as species. A. pseudolwoffii was the predominant species isolated in 25 cases (prevalence 0.26%). Antimicrobial susceptibility was determined for 12 antimicrobials by the disc diffusion method, with A. pseudolwoffii isolates exhibiting the lowest susceptibility to ceftazidime (32%) and co-trimoxazole (60%).

Does the intubation timeline affect the in-hospital mortality of COVID-19 patients? A retrospective cohort study.

Background: Effective strategies for managing coronavirus disease 19 (COVID-19) patients suffering from acute respiratory distress are constantly evolving. The timeline and threshold for transitioning from non-invasive ventilation to intermittent mandatory ventilation in critical cases who develop COVID-19-related respiratory distress are undetermined. The present research intends to investigate if emergency room intubations in COVID-19 patients affect mortality. Methods: Between January 1, 2021 and June 30, 2021, we retrospectively reviewed chart analysis on all patients with confirmed positive COVID-19 screening and who underwent endotracheal intubation. Depending on when the intubation was performed; early in the emergency room or delayed outside the emergency room, patients were separated into two cohorts. In addition to comorbid clinical manifestations, the quick sequential organ failure assessment (qSOFA) score, and in-hospital mortality were all recorded as demographic and clinical information. Results: Fifty-eight of the 224 corona-positive patients who underwent intubation had their intubations performed in the emergency room. Age, sex, alcohol use, and smoking status did not significantly differ between the two categories at the baseline. The mean qSOFA score was higher in the early intubation cohort (3.5; p < 0.000) along with more underlying comorbidities (3.0; p < 0.000). When compared to the late intubation cohort (45.78%), patients treated with early intubation had a significantly greater death rate (67.24%). Conclusion: In summary, we discovered that patients who underwent intubation in the emergency units exhibited a high quick SOFA score as well as maximum co-morbid conditions than patients intubated somewhere else in the hospital. The findings of our investigation imply that intubating patients too early might be risky.

Effectiveness of anti-epidemic measures on ensuring indoor air quality of cleanrooms in a tertiary care hospital.

OBJECTIVES: Maintaining cleanrooms in a hospital is a key part of preventing healthcare-associated infections (HAIs). The aim of this work was to verify the effectiveness of anti-epidemic measures to ensure air quality in cleanrooms in a tertiary care hospital. METHODS: The study was based on the application of anti-epidemic measures and verification of their effectiveness by regular indoor air monitoring between the years 2014-2019. Monitoring was performed according to a recommended procedure based on European standards for cleanrooms. RESULTS: The results demonstrate a reduction in the number of airborne particles and the number of colonies, as well as the elimination of fungi and conditioned pathogens in air samples. CONCLUSIONS: The authors emphasize the importance of established anti-epidemic measures and regular monitoring for the prevention of HAIs.

Genomic Characterization of Cronobacter spp. and Salmonella spp. Strains Isolated From Powdered Infant Formula in Chile.

