scholarly journals Nutrition Related Stress Factors Reduce the Transfer of Extended-Spectrum Beta-Lactamase Resistance Genes between an Escherichia coli Donor and a Salmonella Typhimurium Recipient In Vitro

Biomolecules ◽  
2019 ◽  
Vol 9 (8) ◽  
pp. 324 ◽  
Author(s):  
Eva-Maria Saliu ◽  
Marita Eitinger ◽  
Jürgen Zentek ◽  
Wilfried Vahjen

The transfer of extended spectrum β-lactamase (ESBL)-genes occurs frequently between different bacteria species. The aim of this study was to investigate the impact of nutrition related stress factors on this transfer. Thus, an Escherichia coli donor and a Salmonella Typhimurium recipient were co-incubated for 4 h in media containing different levels of the stress factors’ pH, osmolality, copper, zinc and acetic, propionic, lactic, and n-butyric acid, as well as subtherapeutic levels of cefotaxime, sulfamethoxazole/trimethoprim, and nitrofurantoin. Conjugation frequencies were calculated as transconjugants per donor, recipient, and total bacterial count. A correction factor for the stress impact on bacterial growth was used. Acetic, lactic, and n-butyric, acid, as well as pH, showed no significant impact. In contrast, increasing concentrations of propionate, zinc, copper, and nitrofurantoin, as well as increased osmolality reduced conjugation frequencies. Sulfamethoxazole/trimethoprim and cefotaxime showed increased transconjugants per donor, which decreased after correction for stress. This study showed, for the model mating pair, that conjugation frequencies decreased under different physiological stress conditions, and, thus, the hypothesis that stress factors may enhance conjugation should be viewed with caution. Furthermore, for studies on in vitro gene transfer, it is vital to consider the impact of studied stressors on bacterial growth.

Author(s):  
Alina Karoline Nussbaumer-Pröll ◽  
Sabine Eberl ◽  
Birgit Reiter ◽  
Thomas Stimpfl ◽  
Walter Jäger ◽  
...  

AbstractIn vitro pharmacodynamic models are used to optimize in vivo dosing regimens in antimicrobial drug development. One limiting factor of such models is the lack of host factors such as corpuscular blood components as erythrocytes which have already been shown to impact activity of antibiotics and/or growth of the pathogen. However, the impact of thrombocytes has not previously been investigated. We set out to investigate if the addition of thrombocytes (set to physiological concentrations in blood of healthy human, i.e., 5 × 105 thrombocytes/μL standard growth media Mueller Hinton Broth, MHB) has an influence on bacterial growth and on the efficacy of antibiotics against Gram+ and Gram− bacteria. Growth assays and time-killing-curves (TKC) were performed with ATCC-strains of Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa in triplicate over 24 h. The same approach was followed for 5 clinical isolates of Escherichia coli. Meropenem, ciprofloxacin, and tigecycline were tested as representatives of broad-spectrum antibiotics, and concentrations several-fold above and below the minimal inhibitory concentration (MIC) were simulated. No significant impact of thrombocytes was found on bacterial growth or antimicrobial stability for the investigated agents. Bacteria reduced thrombocyte content to different degree, indicating direct interaction of pathogens and thrombocytes. Impact on bacterial killing was observed but was not fully reproducible when thrombocytes from different donors where used. While interaction of bacteria and thrombocytes was evident in the present study, interaction between antibiotic activity and thrombocytes seems unlikely. Whether variability was caused by different thrombocyte concentrates needs further investigation.


2016 ◽  
Vol 61 (1) ◽  
Author(s):  
G. Mourand ◽  
F. Paboeuf ◽  
M. A. Fleury ◽  
E. Jouy ◽  
S. Bougeard ◽  
...  

