Transfer of antibiotic resistance determinants between lactobacilli isolates from the gastrointestinal tract of chicken

2012 ◽  
Vol 3 (2) ◽  
pp. 137-144 ◽  
Author(s):  
F. Vieira de Souza ◽  
R. Roque ◽  
J.L. Silva Moreira ◽  
M. Resende de Souza ◽  
J.R. Nicoli ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Gastrointestinal Tract ◽  
Broiler Chickens ◽  
Lactobacillus Reuteri ◽  
Multi Drug Resistance ◽  
Bacterial Strains ◽  
Genetic Traits ◽  
In Vivo Experiments

The aim of this study was to assess the potential horizontal transfer of genetic traits for antibiotic resistance between lactobacilli isolated from the chicken gut, both in vitro and in vivo. Thirty-seven Lactobacillus spp. strains isolated from the gizzard, small and large intestines and caeca of free-range broiler chickens showed multi-drug resistance as assessed by disc diffusion assays. The minimum inhibitory concentration (MIC) for vancomycin, tetracycline, erythromycin and chloramphenicol was determined in De Man, Rogosa and Sharpe broth in a microplate assay. Almost all the lactobacilli isolates were resistant to vancomycin (except strains belonging to the Lactobacillus acidophilus group) and to tetracycline (MIC≥128 μg/ml). Only five strains were resistant to erythromycin, and six to chloramphenicol. The transfer rate in filter mating experiments performed using L. acidophilus strain 4M14E (EmR), Lactobacillus vaginalis strain 5M14E (CmR), Lactobacillus salivarius strain 5C14C (EmR), and the 4G14L and 3C14C strains of Lactobacillus reuteri (CmR) showed a frequency of approximately 1×104 cfu/ml of double-resistant transconjugants for the different combinations. The exception was the L. salivarius 5C14C (EmR) and L. vaginalis 5M14E (CmR) mating combination, which produced no transconjugants. In vivo experiments performed in gnotobiotic mice by mating L. acidophilus 4M14E (EmR) with L. reuteri 3C14C (CmR), L. reuteri 4G14L (CmR) or L. vaginalis 5M14E (CmR) resulted in transconjugants at 3.95±0.29, 3.16±0.33, and 4.55±1.52 log10 cfu/g of faeces, respectively. Taken together, these data suggest that genetic exchange may occur between native bacterial strains within the gastrointestinal tract of chickens, which might maintain a dynamic gene pool conferring antibiotic resistance upon indigenous microbiota components, even in the absence of the pathogens. This possibility must be taken into account as a complementary criterion when lactobacilli are screened for probiotic use.

scholarly journals Novel Lytic Phages Protect Cells and Mice against Pseudomonas aeruginosa Infection

2021 ◽  
Author(s):  
Feng Chen ◽  
Xingjun Cheng ◽  
Jianbo Li ◽  
Xiefang Yuan ◽  
Xiuhua Huang ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Mouse Models ◽  
Bacterial Infections ◽  
Phage Therapy ◽  
Inflammatory Responses ◽  
Multi Drug Resistance ◽  
Bacterial Strains ◽  
Genetic Traits

With the fast emergence of serious antibiotic resistance and the lagged discovery of novel antibacterial drugs, phage therapy for pathogenic bacterial infections has acquired great attention in the clinics. However, development of therapeutic phages also faces tough challenges, such as laborious screening and time to generate effective phage drugs since each phage may only lyse a narrow scope of bacterial strains. Identifying highly effective phages with broad host ranges is crucial for improving phage therapy. Here, we isolated and characterized several lytic phages from various environments specific for Pseudomonas aeruginosa by testing their growth, invasion, host ranges, and potential for killing targeted bacteria. Importantly, we identified several therapeutic phages (HX1, PPY9, and TH15) with broad host ranges to lyse laboratory strains and clinical isolates of P. aeruginosa with multi-drug resistance (MDR) both in vitro and in mouse models. In addition, we analyzed critical genetic traits related to the high-level broad host coverages by genome sequencing and subsequent computational analysis against known phages. Collectively, our findings establish that these novel phages may have potential for further development as therapeutic options for patients who fail to respond to conventional treatments. IMPORTANCE Novel lytic phages isolated from various environmental settings were systematically characterized for their critical genetic traits, morphology structures, host ranges against laboratory strains and clinical multi-drug resistant (MDR) Pseudomonas aeruginosa, and antibacterial capacity both in vitro and in mouse models. First, we characterized the genetic traits and compared with other existing phages. Furthermore, we utilized acute pneumonia induced by laboratorial strain PAO1, and W19, an MDR clinical isolate and chronic pneumonia by agar beads laden with FDR1, a mucoid phenotype strain isolated from the sputum of a cystic fibrosis (CF) patient. Consequently, we found that these phages not only suppress bacteria in vitro but also significantly reduce the infection symptom and disease progression in vivo, including lowered bug burdens, inflammatory responses and lung injury in mice, suggesting that they may be further developed as therapeutic agents against MDR P. aeruginosa.

