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Zoonotic origin of antibiotic resistant coliform bacteria has become a great threat to human health. Poultry meat is one of the most common sources of protein that has become the source of many antibiotic resistant bacteria. This investigation aimed to identify the prevalence of antibiotic resistant Escherichia coli strains in poultry meat that are being sold in different market places of Guwahati city. Single disc diffusion method was used to screen 98 E. coli isolates for their susceptibility against 11 different antimicrobial agents, namely, ampicillin, imipenem, cefotaxime, cefpodoxime, ceftazidime, cefoxitin, chloramphenicol, amikacin, streptomycin, nalidixic acid and tetracycline. Double disk diffusion test with cefotaxime and ceftazidime (third generation cephalosporin agents) was done to identify ESBL strains among them. All isolates were resistant to a minimum of 2 antimicrobial agents. Eighty two percent were resistant to broad spectrum antibiotics, such as, tetracycline, 80% to nalidixic acid, 76% to ampicillin. Prevalence of multiple antibiotic resistant or multi-drug resistant and ESBL positive isolates was 86% and 33%, respectively. This study has clearly revelled the risk factor associated with consumption or handling of marketed poultry products and also highlighted the matter of excessive administration of antibiotics in poultry farming practice.

Salmonella is a globally distributed major food-borne pathogen and poultry is one of the predominant sources of salmonellosis in humans. To investigate the presence of motile Salmonella in the poultry hatchery environment, we collected 97 fluff samples from four selected broiler breeder chicken hatcheries from Chattogram, Bangladesh during July-December 2015. To isolate motile Salmonella enterica, we used conventional bacteriological techniques followed by serological verification using anti-Salmonella Poly A-E serum and species confirmation by conventional PCR assay. Antimicrobial susceptibility testing by Kirby-Bauer disc diffusion method for 10 commonly used antibiotics was performed on all isolates. Isolates displaying phenotypic resistance to ampicillin were tested by PCR for blaTEM gene, whereas those resistant to tetracycline were tested for the presence of tetA, tetB and tetC genes. A total of 24 samples (24.7%; 95% CI: 16.5–34.5, N = 97) from 3 hatcheries were positive for motile Salmonella. Of them, 21 (87.5%) and 12 (50.0%) were resistant to ampicillin and tetracycline, respectively, 9 (37.5%) to nalidixic acid and sulphamethoxazole/trimethoprim. No resistance was detected to ceftriaxone, cefoxitin, gentamicin, neomycin, ciprofloxacin and colistin. Ten (42%) of 24 isolates from 2 hatcheries were multi-drug resistant (i.e. resistant to ≥ 3 antimicrobial classes). Six of 21 ampicillin resistant isolates contained blaTEM gene and 10 of 12 tetracycline resistant isolates contained tetA gene. This study highlights the circulation of multi-drug resistant motile Salmonella in the hatchery environment for the first time in Bangladesh. Further epidemiological and molecular studies are therefore needed to identify the serotypes and source of the bacteria in hatcheries.

Salmonella spp. causes foodborne diseases related to the consumption of contaminated foods, especially poultry products. This study aimed to investigate the occurrence of Salmonella spp. serovars in raw eggs from supermarkets and street food markets in southern Brazil, to analyze virulence genes, resistance profiling to antimicrobials and sanitizers, and to determine the susceptibility of the isolates to Butia odorata extract. Among 160 samples analyzed, just two (1.25%) were positive for Salmonella spp.. One positive sample was from egg yolk (S. enterica serovar Gallinarum, isolate S28), and another one was from eggshell (S. enterica serovar Panama, isolate S37). Regarding the virulence genes, the isolate S37 harbored all the genes evaluated (hilA, invA, spvC, sefA, and pefA), while the isolate S28 did not harbor the pefA gene. The isolate S28 was resistant to tobramycin, azithromycin, and trimethoprim, while the isolate S37 showed resistance profile just to nalidixic acid. However, none of the resistance genes evaluated were identified. Both isolates showed resistance to benzalkonium chloride, chlorhexidine digluconate, sodium hypochlorite, and peracetic acid, presenting high MIC values for these sanitizers. In contrast, B. odorata extract showed antimicrobial activity against the isolates S28 and S37, however, more studies are needed to prove its potential as a natural antimicrobial compound. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s13197-021-05113-5) contains supplementary material, which is available to authorized users.

The canary (Serinus canaria) is appreciated for its beautiful song, colors, and docile temperament and drives a lucrative business. However, diseases caused by avian pathogenic Escherichia coli (APEC) compromise the health of canaries, and the inadequate antimicrobial treatment can lead to the emergence of resistant strains. This study aimed to characterize 21 isolates of E. coli obtained from canaries infected with colibacillosis during an outbreak in northern Paraná State, Brazil. APEC and diarrheagenic E. coli (DEC) virulence genes were screened for by polymerase chain reaction (PCR). All isolates were positive for the hlyF, iss, and ompT genes, which are characteristic of APEC. The iroN gene was found in 95.2% of isolates, and none had the iutA gene. The ipaH gene, characteristic of enteroinvasive E. coli (EIEC), was found in 71.4% of isolates, all belonging to the phylogenetic group B1. High genetic similarity (>95%) was found using enterobacterial repetitive intergenic consensus PCR (ERIC-PCR). The isolates belonged to serotypes O117:H4 (71.4%) and O1:H20 (23.8%). This is the first report of a clonal colibacillosis outbreak in canaries caused by APEC. All isolates were resistant to ampicillin, nalidixic acid, ciprofloxacin, enrofloxacin, norfloxacin, and tetracycline. The high rate of multidrug resistance in our study shows the importance of avoiding the inadequate antibiotic treatment. We suggest that further studies should be conducted to contribute to the understanding of colibacillosis in canaries since the health of animals is linked to human and environmental health, as defined by the concept of One Health.

In this work, multi-walled carbon nanotubes were applied as sorbent for extraction, followed by back-extraction and HPLC/UV determination of Nalidixic acid in aqueous solutions. The effects of some parameters such as pH, flow velocity, sorbent amount, and ionic strength were investigated. A little amount of sorbent (5.0 mg) was chosen as its optimum amount which this sorbent was reusable for almost five cycles of adsorption/desorption processes. A preconcentration factor of 500 was obtained. Linear range was 0.19–23.2 mg L−1 (R2 = 0.9979). The limit of detection and RSD for the suggested method were 0.02 mg L−1 and 2.2%, respectively. To assess the method performance for real samples, Nalidixic acid was determined in human blood serum and tablet samples. Advantages of the suggested method for Nalidixic acid extraction and preconcentration are low cost, being less time consuming, simplicity, no usage of organic solvents, and high preconcentration factor and efficiency.

