The comparative anti-oxidant and anti-inflammatory efficacy of postbiotics and probiotics through Nrf-2 and NF-kB pathways in DSS-induced colitis model.

IBD is a disorder which could be caused by oxidative stress. This investigation aims to determine if probiotics and postbiotics can control oxidative stress and inflammation and compare the effectiveness of these two probiotic and postbiotic mixtures of substances. 88 strains of Lactobacillus and Bifidobacterium were tested for antioxidant activity. Male wild-type C57BL/6 mice were divided into four experimental groups, namely high fat diet (HFD) + PBS, HFD + DSS, HFD + DSS + 109 cfu/ml of probiotics, and HFD + DSS + 109 cfu/ml of postbiotics. The phenotypical indices and pathological scores were assessed. The expression of genes related to NF-kB and Nrf2 signaling pathways and enzymes associated with oxidant/anti-oxidant activities, and proinflammatory/inflammatory cytokines were assessed. In contrast to the groups exposed to DSS, mice treated with probiotics mixture and postbiotics mixture alongside DSS displayed alleviation of DSS-induced adverse effects on phenotypical characteristics, as well as molecular indices such as the Nrf2 and NF-kB related genes, with a greater emphasis on the postbiotics component. In accordance with the findings of the present investigation, it can be inferred that even in using a high-fat dietary regimen as an inducer of oxidative stress, the emergence of inflammation can be effectively addressed through the utilization of probiotics and, more specifically, postbiotics.

The effect of novel paraprobiotic cocktail on dextran sodium sulfate induced acute colitis control focusing on autophagy signaling pathway.

PURPOSE: Paraprobiotics are a non-viable form of probiotics that are reported to provide significant health benefits. Nevertheless, little is known about the beneficial effects of paraprobiotics on inflammatory bowel disease. Although probiotics show potential as therapeutic agents for a range of diseases, including inflammatory bowel disease (IBD), there are certain risks associated with their use. These risks include toxin production, hemolytic potential, antibiotic resistance, and the need to analyze metabolic activities. Hence Using paraprobiotic with the lower aforementioned risk would therefore be the preferable option. Here, we conducted an in vivo study to evaluate the preventive effect of our native paraprobiotic cocktail against dextran sulfate sodium (DSS)-induced murine colitis by affecting the autophagy signaling pathway. METHODS: Four-week-old male C57Bl/6 mice were randomly divided into three groups after a two-week acclimation period with normal standard laboratory food diet. Mice were administered PBS (PBS group as control), PBS along with DSS (DSS group, as a control), and a cocktail of paraprobiotics along with DSS (Para group). The severity of colitis, length and histopathology of the colon were evaluated. In addition, the expression of autophagy was assessed using real-time PCR. RESULTS: The results showed that administration of the paraprobiotic cocktail to DSS-treated mice inhibited the severity of colitis symptoms, as evidenced by the inhibition of weight loss and DAI, as well as histopathological scores in the study colon, as well as shortening of colon length caused by DSS. In contrast to the DSS group, the cocktail was able to modulate inflammation through upregulation of autophagy-related genes (becline 1, atg5, atg7, atg12, and atg13). CONCLUSION: Although there are some limitations in our investigation, such as the dosage and duration of treatments, our native paraprobiotic blend effectively prevented the advancement of colitis. This suggests that it plays a vital role in regulating inflammation and preventing colitis by promoting the autophagy mechanism in cases where the consumption of probiotics may have negative consequences.

Investigating the crucial role of selected Bifidobacterium probiotic strains in preventing or reducing inflammation by affecting the autophagy pathway.

