Nalidixic Acid-resistant Strains Research Articles

Effect of a Bacillus subtilis plus Yeast Cell Wall Synbiotic on Salmonella Enteritidis Colonization in Ceca of Layer Pullets

Salmonella Enteritidis (SE) is a major contamination concern in eggs and risk for Salmonellosis in humans. Strains of Bacillus subtilis and yeast cell wall can be used as substitutes for antibiotic substances in feed against Salmonella in poultry. The objective of this study was to assess the effect of BacPack® (Quality Technology International, Inc., Elgin, IL, USA) Q1+1 (BPQ11), a feed additive combination of a Bacillus subtilis strain and Saccharomyces cerevisiae cell wall, on SE cecal colonization in Lohmann LSL pullets. A control group (CON) and a test group (BPQ) were each randomly assigned 100-day-old chicks. CON was fed a corn–soybean meal-based vegetarian mash diet, and BPQ was fed the control diet supplemented with BPQ11 for the duration of the study. At 8 days of age, chicks were orally challenged with a nalidixic acid-resistant SE strain at a dose of 6.3 × 107 colony forming units (CFUs) per bird. At 7, 11, 15, and 19 days post-challenge (DPC), 25 birds per group were euthanized, and their cecal contents were collected and analyzed for SE. SE counts were 6.88, 7.98, 7.79, and 7.50 in CON and 7.18, 7.31, 6.35, and 6.30 log10 CFU/g in BPQ at 7, 11, 15, and 19 DPC, respectively. SE did not differ between CON and BPQ at 7 DPC; however, BPQ had lower (p < 0.0001) SE at 11 (−0.67), 15 (−1.45), and 19 (−1.20 log10 CFU/g) DPC. Results indicate that synbiotic BPQ11 may be a useful dietary pre-harvest tool for SE management in layer birds.

Molecular detection of genes encoding resistance to tetracycline and quinolones among Shigella strains isolated from children with acute diarrhea in southwest Iran.

An increase in the antibiotic resistance of Shigella isolates has caused major global challenges in antimicrobial therapy. Knowledge of local antibiotic resistance trends is essential for selecting appropriate antibiotic treatment regimens. This study aimed to evaluate the frequency of efflux-mediated tetracycline resistance (tet) and plasmid-mediated quinolone resistance (qnr) genes among Shigella isolates. This survey investigated 91 Shigella isolates, obtained from children with acute diarrhea. The isolates were identified using standard biochemical tests and confirmed by polymerase chain reaction (PCR) assay. Besides, the susceptibility of isolates to six selected antibiotics was assessed by the disk diffusion method. All tetracycline-resistant and nalidixic acid and ciprofloxacin resistant strains were screened for tet and qnr genes by a multiplex PCR assay. According to the results of antibiotic susceptibility tests, the highest level of antibiotic resistance was related to tetracycline (80.2%) and doxycycline (78.1%), respectively. All isolates were sensitive to tigecycline. The PCR results showed that 40.6%, 3.1%, 21.8%, 61.6% and 28.7% of the isolates carried qnrA, qnrB, qnrS, tetA, and tetB genes, respectively. None of the isolates contained tetC and tetD genes. The current findings revealed that tetA and qnrA genes might play a key role in conferring tetracycline and quinolone resistance.

In vitro and in vivo evaluation of tannic acid as an antibacterial agent in broilers infected with Salmonella Typhimurium

This study was conducted to evaluate tannic acid (TA) as an antibacterial agent against Salmonella Typhimurium in in vitro and in vivo chicken models. The TA formed an inhibitory zone against Salmonella enterica serotypes including S. Typhimurium, S. Enteritidis, and S. Infantis. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of TA against Salmonella Typhimurium nalidixic acid resistant strain (STNR) were determined as 40 and 700 μg/mL, respectively. Sublethal doses of TA (5, 10, and 20 μg/mL) restricted swimming and swarming motility and biofilm formation of STNR compared to the control group (0 μg/mL) (P < 0.05). The TA-bovine serum albumin (BSA) complex formed at simulated gastric pH (pH 3.75) was hydrolyzed at pH 6.75 and 7.25 (P < 0.05), and the hydrolysis of the TA-BSA complex was stronger at pH 7.25 compared to the pH 6.75 (P < 0.05). The inhibitory zone of the TA-BSA complex against STNR at pH 6.75 was lower than TA without BSA at 30 and 60 min (P < 0.05), but not at 120 min (P > 0.1). The inhibitory zone of the TA-BSA complex against STNR at pH 7.25 was not decreased at 0, 30, and 60 min compared to TA without BSA (P > 0.1). The recovery rate of TA was 83, 54.8, 10.5, and 19.6% in the gizzard, jejunum, ileum, and ceca, respectively, in broiler chickens. The STNR-infected broilers fed 0.25 g/kg of TA had significantly lower unweighted beta diversity distance compared to the sham-challenged control (SCC) and challenged controlled (CC) group on D 21. TA supplementation linearly (P < 0.05) and quadratically (tendency; P = 0.071) reduced relative abundance of the family Peptostreptococcaceae in broilers infected with STNR on D 7. TA supplementation linearly (P < 0.05) and quadratically (tendency; P = 0.06) increased the relative abundance of the family Erysipelotrichaceae in broilers infected with STNR on D 21. Therefore, TA has potential to be used as an antibacterial agent against the S. Typhimurium infection in broilers.