The aim of this study was to investigate the prevalence of thermophilic Campylobacter in the broiler production chain of southern Brazil, by evaluating broiler farms and slaughter line samples, and to determine the genetic diversity, antimicrobial resistance, and virulence genes of the isolates. Of the 140 samples investigated in this study, 75 (53.6%) were positive for thermophilic Campylobacter, and all isolates were identified by phenotypic and molecular tests as C. jejuni. The resistance to nalidixic acid was the most common (74%), followed by resistance to enrofloxacin (67.3%) and ciprofloxacin (37.1%). However, there was no resistance to the macrolides tested which are recommended for the treatment of human campylobacteriosis. The PFGE showed that the isolates were grouped in eight macrorestriction patterns (P1 to P8). A representative isolate of each macrorestriction pattern was investigated for the presence of virulence genes and all isolates carried the cadF, ciaB, cdtA, cdtB, cdtC, and flaA genes. The dnaJ gene was detected in 87.5% (7/8) of the isolates. The flhA and racR genes were detected in 75% (6/8), while the pldA gene was present in 62.5% (5/8) and the wlaN gene in 25% (2/8). The presence of C. jejuni in broiler farms and in the slaughterhouse is a hazard to consumer given that this pathogen can be maintained throughout the broiler production chain and contaminates the final product. Moreover, the presence of the major virulence genes in the isolates demonstrates that they have the ability to develop campylobacteriosis in humans.

A molecularly imprinted polymer micro-solid phase extraction in pipette tip was employed for selective extraction and spectrophotometric determination of acetaminophen and nalidixic acid in pills and seawater samples. Parameters affecting microextraction, including pH of the sample, sample volume, type and elution solvent, number of cycles of extraction and desorption and amount of molecular imprinted polymer, were optimized utilizing both response surface methodology and one-variable-at-a-time methods. Four parameters in three levels were applied for response surface methodology experimental design. The proposed method indicated good linear range of 1.0–150.0 µg L−1 for nalidixic acid and 1.0–100.0 µg L−1 for acetaminophen by low detection limits of 0.2 and 0.3 µg L−1, respectively, for nalidixic acid and acetaminophen. Found recoveries were between 88.7 and 99.8% for real samples, with relative standard deviations less than 5.8%. The suggested protocol was successfully utilized to the separation, preconcentration and determination of nalidixic acid and acetaminophen in commercial pills and seawater samples.

We assessed the quality status of surface and ground waters used for drinking in Ikwo, Southeast Nigeria, using the physicochemical and bacteriological qualities, and antimicrobial susceptibility profiles following standard procedures. Overall. the mean values of the physicochemical qualities of the water samples revealed the following: colour (brown/dark-brown/colourless), odour (objectionable), temperature (28–30 °C), pH (6.30–7.50), electrical conductivity (12.94–12.99 µs cm−1), total dissolved solids (7.74–7.80 mg L−1), alkalinity (0.3–1.4 mg L−1), hardness (19–252 mg L−1), chloride (0.8–3.1 mg L−1), copper (0.01–0.72 mg L−1) and zinc (0.03–1.49 mg L−1). Only nitrate was not detected all through. Likewise, total heterotroph, coliform and Escherichia coli counts ranged from 1.16 to 6.96 × 103 cfu mL−1, 150 to 2400 MPN/100 mL and 45 to 345 CFU/100 mL, respectively. The antimicrobial susceptibility profiles showed that the E. coli isolates were relatively susceptible to gentamycin (58%) and levofloxacin (50%), with high percentages of the isolates displaying resistance against tetracycline (78%), norfloxacin (76%), nalidixic acid (76%), augmentin (68%), ampiclox (62%), doxycycline (62%) and amoxil (52%). While the physicochemical parameters were generally within the permissible limits of the WHO guidelines, reverse is the case for the bacteriological loads. Exceedance of bacteriological water quality criteria and prevalence of multidrug-resistant E. coli indicate high levels of microbial and drug pollutants in the waters, rendering them unfit for direct human ingestion without proper pretreatment.

Aeromonas spp. are opportunistic pathogenic bacteria related to an assembly of infectious diseases in ornamental fish. In the present study, virulence properties and antibiotic susceptibility of 52 guppy-borne Aeromonas spp. were investigated. The isolates were identified as A. veronii (n = 34), A. dhakensis (n = 10), A. hydrophila (n = 3), A. caviae (n = 3) and A. enteropelogenes (n = 2) by gyrB gene sequencing. The gyrB sequence deviation within and among the species ranged from 0 to 2.6% and 2.7-9.2%. Each species formed a distinct group in the unrooted neighbor-joining phylogenetic tree. The phenotypic virulence factors such as β-hemolysis, slime, caseinase, DNase, gelatinase and lipase production were observed in 28 (53.9%), 33 (63.5%), 28 (53.9%), 42 (80.8%), 37 (71.2%) and 42 (80.8%) isolates, respectively. The virulence genes were detected by PCR assay in the following proportions- act (84.6%), hly (80.8%), aer (73.1%), lip (73.1%), gcaT (73.1%), ascV (53.8%), ahyB (53.8%) fla (51.9%), alt (48.1%), ast (36.5%) and ser (34.6%), respectively. The amoxicillin, ampicillin, imipenem, nalidixic acid, oxytetracycline and rifampicin were resistant to more than 70.0% of the isolates in antibiotic susceptibility test. Our study suggests that the ornamental guppy can be a potential reservoir of virulent and multi-drug resistant Aeromonas spp.

Background Salmonella enterica serovar Enteritidis (S. Enteritidis) is one of the most common serovars, associated with human salmonellosis. The food-borne outbreak of this bacterium is mainly related to the consumption of contaminated poultry meat and poultry products, including eggs. Therefore, rapid and accurate detection, besides investigation of virulence characteristics and antimicrobial resistance profiles of S. Enteritidis in poultry and poultry egg samples is essential. A total of 3125 samples (2250 poultry and 875 poultry egg samples), sent to the administrative centers of veterinary microbiology laboratories in six provinces of Iran, were examined for Salmonella contamination, according to the ISO 6579 guideline. Next, duplex PCR was conducted on 250 presumptive Salmonella isolates to detect invA gene for identification of the genus Salmonella and sdf gene for identification of S. Enteritidis. Subsequently, the S. Enteritidis isolates were examined for detection of important virulence genes (pagC, cdtB, msgA, spaN, tolC, lpfC, and spvC) and determination of antibiotic resistance patterns against nalidixic acid, trimethoprim-sulfamethoxazole, cephalothin, ceftazidime, colistin sulfate, and kanamycin by the disk diffusion method. Results Overall, 8.7 and 2.3% of poultry samples and 6.3 and 1.3% of eggs were contaminated with Salmonella species and S. Enteritidis, respectively. The invA and msgA genes (100%) and cdtB gene (6.3%) had the highest and the lowest prevalence rates in S. Enteritidis isolates. The spvC gene, which is mainly located on the Salmonella virulence plasmid, was detected in 50.8% of S. Enteritidis isolates. The S. Enteritidis isolates showed the highest and the lowest resistance to nalidixic acid (87.3%) and ceftazidime (11.1%), respectively. Unfortunately, 27.0% of S. Enteritidis isolates were multidrug-resistant (MDR). Conclusion The rate of contamination with Salmonella in the poultry and egg samples, besides the presence of antimicrobial resistant and MDR Salmonella isolates harboring the virulence genes in these samples, could significantly affect food safety and subsequently, human health. Therefore, continuous monitoring of animal-source foods, enhancement of poultry farm control measures, and limiting the use of antibiotics for prophylactic purposes in food producing animals, are essential for reducing the zoonotic risk of this foodborne pathogen for consumers and also choosing effective antibiotics for the treatment of salmonellosis.