Several studies have shown that probiotics can prevent and reduce inflammation in inflammation-related diseases. However, few studies have focused on the interaction between host and probiotics in modulating the immune system through autophagy. Therefore, we aimed to investigate the preventive and/or therapeutic effects of native potential probiotic breast milk-isolated Bifidobacterium spp. (i.e. B. bifidum, B. longum, and B. infantis) on the inflammatory cascade by affecting autophagy gene expression 24 and 48 h after treatment. Autophagy genes involved in different stages of the autophagy process were selected by quantitative polymerase chain reaction (qPCR). Gene expression investigation was accomplished by exposing the human colorectal adenocarcinoma cell line (HT-29) to sonicated pathogens (1.5 × 108 bacterial CFU ml-1) and adding Bifidobacterium spp. (MOI10) before, after, and simultaneously with induction of inflammation. An equal volume of RPMI medium was used as a control. Generally, our native potential probiotic Bifidobacterium spp. can increase the autophagy gene expression in comparison with pathogen. Moreover, an increase in gene expression was observed with our probiotic strains' consumption in all stages of autophagy. Totally, our selected Bifidobacterium spp. can increase autophagy gene expression before, simultaneously, and after the inflammation induction, so they can prevent and reduce inflammation in an in vitro model of inflammation.

Decoding the genetic structure of conjugative plasmids in international clones of Klebsiella pneumoniae: A deep dive into blaKPC, blaNDM, blaOXA-48, and blaGES genes.

Carbapanem-resistant Klebsiella pneumoniae is a globally healthcare crisis. The distribution of plasmids carrying carbapenemase genes among K. pneumoniae poses a serious threat in clinical settings. Here, we characterized the genetic structure of plasmids harboring major carbapenemases (e.g. blaKPC, blaNDM, blaOXA-48-like, and blaGES) from K. pneumoniae using bioinformatics tools. The plasmids carrying at least one major carbapenemase gene were retrieved from the GenBank database. The DNA length, Inc type, and conjugal apparatus of these plasmids were detected. Additionally, allele types, co-existence, co-occurrence of carbapenemase genes, gene repetition, and sequence types of isolates, were characterized. There were 2254 plasmids harboring carbapenemase genes in the database. This study revealed that blaKPC-2, blaNDM-1, blaOXA-48, and blaGES-5 were the most prevalent allele types. Out of 1140 (50%) plasmids were potentially conjugative. IncFII, IncR, IncX3, and IncL replicon types were predominant. The co-existence analysis revealed that the most prevalent of other resistance genes were blaTEM-1 (related to blaKPC), blaOXA-232 (related to blaOXA-48), bleMBL (related to blaNDM), and aac (6')-Ib4 (related to blaGES). The co-occurrence of carbapenemases was detected in 42 plasmids while 15 plasmids contained carbapenemase gene repetitions. Sequence alignments highlighted that plasmids carrying blaKPC and blaOXA-48-like were more homogeneous whereas the plasmids carrying blaNDM were divergent. It seems that K. pneumoniae utilizes diversity of genetic flexibility and recombination for resistance against carbapenems. The genetic structure of the plasmids showed that class I and III, Tn3 family, Tn5403 family derivatives, and Tn7-like elements were strongly associated with carbapenemases. The mobilizable plasmids carrying carbapenemases play an important role in the spread of these genes. In addition, gene repetition maybe is related to carbapenem heteroresistance. According to MST (minimum spanning tree) results, the majority of plasmids belonged to sequence type (ST) 11, ST14, and ST12. These international clones have a high capacity to acquire the carbapenemase-containing plasmids.

The notable relatedness between ESBL producing Enterobacteriaceae isolated from clinical samples and asymptomatic fecal carriers.