This study characterized five Cronobacter spp. and six Salmonella spp. strains that had been isolated from 155 samples of powdered infant formula (PIF) sold in Chile and manufactured in Chile and Mexico in 2018-2020. Two strains of Cronobacter sakazakii sequence type (ST) ST1 and ST31 (serotypes O:1 and O:2) and one strain of Cronobacter malonaticus ST60 (O:1) were identified. All Salmonella strains were identified as Salmonella Typhimurium ST19 (serotype O:4) by average nucleotide identity, ribosomal multilocus sequence typing (rMLST), and core genome MLST (cgMLST). The C. sakazakii and C. malonaticus isolates were resistant to cephalothin, whereas the Salmonella isolates were resistant to oxacillin and ampicillin. Nineteen antibiotic resistance genes were detected in the C. sakazakii and C. malonaticus isolates; the most prevalent were mcr-9.1, blaCSA , and blaCMA . In Salmonella, 30 genes encoding for aminoglycoside and cephalosporin resistance were identified, including aac(6')-Iaa, ß-lactamases ampH, ampC1, and marA. In the Cronobacter isolates, 32 virulence-associated genes were detected by WGS and clustered as flagellar proteins, outer membrane proteins, chemotaxis, hemolysins, invasion, plasminogen activator, colonization, transcriptional regulator, survival in macrophages, use of sialic acid, and toxin-antitoxin genes. In the Salmonella strains, 120 virulence associated genes were detected, adherence, magnesium uptake, resistance to antimicrobial peptides, secretion system, stress protein, toxin, resistance to complement killing, and eight pathogenicity islands. The C. sakazakii and C. malonaticus strains harbored I-E and I-F CRISPR-Cas systems and carried Col(pHHAD28) and IncFIB(pCTU1) plasmids, respectively. The Salmonella strains harbored type I-E CRISPR-Cas systems and carried IncFII(S) plasmids. The presence of C. sakazakii and Salmonella in PIF is a health risk for infants aged less than 6 months. For this reason, sanitary practices should be reinforced for its production and retail surveillance.

Seroprevalence of Measles Antibodies in the Population of the Olomouc Region, Czech Republic-Comparison of the Results of Four Laboratories.

OBJECTIVES: Although the incidence of measles has decreased globally since the introduction of regular vaccination, its frequency has increased again in recent years. The study is focused on data from the Olomouc Region in the Czech Republic analyzed in four laboratories. The obtained results were compared with already published data. METHODS: The data were provided by individual laboratories in an anonymized form-age at the time of the examination, sex, and result of test. Samples were collected between June 2018 and September 2019 and evaluated on the scale positive-borderline-negative. RESULTS: A total of 7962 sera samples were evaluated using three different methods-two types of ELISA tests and CLIA. Positive result was issued in a total of 62.6 percent of samples, but the results of individual laboratories varied widely from 55.5 to 70.8 percent. However, the same trend with the highest levels of antibodies in people born before beginning of vaccination was observed. CONCLUSIONS: Data show significantly different results depending on the individual laboratories and the detection kits used. The underestimation of the proportion of positive results can cause problems in selecting individuals for revaccination with a live vaccine, which may fail in weakly positive individuals.

COVID-19 vaccine reluctance and possible driving factors: A comparative assessment among pregnant and non-pregnant women.

The coronavirus disease of 2019 (COVID-19) constitutes a serious threat to pregnant women. One of the key strategies for preventing and managing the COVID-19 epidemic is vaccination. Herd immunity is significantly hampered by COVID-19 vaccine reluctance, which poses a potential threat to population health. Therefore, the present work intends to ascertain the incidence and severity of COVID-19 vaccine hesitancy among Pakistani pregnant women, the determinants driving their decision, and a comparative assessment with non-pregnant participants. This cross-sectional survey was carried out from November 2021 to February 2022. The validated vaccination attitude examination (VAX) scale about vaccination reluctance was undertaken by participants, who were also required to indicate whether they would be inclined to acquire the COVID-19 vaccine along with the reasons for reluctance. In comparison to the non-pregnant category with 353 participants, the group of 372 pregnant participants who responded to the questionnaire had a much greater proportion of hesitant respondents. Likewise, contrasted to 31% of non-pregnant participants, about 40% of them attributed their willingness to get vaccinated against coronavirus to social media. They also demonstrated a considerably stronger mean score on all subcategories of the VAX measure. The adjusted odd ratio findings showed that the independent factors for vaccine reluctance appeared to be trusting rumors on social media (adj OR: 2.58), not being afraid of covid-19 (adj OR: 2.01), not believing in COVID-19 existence (adj OR: 2.53), and not believing in vaccines (adj OR: 4.25). Uncertainty about the COVID-19 vaccine is very prevalent among expectant mothers. The investigation accentuates the pressing need to administer COVID-19 vaccination to the general public, including expectant mothers who might be anxious about the vaccine.