Turkey coryza is a major respiratory disease caused by Bordetella avium (B. avium). It occurs in all ages of turkeys and is characterized by high morbidity and low mortality rates. The present study aimed firstly at determination of the prevalence rates of B. avium in turkeys reared in Egypt at different ages using various diagnostic methods including clinical examination, histopathology, enzyme-linked immunosorbent assay (ELISA), bacterial culture, and polymerase chain reaction (PCR). Using PCR, virulence-associated genes were detected in the confirmed B. avium isolates. Furthermore, the antibiotic resistance profiles of the confirmed B. avium isolates were examined. The achieved results indicated isolation and identification of B. avium infection at different ages of turkeys reared in Egypt. The overall PCR-confirmed prevalence rate of B. avium was 22.95%. The identified B. avium strains harbored virulence-associated genes responsible for colonization in the respiratory tract of turkeys including Bordetella virulence gene (100%), fimbriae (71.14%), and filamentous hemagglutinin (85.68%). The isolated B. avium strains showed multidrug resistance profiles. B. avium isolates were resistant to penicillin (92.82%), ceftiofur (85.68%), nalidixic acid (78.54%), and lincomycin (71.40%). The identified B. avium strains showed clear sensitivities to both gentamicin and neomycin, suggesting these as possible antimicrobial candidates for the control of B. avium infection in turkeys.

This article presents the synthesis of a series of amides, based on the interaction of several 3-aminospirohydantoins with nalidixic acid. The target compounds were characterized by physicochemical parameters, IR, 1H and 13C NMR spectral data. The antimicrobial activity of the products obtained was determined against Gram-positive bacteria Staphylococcus aureus and Bacillus subtilis, Gram-negative bacteria Escherichia coli, Pseudomonas aeruginosa and Salmonella abony, the yeasts Candida albicans and Saccharomyces cerevisiae and the molds Penicillium chrysogenum and Aspergillus niger. The relationship between structure and biological activity of the products obtained was discussed. It was found that the most effective compounds are tetralin (5f) and indane (5g) derivatives, which exhibit a pronounced antimicrobial activity against both tested Gram-positive and Gram-negative bacteria.

Exposure of bacteria to a sub-lethal dosage of antibiotic is one the major causes for the onset of antibiotic resistance. Therefore, we aimed to assess the emergence of antibiotic cross-resistance in bacteria after exposure to a sub-lethal dose of veterinary feed directive (VFD) antibiotics, tilmicosin, and florfenicol. The minimum inhibitory concentrations (MICs) of tilmicosin and florfenicol against Salmonella enterica serovar Enteritidis, Klebsiella pneumoniae, Staphylococcus aureus, and Listeria monocytogenes were determined. Next, the pathogens were exposed to a sub-inhibitory concentration of tilmicosin (0.5, 5, 20 µg/ml) and florfenicol (1, 20 µg/ml) for 24 h and 48 h, and acquired cross-resistance to human therapeutic antibiotics was measured by determining the increase in MIC values. MICs of ampicillin, tetracycline, nalidixic acid, and meropenem against Salmonella and Klebsiella were in the range of 20–1000 µg/ml, 5–62.5 µg/ml, 5–125 µg/ml, and 0.05–0.1 µg/ml, respectively, whereas MICs against Staphylococcus and Listeria were 2.5–10 µg/ml, 2.5 µg/ml, 62.5–500 µg/ml, and 0.1–0.2 µg/ml, respectively. Pre-exposure of these bacteria to a sub-inhibitory concentration of tilmicosin and florfenicol, increased cross-resistance against ampicillin, tetracycline, and nalidixic acid from 1.25- to 40-fold compared to the antibiotic unexposed bacteria with the exception of meropenem, which did not show increased resistance. This study could serve as a foundation to understand the mechanisms of acquired cross-resistance to traditional therapeutic antibiotics, and to develop strategies to alleviate such problem by using alternative antimicrobials.

We use genetic assays to suggest that transcription-coupled repair or new origin formation in Escherichia coli involves removal of RNAP to create an RNA primer for DNA synthesis. Transcription factor DksA was shown to play a role in numerous reactions involving RNA polymerase. Some, but not all, of the activities of DksA at promoters or during transcription elongation require (p)ppGpp. In addition to its role during transcription, DksA is also involved in maintaining genome integrity. Cells lacking DksA are sensitive to multiple DNA damaging agents including UV light, ionizing radiation, mitomycin C, and nalidixic acid. Here, we focus on two recent studies addressing the importance of DksA in the repair of double-strand breaks (DSBs), one by Sivaramakrishnan et al. (Nature 550:214-218, 2017) and one originating in our laboratory, Myka et al. (Mol Microbiol 111:1382-1397. https://doi.org/10.1111/mmi.14227 , 2019). It appears that depending on the type and possibly location of DNA damage, DksA can play either a passive or an active role in DSB repair. The passive role relies on exclusion of anti-backtracking factors from the RNAP secondary channel. The exact mechanism of active DksA-mediated DNA repair is unknown. However, DksA was proposed to destabilize transcription complexes, thus clearing the way for recombination and DNA repair. Based on the requirement for DksA, both in repair of DSBs and the R-loop-dependent formation of new origins of DNA replication, we propose that DksA may allow for removal of RNAP without unwinding of the RNA:DNA hybrid, which can then be extended by a DNA polymerase. This mechanism obviates the need for RNAP backtracking to repair damaged DNA.

Background This study aimed to investigate the frequency and the antibiotic resistance patterns of Salmonella species that were isolated from infectious diarrhea samples taken from pediatric patients in central Iran. Methods The study analyzed 230 stool specimens that were cultured on XLD, MacConkey agar and GN broth. Polymerase chain reaction (PCR) assay was used to identify the Salmonella genus. The antibiotic resistance profiles and the frequency of quinolone and integron genes were obtained. Results Out of 230 samples of infectious diarrhea, 21 (9.1%) cases of Salmonella spp. were identified using culture methods. Another 28 (12.1%) samples had positive PCR results, with S. serovar Paratyphi B and C (9/21; 42.8%) and S. Typhi (3/21; 14.3%) being the most recognized. The highest antibiotic resistance rates were found for nalidixic acid (15/21; 71.4%), tetracycline (9/21; 42.8%). However, six (28.5%) of isolates were found resistant to cotrimoxazole, ampicillin and chloramphenicol. Among the plasmid-mediated quinolone resistance (PMQR) determinants, qnrS, qnrA, and qnrB were positive in (9/15; 60%), (6/15; 40%) and (3/15; 20%) of the isolates, respectively. Class 1 and 2 integrons were identified in 15 (71.4%) and 3 (14.3%) isolates, respectively. Conclusion High rates of quinolone resistant and low frequency of MDR Salmonella spp. isolates were identified in central Iran, similar to findings in other parts of Asia. To prevent the spread of these resistant strains, the antimicrobial resistance of Salmonella spp. isolates should be under constant surveillance, and empiric antibiotic therapy should be adapted appropriately.