Evaluation of a Triple Buffered Peptone Broth for Detection of Salmonella in Broiler Feed

The pH of pre-enrichment media containing feed/ingredients can become acidic during incubation due to bacterial utilization of feed carbohydrates. This decrease in pH can result in cell injury or death, negatively impacting the detection of Salmonella. Our objective was to evaluate a new triple buffered peptone (TBP) against buffered peptone water (BPW) and lactose broth (LB) for the recovery of Salmonella from feed. Liquid cultures of nalidixic acid resistant strains of Salmonella (Enteritidis, Heidelberg, Kentucky or Typhimurium) were added to the pre-enrichment media alone, to pre-enrichment media containing feed or to artificially inoculated feed stored 1 or 7 d to evaluate the effect of the medium on the recovery of Salmonella. Three replicates per treatment were conducted. After incubation at 37 °C for 24 h, the pH of the medium was measured prior to plating onto brilliant green sulfa agar plates supplemented with 200 ppm nalidixic acid (BGSNA). Plates were incubated and evaluated for presence of typical Salmonella colonies. The experiment was replicated. TBP was observed to exhibit significantly better buffering capacity than BPW or LB. Additionally, TBP was able to recover Salmonella 100% of the time compared to BPW (97.9%) and LB (61.5%). TBP shows promise to maintain neutral pH during pre-enrichment which may allow for a more accurate detection of Salmonella in feed.

Molecular Detection of gyrA Mutation in Clinical Strains of Klebsiella pneumoniae

Multi-drug resistant (MDR) Klebsiella pneumoniae strains cause the majority of community acquired and life-threatening infections. We aimed to detect the gyrA mutations in the clinical strains of nalidixic acid and ciprofloxacin resistant K. pneumoniae isolated from the patients with urinary tract infections. Bacterial strains were isolated from the patients with urinary tract infections admitted to a major hospital in Tehran, Iran (2017-2018). Bacterial identification was done according to standard microbiological tests. Antimicrobial susceptibility testing for quinolones and fluoroquinolone antibiotics was done using both disc diffusion and minimal inhibitory concentrations (MICs) methods. PCR-RFLP was used to detect the probable mutation in the gyrA gene in nalidixic acid and ciprofloxacin resistant strains. Finally sequencing was performed to detect point mutations in isolated K. pneumoniae strains. One hundred K. pneumonia isolates were recovered from the urine samples of the clinical cases. Antibiotic resistance testing showed that among all K. pneumoniae isolates, 26% and 19% of the strains were resistant to nalidixic acid and ciprofloxacin respectively. MIC value was ≥4 μg/ml for ciprofloxacin resistant isolates. The results of RFLP on gyrA PCR amplicons using HinfI restriction enzyme showed point mutation in this gene in 46% of nalidixic acid and ciprofloxacin resistant K. pneumonia. The data obtained from the sequencing confirmed the RFLP results and indicated the presence of point mutations in codons 83 and 87 in the gyrA gene which leads to the substitution of different amino acids in gyrA protein. Our findings indicated a relative increased rate of resistance against quinolones and fluoroquinolone antibiotics that raised a concern about extensive dissemination of clinical strains of nalidixic acid and ciprofloxacin resistant K. pneumonia. Point mutation of gyrA gene was responsible for the resistance in our strains however to gain more insight into the molecular characterization of quinolone-resistant isolates, other possible mechanisms of the resistance should also be investigated.