BACKGROUND: The investigation of the presence of extended-spectrum beta-lactamase (ESBL) within Enterobacteriaceae in both fecal carriers and patients is an essential matter. Furthermore, the assessment of distinct characteristics exhibited by resistant bacteria obtained from fecal carriers and patients, as well as the comparison of these characteristics between the two groups, could provide a deeper understanding of how the resistant isolates can remain concealed within a dormant reservoir and intensify antimicrobial resistance. The aim of the present study was to concentrate on the comparison of the antimicrobial resistance pattern and molecular features between strains obtained from clinical and carrier sources. MATERIAL AND METHODS: A total of 142 clinical samples and 120 rectal swabs were collected from June to October 2016. ESBL screening was performed using the double-disk synergy test. PCR was done for the detection of ESBL genes. Assessment of biofilm formation, virulence factor genes, and MLVA was performed for K. pneumonae isolates. Phylogroup typing was performed for E. coli isolates. RESULTS: Of 146 samples, 67.6% were E. coli, and 32.4% were K. pneumoniae. The rate of blaCTXM-15 was 89.4%. In K. pneumoniae type D, ompk35 and fimH were the highest. All the K. pneumoniae isolates were classified into 12 mini clusters and the clinical isolates were characterized into 7 mini clusters. The phylogroup B2 in ESBL-EC was the highest (56.2%). DISCUSSION: Comparison of molecular characteristics and clonal relatedness between fecal carriers and patients showed noticeable relatedness and similarity which may indicate that ESBL-KP can be colonized with the same profiles in different settings and, therefore, may be widely distributed in both community and hospital settings. Therefore, implementation of control protocols, including surveillance of the fecal carriers, could impressively reduce silent reservoirs without clinical symptoms as well as patient rates.

Ameliorating inflammation in an in vitro model by screening the anti-inflammatory and immunomodulatory roles of putative probiotics in inflammatory bowel disease.

IBD is considered a relapsing disease with relapsing phases. Probiotics are beneficial microorganisms that modulate inflammatory signaling pathways. Our aim was to identify the precise molecular effects of probiotics on inflammatory signaling pathways during the presence of inflammation. Evaluation of the expression of JAK/STAT and inflammatory genes after treatment of the HT -29 cell line with the sonicated pathogens and probiotics, simultaneously was performed by quantitative real-time polymerase chain reaction (qPCR) assay. The production of IL-6 and IL-1ß after administration of probiotics was conducted by means of cytokine assay. The probiotic cocktail resulted in the downregulation of TIRAP, IRAK4, NEMO, and RIP genes in the NF-кB pathway compared with Sonicat-treated cells. The expression of JAK/STAT genes was various after probiotic treatment. The application of probiotics has been observed to result in a notable decrease in the production of IL-6 and IL-1ß. The investigated probiotic cocktail, especially Bifidobacterium spp. showed anti-inflammatory effects on HT -29 cells via modulation of JAK/STAT and NF-кB signaling pathways. The use of probiotics with the least side effects could be considered a suitable treatment for patients with inflammatory bowel disease, even at the beginning of inflammation.

Investigation of the anti-inflammatory effects of native potential probiotics as supplementary therapeutic agents in an in-vitro model of inflammation.

BACKGROUND: IBD is considered an inflammatory disease with abnormal and exaggerated immune responses. To control the symptoms, different theraputic agents could be used, however, utilizing the agents with the least side effects could be important. Probiotics as beneficial microorganisms are one of the complementory theraputic agents that could be used to modulate inflammatory signaling pathways. In the current study, we aimed to identify the precise molecular effects of potential probiotics on signaling pathways involved in the development of inflammation. METHODS: A quantitative real-time polymerase chain reaction (qPCR) assay was used to analyze the expression of JAK /STAT (JAK1, JAK2, JAK3, TYK2, STAT1, STAT2, STAT3, STAT4, STAT5 and STAT6) and inflammatory genes (NEMO, TIRAP, IRAK, and RIP) after the HT -29 cell line treatment with the sonicated pathogens and potential probiotics. A cytokine assay was also used to evaluate IL -6 and IL -1ß production after potential probiotic treatment. RESULTS: The potential probiotic cocktail downregulated the JAK genes and TIRAP, IRAK4, NEMO, and RIP genes in the NF-kB pathway compared with cells that were treated with sonicated gram negative pathogens. The expression of STAT genes was different after potential probiotic treatment. The production of IL -6 and IL -1ß decreased after potential probiotic treatment. CONCLUSIONS: Considering the importance of controlling the symptoms of IBD to improve the life quality of the patients, using probiotic could be crucial. In the current study the studied native potential probiotic cocktails showed anti-inflammatory effects via modulation of JAK /STAT and NF-kB signaling pathways. This observation suggests that our native potential probiotics consumption could be useful in reducing intestinal inflammation.

Probiotics as functional foods: How probiotics can alleviate the symptoms of neurological disabilities.