Shiga toxin-producing Escherichia coli (STEC) is a pathotype of E. coli that causes enteric and systemic diseases ranging from diarrhoea to severe hemorrhagic colitis (HC) and hemolytic uremic syndrome (HUS). The emergence of multidrug-resistant (MDR) STEC from cattle sources has increased public health risk and limited treatment options. The prevalence of STEC was investigated in 200 raw food samples (milk and beef samples) and 200 diarrheic samples (cattle and human samples) in a matched region. The presence of stx genes (stx1 and stx2), carbapenemase-encoding genes (blaVIM, blaNDM-1, and blaIMP), and extended-spectrum β-lactamase (ESBL)-encoding genes (blaTEM group, blaCTX-M1 group, and blaOXA-1 group) was screened by polymerase chain reaction (PCR). Antibiogram and Enterobacterial repetitive intergenic consensus (ERIC)-PCR were also conducted. STEC isolates were identified in 6.5% (13/200) of food samples [6% (6/100) of milk and 7% (7/100) of beef samples] and in 11% (22/200) of diarrheic cases [12% (12/100) of cattle and 10% (10/100) of human samples]. We found that O26 (4.5%, 18/400) and O111 (1.5%, 6/400) were the most prevalent STEC serovars and were found more commonly in diarrheic samples. STEC strains with both stx genes, stx2 only, and stx1 only genotypes were present in 62.9% (22/35), 20% (7/35), and 17.1% (6/35) of isolates, respectively. Carbapenemase-producing STEC (CP STEC) isolates were found in 1.8% (7/400) of samples [0.5% (1/200) of foods and 3% (6/200) of diarrheic cases]. The blaVIM gene was detected in all CP STEC isolates, and one human isolate carried the blaNDM-1 gene. ESBL-producing STEC strains were detected in 4.3% (17/400) of samples [1.5% (3/200) of food samples and 7% (14/200) of diarrheic cases]. The blaTEM, blaCTX-M1, and blaOXA-1 genes were detected in 42.9% (15/35), 28.6% (10/35), and 2.9% (1/35) of STEC isolates, respectively. Approximately half (51.4%, 18/35) of STEC isolates were MDR STEC; all CP STEC and ESBL-producing STEC were also MDR STEC. The highest antimicrobial resistance rates were found against nalidixic acid (51.4%) and ampicillin (48.6%), whereas the lowest rates were reported against gentamicin (5.7%) and ciprofloxacin (11.4%). MDR STEC strains were 5.3 times more likely to be found in diarrheic cases than in foods (P = 0.009, 95% CI 1.5–18.7). ERIC-PCR was used for genotyping STEC isolates into 27 different ERIC-types (ETs) with a discrimination index of 0.979. Five ETs showed clusters of 2–4 identical isolates that shared the same virulence and antibiotic resistance genetic profile. Human isolates matched food isolates in two of these ET clusters (the O26 CP STEC cluster and the O111 STEC cluster), highlighting the potential cross-species zoonotic transmission of these pathogens and/or their genes in the study region. This is the first detection of CP STEC in milk and diarrheic cattle in Egypt.

Background Helicobacter pullorum commonly colonized in the gastrointestinal tract of poultry and caused gastroenteritis. This bacterium could be transmitted to humans through contaminated food and caused colitis and hepatitis. Currently, the genetic characteristics of the H. pullorum were not recognized enough. In this study, the genomes of 23 H. pullorum strains from different counties were comparatively analyzed. Among them, H. pullorum 2013BJHL was the first isolated and reported in China. Results The genomes of the studied strains were estimated to vary from 1.55 to 2.03 Mb, with a GC content of ~ 34%. 4064 pan genes and 1267 core genes were obtained from the core-pan genome analysis using the Roary pipeline. Core genome SNPs (cg-SNPs) were obtained using Snippy4 software. Two groups were identified with the phylogenetic analysis based on the cg-SNPs. Some adhesion-related, immune regulation, motility-related, antiphagocytosis-related, toxin-related and quorum sensing related genes were identified as virulence factors. APH(3′)-IIIa, APH(2′’)-If, and AAC(6′)-Ie-APH(2′’)-Ia were identified as antibiotic resistance genes among the H. pullorum genomes. cat, SAT-4 and tetO genes were only identified in 2013BJHL, and tet(C) was identified in MIT98-5489. MIC determination revealed that the 2013BJHL showed acquired resistance to ciprofloxacin, nalidixic acid, tetracycline, gentamicin, streptomycin and erythromycin, only sensitive to ampicillin. The antibiotic resistance genetic determinants on the 2013BJHL genome correlate well with observed antimicrobial susceptibility patterns. Two types of VI secretion system (T6SS) were identified in 52.2% (12/23) the studied strains. Conclusion In this study, we obtained the genetic characteristics of H. pullorum from different sources in the world. The comprehensive genetic characteristics of H. pullorum were first described. H. pullorum showed highly genetic diversity and two sub-types of T6SSs were first identified in H. pullorum. 2013BJHL was found to be multidrug resistant as it was resistant to at least three different antibiotic classes.

Background Riemerella anatipestifer is one of the most serious infectious disease-causing pathogens in the duck industry. Drug administration is an important method for prevention and treatment of infection in duck production, leading to widespread drug resistance in R. anatipestifer. Methods For a total of 162 isolates of R. anatipestifer, the MICs were determined for a quinolone antimicrobial agent, namely, nalidixic acid, and three fluoroquinolones, namely, ciprofloxacin, enrofloxacin and ofloxacin. The gyrA, parC, and parE gene fragments were amplified by PCR to identify the mutation sites in these strains. Site-directed mutants with mutations that were detected at a high frequency in vivo were constructed (hereafter referred to as site-directed in vivo mutants), and the MICs of these four drugs for these strains were determined. Results In total, 100, 97.8, 99.3 and 97.8% of the 137 R. anatipestifer strains isolated between 2013 and 2018 showed resistance to nalidixic acid, ciprofloxacin, enrofloxacin, and ofloxacin, respectively. The high-frequency mutation sites were detected in a total of 162 R. anatipestifer strains, such as Ser83Ile and Ser83Arg, which are two types of substitution mutations of amino acid 83 in GyrA; Val799Ala and Ile811Val in ParC; and Val357Ile, His358Tyr, and Arg541Lys in ParE. MIC analysis results for the site-directed in vivo mutants showed that the strains with only the Ser83Ile mutation in GyrA exhibited an 8–16-fold increase in MIC values, and all mutants showed resistance to ampicillin and ceftiofur. Conclusions The resistance of R. anatipestifer to quinolone agents is a serious problem. Amino acid 83 in GyrA is the major target mutation site for the fluoroquinolone resistance mechanism of R. anatipestifer.