Oxacillin (Methicillin) Resistant Staphylococci in Domestic Animals in the Czech Republic.

The aim of this study was to describe the prevalence of different Staphylococcus species isolated from pathological processes and lesions in domestic animals in the Czech Republic and to detect and describe oxacillin (methicillin)-resistant strains (MRS). During the years 2019-2020, a total of 5218 veterinary clinical samples from the Czech Republic were tested. Testing was performed by culture methods and typing by molecular phenotypic methods MALDI-TOF MS and PCR. Antimicrobial susceptibility testing of the strains was performed by the disk diffusion method. A total of 854 staphylococci strains were identified (16.37% prevalence), out of which 43 strains of 6 species of staphylococci were MRS (n = 43; 0.82% prevalence). Of the MRS strains, the most prevalent species were Staphylococcus pseudintermedius (n = 24; 0.46% prevalence) and Staphylococcus aureus (n = 7; 0.13% prevalence). Susceptibility testing showed resistance to beta-lactam antibiotics and, depending on the species, also to trimethoprim/sulfamethoxazole, gentamicin, tetracycline, erythromycin, clindamycin, and enrofloxacin. For further characterization of MRS, PCR assay for virulence factor genes was performed. Seven of the 14 target genes were observed only in S. aureus, except for the eno gene encoding laminin-binding protein, which was also detected in other staphylococci. It is necessary to emphasize the issue of correct using of antimicrobials in practice and antibiotic policy in university teaching and to create stricter legislation that would prevent the widespread use of antimicrobials in veterinary medicine, especially in livestock to reduce the emergence and spread of antimicrobial resistance.

Draft Genome Sequences of Seven Cronobacter sakazakii Strains Carrying the mcr 9.1 Gene Isolated in Chile.

Cronobacter sakazakii is a pathogen that causes severe diseases such as meningitis and necrotizing enterocolitis in infants under 12 months, associated with the consumption of contaminated rehydrated powdered infant formula (PIF). We present seven C. sakazakii genome sequences isolated from PIF and dairy products in Chile in 2017.

Profiling the Virulence and Antibiotic Resistance Genes of Cronobacter sakazakii Strains Isolated From Powdered and Dairy Formulas by Whole-Genome Sequencing.

Cronobacter sakazakii is an enteropathogen that causes neonatal meningitis, septicemia, and necrotizing enterocolitis in preterm infants and newborns with a mortality rate of 15 to 80%. Powdered and dairy formulas (P-DF) have been implicated as major transmission vehicles and subsequently the presence of this pathogen in P-DF led to product recalls in Chile in 2017. The objective of this study was to use whole genome sequencing (WGS) and laboratory studies to characterize Cronobacter strains from the contaminated products. Seven strains were identified as C. sakazakii, and the remaining strain was Franconibacter helveticus. All C. sakazakii strains adhered to a neuroblastoma cell line, and 31 virulence genes were predicted by WGS. The antibiograms varied between strains. and included mcr-9.1 and bla CSA genes, conferring resistance to colistin and cephalothin, respectively. The C. sakazakii strains encoded I-E and I-F CRISPR-Cas systems, and carried IncFII(pECLA), Col440I, and Col(pHHAD28) plasmids. In summary, WGS enabled the identification of C. sakazakii strains and revealed multiple antibiotic resistance and virulence genes. These findings support the decision to recall the contaminated powdered and dairy formulas from the Chilean market in 2017.

Virulence and Antibiotic Resistance Genes in Listeria monocytogenes Strains Isolated From Ready-to-Eat Foods in Chile.