ABSTRACT Four trials were conducted to evaluate the impact of Escherichia coli probiotic strain ED1a administration to pigs on the gut carriage or survival in manure of extended-spectrum-β-lactamase-producing E. coli. Groups of pigs were orally inoculated with strain E. coli M63 carrying the bla CTX-M-1 gene (n = 84) or used as a control (n = 26). In the first two trials, 24 of 40 E. coli M63-inoculated pigs were given E. coli ED1a orally for 6 days starting 8 days after oral inoculation. In the third trial, 10 E. coli M63-inoculated pigs were given either E. coli ED1a or probiotic E. coli Nissle 1917 for 5 days. In the fourth trial, E. coli ED1a was given to a sow and its 12 piglets, and these 12 piglets plus 12 piglets that had not received E. coli ED1a were then inoculated with E. coli M63. Fecal shedding of cefotaxime-resistant Enterobacteriaceae (CTX-RE) was studied by culture, and bla CTX-M-1 genes were quantified by PCR. The persistence of CTX-RE in manure samples from inoculated pigs or manure samples inoculated in vitro with E. coli M63 with or without probiotics was studied. The results showed that E. coli M63 and ED1a were good gut colonizers. The reduction in the level of fecal excretion of CTX-RE in E. coli ED1a-treated pigs compared to that in nontreated pigs was usually less than 1 log10 CFU and was mainly observed during the probiotic administration period. The results obtained with E. coli Nissle 1917 did not differ significantly from those obtained with E. coli ED1a. CTX-RE survival did not differ significantly in manure samples with or without probiotic treatment. In conclusion, under our experimental conditions, E. coli ED1a and E. coli Nissle 1917 could not durably prevent CTX-RE colonization of the pig gut.


2000 ◽  
Vol 118 (1) ◽  
pp. 21-29 ◽  
Author(s):  
Ulysses Fagundes-Neto ◽  
Isabel Cristina Affonso Scaletsky

Diarrheal disease is still the most prevalent and important public health problem in developing countries, despite advances in knowledge, understanding, and management that have occurred over recent years. Diarrhea is the leading cause of death in children under 5 years of age. The impact of diarrheal diseases is more severe in the earliest periods of life, when taking into account both the numbers of episodes per year and hospital admission rates. This narrative review focuses on one of the major driving forces that attack the host, namely the enteropathogenic Escherichia coli (EPEC) and the consequences that generate malnutrition in an early phase of life. EPEC serotypes form dense microcolonies on the surface of tissue-culture cells in a pattern known as localized adherence (LA). When EPEC strains adhere to epithelial cells in vitro or in vivo they cause characteristic changes known as Attaching and Effacement (A/E) lesions. Surface abnormalities of the small intestinal mucosa shown by scanning electron microscopy in infants with persistent diarrhea, although non-specific, are intense enough to justify the severity of the clinical aspects displayed in a very young phase in life. Decrease in number and height of microvilli, blunting of borders of enterocytes, loss of the glycocalyx, shortening of villi and presence of a mucus pseudomembrane coating the mucosal surface were the abnormalities observed in the majority of patients. These ultrastructural derangements may be due to an association of the enteric enteropathogenic agent that triggers the diarrheic process and the onset of food intolerance responsible for perpetuation of diarrhea. An aggressive therapeutic approach based on appropriate nutritional support, especially the utilization of human milk and/or lactose-free protein hydrolyzate-based formulas and the adequate correction of the fecal losses, is required to allow complete recovery from the damage caused by this devastating enteropathogenic agent.


2007 ◽  
Vol 51 (4) ◽  
pp. 1481-1486 ◽  
Author(s):  
C. Andrew DeRyke ◽  
Mary Anne Banevicius ◽  
Hong Wei Fan ◽  
David P. Nicolau

ABSTRACT The purpose of this study was to examine the in vivo efficacies of meropenem and ertapenem against extended-spectrum-β-lactamase (ESBL)-producing isolates with a wide range of MICs. Human-simulated dosing regimens in mice were designed to approximate the free drug percent time above the MIC (fT>MIC) observed for humans following meropenem at 1 g every 8 h and ertapenem at 1 g every 24 h. An in vivo neutropenic mouse thigh infection model was used to examine the bactericidal effects against 31 clinical ESBL Escherichia coli and Klebsiella pneumoniae isolates and 2 non-ESBL isolates included for comparison at a standard 105 inoculum. Three isolates were examined at a high 107 inoculum as well. Meropenem displayed greater in vitro potency, with a median MIC (range) (μg/ml) of 0.125 (0.03 to 32), than did ertapenem, with 0.5 (0.012 to 128). Seven of the 31 ESBL isolates were removed from the efficacy analysis due to their inability to establish infection in the mouse model. When MICs were ≤1.5 μg/ml for ertapenem (≤0.5 μg/ml for meropenem), similar reductions in CFU (≈ 2-log kill) were observed for both ertapenem (fT>MIC ≥ 23%) and meropenem (fT>MIC ≥ 75%). Ertapenem showed bacterial regrowth for seven of eight isolates, with MICs of ≥2 μg/ml (fT>MIC ≤ 20%), while meropenem displayed antibacterial potency that varied from a static effect to a 1-log bacterial reduction in these isolates (fT>MIC = 30 to 65%). At a 107 inoculum, both agents eradicated bacteria due to adequate exposures (fT>MIC = 20 to 45%). Due to low MICs, no difference in bacterial kill was noted for the majority of ESBL isolates tested. However, for isolates with raised ertapenem MICs of ≥2 μg/ml, meropenem displayed sustained efficacy due to its greater in vitro potency and higher resultant fT>MIC.