Antimicrobial and immunomodulatory effects of tannic acid supplementation in broilers infected with Salmonella Typhimurium

Infection by Salmonella Typhimurium, a food-borne pathogen, can reduce the poultry production efficiency. The objective of this study was to investigate the effects of tannic acid (TA) supplementation on growth performance, Salmonella colonization, gut barrier integrity, serum endotoxin levels, antioxidant capacity, gut health, and immune function in broilers infected with the Salmonella enterica serovar Typhimurium nalidixic acid resistant strain (STNR). A total of 546 one-day-old broilers were arbitrarily allocated into 6 treatments including 1) Sham-challenged control (SCC; birds fed a basal diet and administrated peptone water); 2) Challenged control (CC; birds fed a basal diet and inoculated with 108 STNR); 3) Tannic acid 0.25 (TA0.25; CC + 0.25 g/kg TA); 4) TA0.5 (CC + 0.5 g/kg TA); 5) TA1 (CC + 1 g/kg TA); and 6) TA2 (CC + 2 g/kg TA). On D 7, supplemental TA linearly reduced STNR colonization in the ceca (P < 0.01), and TA1 and TA2 group had significantly lower reduced STNR colonization in the ceca (P < 0.01). On D 7 to 21, average daily gain tended to be linearly increased by supplemental TA (P = 0.097). The serum endotoxin levels were quadratically decreased by supplemental TA on D 21 (P < 0.05). Supplemental TA quadratically increased ileal villus height (VH; P < 0.05), and the TA0.25 group had higher ileal VH compared to the CC group (P < 0.05). Supplemental TA linearly increased percentage of peripheral blood CD8+ T cells on D 18 (P < 0.01). The TA0.5 group had significantly lower lymphocyte numbers compared to the CC groups (P < 0.05). The abundance of monocytes linearly increased with TA supplementation (P < 0.01). Therefore, broilers fed TA had reduced STNR colonization, increased growth performance, decreased serum endotoxin levels, enhanced gut health in the broilers, and stimulated the immune system in broilers infected with STNR. Supplementation of TA (1–2 g/kg) enhanced growth performance and gut health via antimicrobial and immunostimulatory effects in broilers infected with STNR.

Molecular Characterization of Antibiotic Resistance and Genetic Diversity of Klebsiella pneumoniae Strains.

The aims of this study were the molecular characterization of antibiotic resistance and genotyping of Klebsiella pneumoniae strains isolated from clinical cases in Tehran, Iran. A total of 100 different types of clinical human samples were collected from a major teaching hospital in Tehran, Iran. Bacterial isolates were identified using standard microbiological tests. Antimicrobial susceptibility testing was done according to the latest CLSI guidelines. PCR was used to amplify the gyrA gene in quinolone-resistant isolates and sequencing was performed for the detection of probable mutations between the isolates. The occurrence of plasmid-mediated quinolone resistance genes (qnrA, qnrB, and qnrS) was also investigated by PCR. Finally, genotyping of the strains was performed by PFGE in a standard condition. The susceptibility pattern revealed a high and low level of resistance against meropenem (20%) and trimethoprim (37%), respectively. PCR and sequencing detected mutation in the gyrA gene in 51% of quinolone-resistant K. pneumoniae. According to the susceptibility report, among nalidixic acid-resistant strains, 60.5%, 50%, and 42.9% of isolates contained qnrA, qnrB, and qnrS, respectively. Among ciprofloxacin-resistant strains, qnrA was the most frequent PMQR gene. The PFGE differentiated the strains into 31 different genetic clusters so that the highest number (7/66) was in category A. Our results indicated that the frequency of resistance to various antibiotics particularly trimethoprim, nalidixic acid, and cefoxitin are increasing. The presence of qnr (S and A) genes and point mutation of the gyrA gene were likely to be responsible for the resistance toward nalidixic acid and ciprofloxacin in our strains. Also, the results obtained from genotyping indicated that the K. pneumoniae strains isolated in this study belonged to the diverse clones.