Listeria monocytogenes is causing listeriosis, a rare but severe foodborne infection. Listeriosis affects pregnant women, newborns, older adults, and immunocompromised individuals. Ready-to-eat (RTE) foods are the most common sources of transmission of the pathogen This study explored the virulence factors and antibiotic resistance in L. monocytogenes strains isolated from ready-to-eat (RTE) foods through in vitro and in silico testing by whole-genome sequencing (WGS). The overall positivity of L. monocytogenes in RTE food samples was 3.1% and 14 strains were isolated. L. monocytogenes ST8, ST2763, ST1, ST3, ST5, ST7, ST9, ST14, ST193, and ST451 sequence types were identified by average nucleotide identity, ribosomal multilocus sequence typing (rMLST), and core genome MLST. Seven isolates had serotype 1/2a, five 1/2b, one 4b, and one 1/2c. Three strains exhibited in vitro resistance to ampicillin and 100% of the strains carried the fosX, lin, norB, mprF, tetA, and tetC resistance genes. In addition, the arsBC, bcrBC, and clpL genes were detected, which conferred resistance to stress and disinfectants. All strains harbored hlyA, prfA, and inlA genes almost thirty-two the showed the bsh, clpCEP, hly, hpt, iap/cwhA, inlA, inlB, ipeA, lspA, mpl, plcA, pclB, oat, pdgA, and prfA genes. One isolate exhibited a type 11 premature stop codon (PMSC) in the inlA gene and another isolate a new mutation (deletion of A in position 819). The Inc18(rep25), Inc18(rep26), and N1011A plasmids and MGEs were found in nine isolates. Ten isolates showed CAS-Type II-B systems; in addition, Anti-CRISPR AcrIIA1 and AcrIIA3 phage-associated systems were detected in three genomes. These virulence and antibiotic resistance traits in the strains isolated in the RTE foods indicate a potential public health risk for consumers.

Molecular Characterization of Cronobacter sakazakii Strains Isolated from Powdered Milk.

Cronobacter spp. are opportunistic pathogens of the Enterobacteriaceae family. The organism causes infections in all age groups, but the most serious cases occur in outbreaks related to neonates with meningitis and necrotizing enterocolitis. The objective was to determine the in silico and in vitro putative virulence factors of six Cronobacter sakazakii strains isolated from powdered milk (PM) in the Czech Republic. Strains were identified by MALDI-TOF MS and whole-genome sequencing (WGS). Virulence and resistance genes were detected with the Ridom SeqSphere+ software task template and the Comprehensive Antibiotic Resistance Database (CARD) platform. Adherence and invasion ability were performed using the mouse neuroblastoma (N1E-115 ATCCCRL-2263) cell line. The CRISPR-Cas system was searched with CRISPRCasFinder. Core genome MLST identified four different sequence types (ST1, ST145, ST245, and ST297) in six isolates. Strains 13755-1B and 1847 were able to adhere in 2.2 and 3.2 × 106 CFU/mL, while 0.00073% invasion frequency was detected only in strain 1847. Both strains 13755-1B and 1847 were positive for three (50.0%) and four virulence genes, respectively. The cpa gene was not detected. Twenty-eight genes were detected by WGS and grouped as flagellar or outer membrane proteins, chemotaxis, hemolysins, and invasion, plasminogen activator, colonization, transcriptional regulator, and survival in macrophages. The colistin-resistance-encoding mcr-9.1 and cephalothin-resis-encoding blaCSA genes and IncFII(pECLA) and IncFIB(pCTU3) plasmids were detected. All strains exhibited CRISPR matrices and four of them two type I-E and I-F matrices. Combined molecular methodologies improve Cronobacter spp. decision-making for health authorities to protect the population.

PCR Detection of Oxacillinases in Bacteria.