Objective Food-borne pathogens are a major cause of illnesses, death and expenses. Their occurrence in meat and other food is considered a global health problem. The burden of food-borne disease is increasing due to antimicrobial resistance which represents a greater risk of treatment failure. However, very little is known about the antibiotic resistance profile of food-borne pathogens in Nepal. This study was conducted to examine the antibiotic resistance profile of common food-borne bacterial pathogens isolated from raw meat sold in Nepal. A total of 83 meat samples were collected from the market and analyzed. Results The prevalence of Staphylococcus aureus, Escherichia coli, Salmonella, Shigella, and Vibrio were 68%, 53%, 35%, 6%, and 6% respectively. The resistance of Salmonella was most frequently observed to amoxicillin (100%), tetracycline (24%), chloramphenicol (11%), and nalidixic acid (11%). S. aureus was resistant to amoxicillin (100%) followed by tetracycline (63%), nalidixic acid (17%), and cefotaxime (13%) respectively. Vibrio isolates resisted amoxicillin (100%), tetracycline (40%) and chloramphenicol (20%). Shigella expressed the highest resistance to amoxicillin (100%), followed by chloramphenicol (80%), tetracycline (60%) and nalidixic acid (20%). E. coli exhibited the highest resistance to amoxicillin (100%), followed by tetracycline (93%), nalidixic acid (25%) and cefotaxime (19%).

Background Bacterial infection of the urinary tract is among the common reasons for seeking medical attention in the community. Rapidly increasing antibiotic resistance of uropathogens is resulting in limited treatment options. Therefore, knowledge of the current uropathogens and their antibiotic susceptibility is important for better treatment of urinary tract infection. Methods A cross-sectional study design was conducted from February to September thirty, 2017 among students who came to Mekelle University student’s clinics with symptomatic urinary tract infection during the study period.. Mid-stream urine specimens were collected from 341individuals with suspected urinary tract infection for bacteriological identification and antimicrobial susceptibility testing. Data on socio-demographic, clinical and risk factors were also collected using a structured questionnaire. Results Among the 341 study participants, 72(21.1%) showed significant bacteriuria. Escherichia coli (48.6%), Coagulase-negative staphylococci (23%), Staphylococcus aureus (13.5%), and Klebsiella spp. (8.1%) were common bacterial isolates. Resistance to ampicillin (81–100%), amoxicillin/clavulanic acid (77–93.6%), co- trimoxazole (55 72.3%), nalidixic acid (57.4%) and tetracycline (46–55.5%) was seen by most isolates. Multidrug resistance was observed in 73% of the bacterial isolates, and 25.5% of the Gram-negative isolates were extended-spectrum beta-lactamase producers. Being female, a history of urinary tract infection, a history of catheterization and frequent sexual activity were found to be statistically associated with urinary tract infection. Conclusion Urinary tract infection is a problem among university students with a prevalence of 21.1%. All isolates have developed resistance to most of the commonly used antibiotics. Therefore, health education on the transmission and causes of urinary tract infection are recommended for the students.

The host–guest inclusion complex structure and binding ability of two different quinolones with γ-cyclodextrin (γ-CD) were investigated in solution by means of UV–Vis and 1H NMR spectroscopy. Competition of oxolinic and nalidixic acid molecules for the γ-CD cavity was evaluated by determination of association constants. Both quinolones form 1:1 inclusion complexes, their binding constants at room temperature (25 °C) under acidic and basic conditions were calculated using Benesi–Hildebrand equation. The stability of the complexes was dependent on the structure of the quinolone. In general, the weaker binding constants were observed for oxolinic acid-γ-CD complexes (1616 and 1765 M−1) and the larger binding constants were obtained for nalidixic acid-γ-CD complexes (3760 and 3840 M−1). 1H NMR studies in D2O were performed to elucidate the structure of each inclusion complex, nalidixic acid molecule penetrates more deeply into the γ-CD cavity and an intermolecular hydrogen bond is formed. Knowledge about structure and relative stability of quinolone-γ-CD complexes will be useful for future applications of these antimicrobial agents in medicinal chemistry.

The present study aims at evaluating the potentiating ability of Lantana camara leaves and flower extracts in enhancing the antibacterial efficacy of antibiotics against pathogenic bacterial species. Methanolic extracts of L. camara leaves and flowers (pink turn yellow) were prepared by the solvent extraction method. The leaves and flower extracts alone and their combination with five different antibiotics; Ciprofloxacin, Streptomycin, Tetracycline, Nalidixic acid and Norfloxacin were assessed for their synergistic antibacterial potential against Escherichia coli MTCC 1698, Staphylococcus aureus MTCC 3160, Enterobacter fecalis MTCC 439, Proteus vulgaris MTCC 1771 and Salmonella typhi MTCC 733, using an agar disc diffusion method and checkerboard antibacterial susceptibility assay. The minimum inhibitory concentration of Lantana extracts and antibiotics was assessed using broth micro-dilution assay. In agar disc diffusion assay, the combination of the flower extract and streptomycin depicted 5.26–25% increase in ZOI against S. aureus, and S. typhi. In the checkerboard antibacterial assay, the Ciprofloxacin and flower extract showed a synergistic inhibitory effect against E. coli, S. aureus, E. fecalis, P. vulgaris and S. typhi (FIC index, ranging from 0.39 to 0.14), while the combination of leaves extract and streptomycin exerted additive effect against S. typhi and E. coli (FICI = 0.46 and 0.48 respectively). The MICs of Lantana leaves and flower extracts against the test bacterial strains were depicted between 31.25 μg/ml and 250 µg/ml. L. camara leaves and flower extracts enhance the antibacterial potency of antibiotics and their synergistic combination could emerge as a novel therapeutics against pathogenic bacterial species.