Molecular Characterization of Quinolone Resistance Determinants in Non-Typhoidal Salmonella Strains Isolated in Tehran, Iran

Background: Quinolone resistant Salmonella serotypes have been reported in recent years and have become increasingly widespread worldwide. Objectives: We evaluated the molecular mechanism of quinolone resistance in non-typhoidal Salmonella strains isolated from clinical samples in Tehran, Iran. Methods: The present study included the Salmonella isolates originated from hospitalized individuals and outpatients in Tehran, Iran. Serotyping of nalidixic acid-resistant Salmonella isolates was done by slide agglutination method. Then, the quinolone resistance-determining region (QRDR) of topoisomerase gene gyrA and the plasmid-mediated quinolone resistance (PMQR) determinants were detected using the polymerase chain reaction (PCR) method. Restriction fragment length polymorphism (RFLP) analysis was also employed to determine the possible mutation in the gyrA gene of those strains. Mutant strains were detected by enzymatic digestion, and their PCR products were sequenced immediately. Results: Amongst 141 isolates, 60% showed nalidixic acid resistance, whereas none of them were ciprofloxacin-resistant. The commonly prevalent serotypes were S. Enteritidis and S. Infantis. Of 85 nalidixic acid-resistant strains, 17 (20%) isolates harbored the qnrS gene. However, PCR analysis of the quinolone-resistant strains did not detect qnrA and qnrB genes. PCR-RFLP and sequencing analysis of the QRDRs of the gyrA gene indicated that 16 (18.8%) isolates had mutant patterns, and the most common point mutation was serine to phenylalanine at position 83. Conclusions: Our results demonstrated that point mutations in gyrA and the existence of plasmid-mediated gene qnrS were important mechanisms of quinolone resistance in non-typhoidal Salmonella strains isolated from human origin. Other alternative mechanisms of resistance, such as alterations in the expression of efflux pumps, should be studied to provide greater insight into the molecular mechanism of quinolone-resistant non-typhoidal Salmonella isolates.

The effect of crisping, misting, and storage temperature on the survival or growth of Listeria monocytogenes and natural psychrotrophic bacteria on romaine lettuce.

L. monocytogenes has been linked to fresh produce and detected in the retail environment. This study simulated the retail practices (crisping, misting, and storage) of unbagged whole heads of romaine lettuce to determine the growth of L. monocytogenes and natural psychrotrophic microflora. Three nalidixic acid-resistant strains of L. monocytogenes strains were inoculated to each head of lettuce (≈5 log10 CFU/g). For crisping, 24 heads of romaine lettuce were immersed in tap water or electrolyzed water (EW; free chlorine: 55 ppm) for 5 min, followed by holding at 5 °C for 2 h. The water-crisped (WC), EW crisped (EWC), or non-crisped (NC) lettuces were placed in a commercial refrigerated cabinet for misting at 5 °C. After 24-h misting, heads of lettuce were placed in perforated drain boxes with cover at 5 °C or 15 °C. The tap water and EW crisping achieved 1.3 and 2.9 log10 CFU/g reduction of L. monocytogenes, respectively. Approximately 1 log additional reduction of L. monocytogenes in the non-crisped lettuce was shown after misting (p < 0.05), but no significant effect of misting on the population of L. monocytogenes was observed on WC or EWC lettuces (p > 0.05). Regardless of the storage temperature or misting, L. monocytogenes populations remained significantly (p < 0.05) lower on EWC lettuce than NC and WC lettuce. On days 4 and 7 of storage, the natural psychrotrophic bacteria on lettuce stored at 5 °C was significantly lower than stored at 15 °C, and its population was not affected by crisping and misting. These provide insight into the influence of retail lettuce handling practices on the risk of L. monocytogenes.

Effects of a synbiotic combination of Bacillus subtilis and yeast cell wall-derived glucomannan on cecal colonization of Salmonella Enteritidis in layer chickens

Salmonella Enteritidis ( SE ) has been a major foodborne concern associated with poultry and poultry products. Direct-fed microbials ( DFM ), such as Bacillus subtilis strains , and mannan-oligosaccharides ( MOS ) added into poultry feed have emerged as an alternative approach to antimicrobials to reduce Salmonella colonization in chickens. This study was conducted to evaluate the effect of a synbiotic feed additive product (BacPack Q1+1) combining a selected Bacillus subtilis strain and yeast cell wall-derived MOS on SE colonization in the ceca of Hy-line W36 pullets. Day-old chicks were raised in cage units and orally challenged at 3 wk-of-age with a nalidixic acid resistant SE strain at a dose of 1.6 × 10 9 colony forming units ( CFU ) per bird. SE colonization was assessed by enumeration of SE in cecal pouches at 7, 10 and 13 days postinfection ( dpi ). SE was not detectable in the ceca of 68% of synbiotic supplemented pullets compared to 36% of the controls at 13 dpi ( P = 0.049). The findings of this study suggest that synbiotic combinations of DFM and prebiotics could be a useful tool for reducing the prevalence of Salmonella in layer chickens.

