Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

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    We will mail you new results for this query: keywords==Antibiotics
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Multiclass screening in urine by comprehensive two-dimensional liquid chromatography time of flight mass spectrometry for residues of sulphonamides, beta-agonists and steroids
Blokland, M.H. ; Zoontjes, P.W. ; Ginkel, L.A. van; Schans, M.G.M. van de; Sterk, S.S. ; Bovee, T.F.H. - \ 2018
Food Additives & Contaminants. Pt. A, Chemistry, Analysis, Control, Exposure & Risk Assessment 35 (2018)9. - ISSN 1944-0049 - p. 1703 - 1715.
Antibiotics - comprehensive 2D-LC - growth promoters - LC x LC - mass spectrometry - residues

Nowadays routine residue monitoring involves the analysis of many compounds from different classes, mainly in urine. In the past two decades, developments heavily focused on the use of mass spectrometers (MS) and faster and more sensitive MS detectors have reached the market. However, chromatographic separation (CS) was rather ignored and the cognate developments in CS were not in line. As a result, residue analysis did not improve to the extent anticipated. CS by LC x LC is a promising technique and will enable a further increase in the range of compounds and compound classes that can be detected in a single run. In the present study, a self-built LC x LC system, using a 10 port valve, was connected to a single quadrupole MS with electrospray interface. Standards containing a mixture of sulphonamides, β-agonists and (steroid) hormones, 53 compounds, in total, were analysed. Results demonstrated that these compounds were well separated and could be detected at low levels in urine, i.e. limit of detection (LOD) from 1 µg L−1 for most β-agonists to 10 µg L−1 for some sulphonamides and most hormones. To enhance the sensitivity, optimisation was performed on an advanced commercial LC x LC system connected to a full scan accurate MS. This ultimately resulted in a fast high throughput untargeted method, including a simple sample clean-up in a 96-well format, for the analysis of urine samples.

Impact of lincosamides antibiotics on the composition of the rat gut microbiota and the metabolite profile of plasma and feces
Behr, C. ; Ramírez-Hincapié, S. ; Cameron, H.J. ; Strauss, V. ; Walk, T. ; Herold, M. ; Beekmann, K. ; Rietjens, I.M.C.M. ; Ravenzwaay, B. van - \ 2018
Toxicology Letters 296 (2018). - ISSN 0378-4274 - p. 139 - 151.
Antibiotics - Gut microbiome - Metabolomics - Microbiome-related metabolites - Repeated dose oral toxicity study - Taxonomic profiling

The importance of the gut microorganisms and their wide range of interactions with the host are well-acknowledged. In this study, lincomycin and clindamycin were used to modulate microbial communities of Wistar rats to gain a comprehensive understanding of the implications of microbiome alterations. A metabolomics approach and taxonomic profiling were applied to characterize the effects of these antibiotics on the functionality of the microbiome and to identify microbiome-related metabolites. After treatment, the diversity of the microbial community was drastically reduced. Bacteroidetes and Verrucomicrobia were drastically reduced, Tenericutes and Deferribacteres completely disappeared, while abundance of Firmicutes and Proteobacteria were highly increased. Changes in plasma and feces metabolites were observed for metabolites belonging mainly to the class of complex lipids, fatty acids and related metabolites as well as amino acids and related compounds. Bile acid metabolism was markedly affected: taurocholic acid, glycochenodeoxycholic acid and cholic acid presented abrupt changes showing a specific metabolite pattern indicating disruption of the microbial community. In both plasma and feces taurocholic acid was highly upregulated upon treatment whereas glycochenodeoxycholic acid was downregulated. Cholic acid was upregulated in feces but downregulated in plasma. These results show that changes in the gut microbial community lead to alterations of the metabolic profile in blood and feces of the host and can be used to identify potentially microbiome-related metabolites. This implies that metabolomics could be a suitable tool to estimate the extent of changes induced in the intestinal microbiome with respect to consequences for the host.