Prevalence and distribution of Shiga toxin-producing Escherichia coli (STEC) serogroups from the faecal samples of cattle and camels slaughter in Maiduguri abattoir and their antibiotic resistance profile of the isolates were determined. The highest prevalence (24%) was recorded in the month of September and more STEC isolates came from cattle than the camels. There was significant (P

Specific features for the development of resistance in Campylobacter jejuni strains were studied after treatment with antibiotics of 6 pharmacological groups. Populations of 18 native strains of C. jejuni (isolated from raw poultry products) and their subcultures (obtained after 2-3-fold stress exposures to antimicrobial agents in subinhibitory doses) were examined to evaluate the expression of phenotypic antibiotic resistance. Genotypic properties of strains were studied by the PCR with primers that detect the presence of genes for resistance to aminoglycosides (aphA-1, aphA-3, and aphA-7), tetracyclines (tetO), and quinolones (GZgyrA). The majority of test strains of C. jejuni exhibited a high resistance to nalidixic acid, ciprofloxacin, and tetracycline, which reached the maximum value after numerous passages. The expression of antibiotic resistance was greatest in the presence of nalidixic acid and tetracycline. Ciprofloxacin resistance of 33% strains, which were initially resistant to this antibiotic, was increased after 2-3-fold treatment. We revealed a high degree of correspondence between phenotypic and genotypic profiles of antibiotic resistance in food isolates of Campylobacter. One, two, or more genes of aphA were identified in 85% strains phenotypically resistant to aminoglycosides. The tetO gene was found nearly in all strains resistant to tetracycline. Studying the biofilm matrix in C. jejuni after culturing with antibiotics in subinhibitory doses showed that quinolones (particularly nalidixic acid) and tetracyclines potentiate the formation of biofilms and increase the tolerance of Campylobacter to stress exposures. The intensity of biofilm growth was shown to depend little on the effect of macrolides and aminoglycosides. Therefore, the presence of these agents in residual concentrations is associated with a lower risk for the development of antibiotic resistance in C. jejuni populations.

Emulsifiers like polysorbate family, carboxymethyl cellulose are widely used in the preparation of drugs, vaccines, food, cosmetics, and skin care products. In this study, strain SP2 is isolated for the high-efficiency utilization of emulsifier as its sole carbon and energy source. Strain grows on the polysorbate family with generation time ranging from 0.8 to 2.2 h. It is sensitive to ampicillin, gentamicin, imipenem, penicillin, and vancomycin, but resistant to amikacin, cefoxitin, erythromycin, kanamycin, nalidixic acid, and tetracycline. Based on 16S rRNA gene, strain is identified as Acinetobacter soli. Compared to M. pamicella, which is capable of degrading 50% polysorbate 80 within 3 days, isolate PS2 could dispose 51.6% polysorbate 80 within only 8 h. Using cell crude extract from strain SP2, the liberation of free fatty acid in the reaction mixture containing polysorbate 80 suggests that the mechanism for polysorbate utilization belongs to the β-oxidation.

Made available in DSpace on 2018-12-11T16:51:52Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-06-01 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Solid-state [Ln(nal)3]·nH2O compounds, where Ln represents light trivalent lanthanide (La to Sm, except Pm), nal is nalidixate (C12H11N2O3), and n is the number of water molecules, were synthesized. Characterization and investigation were made by means of complexometry, elemental analysis, powder X-ray diffraction, Fourier transform infrared, near infrared (NIR) and diffuse reflectance spectroscopies, simultaneous thermogravimetry and differential thermal analysis, and differential scanning calorimetry. The dehydration process occurs in a single step up to 453 K. The final temperature of dehydration decreases from Pr(III) to Sm(III), and the thermal stability of anhydrous compounds increases from La(III) to Sm(III) (except Ce), and these behaviors are related to the decrease in the ionic radius of the lanthanide ion. The thermal decomposition of the anhydrous compounds occurs in three, four, or five consecutive steps, with formation of the respective oxides CeO2, Pr6O11, and Ln2O3 (Ln = La, Nd, and Sm) as final residues. The results also provide information concerning the composition and thermal behavior of these compounds. Spectroscopic studies in the UV–Vis and NIR regions provide information about the characteristics 4f–4f transitions for the Pr(III), Nd(III), and Sm(III) compounds, as well as the NIR region characteristic overtone and combination bands of the compounds. Faculdade de Engenharia Universidade Estadual Paulista (UNESP) Instituto de Química Universidade Estadual de Campinas (UNICAMP) Faculdade de Ciências Universidade Estadual Paulista (UNESP) Faculdade de Engenharia Universidade Estadual Paulista (UNESP) Faculdade de Ciências Universidade Estadual Paulista (UNESP) CNPq: 150759/2009 FAPESP: 2006/06951-3 CAPES: 472233/2008-4

Marrubium vulgare L. is used in folk medicine of several countries. Many researches showed that it is used for treatment of variety of diseases, for example as remedy for asthma and diabetes, duo to its antibacterial effects. The present study aimed to access the effect of the sampling location on the antibacterial activity of the flavonoids extracted from Marrubium vulgare L. leaves, against three pathogenic bacteria; Escherichia coli, Bacillus cereus, and Proteus mirabilis. The leaves were collected from three different sampling locations belonging northwest Algeria: Tessala Mountain, M’sila forest and Ain Skhouna. The flavonoid extraction was carried out using organic solvents with increasing polarity. The bacterial susceptibility testing by diffusion agar method showed that the diameters zones varied significantly (P

We attempted to isolate lactic acid bacteria (LAB) from the marine oyster (Crassostrea gigas) and selected several environmental stress-resistant isolates for the development of a future probiotic adjuvant for marine aquaculture. Twenty-six presumptive LAB isolates were extracted from oysters and screened (by an agar diffusion assay) for antimicrobial activity toward various pathogens: Vibrio parahaemolyticus, Streptococcus iniae, and Edwardsiella tarda. Eight isolates had an antibacterial activity toward V. parahaemolyticus; in particular, 6 isolates showed a growth-inhibitory activity, with inhibition zone diameters > 15 mm. Of these, 5 isolates (JL17, JL18, JL28, HL7, and HL32) were also active against S. iniae and E. tarda. Enterococcus faecium HL7 was selected as the isolate most resistant to environmental stressors: the minimum NaCl, ethanol, and hydrogen peroxide concentrations at which HL7 cells lost their viability were 1.9 M, 11%, and 0.013%, respectively. When an antibiotic sensitivity test was performed on E. faecium HL7, this isolate was found to be resistant to trimethoprim/sulfamethoxazole, cephalothin, ampicillin, rifampin, gentamicin, cefotaxime, cefepime, cefotetan, nalidixic acid, and kanamycin. While the oyster model studies provided indication that E. faecium HL7 could be a good candidate as biocontrol agent against V. vulnificus, further optimization is needed in the actual animal rearing situation.

1-Ethyl-7-methyl-4-oxo-[1,8]naphthyridine-3-carboxylic acid (nalidixic acid) is one of the most important and highly-consumed quinolone antibiotic drugs. In the present study, we prepared and characterized the nalidixic acid (NA)-layered zinc hydroxide (LZH) nanohybrid composites. The two subsequent ratios of the nanocomposites showed a basal spacing of 10.56 and 11.18 A. These prepared nanocomposites were analyzed by PXRD, FTIR, TGA/DTA, BET, EDS, and FESEM, all representing the successful intercalation of NA into LZH nanohybrid gallery spaces. Accordingly, by the PXRD analysis, the average particle size and micro-strain values of the LZH and the two ratios of the nanocomposites were calculated and compared. The biopolymer NA-chitosan and NA-LZH-chitosan nanocomposites were prepared. Furthermore, drug release studies from LZH and chitosan substrates were measured at pH = 4.8 and 7.4 and acetate-buffered and phosphate-buffered saline solutions, respectively.