Neurological disorders are diseases of the central nervous system with progressive loss of nervous tissue. One of the most difficult problems associated with neurological disorders is that there is no clear treatment for these diseases. In this review, the physiopathology of some neurodegenerative diseases, etiological causes, drugs used and their side effects, and finally the role of probiotics in controlling the symptoms of these neurodegenerative diseases are presented. Recently, researchers have focused more on the microbiome and the gut-brain axis, which may play a critical role in maintaining brain health. Probiotics are among the most important bacteria that have positive effects on the balance of homeostasis via influencing the microbiome. Other important functions of probiotics in alleviating symptoms of neurological disorders include anti-inflammatory properties, short-chain fatty acid production, and the production of various neurotransmitters. The effects of probiotics on the control of abnormalities seen in neurological disorders led to probiotics being referred to as "psychobiotic. Given the important role of the gut-brain axis and the imbalance of the gut microbiome in the etiology and symptoms of neurological disorders, probiotics could be considered safe agents that positively affect the balance of the microbiome as complementary treatment options for neurological disorders.

Determination of seroprevalence of brucellosis in livestock and high-risk population in Kurdistan, Western Iran.

BACKGROUND: Brucellosis is one of the most prevalent zoonotic diseases. Using serological tests are valid and rapid methods that could be used in the detection of the history of getting brucellosis. Considering that Iran is an endemic country for brucellosis, we aimed to investigate the rate of seroprevalence of brucellosis among livestock and human in Kurdistan province. MATERIAL AND METHOD: Serum sampling was performed from 51 slaughterhouse workers, veterinarians, and husbandry workers, along with 260 livestock (80 cattle, 120 sheep, and 60 goats). Rose Bengal plate test (RBPT), and Enzyme-linked immunosorbent assay (ELISA) were used for livestock and the anti-Brucella IgG antibody was evaluated in human participants. RESULTS: The seroprevalence (based on ELISA assay) in sheep, goats, and cows was 5.8%, 5%, and 1.2%, respectively. Also, the rate of anti-Brucella IgG was 3.9% among human participants. DISCUSSION: the current study, provided some valuable information on the seroprevalence of brucellosis in animal and human participants from the west of Iran. Considering the effects of brucellosis on causing reproductive disorders, including abortion, placental retention, andendometritis controlling the infection could have a significant impact on terms of economy.

Antibiotic resistance and the alternatives to conventional antibiotics: The role of probiotics and microbiota in combating antimicrobial resistance.

From the introduction of the first antibiotic to the present day, the emergence of antibiotic resistance has been a difficult problem for medicine. Regardless of the type of antibiotic resistance, the presence of resistant isolates in clinical and even asymptomatic fecal carriers becomes a difficult public health problem. Therefore, the use of new antimicrobial combination therapies or alternative agents with antimicrobial activity that have the least side effects, including plant-, metal-, and nanoparticle-based agents, could be crucial and useful. Recently, the use of probiotics as a hypothetical candidate to combat infectious disease control and antimicrobial resistance has received notable attention. Considering the alteration of the microbiota in fecal carriers and also in patients with resistant bacterial isolates, the use of probiotics could have an appropriate effect on the balance of the microbial population. In this review, we have attempted to discuss the history of antimicrobial resistance and provide an overview of microbiota change and the use of probiotics as new agents with antimicrobial activity associated with the emergence of resistant isolates.

The Role of Combining Probiotics in Preventing and Controlling Inflammation: A Focus on the Anti-Inflammatory and Immunomodulatory Effects of Probiotics in an In Vitro Model of IBD.