Reduction of Salmonella Infantis on skin-on, bone-in chicken thighs by cetylpyridinium chloride application and the impact on the skin microbiota

Salmonella Infantis has been the etiological agent of numerous foodborne outbreaks of nontyphoidal Salmonella. Consequently, there is an emergent need to mitigate Salmonella Infantis among poultry. Thus, this study evaluated the efficacy of cetylpyridinium chloride (CPC) versus peroxyacetic acid (PAA), on bone-in, skin-on chicken thighs for the reduction of Salmonella and changes in the microbiota. Exactly 100 skin-on, bone-in chicken thighs (2 trials, 0 and 24 h, k = 5, n = 5, N = 50) were inoculated with 108 CFU/mL of a nalidixic acid resistant strain of S. Infantis for an attachment of 106 CFU/g. Thighs were treated with 20 s part dips (350 mL): a no inoculum, no treatment control (NINTC); no treatment control (NTC); tap water (TW); TW+CPC; TW+PAA. Following treatment, thighs were rinsed in 150 mL of nBPW, and rinsates were collected. Rinsates were spot plated for Salmonella and aerobic bacteria (APC). Log10 transformed counts were analyzed using a mixed-effects model (random effect = trial) with means separated using Tukey's HSD (P ≤ 0.05). The genomic DNA of rinsates was extracted, and the 16S rDNA was sequenced on an Illumina MiSeq. Microbiota data were analyzed using QIIME2, with data considered significant at P ≤ 0.05 (main effects) and Q≤0.05 (pairwise differences). Treatment × time interactions were observed for both Salmonella and APC (P < 0.05). The treatment of thighs with PAA and CPC reduced Salmonella and APC in respect to the controls. Numerically, thighs treated with CPC had less Salmonella (4.29 log10CFU/g) and less APC (4.56 log10CFU/g) at 24 h than all other treatments (P > 0.05). Differences in diversity metrics were not consistently observed between treatments; however, in trial 2, the NTC treated thighs were different than those treated with CPC (P < 0.05; Q < 0.05). In both trials, ANCOM, the analysis of microbiome compositional profiles, revealed shifts at both the phylum and order levels with thighs being different in the relative abundances of Proteobacteria (P < 0.05). In conclusion, treatment of skin-on poultry parts with CPC may reduce the risk of foodborne outbreaks caused by Salmonella Infantis.

Combination of probiotic and prebiotic impacts Salmonella Enteritidis infection in layer hens

Mitigation of food pathogens such as Salmonella continues to be a focus in poultry production, whether on shell eggs or meat products. No single approach is fully able to eliminate the risk, but rather comprehensive programs are used in a multifaceted approach. Targeted in-feed additives are a component of a food safety program and may help to reduce Salmonella infection. This study was aimed to evaluate the efficacy of a probiotic and prebiotic combination feed-additive product (Fulfill Plus) to reduce Salmonella Enteritidis ( S.E. ) infection in layer hens. One-hundred and forty-four (144) 1-day-old Hy-line pullets were raised on new litter until 10 wk of age when they were moved into individual cages. There were 3 treatments, 1) Untreated 2) probiotic and prebiotic combination given D 0 to termination, and 3) probiotic and prebiotic combination given Wk 10 to termination. At 17 weeks-of-age each bird was orally challenged with 3 × 10 6 CFU/bird nalidixic acid resistant strain of S.E. Seven (7) and 14 d postchallenge, ceca and ovaries were aseptically collected and cultured for S.E. Significant differences ( P < 0.05) in prevalence and mean cecal S.E. log 10 MPN/g were observed between 7 d postinfection compared with 14 d postinfection. Addition of probiotic and prebiotic combination at wk 10 (Treatment 3) significantly ( P < 0.05) reduced S.E. numbers in the ceca of these pullets compared to probiotic and prebiotic combination given at D 0 (Treatment 2). Significant differences ( P < 0.05) in prevalence and mean ovary S.E. log10 MPN/g were observed with Treatment 2 being significantly lower than Treatment 3.