Microbiome-related metabolite changes in gut tissue, cecum content and feces of rats treated with antibiotics
Behr, C. ; Sperber, S. ; Jiang, X. ; Strauss, V. ; Kamp, H. ; Walk, T. ; Herold, M. ; Beekmann, K. ; Rietjens, I.M.C.M. ; Ravenzwaay, B. van - \ 2018
Toxicology and Applied Pharmacology 355 (2018). - ISSN 0041-008X - p. 198 - 210.
Antibiotics - Gut content and tissue - Gut microbiome - Metabolite profiling - Metabolomics - Repeated dose oral toxicity study

The metabolic functionality of the gut microbiota contributes to the metabolism and well-being of its host, although detailed insight in the microbiota's metabolism is lacking. Omics technologies could facilitate unraveling metabolism by the gut microbiota. In this study, we performed metabolite profiling of different matrices of the gut, after antibiotic treatment of rats in order to evaluate metabolite changes observed at different dose levels and in different sexes, and to identify the best tissue matrix for further investigations regarding an assessment of metabolic effects of new compounds with antibiotic activity. Three different antibiotics (vancomycin, streptomycin and roxithromycin) were administered orally to rats for 28 days according to the OECD 407 guideline with a subsequent metabolic profiling in feces, cecum content and gut tissue (jejunum, ileum, cecum, colon and rectum). The data were analyzed in the MetaMap®Tox database. Treatment-related effects could be observed in the metabolite profile of feces and cecum content, but not of the different gut tissues. The metabolite profile showed compound specific effects on the microbiome. In line with the activity spectra of the antibiotics tested, vancomycin showed the largest effects, followed by roxithromycin and then by streptomycin for which changes were modest. In general, for all antibiotics the largest changes were observed for the classes of lipids (increase up to 94-fold), bile acids (increase up to 33-fold), amino acids (increase up to 200-fold) and amino acid related (increase up to 348-fold). The most relevant changes in metabolite values were similar in feces and cecum content and among sexes. The results of this targeted analysis indicate that the metabolic profiles of male and female animals in the gut microbiome are comparable. Concluding, taking other samples than feces does not add any extra information. Thus, as a non-invasive sampling method, feces provide a suitable matrix for studies on metabolism by the gut microbiota.

The persistence of a broad range of antibiotics during calve, pig and broiler manure storage
Berendsen, B.J.A. ; Lahr, J. ; Nibbeling, C. ; Jansen, L.J.M. ; Bongers, I.E.A. ; Wipfler, E.L. ; Schans, M.G.M. van de - \ 2018
Chemosphere 204 (2018). - ISSN 0045-6535 - p. 267 - 276.
Antibiotics - Dissipation - Environment - Fate - Manure - Persistence - Reviewer suggestions

After administration to livestock, a large fraction of antibiotics are excreted unchanged via excreta and can be transferred to agricultural land. For effective risk assessment a critical factor is to determine which antibiotics can be expected in the different environmental compartments. After excretion, the first relevant compartment is manure storage. In the current study, the fate of a broad scope of antibiotics (n = 46) during manure storage of different livestock animals (calves, pigs, broilers) was investigated. Manure samples were fortified with antibiotics and incubated during 24 days. Analysis was carried out by LC-MS. The dissipation of the antibiotics was modelled based on the recommendations of FOCUS working group. Sulphonamides relatively quickly dissipate in all manure types, with a DT90 of in general between 0.2 and 30 days. Tetracyclines (DT90 up to 422 days), quinolones (DT90 100–5800 days), macrolides (DT90 18–1000 days), lincosamides (DT90 135–1400 days) and pleuromutilins (DT90 of 49–1100 days) are in general much more persistent, but rates depend on the manure type. Specifically lincomycin, pirlimycin, tiamulin and most quinolones are very persistent in manure with more than 10% of the native compound remaining after a year in most manure types. For all compounds tested in the sub-set, except the macrolides, the dissipation was an abiotic process. Based on the persistence and current frequency of use, oxytetracycline, doxycycline, flumequine and tilmicosin can be expected to end up in environmental compartments. Ecotoxicological data should be used to further prioritize these compounds.