Oxacillinases (OXA) have been mostly described in Enterobacteriaceae, Acinetobacter, and Pseudomonas species. Recent years have witnessed an increased prevalence of intrinsic and/or acquired ß-lactamase-producing Acinetobacter in food-producing animals. This study was conducted to assess the prevalence of OXA among selected bacterial species and to characterize these enzymes by in silico analysis. Screening of OXA was performed by PCR amplification using specific pairs of oligonucleotides. Overall, 40 pairs of primers were designed, of which 6 were experimentally tested in vitro. Among 49 bacterial isolates examined, the presence of blaOXA-1-like genes was confirmed in 20 cases (41%; 19 times in Klebsiella pneumoniae and once in Enterobacter cloacae). No OXA were found in animal isolates. The study results confirmed the specificity of the designed oligonucleotide pairs. Furthermore, the designed primers were found to possess the ability to specifically detect 90.2% of all OXA. These facts suggest that the in silico and in vitro tested primers could be used for single or multiplex PCR to screen for the presence of OXA in various bacteria, as well as to monitor their spread. At the same time, the presence of conserved characteristic amino acids and motifs was confirmed by in silico analysis of sequences of representative members of OXA.

Microbiological Quality of Powdered Infant Formula in Latin America.

ABSTRACT: Cronobacter is a bacterial genus that includes seven species, and the species Cronobacter sakazakii is most related to meningitis and septicemia in infants associated with powdered infant formula (PIF). The objectives of this study were to evaluate the presence of C. sakazakii and to determine the microbiological quality of PIF for infant consumption. To do this, a total of 128 PIF samples were analyzed in four brands and countries (Chile, Mexico, Holland, and Brazil), considering three types of PIF: premature (PIF1), infant (PIF2), and follow-up (PIF3). Aerobic plate counts (APC) and Enterobacteriaceae (ENT) were assessed in accordance with Chilean official standards. The outer membrane protein A (ompA) gene was amplified to detect Cronobacter spp. and the fusA gene was amplified to identify C. sakazakii by using the PubMLST Web site and BLAST (NCBI). The antibiotic resistance profile was performed according to the Clinical and Laboratory Standards Institute standards. The pathogen was quantified by the most probable number (MPN). The results showed that APC median values for PIF1, PIF2, and PIF3 were 3.2, 4.9, and 4.8 log CFU g-1, respectively. The APC were higher in PIF2 (P < 0.01) from Holland (P < 0.01) in the commercial brand 4 (P < 0.01). The ENT median values in PIF1, PIF2, and PIF3 were 1.8, 1.5, and 1.7 log CFU g-1, respectively. Five strains of C. sakazakii and one strain of Cronobacter malonaticus were identified as having values between 0.023 and 2.3 MPN/g. All strains (100%) harbored the ompA, plasminogen activator (cpa), and hemolysin (hly) virulence genes. To conclude, C. sakazakii was found in four PIF samples from four Chilean products and one from Mexico, which is distributed throughout America. C. sakazakii strains exhibit virulence factors and resistance to ampicillin, thus posing a risk when PIFs are consumed by infants.

Antibiotic Susceptibility of Cronobacter spp. Isolated from Clinical Samples.