Objective: IBD is an inflammatory disease with abnormalities such as dysbiosis and abnormal immune system activity. Probiotics, as live beneficial microorganisms, play a role in maintaining health through various mechanisms, including the modulation of the immune system and the control of inflammation. Here, we aimed to investigate the efficacy of a probiotic mixture of Lactobacillus spp. and Bifidobacterium spp. in modulating JAK/STAT and NF-kB inflammatory signaling pathways. Method: A quantitative real-time polymerase chain reaction (qPCR) assay was conducted to analyze the expression of JAK/STAT and inflammatory genes after treatment with the probiotic mixture before, after, and simultaneously with the sonicated pathogen in the HT-29 cell line. The production of IL-6 and IL-1ß after probiotic treatment was investigated via cytokine assay. Results: Treatment with probiotics resulted in downregulation of TIRAP, IRAK4, NEMO, and RIP genes in the NF-kB pathway and JAK/STAT genes compared with sonicat-treated cells as inflammation inducers. The production of IL-6 and IL-1 decreased after probiotic treatment. Conclusions: The probiotic mixture of Lactobacillus spp. and Bifidobacterium spp. showed anti-inflammatory effects by modulating JAK/STAT and NF-kB signaling pathways. The use of probiotics could be considered as an appropriate complementary treatment for patients with inflammatory bowel disease.

Anti-inflammatory and immunomodulatory effects of Lactobacillus spp. as a preservative and therapeutic agent for IBD control.

BACKGROUND: Probiotics have a beneficial effect on inflammatory responses and immune regulation, via Janus kinase/signal transduction and activator of transcription (JAK/STAT) and NF-κB signaling pathways. To evaluate the precise effects of Lactobacillus spp. as a protective and therapeutic agent, we aimed to investigate the efficacy of Lactobacillus spp. in modulating JAK/STAT and nuclear factor kappa B (NF-κB) inflammatory signaling pathways. METHODS: A quantitative real-time polymerase chain reaction (qPCR) assay was used to analyze the expression of JAK/STAT and inflammatory genes (TIR-associated Protein [TIRAP], Interleukin 1 Receptor Associated Kinase[IRAK4], Nuclear factor-kappa B Essential Modulator [NEMO], and receptor interacting protein [RIP]) followed by treatment of the HT-29 cell line with sonicated pathogens before, after, and simultaneously with Lactobacillus spp. A cytokine assay was also used to evaluate interleukin (IL)-6 and IL-1ß production after treatment with Lactobacillus spp. RESULTS: Lactobacillus spp. downregulated JAK and TIRAP, IRAK4, NEMO, and RIP genes in the NF-κB pathway compared to sonicate-treated cells. The expression of STAT genes was different after treatment with probiotics. The production of IL-6 and IL-1ß decreased after probiotic treatment. CONCLUSIONS: Our Lactobacillus spp. cocktail showed anti-inflammatory effects on HT-29 cells by modulating JAK/STAT and NF-κB signaling pathways in all three treatment variants. Therefore, Lactobacillus spp. as a dietary supplement can both prevent and reduce inflammation-related diseases such as inflammatory bowel disease.

High heterogeneity of fecal carriage extended-spectrum beta-lactamase-producing E. coli isolated from iranian community and clinical settings.

BACKGROUND: Extended-spectrum beta-lactamase-producing enterobacteria (ESBL-PE) in carriers have become a global health problem. Using molecular typing techniques, including PFGE, could be useful to determine the source of bacterial dissemination. The current study aimed to investigate the intestinal carriage of ESBL-producing E. coli (ESBL-EC) and clonal relatedness among ESBL-EC isolated from hospitalized and outpatient fecal carriers in Iran. METHODS: A total of 120 rectal swabs were collected; 50.8% (61/120) from intensive care unit (ICU) inpatients and 49.2% (59/120) from outpatients. MacConkey agar enriched with cefotaxime was used to screen the ESBL-EC. PCR assays were performed to detect ESBL and carbapenemase genes. Pulse-fields gel electrophoresis (PFGE) was performed to assess clonal relatedness. RESULTS: Totally, 60.0% (72/120) were carrier for ESBL-EC. The rates of resistance against ceftazidime and cefepime were 90.2% (65/72) and 93.0% (67/72), respectively. The rates of blaCTX-M-15, blaTEM, blaSHV, blaNDM-1, blaOXA-48 and blaIMP was 90.2% (65/72), 50.0% (36/72), 5.5% (4/72), 4.1% (3/72), 4.1% (3/72) and 1.3% (1/72), respectively. Based on a cut-off 80%, 69 ESBL-EC isolates could be categorized in 10 mini-cluster and 47 isolates were considered as singletons. DISCUSSION: High heterogeneity among isolates from ESBL-EC suggests that this bacterium probably has a different source of dissemination. Screening of carriers in hospitals and communities could help the infection control program in public health.