Loss of angiopoietin-like 4 (ANGPTL4) in mice with diet-induced obesity uncouples visceral obesity from glucose intolerance partly via the gut microbiota
Janssen, Aafke W.F. ; Katiraei, Saeed ; Bartosinska, Barbara ; Eberhard, Daniel ; Willems van Dijk, Ko ; Kersten, Sander - \ 2018
Diabetologia 61 (2018)6. - ISSN 0012-186X - p. 1447 - 1458.
Angiopoietin-like 4 - Antibiotics - Glucose tolerance - Gut microbiota - Insulin secretion - White adipose tissue
Aims/hypothesis: Angiopoietin-like 4 (ANGPTL4) is an important regulator of triacylglycerol metabolism, carrying out this role by inhibiting the enzymes lipoprotein lipase and pancreatic lipase. ANGPTL4 is a potential target for ameliorating cardiometabolic diseases. Although ANGPTL4 has been implicated in obesity, the study of the direct role of ANGPTL4 in diet-induced obesity and related metabolic dysfunction is hampered by the massive acute-phase response and development of lethal chylous ascites and peritonitis in Angptl4−/− mice fed a standard high-fat diet. The aim of this study was to better characterise the role of ANGPTL4 in glucose homeostasis and metabolic dysfunction during obesity. Methods: We chronically fed wild-type (WT) and Angptl4−/− mice a diet rich in unsaturated fatty acids and cholesterol, combined with fructose in drinking water, and studied metabolic function. The role of the gut microbiota was investigated by orally administering a mixture of antibiotics (ampicillin, neomycin, metronidazole). Glucose homeostasis was assessed via i.p. glucose and insulin tolerance tests. Results: Mice lacking ANGPTL4 displayed an increase in body weight gain, visceral adipose tissue mass, visceral adipose tissue lipoprotein lipase activity and visceral adipose tissue inflammation compared with WT mice. However, they also unexpectedly had markedly improved glucose tolerance, which was accompanied by elevated insulin levels. Loss of ANGPTL4 did not affect glucose-stimulated insulin secretion in isolated pancreatic islets. Since the gut microbiota have been suggested to influence insulin secretion, and because ANGPTL4 has been proposed to link the gut microbiota to host metabolism, we hypothesised a potential role of the gut microbiota. Gut microbiota composition was significantly different between Angptl4−/− mice and WT mice. Interestingly, suppression of the gut microbiota using antibiotics largely abolished the differences in glucose tolerance and insulin levels between WT and Angptl4−/− mice. Conclusions/interpretation: Despite increasing visceral fat mass, inactivation of ANGPTL4 improves glucose tolerance, at least partly via a gut microbiota-dependent mechanism.
Prevalence of antibiotics and antibiotic resistance genes in a wastewater effluent-receiving river in the Netherlands
Sabri, N.A. ; Schmitt, H. ; Zaan, B. Van der; Gerritsen, H.W. ; Zuidema, T. ; Rijnaarts, H.H.M. ; Langenhoff, A.A.M. - \ 2018
Journal of Environmental Chemical Engineering (2018). - ISSN 2213-2929
Antibiotic resistance genes - Antibiotics - River - Wastewater treatment plant
Antibiotics are being used intensively for humans and livestock worldwide and have led to the presence of antibiotic resistance bacteria (ARB) and antibiotic resistance genes (ARGs) in the environment. Wastewater treatment plants (WWTPs) have been identified as a point source for ARB&Gs, and water catchments consequently are potential receptors of ARB&Gs. The objective of this study was to investigate the occurrence of antibiotics (macrolides, sulfonamides, tetracyclines), ARGs (ermB, sul1, sul2, tetW), and class 1 integron (targeting the integrase gene), in a Dutch river that receives wastewater treatment plant effluent. Sediment and water samples were collected during one year along the river. The WWTP significantly increased the amounts of antibiotics and ARGs in the river as compared to the upstream samples, of which the antibiotics decreased once they entered the river. ARGs were persistent in the water and sediment from the WWTP effluent discharge point until 20 km downstream. This study provides insight in the prevalence of antibiotics and ARGs in a wastewater effluent-receiving river system in the Netherlands. Even though human antibiotic usage is low in the Netherlands, antibiotics, residues of antibiotics, and ARGs are detected in the river surface water-sediment system, which shows that a river has the potential to act as a reservoir of ARGs.
Effects of temperature, genetic variation and species competition on the sensitivity of algae populations to the antibiotic enrofloxacin
Rico, Andreu ; Zhao, Wenkai ; Gillissen, Frits ; Lürling, Miquel ; Brink, Paul J. van den - \ 2018
Ecotoxicology and Environmental Safety 148 (2018). - ISSN 0147-6513 - p. 228 - 236.
Antibiotics - Cyanobacteria - Green algae - Species competition - Temperature-dependent sensitivity