Cronobacter spp. have been recognized as causative agents of various severe infections in pre-term or full-term infants as well as elderly adults suffering from serious underlying disease or malignancy. A surveillance study was designed to identify antibiotic resistance among clinical Cronobacter spp. strains, which were isolated from patients of two hospitals between May 2007 and August 2013. Altogether, 52 Cronobacter spp. isolates were analyzed. Although MALDI-TOF mass spectrometry recognized all Cronobacter sakazakii and Cronobacter malonaticus strains, it could not identify Cronobacter muytjensii strain. Nevertheless, all strains were identified as Cronobacter spp. using multilocus sequence typing (MLST). Strains were tested against 17 types of antibiotics, using the standard microdilution method according to the 2018 European Committee on Antimicrobial Susceptibility Testing criteria. Three Cronobacter species were identified as C. sakazakii (n = 33), C. malonaticus (n = 18), and C. muytjensii (n = 1); all isolates were susceptible to all tested antibiotics. All strains were PCR-negative for bla TEM, bla SHV, and bla CTX-M ß-lactamase genes, as well. Even though the results of this study showed that Cronobacter spp. isolates were pan-susceptible, continued antibiotic resistance surveillance is warranted.Cronobacter spp. have been recognized as causative agents of various severe infections in pre-term or full-term infants as well as elderly adults suffering from serious underlying disease or malignancy. A surveillance study was designed to identify antibiotic resistance among clinical Cronobacter spp. strains, which were isolated from patients of two hospitals between May 2007 and August 2013. Altogether, 52 Cronobacter spp. isolates were analyzed. Although MALDI-TOF mass spectrometry recognized all Cronobacter sakazakii and Cronobacter malonaticus strains, it could not identify Cronobacter muytjensii strain. Nevertheless, all strains were identified as Cronobacter spp. using multilocus sequence typing (MLST). Strains were tested against 17 types of antibiotics, using the standard microdilution method according to the 2018 European Committee on Antimicrobial Susceptibility Testing criteria. Three Cronobacter species were identified as C. sakazakii (n = 33), C. malonaticus (n = 18), and C. muytjensii (n = 1); all isolates were susceptible to all tested antibiotics. All strains were PCR-negative for bla TEM, bla SHV, and bla CTX-M ß-lactamase genes, as well. Even though the results of this study showed that Cronobacter spp. isolates were pan-susceptible, continued antibiotic resistance surveillance is warranted.

Antibiotic susceptibility and production of endotoxin by Ochrobactrum anthropi isolated from environment and from patients with cystic fibrosis.

The aim of this work was to compare production of endotoxin and to determine susceptibility to antibiotics in two groups of specimens-wild-type strains Ochrobactrum anthropi isolated from the environment and the strains isolated from patients with cystic fibrosis. The determination of the endotoxin produced by the test strains was carried on by using a limulus amebocyte lysate test (LAL test). Determination of ATB sensitivity was accomplished by means of a broth dilution method in a microtiter plate (MIC). No significant difference was found between the group of ochrobacters isolated from the environment and the group of ochrobacters isolated from cystic fibrosis patients. Antibiotic sensitivity testing has indicated that the resistance to tigecycline, trimethoprim/sulfamethoxazole, and gentamicin was slightly higher in strains isolated from cystic fibrosis patients in comparison with strains isolated from the environment. In general, most of the test strains were sensitive to most of the antibiotics tested. Significant resistance has been demonstrated for cefotaxime. Resistance was also found for gentamicin in strains number 4 and 7. The MIC was equal to the breakpoint for this antibiotic (8000 mg/L).

Characterization of the pathogenicity of clinical Cronobacter malonaticus strains based on the tissue culture investigations.

Cronobacter malonaticus is a member of the genus Cronobacter which is considered an opportunistic pathogen. The significance of C. malonaticus has recently increased since it was documented to be involved in several serious neonatal infections. However, the virulence factors of C. malonaticus including their ability to adhere, invade and overcome host barriers have not been studied before. Unlike previous Cronobacter research, this study is mainly focused on C. malonaticus and is aimed to investigate its virulence characteristics that enable this species to cause adult and neonatal infections. Altogether, 20 strains were included in this study (19 clinical and one environmental strain). Our data showed that the clinical C. malonaticus has an ability to adhere and invade Caco-2, HBMEC, A549 and T24 cell lines. Moreover, the result showed that certain strains of C. malonaticus (including 1827 and 2018) were able to persist well in macrophages. However, ST7 strains 1827 and 2018 proved to be the most invasive strains among all used strains. The CDC strain 1569 (ST307) which was isolated from the blood of a fatal neonatal case showed also significant results in this study as it was able to invade all used human cells and survive and replicate within microphages. Finally, the findings of this study confirm the potential ability of C. malonaticus to cause serious infections in neonates or adults such as necrotising enterocolitis, meningitis, bacteraemia, pneumonia and urinary tract infection.