Food supplement “carrageenan” and its effect on the organism

2020 ◽  
Author(s):  
O.E. Luneva ◽  
Keyword(s):  
Gastrointestinal Tract ◽  
Food Additives ◽  
Food Supplement ◽  
The Body ◽  
Laboratory Rats ◽  
Experimental Part ◽  
In Vivo Experiments ◽  
Physiological Processes

Food additives are positioned as harmless, although, their components affectthe physiological processes associated with the permeability of the wall of the gastrointestinal tract (GIT) and intestinal microbiota. This article describes thecarrageenan supplement and its effects on the body in in vitro and in vivo experiments. The experimental part is devoted to analysis of the intestinalmicrobiota of laboratory rats with the consumption of the carrageenan dietary supplement in the amount of about 4,4 % of the standard feed.

scholarly journals Curcumin Analogues as a Potential Drug against Antibiotic Resistant Protein, β-Lactamases and L, D-Transpeptidases Involved in Toxin Secretion in Salmonella typhi: A Computational Approach

2021 ◽  
Vol 2 (1) ◽  
pp. 77-100
Author(s):  
Tanzina Akter ◽  
Mahim Chakma ◽  
Afsana Yeasmin Tanzina ◽  
Meheadi Hasan Rumi ◽  
Mst. Sharmin Sultana Shimu ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Typhoid Fever ◽  
Homology Modeling ◽  
Salmonella Typhi ◽  
Multidrug Resistant ◽  
Drug Candidate ◽  
In Vivo Experiments ◽  
Curcumin Analogs

Typhoid fever caused by the bacteria Salmonella typhi gained resistance through multidrug-resistant S. typhi strains. One of the reasons behind β-lactam antibiotic resistance is -lactamase. L, D-Transpeptidases is responsible for typhoid fever as it is involved in toxin release that results in typhoid fever in humans. A molecular modeling study of these targeted proteins was carried out by various methods, such as homology modeling, active site prediction, prediction of disease-causing regions, and by analyzing the potential inhibitory activities of curcumin analogs by targeting these proteins to overcome the antibiotic resistance. The five potent drug candidate compounds were identified to be natural ligands that can inhibit those enzymes compared to controls in our research. The binding affinity of both the Go-Y032 and NSC-43319 were found against β-lactamase was −7.8 Kcal/mol in AutoDock, whereas, in SwissDock, the binding energy was −8.15 and −8.04 Kcal/mol, respectively. On the other hand, the Cyclovalone and NSC-43319 had an equal energy of −7.60 Kcal/mol in AutoDock, whereas −7.90 and −8.01 Kcal/mol in SwissDock against L, D-Transpeptidases. After the identification of proteins, the determination of primary and secondary structures, as well as the gene producing area and homology modeling, was accomplished. The screened drug candidates were further evaluated in ADMET, and pharmacological properties along with positive drug-likeness properties were observed for these ligand molecules. However, further in vitro and in vivo experiments are required to validate these in silico data to develop novel therapeutics against antibiotic resistance.

scholarly journals Identification of Lactobacillus reuteri Genes Specifically Induced in the Mouse Gastrointestinal Tract

2003 ◽  
Vol 69 (4) ◽  
pp. 2044-2051 ◽  
Author(s):  
Jens Walter ◽  
Nicholas C. K. Heng ◽  
Walter P. Hammes ◽  
Diane M. Loach ◽  
Gerald W. Tannock ◽  
...  
Keyword(s):  
Gastrointestinal Tract ◽  
Reporter Gene ◽  
Stress Responses ◽  
Lactobacillus Reuteri ◽  
Xylose Isomerase ◽  
Methionine Sulfoxide ◽  
Methionine Sulfoxide Reductase ◽  
Chromosomal Dna