Primary producers are amongst the most sensitive organisms to antibiotic pollution in aquatic ecosystems. To date, there is little information on how different environmental conditions may affect their sensitivity to antibiotics. In this study we assessed how temperature, genetic variation and species competition may affect the sensitivity of the cyanobacterium Microcystis aeruginosa and the green-algae Scenedesmus obliquus to the antibiotic enrofloxacin. First, we performed single-species tests to assess the toxicity of enrofloxacin under different temperature conditions (20 °C and 30 °C) and to assess the sensitivity of different species strains using a standard temperature (20 °C). Next, we investigated how enrofloxacin contamination may affect the competition between M. aeruginosa and S. obliquus. A competition experiment was performed following a full factorial design with different competition treatments, defined as density ratios (i.e. initial bio-volume of 25/75%, 10/90% and 1/99% of S. obliquus/M. aeruginosa, respectively), one 100% S. obliquus treatment and one 100% M. aeruginosa treatment, and four different enrofloxacin concentrations (i.e. control, 0.01, 0.05 and 0.10 mg/L). Growth inhibition based on cell number, bio-volume, chlorophyll-a concentration as well as photosynthetic activity were used as evaluation endpoints in the single-species tests, while growth inhibition based on measured chlorophyll-a was primarily used in the competition experiment. M. aeruginosa photosynthetic activity was found to be the most sensitive endpoint to enrofloxacin (EC50–72 h =0.02 mg/L), followed by growth inhibition based on cell number. S. obliquus was found to be slightly more sensitive at 20 °C than at 30 °C (EC50–72 h cell number growth inhibition of 38 and 41 mg/L, respectively), whereas an opposite trend was observed for M. aeruginosa (0.047 and 0.037 mg/L, respectively). Differences in EC50–72 h values between algal strains of the same species were within a factor of two. The competition experiment showed that M. aeruginosa growth can be significantly reduced in the presence of S. obliquus at a density ratio of 75/25% M. aeruginosa/S. obliquus, showing a higher susceptibility to enrofloxacin than in the single-species test. The results of this study confirm the high sensitivity of cyanobacteria to antibiotics and show that temperature and inter-strain genetic variation may have a limited influence on their response to them. The results of the competition experiment suggest that the structure of primary producer communities can be affected, at least temporarily, at antibiotic concentrations close to those that have been measured in the environment.

Interaction networks, ecological stability, and collective antibiotic tolerance in polymicrobial infections
Vos, Marjon G.J. de; Zagorski, Marcin ; McNally, Alan ; Bollenbach, Tobias - \ 2017
Proceedings of the National Academy of Sciences of the United States of America 114 (2017)40. - ISSN 0027-8424 - p. 10666 - 10671.
Antibiotics - Infection - Microbiology - Systems biology

Polymicrobial infections constitute small ecosystems that accommodate several bacterial species. Commonly, these bacteria are investigated in isolation. However, it is unknown to what extent the isolates interact and whether their interactions alter bacterial growth and ecosystem resilience in the presence and absence of antibiotics. We quantified the complete ecological interaction network for 72 bacterial isolates collected from 23 individuals diagnosed with polymicrobial urinary tract infections and found that most interactions cluster based on evolutionary relatedness. Statistical network analysis revealed that competitive and cooperative reciprocal interactions are enriched in the global network, while cooperative interactions are depleted in the individual host community networks. A population dynamics model parameterized by our measurements suggests that interactions restrict community stability, explaining the observed species diversity of these communities. We further show that the clinical isolates frequently protect each other from clinically relevant antibiotics. Together, these results highlight that ecological interactions are crucial for the growth and survival of bacteria in polymicrobial infection communities and affect their assembly and resilience.