The effects of the probiotic cocktail on modulation of the NF-kB and JAK/STAT signaling pathways involved in the inflammatory response in bowel disease model.

BACKGROUND: Probiotics positively affect inflammatory responses, in part, through Janus kinase/signal transduction and activator of transcription (JAK/STAT) and inflammatory signaling pathways. To evaluate the precise effects of probiotics as protective treatment, we aimed to investigate the effectiveness of Lactobacillus spp., Bifidobacterium spp., and a mixture of these probiotics in modulating the JAK/STAT and inflammatory signaling pathways. METHODS: A quantitative real-time polymerase chain reaction (qPCR) assay was used to analyze the expression of JAK/STAT and inflammatory genes (TIRAP, IRAK4, NEMO, and RIP) following HT-29 cell line treatment with sonicated pathogens Lactobacillus spp., Bifidobacterium spp., and a mixed cocktail. A cytokine assay was also used to evaluate the IL-6 and IL-1ß production following the probiotic treatment. RESULTS: The probiotic cocktail downregulated the JAK genes and TIRAP, IRAK4, NEMO, and RIP genes in the NF-kB pathway compared to sonicate pathogen treatment cells. The expression of STAT genes was variable following probiotic treatment. The IL-6 and IL-1ß production decreased after probiotic treatment. CONCLUSIONS: Our probiotic cocktail showed anti-inflammatory effects on HT-29 cells by modulating JAK/STAT and NF-kB pathways. Therefore, Lactobacillus spp. and Bifidobacterium spp. probiotics as nutritional supplements may reduce inflammation-associated diseases such as inflammatory bowel disease (IBD).

Seroepidemiology of leptospirosis in livestock and workers of high-risk occupation in Kurdistan, Iran.

BACKGROUND: Leptospirosis is one of the major zoonotic infectious diseases which could cause disease in both animals and humans. Using ELISA is one of the serological tests that could be used in the detection of leptospirosis. Based on the different reports about the prevalence of leptospirosis in different parts of our country, we aimed to investigate the rate of Leptospira spp, among livestock and human in Kurdistan province. MATERIAL AND METHOD: ELISA assay was performed by ELISA kit (Novatec, Germany) for quantitative detection of anti-Leptospira IgG in human and IgM antibody and total antibodies (IgM and IgG) in serum samples of livestock. RESULTS: In the present study, the seroprevalence in sheep, goats, and cows was (2/30) 6.7% [95% CI 0.8%,22.1%], (1/31) 3.2% [ 95% CI 0.08%,16.7%] and 0%, respectively. Also, the rate of anti-Leptospira was (1/51) 1.9% [95% CI 0.05%,10.4%] among 51 human participants. DISCUSSION: the current study, provided some valuable information on the rate of leptospirosis in animal and human participants from west of Iran, which can be useful in terms of monitoring the disease in the area and helping the health care system to control the roots of bacterial transmission.

The monitoring of Francisella tularensis in surface water of East Azerbaijan province, Iran.

BACKGROUND: Francisella tularensis could be disseminated through arthropod bites and exposure to infected animals, water, and aerosols. Water sources that are contaminated with rodent excrement could be a source of contamination; therefore, an analysis of water samples is an appropriate method to investigate the routes of dissemination. Since an outbreak occurred in one of the villages in East Azerbaijan. The current study aimed to investigate the Francisella isolation in the different water samples from East Azerbaijan, Iran. Sampling was carried out in East Azerbaijan province. Forty-six specimens of surface water were collected. Filtration, culture, and inoculation of the water sample into NMRI (Naval Medical Research Institute) inbreed mice were performed. DNA was extracted from filtered water samples, different organs of inoculated mice, and bacterial isolates and was tested by TaqMan real-time PCR by targeting ISFtu2 and fopA genes. Despite the unsuccessfulness in isolation of F. tularensis, molecular test results indicate the presence of bacteria in surface water. The highest rate of F. tularensis (ten from 46 water samples, 21.7%) was detected from injected mice based on molecular methods. Despite the high efforts of researchers to isolate Francisella spp. in Iran, in recent years, and also the evidence that shows the presence of this bacterium in different parts of the country, the culture was not successful again in this study and the molecular method still is recommended to identify the possible sources of Francisella spp.