Background Enteric bacterial pathogens are the major causes of food-borne gastroenteritis in humans and remain important public health problems worldwide. The emergence of antimicrobial resistance is a global concern, particularly in developing countries. The aim of this study was to determine the prevalence of enteric bacteria pathogens and their antimicrobial susceptibility patterns among food handlers in Gondar town, Northwest Ethiopia. Methods A cross-sectional study was conducted from February 4 to April 16, 2018. A total of 257 food handlers were selected using a multistage sampling technique. Data on socio-demographic characteristics were collected using a structured questionnaire. Stool samples were collected and inoculated into appropriate media. Enteric bacterial pathogens were identified using standard microbiological methods. Antimicrobial susceptibility tests were performed using the disk diffusion technique as per the standard Kirby-Bauer method. Data were entered and analyzed using SPSS version 20 software. Results The overall prevalence of enteric bacteria was 34/257 (13. 2%, [95% CI, 8.9–17.5%]). Shigella species was the leading isolate that accounted for 26/257 (10.1%) followed by Enterohemorrhagic Escherichia coli (EHEC) O157: H7 5/257 (1.9%) and Salmonella species 3/257 (1.2%). Shigella spp. was susceptible to ciprofloxacin 26 (100%), ceftriaxone 25 (96.1%), chloramphenicol 24 (92.3%), nalidixic acid 24 (92.3%), and gentamicin 20 (76.9%). Escherichia coli O157: H7 and Salmonella spp. showed the maximum (100%) susceptibility results to ceftriaxone, chloramphenicol, ciprofloxacin, and gentamicin. The overall prevalence of Multidrug resistance (MDR) in the current study was 14/34 (41.2%). Conclusion Our study showed high prevalence of enteric bacterial pathogens among food handlers. All isolates were susceptible to ciprofloxacin. However, a substential number of isolates were resistant to commonly prescribed antibiotics and the prevalence of MDR was high.

The aim of the present study was to assess the antibiotic susceptibility profile of Pediococcus strains from diverse sources. From a total of 115 dairy and non-dairy samples, 40 Pediococcus strains were isolated. Their biochemical and molecular characterization confirmed them as P. pentosaceus and P. acidilactici. All the 40 identified isolates were evaluated for antibiotic susceptibility using disc diffusion assay against a total of 20 antibiotics. The isolates exhibited varied range of responses towards the antibiotics depending on the strain type, source and location of isolation. All the isolates were either sensitive or intermediate resistant to amoxycillin, erythromycin, ceftriaxone, cloxacillin, cefoperazone, penicillin, netillin, gentamycin and chloramphenicol. Resistance towards vancomycin and nalidixic acid was exhibited by most of the isolates. A total of 16 strains belonging to dosa batter (n = 4; n = number of isolates), fermented vegetables (n = 4), fermented grape juice (n = 4), idly batter (n = 3) and the only isolate from butter milk exhibited sensitivity/intermediate towards 80–90% of the studied antibiotics. No considerable difference in susceptibility pattern was observed between the two Pediococcus species, i.e., P. pentosaceus and P. acidilactici. Overall, the maximum resistance was exhibited by isolates belonging to silage (Sil-2; 50%) followed by cow milk (PD-41), dosa batter (8-PD) and human isolate (PD-45) which showed resistance towards 40% of studied antibiotics. The susceptibility profiling of Pediococcus strains will be helpful in their safer selection for future food and feed applications.

The aims of the present study were to determine (i) the profiles of phylogroup and (ii) the antimicrobial susceptibility of pathogenic Escherichia coli strains isolated from calves, and of Salmonella spp. strains isolated from calves and pigs in Minas Gerais State, Brazil. Sixty-one pathogenic E. coli strains and Salmonella spp. (n = 24) strains isolated from fecal samples of calves and Salmonella spp. (n = 39) strains previously isolated from fecal samples of growing/finishing pigs were tested. The minimum inhibitory concentration (MIC) using the agar dilution method was determined for nalidixic acid, amikacin, amoxicillin, ampicillin, cefoxitin, norfloxacin, gentamicin, tetracycline, and trimethoprim-sulfamethoxazole. All E. coli isolates were susceptible to amikacin. Tetracycline was the antimicrobial that presented the higher frequency of resistance among E. coli strains, followed by ampicillin, trimethoprim-sulfamethoxazole, amoxicillin, nalidixic acid, norfloxacin, gentamicin, and cefoxitin. E. coli (n = 61) strains isolated from calves belonged to different phylogroup namely, phylogroup A (n = 26), phylogroup B1 (n = 31), phylogroup E (n = 3), and phylogroup F (n = 1). Phylogroups B2, C, and D were not identified among the E. coli in the present study. All Salmonella spp. (n = 24) strains isolated from fecal samples of calves were susceptible to amikacin, amoxicillin, ampicillin, norfloxacin, gentamicin, tetracycline, and trimethoprim-sulfamethoxazole. Resistance to nalidixic acid and cefoxitin was detected in 16.66 and 8.33 % of the Salmonella spp. strains, respectively. Among the Salmonella spp. (n = 39) strains isolated from fecal samples of pigs, the higher frequency of resistance was observed to tetracycline, followed by amoxicillin, gentamicin, ampicillin, trimethoprim-sulfamethoxazole, nalidixic acid, cefoxitin, and norfloxacin. All strains were susceptible to amikacin. Forty-eight (78.68 %) of the E. coli strains were classified as multidrug-resistant, whereas among Salmonella spp. strains, the percentage of multidrug resistance was 57.14 %, being all multidrug-resistant strains isolated from pigs (92.30 %). The results from the present study indicate a high frequency of antimicrobial resistance among pathogenic E. coli strains isolated from calves and Salmonella spp. strains isolated from pigs and a high rate of susceptibility to most antimicrobials tested among Salmonella spp. strains isolated from calves. Our study highlights the presence of multidrug-resistant strains of E. coli and Salmonella spp. isolated from food-producing animals in Minas Gerais, Brazil. Electronic supplementary material The online version of this article (doi:10.1007/s11250-016-1152-0) contains supplementary material, which is available to authorized users.