Effect of dietary direct-fed microbial and yeast cell walls on cecal digesta microbiota of layer chicks inoculated with nalidixic acid resistant Salmonella Enteritidis

Salmonella enterica serovar Enteritidis (SE) has consistently been the most common serotype associated with the foodborne Salmonellosis worldwide. In this study, the effect of a dietary direct-fed microbial (DFM) and yeast cell walls (YCW) under a challenge of nalidixic acid resistant SE strain using layer chicks has been investigated. A total of 160 newly hatched Dekalb White female chicks were randomly assigned into 2 experimental groups (80 birds/treatment), control group (CON) and treatment group (DY). Chicks were fed ad libitum a non–medicated-corn-soy based diet and DY was supplemented with the combination of DFM and YCW. At 8 days of age, 2.1 × 109 CFU/bird of the SE was given to all chicks by oral administration. On 3 days postinoculation (dpi), 20 chicks/group were euthanized and all cecal contents were collected for analysis. On 6, 10, and 14 dpi, the cecal contents were sampled from 16 chicks per group. The number of SE in the cecal contents was counted using culture-based methods. A 16S rRNA-based microbiota analysis was performed for additional microbial profiling. The CON and DY showed difference (P ≤ 0.05) in β diversity throughout the trial. Prevalence of SE in cecal contents was lower (P ≤ 0.05) in DY across all time-points. Lower abundance of Salmonella spp. was also shown in DY by liner discriminant analysis effect size (LEfSe). DY increased (P ≤ 0.05) diversity of bacterial species in the cecal contents in DY at 10 and 14 dpi. For the SE challenged birds, SE reduction in DY was observed at 3 dpi and until the end of the trial at 14 dpi confirming a numerically larger difference between groups as well as an increase in bacterial species diversity in DY. It could be hypothesized that the SE reduction shown immediately after the challenge and the greater SE reduction shown after 10 dpi may be the synergistic effect of the combined feed additives.

The use of roller swabs for Salmonella detection in poultry litter

Litter sampling is utilized as a noninvasive and practical method to determine broiler flock Salmonella status. The common methods include boot cover/sock ( BC ), drag swab ( DS ), or litter grab sampling ( LG ). Roller swabs are a new research method that can be used to sample litter without entering research pens. This study aimed to assess the use of roller swabs ( RS ) for Salmonella ( S ) detection in comparison to BC, DS, and LG. For Experiment 1, litter was sampled for two weeks following a broiler flock that was challenged at 6 days of age with 1 × 10 7 cfu of a nalidixic acid resistant strain of Salmonella Enteritidis to establish a high litter prevalence of S. In Experiment 2, sampling occurred after a subsequent flock was raised on the same litter. In Experiment 1, S was detected by RS less frequently (81%) than DS (95%), BC (97%), and LG (98%; P < 0.0001). In Experiment 2, S detection using RS (23%) was similar to LG (17%), higher than DS (6%, P = 0.022), and lower than BC (55%, P < 0.0001). Although RS were a less sensitive sampling method when the litter prevalence of S was high, RS were equivalent to or better than LG and DS methods when S prevalence was low. The use of roller swabs allows for sampling of litter without entering each pen and has the potential to be utilized for Salmonella detection in research pen trials.

The Role of Aquatic Ecosystems (River Tua, Portugal) as Reservoirs of Multidrug-Resistant Aeromonas spp.

The inappropriate use of antibiotics, one of the causes of the high incidence of antimicrobial-resistant bacteria isolated from aquatic ecosystems, represents a risk for aquatic organisms and the welfare of humans. This study aimed to determine the antimicrobial resistance rates among riverine Aeromonas spp., taken as representative of the autochthonous microbiota, to evaluate the level of antibacterial resistance in the Tua River (Douro basin). The prevalence and degree of antibiotic resistance was examined using motile aeromonads as a potential indicator of antimicrobial susceptibility for the aquatic environment. Water samples were collected from the middle sector of the river, which is most impacted area by several anthropogenic pressures. Water samples were plated on an Aeromonas-selective agar, with and without antibiotics. The activity of 19 antibiotics was studied against 30 isolates of Aeromonas spp. using the standard agar dilution susceptibility test. Antibiotic resistance rates were fosfomycin (FOS) 83.33%, nalidixic acid (NA) 60%, cefotaxime (CTX) 40%, gentamicin (CN) 26.67%, tobramycin (TOB) 26.67%, cotrimoxazole (SXT) 26.67%, chloramphenicol (C) 16.67%, and tetracycline (TE) 13.33%. Some of the nalidixic acid-resistant strains were susceptible to fluoroquinolones. Multiple resistance was also observed (83.33%). The environmental ubiquity, the natural susceptibility to antimicrobials and the zoonotic potential of Aeromonas spp. make them optimal candidates for studying antimicrobial resistance (AMR) in aquatic ecosystems. Aquatic environments may provide an ideal setting for the acquisition and dissemination of antibiotic resistance because anthropogenic activities frequently impact them. The potential risk of multi- and pan-resistant bacteria transmission between animals and humans should be considered in a “One Health—One World” concept.