First Report of Extended-Spectrum Betalactamase-Producing Klebsiella pneumoniae Among Fecal Carriage in Iran: High Diversity of Clonal Relatedness and Virulence Factor Profiles.

Increasing rate of silent intestinal carriers with extended-spectrum betalactamase (ESBL)-producing Klebsiella pneumonia (ESBL-KP) has given rise to a serious healthcare problem in clinical settings. Various epidemiological studies are being conducted to determine clonal relatedness among carriers. In this study, we investigated the intestinal carriage of ESBL-KP and clonal relatedness among ESBL-KP isolated from fecal carriage in Iran for the first time. A total of 120 rectal swabs (RSs) were collected including 61 from inpatients of intensive care unit and 59 from outpatients. ESBL-KP screening was performed using MacConkey agar supplemented with cefotaxime. PCR was done for detection of ESBL, carbapenemase, and virulence factor genes. Conjugation experiments and PCR-based replicon typing were performed. Clonal relatedness was investigated by multilocus sequence typing (MLST) and multiple locus variable number tandem repeat analysis (MLVA). Out of a total of 120 RSs, 18.3% (22/120) ESBL-KP were isolated. The rate of blaCTXM-15 was 81%. ompk35 was the most prevalent virulence gene detected in 86.3% of the isolates. In conjugation experiments, three out of five tested isolates had conjugative plasmids. The most prevalent plasmid types belonged to IncL/M, IncA/C, and Inc FII. The MLST analysis showed that the main sequence types (STs) identified among ESBL-KP isolates were ST147, ST15, and ST16. The isolates were characterized into 4 miniclusters and 11 singletons using MLVA. High heterogeneity among ESBL-KP isolates indicated that this bacterium could be colonized in different sites and easily transferred. Screening of carriers in hospitals and community could help in controlling of infection in the healthcare and community settings.

Considerable rate of putative virulent phylo-groups in fecal carriage of extended-spectrum ß-lactamase producing Escherichia coli.

Extended-Spectrum Beta-lactamase producing Enterobacteriales (ESBL-PE) in fecal carriage have become a global health concern. Detection of putative virulent ESBL-producing E.coli (ESBL-EC) isolates among asymptomatic carriers is a threatening issue in public health. The aim of this study was to investigate the intestinal carriage of ESBL-EC, phylo-groups and clonal relatedness among putative virulent groups of ESBL-EC isolated from fecal carriages. A total of 120 rectal swabs; 50.8% (61/120) from inpatients of intensive care unit (ICU) and 49.2% (59/120) from outpatients were collected. The ESBL-EC screening was performed by using MacConkey agar supplemented with cefotaxime. PCR assays were applied for determination of phylo-groups, detection of ESBL and carbapenemase genes. Conjugation experiment, plasmid replicon typing and Multilocus Sequence Typing (MLST) were performed for putative virulent phylo-groups. Totally, of 120 studied individuals, 60.0% (72/120) were carrier for ESBL-EC. The rate of blaCTX-M-15, blaTEM, blaSHV was 90.2% (65/72), 50.0% (36/72) and 5.5% (4/72), respectively. The frequency of phylo-groups A, B1, B2, C, D, and F were 20.8% (15/72), 6.9% (5/72), 20.8% (15/72), 2.7% (2/72), 13.8 (10/72) and 12.5% (9/72), respectively. In conjugation experiments, of 6 tested isolates, 5 had conjugative plasmids. The most prevalent plasmid types belonged to IncF incompatibility groups. The MLST analysis showed that the main sequence types among ESBL-EC isolates were ST769 and ST472. The current study provides novel information about the presence of the ESBL-EC isolates, particularly putative virulent phylo-groups among fecal carriages in Iran. Our data revealed that there was almost high ST heterogeneity among putative ESBL-EC isolates. In order to implementation of effective infection control program, detection of fecal carriage in appropriate time typically at the beginning of admission to the hospital is recommended.