A probabilistic approach to assess antibiotic resistance development risks in environmental compartments and its application to an intensive aquaculture production scenario
Rico, Andreu ; Jacobs, Rianne ; Brink, Paul J. Van den; Tello, Alfredo - \ 2017
Environmental Pollution 231 (2017). - ISSN 0269-7491 - p. 918 - 928.
Antibiotic resistance - Antibiotics - Aquaculture - Environment - Risk assessment
Estimating antibiotic pollution and antibiotic resistance development risks in environmental compartments is important to design management strategies that advance our stewardship of antibiotics. In this study we propose a modelling approach to estimate the risk of antibiotic resistance development in environmental compartments and demonstrate its application in aquaculture production systems. We modelled exposure concentrations for 12 antibiotics used in Vietnamese Pangasius catfish production using the ERA-AQUA model. Minimum selective concentration (MSC) distributions that characterize the selective pressure of antibiotics on bacterial communities were derived from the European Committee on Antimicrobial Susceptibility Testing (EUCAST) Minimum Inhibitory Concentration dataset. The antibiotic resistance development risk (RDR) for each antibiotic was calculated as the probability that the antibiotic exposure distribution exceeds the MSC distribution representing the bacterial community. RDRs in pond sediments were nearly 100% for all antibiotics. Median RDR values in pond water were high for the majority of the antibiotics, with rifampicin, levofloxacin and ampicillin having highest values. In the effluent mixing area, RDRs were low for most antibiotics, with the exception of amoxicillin, ampicillin and trimethoprim, which presented moderate risks, and rifampicin and levofloxacin, which presented high risks. The RDR provides an efficient means to benchmark multiple antibiotics and treatment regimes in the initial phase of a risk assessment with regards to their potential to develop resistance in different environmental compartments, and can be used to derive resistance threshold concentrations.
ANGPTL4 promotes bile acid absorption during taurocholic acid supplementation via a mechanism dependent on the gut microbiota
Janssen, Aafke W.F. ; Dijk, Wieneke ; Boekhorst, Jos ; Kuipers, Folkert ; Groen, Albert K. ; Lukovac, Sabina ; Hooiveld, Guido J.E.J. ; Kersten, Sander - \ 2017
Biochimica et Biophysica Acta. Molecular and Cell Biology of Lipids 1862 (2017)10. - ISSN 1388-1981 - p. 1056 - 1067.
Angiopoietin-like 4 - Antibiotics - Bile acids - Gut microbiota - Lipoprotein lipase - Triglycerides
Angiopoietin-like 4 (ANGPTL4) raises plasma triglyceride levels by inhibiting lipoprotein lipase. A set of compounds that are able to reduce plasma triglyceride levels are bile acids (BA). Because BA have been shown to decrease ANGPTL4 secretion by intestinal cells, we hypothesized that BA lower plasma triglycerides (partly) via ANGPTL4. To test that hypothesis, wild-type and Angptl4−/− mice were fed chow supplemented with taurocholic acid (TCA) for seven days. TCA supplementation effectively lowered plasma triglycerides in wild-type and Angptl4−/− mice, indicating that ANGPTL4 is not required for plasma triglyceride-lowering by BA. Intriguingly, however, plasma and hepatic BA concentrations were significantly lower in TCA-supplemented Angptl4−/− mice than in TCA-supplemented wild-type mice. These changes in the Angptl4−/− mice were accompanied by lower BA levels in ileal scrapings and decreased expression of FXR-target genes in the ileum, including the BA transporter Slc10a2. By contrast, faecal excretion of specifically primary BA was higher in the Angptl4−/− mice, suggesting that loss of ANGPTL4 impairs intestinal BA absorption. Since the gut microbiota converts primary BA into secondary BA, elevated excretion of primary BA in Angptl4−/− mice may reflect differences in gut microbial composition and/or functionality. Indeed, colonic microbial composition was markedly different between Angptl4−/− and wild-type mice. Suppression of the gut bacteria using antibiotics abolished differences in plasma, hepatic, and faecal BA levels between TCA-supplemented Angptl4−/− and wild-type mice. In conclusion, 1) ANGPTL4 is not involved in the triglyceride-lowering effect of BA; 2) ANGPTL4 promotes BA absorption during TCA supplementation via a mechanism dependent on the gut microbiota.
The analysis of tetracyclines, quinolones, macrolides, lincosamides, pleuromutilins, and sulfonamides in chicken feathers using UHPLC-MS/MS in order to monitor antibiotic use in the poultry sector
Jansen, Larissa J.M. ; Bolck, Yvette J.C. ; Rademaker, Janneau ; Zuidema, Tina ; Berendsen, Bjorn J.A. - \ 2017
Analytical and Bioanalytical Chemistry 409 (2017)21. - ISSN 1618-2642 - p. 4927 - 4941.
Antibiotics - Feathers - LC-MS/MS - Monitoring - Validation
In The Netherlands, all antibiotic treatments should be registered at the farm and in a central database. To enforce correct antibiotic use and registration, and to enforce prudent use of antibiotics, there is a need for methods that are able to detect antibiotic treatments. Ideally, such a method is able to detect antibiotic applications during the entire lifespan of an animal, including treatments administered during the first days of the animals’ lives. Monitoring tissue, as is common practice, only provides a limited window of opportunity, as residue levels in tissue soon drop below measurable quantities. The analysis of feathers proves to be a promising tool in this respect. Furthermore, a qualitative confirmatory method was developed for the analyses of six major groups of antibiotics in ground chicken feathers, aiming for a detection limit as low as reasonably possible. The method was validated according to Commission Decision 2002/657/EC. All compounds comply with the criteria and, as a matter of fact, 58% of the compounds could also be quantified according to regulations. Additionally, we demonstrated that a less laborious method, in which whole feathers were analyzed, proved successful in the detection of applied antibiotics. Most compounds could be detected at levels of 2 μg kg−1 or below with the exception of sulfachloropyridazine, tylosin, and tylvalosin. This demonstrates the effectiveness of feather analysis to detect antibiotic use to allow effective enforcement of antibiotic use and prevent the illegal, off-label, and nonregistered use of antibiotics.
Gut microbiome-related metabolic changes in plasma of antibiotic-treated rats
Behr, C. ; Kamp, H. ; Fabian, E. ; Krennrich, G. ; Mellert, W. ; Peter, E. ; Strauss, V. ; Walk, T. ; Rietjens, I.M.C.M. ; Ravenzwaay, B. van - \ 2017
Archives of Toxicology 91 (2017)10. - ISSN 0340-5761 - p. 3439 - 3454.
Antibiotics - Metabolomics - Microbiome - Plasma metabolite profiling - Repeated dose oral toxicity study