Effects of a mannan-rich yeast cell wall-derived preparation on cecal concentrations and tissue prevalence of Salmonella Enteritidis in layer chickens.

Salmonella Enteritidis (SE) has been the most common Salmonella serotype associated with foodborne infections in the last several years. Dietary applications of yeast-based preparations in feed have shown to reduce Salmonella colonization in chickens augmenting SE control strategies. This study was conducted to evaluate the effects of a mannan-rich yeast cell wall-derived preparation (Actigen®) administered in feed at a rate of 400 g/ton on SE colonization in the cecum and internal organs of commercial layer chickens. Sixteen week-old layer pullets were orally challenged with a selected nalidixic acid resistant SE strain at a dose of 1.7×10^9 colony forming units (CFU) per bird. SE colonization was assessed by evaluating isolation rates from ovary and pooled liver/spleen samples as well as enumeration of SE in cecal pouches one week post-challenge. Recovery rates of SE from the ovaries of directly challenged birds receiving Actigen® were significantly lower (P <0.02) than those in directly challenged birds fed an unsupplemented control diet. Recovery rates of SE from pooled liver/spleen samples were not significantly different between Actigen®-treated pullets and controls (P = 0.22). Using direct plate count methods, cecal SE concentrations were 1 log10 lower (P <0.001) in challenged pullets in the Actigen®-supplemented group than in the challenged controls. The SE concentration distributions in the ceca were similar in groups testing positive and groups testing negative for SE in the ovaries and liver/spleens tissues. As a result, SE concentrations in the ceca could not be directly related to the occurrence or prevalence of SE in these tissues. In conclusion, Actigen® supplementation appears to decrease the prevalence of SE in ovarian tissue and concentrations of SE in cecal contents and may be useful as a tool for reducing the risk of eggshell contamination and transovarian transmission of SE in eggs.

Green Synthesis of Silver Nanoparticles with Antimicrobial Properties Using Phyla dulcis Plant Extract.

Foodborne illnesses caused by the consumption of food contaminated with foodborne pathogens at infectious doses are becoming a common health issue throughout the world. Metal nanoparticles with potential antimicrobial properties are an area that can be explored to discover novel antimicrobial agents. The traditional synthesis methods of metal nanoparticles involve the use of toxic chemicals and the generation of harmful byproducts. In this study, a greener method to synthesize silver nanoparticles (AgNPs) with potential antimicrobial properties was investigated. The aqueous extract of the medicinal plant Phyla dulcis Trev. (verbenaceae) was used as the reducing and stabilizing agent to synthesize AgNPs using microwave irradiation. The formation of AgNPs was confirmed using ultraviolet-visible spectroscopy by the appearance of characteristic surface plasmon resonance peaks in the 430-440 nm wavelength range. The size and stability of the AgNPs were studied using Zetasizer nano-series for 5 weeks after synthesis. The average particle size remained between 63 and 76 nm during the first 4 weeks and increased to 114 nm in the fifth week showing possible aggregation after the fourth week. The zeta potential remained between -20 and -24 mV throughout the 5 weeks showing relatively good stability. Scanning electron microscopy/energy dispersive X-ray spectroscopy showed the association of phytoconstituents with the AgNPs. X-ray photoelectron spectroscopy analysis confirmed the formation of metallic nanoparticles starting from silver nitrate. Finally, the AgNPs were tested to be effective against Escherichia coli O157:H7 (ATCC 43888), Salmonella Typhimurium (novobiocin and nalidixic acid resistant strain), Listeria monocytogenes (4b; ATCC 19115), and Staphylococcus aureus (ATCC 6538) strains, which are known to be common foodborne pathogens.

Microbial virulence, molecular epidemiology and pathogenic factors of fluoroquinolone-resistant Haemophilus influenzae infections in Guangzhou, China