Molecular epidemiology of NDM-1- and OXA-48-producing Klebsiella pneumoniae in an Iranian hospital: clonal dissemination of ST11 and ST893.

Objectives: Despite the fact that the blaOXA-48 and blaNDM-1 genes have successfully disseminated among Klebsiella pneumoniae isolates worldwide, outbreaks remain unidentified in Iran. Here we examined the molecular epidemiology of 96 carbapenem-resistant K. pneumoniae recovered from an Iranian hospital. Methods: A total of 96 non-replicate carbapenem-resistant K. pneumoniae were recovered from clinical specimens in a university hospital. Detection of ESBLs and carbapenemases produced by studied strains was performed using PCR and DNA sequencing. The bacterial isolates were assigned to clonal lineages by PFGE and MLST. In addition, plasmids were analysed by PCR-based replicon typing and conjugation assays. Results: All isolates harboured blaOXA-48 and blaNDM-1 genes together or alone. Almost all strains also carried ESBL genes. Eighty-seven isolates of K. pneumoniae were categorized into seven pulsotypes. The predominant strain clusters/pulsotypes associated with the outbreak corresponded to ST11 (48/96) and ST893 (31/96). Plasmids carrying blaOXA-48 and blaNDM-1 were successfully transferred to Escherichia coli K12 as the recipient strain. blaOXA-48 was located on IncL/M plasmids of ∼39 kb, while blaNDM-1 was carried by either an IncFII plasmid of ∼50 kb or an untypeable plasmid of ∼4 or 10 kb. Conclusions: We describe two separate outbreaks of blaOXA-48- and blaNDM-1-carrying K. pneumoniae strains associated with dissemination of the ST11 and ST893 clones, with the ICU acting as the epicentre. The spread of plasmids carrying carbapenemase genes resulting in fulminant antimicrobial resistance is a severe concern.

Emergence of carbapenem resistant Escherichia coli isolates producing blaNDM and blaOXA-48-like carried on IncA/C and IncL/M plasmids at two Iranian university hospitals.

The emergence of carbapenem resistance among Escherichia coli is a serious threat to public health. The objective of this study was to investigate resistance genes and clonality of carbapenem resistant E. coli in Iran. Between February 2015 and July 2016, a total of 32 non-duplicate E. coli isolates that were ertapenem resistant or intermediate (R/I-ETP) were collected from patient clinical or surveillance cultures (rectal swabs) at two university hospitals. Resistance genes were identified by PCR and sequencing. Conjugation experiments, PCR-based replicon typing, PFGE and multilocus sequence typing (MLST) were performed. PCR assays showed, among the 32 isolates, twenty-nine strains produced carbapenemase genes. The predominant carbapenemase was blaOXA-48 (82.8%), followed by blaNDM-1 (31%), blaNDM-7 (6.9%) and blaOXA-181 (3.4%). Seven of the blaNDM positive isolates co-harbored blaOXA-48 carbapenemases. The blaNDM and blaOXA-48 were found in IncA/C and IncL/M conjugative plasmids, respectively. The blaCTX-M-15, qnrA and intI1 genes were also present in most isolates. The PFGE revealed genetic diversity among the 28 E. coli isolates, which belonged to six minor PFGE clusters and 14 isolates were singletons. The 26 isolates were distributed into 18 STs, of which two were dominant (ST648 and ST167). We identified one blaNDM-1-positive ST131 E. coli isolates that harbor the blaCTX-M-15 and blaTEM genes. Horizontal transfer of IncA/C and IncL/M plasmids has likely facilitated the spread of the blaOXA-48 and blaNDM genes among E. coli. Their clonal diversity and the presence of faecal carriers in isolates suggest an endemic spread of OXA-48 and NDM. Therefore, it emphasizes the critical importance of monitoring and controlling the spread of carbapenem resistant E. coli.