1991 ◽  
Vol 54 (7) ◽  
pp. 496-501 ◽  
Author(s):  
ARTHUR HINTON ◽  
GEORGE E. SPATES ◽  
DONALD E. CORRIER ◽  
MICHAEL E. HUME ◽  
JOHN R. DELOACH ◽  
...  

A Veillonella species and Enterococcus durans were isolated from the cecal contents of adult broilers. Mixed cultures of Veillonella and E. durans inhibited the growth of Salmonella typhimurium and Escherichia coli 0157:H7 on media containing 2.5% lactose (w/v). The growth of S. typhimurium or E. coli 0157:H7 was not inhibited by mixed cultures containing Veillonella and E. durans on media containing only 0.25% lactose or by pure cultures of Veillonella or E. durans on media containing either 0.25% or 2.5% lactose. The mixed cultures of Veillonella and E. durans produced significantly (P<0.05) more acetic, propionic, and lactic acids in media containing 2.5% lactose than in media containing 0.25% lactose. The inhibition of the enteropathogens was related to the production of lactic acid from lactose by the E. durans and the production of acetic and propionic acids from lactic acid by the Veillonella.


2015 ◽  
Vol 59 (4) ◽  
pp. 1962-1968 ◽  
Author(s):  
Sun Hee Park ◽  
Su-Mi Choi ◽  
Dong-Gun Lee ◽  
Sung-Yeon Cho ◽  
Hyo-Jin Lee ◽  
...  

ABSTRACTExtended-spectrum β-lactamase-producingEscherichia coli(ESBL-EC) is increasingly identified as a cause of acute pyelonephritis (APN) among patients without recent health care contact, i.e., community-associated APN. This case-control study compared 75 cases of community-associated ESBL-EC APN (CA-ESBL) to 225 controls of community-associated non-ESBL-EC APN (CA-non-ESBL) to identify the risk factors for ESBL-EC acquisition and investigate the impact of ESBL on the treatment outcomes of community-associated APN (CA-APN) caused byE. coliat a Korean hospital during 2007 to 2013. The baseline characteristics were similar between the cases and controls; the risk factors for ESBL-EC were age (>55 years), antibiotic use within the previous year, and diabetes with recurrent APN. The severity of illness did not differ between CA-ESBL and CA-non-ESBL (Acute Physiology and Chronic Health Evaluation [APACHE] II scores [mean ± standard deviation], 7.7 ± 5.9 versus 6.4 ± 5.3;P= 0.071). The proportions of clinical (odds ratio [OR], 1.76; 95% confidence interval [CI], 0.57 to 5.38;P= 0.323) and microbiological (OR, 1.16; 95% CI, 0.51 to 2.65;P= 0.730) cures were similar, although the CA-ESBL APN patients were less likely to receive appropriate antibiotics within 48 h. A multivariable Cox proportional hazards analysis of the prognostic factors for CA-APN caused byE. colishowed that ESBL production was not a significant factor for clinical (hazard ratio [HR], 0.39; 95% CI, 0.12 to 1.30;P= 0.126) or microbiological (HR, 0.49; 95% CI, 0.21 to 1.12;P= 0.091) failure. The estimates did not change after incorporating weights calculated using propensity scores for acquiring ESBL-EC. Therefore, ESBL production did not negatively affect treatment outcomes among patients with community-associatedE. coliAPN.


2016 ◽  
pp. 39-44
Author(s):  
Ifra Tun Nur ◽  
Jannatun Tahera ◽  
Md Sakil Munna ◽  
M Majibur Rahman ◽  
Rashed Noor