ABSTRACT Lactobacilli are common inhabitants of the gastrointestinal tracts of mammals and have received considerable attention due to their putative health-promoting properties. Little is known about the traits that enhance the ability of these bacteria to inhabit the gastrointestinal tract. In this paper we describe the development and application of a strategy based on in vivo expression technology (IVET) that enables detection of Lactobacillus reuteri genes specifically induced in the murine gut. A plasmid-based system was constructed containing ′ermGT (which confers lincomycin resistance) as the primary reporter gene for selection of promoters active in the gastrointestinal tract of mice treated with lincomycin. A second reporter gene, ′bglM (β-glucanase), allowed differentiation between constitutive and in vivo inducible promoters. The system was successfully tested in vitro and in vivo by using a constitutive promoter. Application of the IVET system with chromosomal DNA of L. reuteri 100-23 and reconstituted lactobacillus-free mice revealed three genes induced specifically during colonization. Two of the sequences showed homology to genes encoding xylose isomerase (xylA) and peptide methionine sulfoxide reductase (msrB), which are involved in nutrient acquisition and stress responses, respectively. The third locus showed homology to the gene encoding a protein whose function is not known. Our IVET system has the potential to identify genes of lactobacilli that have not previously been functionally characterized but which may be essential for growth of these bacteria in the gastrointestinal ecosystem.

Bioavailability and Speciation of Mineral Micronutrients: The Enzymolysis Approach

1997 ◽  
Vol 80 (4) ◽  
pp. 920-927 ◽  
Author(s):  
Pierre Hocquellet ◽  
Marie-Dominique L'Hotellier
Keyword(s):  
Gastrointestinal Tract ◽  
Absorption Spectrometry ◽  
Insoluble Residue ◽  
In Vivo Experiments ◽  
Treatment Data ◽  
Dietary Components ◽  
Vitro Model ◽  
Good Agreement

Abstract Speciation analyses are essential to investigate the effects of dietary components on bioavailability of mineral micronutrients. Enzymolysis was used. An in vitro model simulating enzymatic activity in the gastrointestinal tract of monogastric species was developed and used to assess availability of Fe, Cu, Mn, and Zn in some foodstuffs. The solubility of each element in samples was measured by atomic absorption spectrometry after enzymatic treatment. Data are in good agreement with information obtained from earlier, more expensive nutritional surveys or in vivo experiments and, therefore, allow prediction of the tendency of a particular food to induce mineral deficiency. In addition, ligands responsible for inhibiting intestinal absorption were identified by determining the amount of metal released after treatment of the insoluble residue with an appropriate enzyme such as cellulase and phytase, used respectively to study fiber and phytate interactions. Enzymolysis may be useful for optimizing mineral supplementations though its nutritional significance is somewhat limited by the fact that it does not take into account the dynamic changes in the gastrointestinal tract. Enzymolysis is a prerequisite for further speciation studies of complex systems and in some instances is the only way for specifying physicochemical forms of elements.

scholarly journals In Vitro and In Vivo Study of the Gastrointestinal Absorption and Metabolisation of Hymenocardine, a Cyclopeptide Alkaloid

Planta Medica ◽  
2017 ◽  
Vol 83 (09) ◽  
pp. 790-796 ◽  
Author(s):  
Emmy Tuenter ◽  
Sebastiaan Bijttebier ◽  
Kenn Foubert ◽  
Annelies Breynaert ◽  
Sandra Apers ◽  
...  
Keyword(s):  
Gastrointestinal Tract ◽  
Rat Plasma ◽  
In Vivo Study ◽  
Root Bark ◽  
In Vivo Experiments ◽  
Cyclopeptide Alkaloid ◽  
Blood And Urine Samples ◽  
The Stability

AbstractHymenocardine is a cyclopeptide alkaloid present in the root bark of Hymenocardia acida. In traditional African medicine, the leaves and roots of this plant are used to treat malaria, and moderate in vitro antiplasmodial activity has been reported for hymenocardine. However, in view of its peptide-like nature, potential metabolisation after oral ingestion has to be taken into account when considering in vivo experiments. In this study, the stability and small intestinal absorption of hymenocardine was assessed using an in vitro gastrointestinal dialysis model. In addition, potential liver metabolisation was investigated in vitro by incubation with a human S9 fraction. Moreover, hymenocardine was administered to rats per os, and blood and urine samples were collected until 48 and 24 h after oral administration, respectively. All samples resulting from these three experiments were analyzed by LC-MS. Analysis of the dialysate and retentate, obtained from the gastrointestinal dialysis model, indicated that hymenocardine is absorbed unchanged from the gastrointestinal tract, at least in part. After S9 metabolisation, several metabolites of hymenocardine could be identified, the major ones being formed by the reduction and/or the loss of an N-methyl group. The in vivo study confirmed that hymenocardine is absorbed from the gastrointestinal tract unchanged, since it could be identified in both rat plasma and urine, together with hymenocardinol, its reduction product.

scholarly journals The Use of Probiotics in the Reduction of Campylobacter spp. Prevalence in Poultry