BackgroundFluoroquinolone-resistant Haemophilus influenzae (FRHI) has been reported worldwide but remain unclear in China.MethodsA total of 402 H. influenzae isolates collected from 2016 to 2017 were included. Antimicrobial susceptibility on 10 antibiotics was performed, and minimum inhibitory concentration of ciprofloxacin- and nalidixic acid-resistant strains were further determined by E-test strips, with risk factors also evaluated. Strains with resistance or reduced susceptibility to ciprofloxacin were subjected to sequencing of the quinolone resistance-determining regions (QRDR) and plasmid-mediated quinolone resistance genes by sequencing, with multi-locus sequence typing.Results2.2% of H. influenzae strains were non-susceptible (7/402, 1.7%) or susceptible (2/402, 0.5%) to ciprofloxacin but NAL-resistant by E-test, and multidrug resistance was more common in fluoroquinolones non-susceptible H. influenzae group (p = 0.000). Infection risk factors included invasive procedure (p = 0.011), catching cold/previous contact with someone who had a cold (p = 0.019), fluoroquinolones use during previous 3 months (p = 0.003). With none of mutations obtained in gyrB, parE and other plasmid-mediated quinolone resistance genes, 7 and 4 strains were found for Ser-84-Leu substitutions in gyrA and one amino acid substitution in the QRDR of gyrA linked with one amino acid substitution in the QRDR of parC, respectively. In addition, five sequence types (ST) were identified, with ST1719 firstly found.ConclusionsFor the first time, this study has reported the incidence, risk factors, molecular determinants on fluoroquinolones resistance and ST of FRHI strains in mainland China, representing the first evidence of mutation of gyrA and parC in China and the new ST1719 worldwide.

A Comparative Study Between Cefixime and Ofloxacin in The Treatment of Uncomplicated Typhoid Fever Attending A Tertiary Care Teaching Hospital

Background: Typhoid fever is a gastrointestinal infection caused by Gram-negative bacterium Salmonella enterica serovar typhi (S.typhi). Typhoid fever continues to be one of the major public health problems in developing countries. Antimicrobial therapy is critical for the clinical management of enteric fever. Incidences of multi-drug resistance to S. typhi (MDRST) and nalidixic acid resistant (NAR) strain have limited treatment options. Resistance pattern and time to fever clearance vary in different geographical areas and overtimes. Hence, this study was conducted to compare efficacy and safety profile of Cefixime and Ofloxacin in uncomplicated typhoid fever in this region.Method: 50 adults proven cases of typhoid fever of the age group of 18-57 years of either sex were included in the study. Group I was treated with Cefixime 200 mg twice a day for 7 days and CG I was treated with Ofloxacin 200 mg twice a day for 7 days. Patients were clinically and bacteriologically evaluated during the study period and follow-up.Result: So both study groups were found comparable in terms of mean fever clearance time. 96% cure rates were observed in both groups. No relapse was recorded.Conclusion: Both Cefixime and Ofloxacin are equally efficacious and safe in the treatment of uncomplicated typhoid fever.Med Phoenix Vol.2(1) July 2017, 3-7

Nalidixic acid screening test in detection of decreased fluoroquinolone susceptibility in Salmonella typhi isolated from blood.

To determine the validity of nalidixic acid screening test in the detection of high MICs of fluoroquinolone against Salmonella(S.) typhi isolated from blood and correlate zone diameters of ofloxacin with that of MIC value for nalidixic acid sensitive and resistant strains. Cross-sectional analytical study. Clinical Microbiology Laboratory of the Aga Khan Hospital, Karachi from January 2002 to December 2003. Two hundred S. typhi isolates from blood were included for nalidixic acid screening and ofloxacin susceptibility. Antibiotic susceptibilities for both the antibiotics were obtained by disc diffusion method whereas MICs were determined by standard agar dilution method as recommended by NCCLS guidelines. Sensitivity, specificity and correlation between both antimicrobial susceptibility methods were calculated and results expressed as scattergrams. The results broadly classify S. typhi isolates into nalidixic acid resistant strains with no zone of inhibition around 30 mug nalidixic acid disc and nalidixic acid sensitive strains with mean zone of inhibition of 24.9 mm. All S. typhi isolates with ofloxacin MIC of capital ZHE, Cyrillic 0.125 microg/ml were found to be nalidixic acid resistant (MIC capital ZHE, Cyrillic32 microg/ml) whereas the isolates with ofloxacin MIC 0.06 microg/ml were nalidixic acid sensitive (MIC 8 microg/ml). Screening for nalidixic acid resistance was found to be 100% sensitive and 97% specific in identifying S. typhi strains with reduced susceptibility to fluoroquinolone (MIC capital ZHE, Cyrillic 0.125 microg/ml). Nalidixic acid resistance as a screening method is proved to be significant in identifying S. typhi isolates with reduced susceptibility to fluoroquinolones. It is also suggested that inhibition zone of 25 mm around 5 microg ofloxacin disc is appropriate as a selection criterion to detect S. typhi isolates with reduced susceptibility to fluoroquinolones.