With a previous observation of Escherichia coli growth cessation along with temperature variation within three different bacteriological culture media (nutrient agar, Luria-Bertani agar and minimal agar), current investigation further depicted on the possible growth dynamics of Escherichia coli (SUBE01) and Salmonella (SUBS01) growth and viability upon supplementation of different carbon sources (dextrose, sucrose, lactose, glycerol and tween 20) at 37°C under the aeration of 100 rpm. Viability of the tested bacterial species was assessed through the enumeration of the colony forming unit (cfu) appeared upon prescribed incubation for 12-24 hours on different agar plates consisting of the above mentioned carbon sources. Besides, to inspect the cellular phenotypic changes, morphological observations were conducted under the light microscope. Variations in bacterial growth (either growth acceleration or cessation) were further noticed through the spot tests on the agar plates. Considerable shortfalls in the culturable cells of E. coli and Salmonella spp. were noted in the minimal media separately consisting of sucrose, lactose, glycerol or tween 20 while an opposite impact of accelerated growth was noticed in the media supplied with dextrose. The data revealed a hierarchy of consequence of carbon sources as nutrient generators whereby the favourable bacterial growth and survival order of the carbon sources was estimated as dextrose > glycerol > lactose > tween 20 > sucrose.Bangladesh J Microbiol, Volume 32, Number 1-2,June-Dec 2015, pp 39-44


2012 ◽  
Author(s):  
Νίκη Λάμπρη

Εισαγωγή. Η παραγωγή β-λακταμασών ευρέως φάσματος Extended-spectrum b-lactamases (ESBLs) από ουροπαθογόνα της τάξεως των Εντεροβακτηριακών αποτελεί ένα αναδυόμενο και συχνά δύσκολο στην αντιμετώπισή του θεραπευτικό πρόβλημα, που αφορά πρωτίστως λοιμώξεις που αποκτώνται ενδονοσοκομειακά άλλα εσχάτως και σε λοιμώξεις εκ της κοινότητας. Η κυριότερη θεραπευτική πρόκληση για τον ιατρό της κοινότητας είναι η έλλειψη αποτελεσματικών αντιμικροβιακών για από του στόματος εξωνοσοκομειακή χορήγηση εφόσον η βαρύτητα της λοίμωξης το επιτρέπει. Σκοπός της παρούσας διατριβής ήταν να διερευνήσει την in vitro δραστικότητα του συνδυασμού δυο αντιμικροβιακών με δυνατότητα εξωνοσοκομειακής χορήγησης, της μεσιλλινάμης με το κλαβουλανικό οξύ έναντι στελεχών ουροπαθογόνων εκ της κοινότητας με παραγωγή ESBL. Υλικό και μέθοδος. Στα πλαίσια μιας μεγάλης πολυκεντρικής επιδημιολογικής μελέτης αντοχής των ουροπαθογόνων από ασθενείς που αντιμετωπίζονται εξωνοσοκομειακά, στελέχη Escherichia coli που απομονώθηκαν κατά τα έτη 2005-2006 και παρήγαγαν ESBL ταυτοποιήθηκαν μοριακά και μελετήθηκε η ευαισθησία τους in vitro στην μεσιλλινάμη και στο συνδυασμό μεσιλλινάμης με κλαβουλανικό οξύ. H μελέτη της in vitro δραστικότητας της μεσιλλινάμης σε συνέργεια με το κλαβουλανικό οξύ έγινε με τα ακόλουθα βήματα: 1) Προσδιορισμό της MIC της μεσιλλινάμης έναντι ουροπαθογόνων E.coli στελεχών, απομονωθέντων από εξωτερικούς ασθενείς 2) Επιλογή στελεχών με φαινότυπο παραγωγής ESBL 3) Επιβεβαίωση και χαρακτηρισμό του είδους της β-λακταμάσης με την τεχνική ισοηλεκτρικής εστίασης (isoelectric focusing) και αντίδραση αλυσιδωτής πολυμεράσης (PCR) 4) Μελέτη της συνέργειας της μεσιλλινάμης με το κλαβουλανικό οξύ με τη μέθοδο διάχυσης στο άγαρ 5) Μελέτη της βακτηριοκτόνου δράσης της μεσιλλινάμης έναντι στελεχών που παράγουν ESBL με τη μέθοδο της καμπύλης θανάτωσης στο χρόνο (killing curves) και μελέτη της συνέργειας της μεσιλλινάμης με το κλαβουλανικό οξύ με την ίδια μέθοδο. Για τις in vitro αλληλεπιδράσεις της μεσιλλινάμης με το κλαβουλανικό (σε σταθερή συγκέντρωση 4 mg/L) χρησιμοποιήθηκαν δύο πυκνότητες βακτηριακού εναιωρήματος (104 και 106 cfu/ίχνος) τόσο στη μέθοδο διάχυσης στο άγαρ όσο και στη μέθοδο της καμπύλης θανάτωσης στο χρόνο. Αποτελέσματα. Από τα 48 ανευρεθέντα ουροπαθογόνα με παραγωγή ESBL τα 47 (ποσοστό 97,9%) ήταν ευαίσθητα in vitro στη μεσιλλινάμη. Τα ένζυμα τύπου CTX-M ήταν κυρίαρχα (87,2%), με συχνότερη την κεφοταξιμάση CTX-M-3. SHV, VEB, TEM και τύπου- OXA ένζυμα ταυτοποιήθηκαν σε ποσοστό 10,6%, 2,1%, 19,2% και 12,8% αντίστοιχα. Συνέργεια με το κλαβουλανικό καταδείχθηκε σε ποσοστό 60,4% με τη μέθοδο της διάχυσης σε άγαρ και σε ποσοστό 43,8% με τη μέθοδο καμπύλης θανάτωσης στο χρόνο. Στο 64,6% των στελεχών παρατηρήθηκε φαινόμενο επίδρασης του βακτηριακού εναιωρήματος, το οποίο όμως αντιστράφηκε με τη συνέργεια που καταδείχθηκε με το καλβουλανικό στο 85,4% των μελετηθέντων στελεχών. Όταν χρησιμοποιήθηκε πυκνό βακτηριακό εναιώρημα, 60,4% των μικροβίων (29/48) ήταν ανθεκτικά στη μεσιλλινάμη αλλά με την προσθήκη κλαβουλανικού το 97,9% εξ’αυτών (47/48) ήταν ευαίσθητα στο συνδυασμό. Συμπεράσματα. Ο επιπολασμός ESBL ενζύμων στα ουροπαθογόνα E.coli από τον ελληνικό χώρο κατά τα έτη 2005-2006 ήταν σχετικά χαμηλός (1.9%) και οι κεφοταξιμάσες (CTX-M) ήταν οι συχνότερες ESBL που ανιχνεύθηκαν. Μεταξύ μεσιλλινάμης και κλαβουλανικού διαπιστώθηκε συνέργεια σε 60% και αδιαφορία σε 40% των E.coli που παρήγαγαν ESBL. Όλα τα στελέχη με MIC ≥4 μg/ml παρουσίασαν συνέργεια. Η μεσιλλινάμη δυνατόν να αποτελεί μια δόκιμη θεραπεία για ανεπίπλεκτες λοιμώξεις του ουροποιητικού από ουροπαθογόνα που παράγουν ESBL με χαμηλές ελάχιστες ανασταλτικές πυκνότητες. Η προσθήκη κλαβουλανικού οξέος αυξάνει και επεκτείνει τη δραστικότητά της έναντι των ESBL-παραγωγών ουροπαθογόνων ακόμη και σε υψηλές βακτηριακές συγκεντρώσεις. Κατά συνέπεια, ο συνδυασμός μεσιλλινάμης με κλαβουλανικό οξύ χρήζει περαιτέρω μελέτης σε κλινικό επίπεδο.