Animals ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 1355
Author(s):  
Marcin Śmiałek ◽  
Joanna Kowalczyk ◽  
Andrzej Koncicki
Keyword(s):  
Gastrointestinal Tract ◽  
Broiler Chickens ◽  
Poultry Meat ◽  
In Vivo Studies ◽  
Population Count ◽  
Sources Of Infection ◽  
Carcass Contamination ◽  
Campylobacter Spp

Campylobacter spp. are widely distributed microorganisms, many of which are commensals of gastrointestinal tract in multiple animal species, including poultry. Most commonly detected are C. jejuni and C. coli. Although infections are usually asymptomatic in poultry, poultry meat and products represent main sources of infection with these bacteria to humans. According to recent EFSA report, campylobacteriosis is the most commonly reported zoonotic disease. In 2018, EFSA Panel on Biological Hazards indicated that use of feed and water additives is the second most likely strategy that can be successful in minimizing Campylobacter spp. colonization rate in broiler chickens. One of those feed and water additives are probiotics. From numerous research papers it can be concluded that probiotics exhibit plenty of mechanisms of anti-Campylobacter activity, which were evaluated under in vitro conditions. These results, to some extent, can explain the efficacy of probiotics in in vivo studies, although different outcome can be observed under these two laboratory conditions. Probiotics are capable of reducing Campylobacter spp. population count in poultry gastrointestinal tract and they can reduce carcass contamination. Potential modes of anti-Campylobacter activity of probiotics, results of in vivo studies and studies performed at a farm level are widely discussed in the paper.

scholarly journals In Vitro and In Vivo Survival and Transit Tolerance of Potentially Probiotic Strains Carried by Artichokes in the Gastrointestinal Tract

2006 ◽  
Vol 72 (4) ◽  
pp. 3042-3045 ◽  
Author(s):  
Francesca Valerio ◽  
Palmira De Bellis ◽  
Stella Lisa Lonigro ◽  
Lorenzo Morelli ◽  
Angelo Visconti ◽  
...  
Keyword(s):  
Gastrointestinal Tract ◽  
Lactobacillus Plantarum ◽  
Bacterial Strains ◽  
Gastrointestinal Digestion ◽  
Feeding Study ◽  
Simulated Gastrointestinal Digestion ◽  
Probiotic Strains ◽  
Transit Tolerance

ABSTRACT The ability of potentially probiotic strains of Lactobacillus plantarum and Lactobacillus paracasei to survive on artichokes for at least 90 days was shown. The anchorage of bacterial strains to artichokes improved their survival in simulated gastrointestinal digestion. L. paracasei IMPC2.1 was further used in an artichoke human feeding study involving four volunteers, and it was shown that the organism could be recovered from stools.

scholarly journals Detection, Characterization, and In Vitro and In Vivo Expression of Genes Encoding S-Proteins in Lactobacillus gallinarum Strains Isolated from Chicken Crops

2005 ◽  
Vol 71 (11) ◽  
pp. 6633-6643 ◽  
Author(s):  
Karen E. Hagen ◽  
Le Luo Guan ◽  
Gerald W. Tannock ◽  
Doug R. Korver ◽  
Gwen E. Allison
Keyword(s):  
Broiler Chickens ◽  
Surface Proteins ◽  
Specific Gene ◽  
Published Data ◽  
Bacterial Strains ◽  
S Protein ◽  
Genes Encoding ◽  
S Proteins

ABSTRACT Thirty-eight isolates of Lactobacillus gallinarum cultured from the crops of broiler chickens were screened for the presence of genes encoding S-layer proteins. All of the isolates had two S-protein genes, which were designated Lactobacillus gallinarum S-protein (lgs) genes. One gene in each isolate was either lgsA or lgsB. The Lactobacillus isolates were further characterized by pulsed-field gel electrophoresis of DNA digests, which grouped the isolates into 17 genotypes (strains). The second gene in each of eight representative strains was sequenced and shown to differ among strains (lgsC, lgsD, lgsE, lgsF, lgsG, lgsH, and lgsI). The genome of each strain thus encoded a common S-protein (encoded by either lgsA or lgsB) and a strain-specific S-protein. The extraction of cell surface proteins from cultures of the eight strains showed that each strain produced a single S-protein that was always encoded by the strain-specific lgs gene. Two of the strains were used to inoculate chickens maintained in a protected environment which were Lactobacillus-free prior to inoculation. DNAs and RNAs extracted from the digesta of the chickens were used for PCR and reverse transcription-PCR, respectively, to demonstrate the presence and transcription of lgs genes in vivo. In both cases, only the strain-specific gene was transcribed. Both of the strains adhered to the crop epithelium, consistent with published data predicting that S-proteins of lactobacilli are adhesins. The results of this study provide a basis for the investigation of gene duplication and sequence variation as mechanisms by which bacterial strains of the same species can share the same habitat.

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