The intestinal microbiota contributes to the metabolism of its host. Adequate identification of the microbiota’s impact on the host plasma metabolites is lacking. As antibiotics have a profound effect on the microbial composition and hence on the mammalian-microbiota co-metabolism, we studied the effects of antibiotics on the “functionality of the microbiome”—defined as the production of metabolites absorbed by the host. This metabolomics study presents insights into the mammalian-microbiome co-metabolism of endogenous metabolites. To identify plasma metabolites related to microbiome changes due to antibiotic treatment, we have applied broad-spectrum antibiotics belonging to the class of aminoglycosides (neomycin, gentamicin), fluoroquinolones (moxifloxacin, levofloxacin) and tetracyclines (doxycycline, tetracycline). These were administered orally for 28 days to male rats including blood sampling for metabolic profiling after 7, 14 and 28 days. Fluoroquinolones and tetracyclines can be absorbed from the gut; whereas, aminoglycosides are poorly absorbed. Hippuric acid, indole-3-acetic acid and glycerol were identified as key metabolites affected by antibiotic treatment, beside changes mainly concerning amino acids and carbohydrates. Inter alia, effects on indole-3-propionic acid were found to be unique for aminoglycosides, and on 3-indoxylsulfate for tetracyclines. For each class of antibiotics, specific metabolome patterns could be established in the MetaMap®Tox data base, which contains metabolome data for more than 550 reference compounds. The results suggest that plasma-based metabolic profiling (metabolomics) could be a suitable tool to investigate the effect of antibiotics on the functionality of the microbiome and to obtain insight into the mammalian-microbiome co-metabolism.