2021 ◽  
Vol 31 (4) ◽  
pp. 2
Author(s):  
IDSAP Peramiarti

Diarrhea is defecation with a frequency more often than usual (three times or more) a day (10 mL/kg/day) with a soft or liquid consistency, even in the form of water alone. Pathogenic bacteria, such as Escherichia coli, Salmonella typhimurium, and Shigella sp., play a role in many cases, to which antibiotics are prescribed as the first-line therapy. However, since antibiotic resistance cases are often found, preventive therapies are needed, such as consuming yogurt, which is produced through a fermentation process by lactic acid bacteria (LAB). This research aimed to determine the activity of lactic acid bacteria (Liactobacillus bulgaricus and Streptococcus thermophilus) in yogurt in inhibiting the growth of the pathogenic bacteria E. coli, S. typhimurium, and Shigella sp. The research applied in vitro with the liquid dilution test method and the true experimental design research method with post-test-only and control group design. The design was used to see the inhibitory effect of yogurt LAB on the growth of E. coli, S. typhimurium, and Shigell sp. to compare the effect of several different yogurt concentrations, namely 20%, 40%, 60%, and 80%. The results of the Least Significance Different analysis showed that there was a significant difference between yogurt with a concentration of 0% and that with various concentrations in inhibiting the growth of E. coli, S. typhimurium, and Shigella sp. with a p-value of &lt;0.05. Whereas, there was no significant difference in the various concentrations of yogurt in inhibiting the growth of the three kinds of bacteria with a p-value of &gt; 0.05.<p class="Default" align="center"> </p>


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