Critically ill patients demonstrate large interpersonal variation in intestinal microbiota dysregulation : a pilot study
Lankelma, Jacqueline M. ; Vught, Lonneke A. van; Belzer, Clara ; Schultz, Marcus J. ; Poll, Tom van der; Vos, Willem M. de; Wiersinga, W.J. - \ 2017
Intensive Care Medicine 43 (2017)1. - ISSN 0342-4642 - p. 59 - 68.
Antibiotics - Critically ill - Gut microbiota - Intensive care unit - Sepsis

Purpose: The intestinal microbiota has emerged as a virtual organ with essential functions in human physiology. Antibiotic-induced disruption of the microbiota in critically ill patients may have a negative influence on key energy resources and immunity. We set out to characterize the fecal microbiota composition in critically ill patients both with and without sepsis and to explore the use of microbiota-derived markers for clinical outcome measurements in this setting. Methods: In this prospective observational cohort study we analyzed the fecal microbiota of 34 patients admitted to the intensive care unit. Fifteen healthy subjects served as controls. The fecal microbiota was phylogenetically characterized by 16S rRNA gene sequencing, and associations with clinical outcome parameters were evaluated. Results: A marked shift in fecal bacterial composition was seen in all septic and non-septic critically ill patients compared with controls, with extreme interindividual differences. In 13 of the 34 patients, a single bacterial genus made up >50% of the gut microbiota; in 4 patients this was even >75%. A significant decrease in bacterial diversity was observed in half of the patients. No associations were found between microbiota diversity, Firmicutes/Bacteroidetes ratio, or Gram-positive/Gram-negative ratio and outcome measurements such as complications and survival. Conclusions: We observed highly heterogeneous patterns of intestinal microbiota in both septic and non-septic critically ill patients. Nevertheless, some general patterns were observed, including disappearance of bacterial genera with important functions in host metabolism. More detailed knowledge of the short- and long-term health consequences of these major shifts in intestinal bacterial communities is needed.

Long-term effects of early life microbiota disturbance on adaptive immunity in laying hens
Simon, K. ; Verwoolde, M.B. ; Zhang, J. ; Smidt, H. ; Vries Reilingh, G. De; Kemp, B. ; Lammers, A. - \ 2016
Poultry Science 95 (2016)7. - ISSN 0032-5791 - p. 1543 - 1554.
Antibiotics - Chicken - Immune response - Microbiota

Due to an interplay between intestinal microbiota and immune system, disruption of intestinal microbiota composition during immune development may have consequences for immune responses later in life. The present study investigated the effects of antibiotic treatment in the first weeks of life on the specific antibody response later in life in chickens. Layer chicks received an antibiotic cocktail consisting of vancomycin, neomycin, metronidazole, and amphotericin-B by oral gavage every 12 h, and ampicillin and colistin in drinking water for the first week of life. After the first week of life, chicks received ampicillin and colistin in drinking water for two more weeks. Control birds received no antibiotic cocktail and plain drinking water. Fecal microbiota composition was determined during antibiotic treatment (d 8 and 22), two weeks after cessation of antibiotic treatment (d 36), and at the end of the experimental period at d 175 using a 16S ribosomal RNA gene targeted microarray, the Chicken Intestinal Tract Chip (ChickChip). During antibiotic treatment fecal microbiota composition differed strongly between treatment groups. Fecal microbiota of antibiotic treated birds consisted mainly of Proteobacteria, and in particular E.coli, whereas fecal microbiota of control birds consisted mainly of Firmicutes, such as lactobacilli and clostridia. Two weeks after cessation of antibiotic treatment fecal microbiota composition of antibiotic treated birds had recovered and was similar to that of control birds. On d 105, 12 weeks after cessation of antibiotic treatment, chicks of both treatment groups received an intra-tracheal lipopolysaccharide (LPS)/human serum albumin (HuSA) challenge. Antibody titers against LPS and HuSA were measured 10 days after administration of the challenge. While T cell independent antibody titers (LPS) were not affected by antibiotic treatment, antibiotic treated birds showed lower T cell dependent antibody titers (HuSA) compared with control birds. In conclusion, intestinal microbial dysbiosis early in life may still have effects on the specific antibody response months after cessation of antibiotic treatment and despite an apparent recovery in microbiota composition.

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