The bacterial isolate Bacillus thuringiensis TS110 was isolated from the rice field soil of Burdwan district, West Bengal, India. Bioassay test of the bacteria TS110 against 3rd, 4th and 5th instar larvae of Cnaphalocrocis medinalis was carried out. Cut leaf assay, potted plant assay and field assay were done. During filed assay, it has been observed that the LC50 (×107) values of TS110 against 3rd, 4th and 5th instar larvae of C. medinalis were 3.77, 5.29, 4.83 and 4.93, 4.42, 4.72 in dry and wet season, respectively. The morphological, biochemical and phylogenetic analysis of the isolate TS110 were done. TS110 was positive for catalase, nitrate reductase, methyl red, voges-proskauer, oxidase, urease, indole, citrate utilization, arginine dihydrolase test, starch, lipid, gelatin, casein, and lecithin hydrolysis test. TS110 showed fermentation test positive for glucose, fructose, mannose, arabinose and trehalose in nutrient broth medium. The antibiotic sensitivity test showed that the bacterial isolate was sensitive to kanamycin (30 µg/disc), nalidixic acid (30 µg/disc), rifampicin (5 μg/disc), doxycycline (30 µg/disc), gatifloxacin (10 µg/disc), vancomycin (30 µg/disc), gentamycin (10 µg/disc), ampicillin (10 µg/disc), ofloxacin (5 μg/disc), levofloxacin (5 μg/disc), streptomycin (10 µg/disc), gentamycin (10 µg/disc). The phylogenetic analysis revealed that TS110 was closely related to different species of B. thuringiensis submitted to the GenBank. On the basis of morpho-physiological and molecular characterization, the bacterial isolate was identified as B. thuringiensis.

Objective: To provide the epidemiology, resistance pattern, and characterization of integrons in Shigella flexneri isolated between 2001 and 2011 in Jiangsu Province. Method: A total of 624 strains of S. flexneri were collected from both outpatients and inpatients in hospitals in Jiangsu Province from January 2001 to December 2011. The Kirby–Bauer disk diffusion method was used to perform the antimicrobial susceptibility test. Polymerase chain reaction (PCR) was used in the detection of integrons. Pulsed-field gel electrophoresis (PFGE) was applied in the homology studies. Result: Serotype 2a accounted for the largest proportion in S. flexneri, namely 26.4 %. Notably, an increasing trend was detected in the resistance to common antimicrobial agents during the period 2001–2011. In recent years, more than 80.0 % isolates of S. flexneri have proved to be resistant to ampicillin, nalidixic acid, and tetracycline. The positive rates of class 1, class 2, and the atypical class 1 integrons in S. flexneri are 69.3 %, 87.8 %, and 89.2 % respectively. Most integrons detected in our research carry genes encoding resistance to trimethoprim and streptomycin. Conclusion: Antimicrobial resistance in S. flexneri has demonstrated a continuous rising trend in Jiangsu Province. A high prevalence of integrons and gene cassettes play an important role in the transmission of drug resistance in S. flexneri. Effective measures are urgently needed to control the spread of multi-drug-resistant S. flexneri, and more continuing active surveillance of antimicrobial resistance should be established worldwide, especially in developing countries.

This study aimed to evaluate the use of nalidixic acid-adapted strains of three major Shiga toxin-producing Escherichia coli (STEC) and non-pathogenic E. coli for the use of radio frequency (RF) heating using phosphate buffer saline (PBS). The effectiveness of RF was evaluated on cocktails of various STEC serotypes (O157:H7, O26:H11, O111) and non-pathogenic E. coli at different endpoint temperatures (55, 60, and 65 °C). All strains were successfully adapted to nalidixic acid (Nal). In general, the results indicated that Nal-adapted strains were not significantly different from Nal-sensitive strains evaluated either before treatment or at the endpoint temperatures. Nal-adapted strains were, therefore, confirmed to be effective as marker organisms in studies involving the use of RF in buffer. Results also showed that the thermal inactivation of strains was more effective as the treatment temperature increased, particularly at 65 °C, which showed a 6.0 log CFU/ml reduction. The results of the present study serve as a baseline to study RF as a potential intervention technology for non-intact beef products.

We present the MilliDrop Analyzer (MDA), a droplet-based millifluidic system for digital antimicrobial susceptibility testing (D-AST), which enables us to determine minimum inhibitory concentrations (MICs) precisely and accurately. The MilliDrop technology was validated by using resazurin for fluorescence readout, for comparison with standard methodology, and for conducting reproducibility studies. In this first assessment, the susceptibility of a reference Gram-negative strain Escherichia coli ATCC 25922 to gentamicin, chloramphenicol, and nalidixic acid were tested by the MDA, VITEK®2, and broth microdilution as a reference standard. We measured the susceptibility of clinically relevant Gram-positive strains of Staphylococcus aureus to vancomycin, including vancomycin-intermediate S. aureus (VISA), heterogeneous vancomycin-intermediate S. aureus (hVISA), and vancomycin-susceptible S. aureus (VSSA) strains. The MDA provided results which were much more accurate than those of VITEK®2 and standard broth microdilution. The enhanced accuracy enabled us to reliably discriminate between VSSA and hVISA strains.

Resistance to quinolone antibiotics has been a serious problem ever since nalidixic acid was introduced into clinical medicine. Over time, resistance of pathogenic microbes to nalidixic acid led to the design of novel variants to revive its potential application. In the present work, a series of eight nanoformulations of nalidixic acid-based diacyl and sulfonyl acyl hydrazine derivatives were prepared. All nanoformulations were found to be stable at different storage temperatures. Antibacterial and anticandida activity of the eight nanoformulations presented encouraging results when compared with their non-nano parent counterparts. The nanoformulations of chloro, furanyl, and sulfonyl acyl substituted derivatives of nalidixic acid displayed most promising results (MIC ranging from 50 to 100 μg mL−1) against the tested bacteria and yeast. Among the screened bacteria, Acinetobacter baumannii displayed maximum sensitivity to the above nanoformulations. Biosafety study on the mammalian model—wax moth, Galleria mellonella—showed that all eight prepared nanoformulations were absolutely nontoxic to the larvae and subsequent pupae and hence may likely have no or low toxicity against mammalian systems. Graphical Abstract Electronic supplementary material The online version of this article (10.1186/s11671-018-2718-8) contains supplementary material, which is available to authorized users.

Background Diarrhoeal disease is the second leading cause of death among children aged below 5 years. Even though, both preventable and treatable diseases, globally there are nearly 1.7 billion cases of childhood diarrhoeal disease and responsible for killing around 525,000 children every year. Shigella and Salmonella species were the leading cause of etiologic agents for diarrhoea associated deaths. The aim of this study was to determine the prevalence and antibiotic susceptibility patterns of Shigella and Salmonella isolated from children aged below 5 years with diarrhoea attending Nigist Eleni Mohammed Memorial Hospital, Hossana, South Ethiopia. Methods A cross sectional study was conducted from June 02 to September 24, 2017. Two hundred four children aged below 5 years with diarrhoea were enrolled consecutively using convenience sampling technique. Stool specimens were processed in accordance with the standard bacteriological methods and antibiotic susceptibility pattern of the isolates was determined using disc diffusion method. Data were analyzed using SPSS version 20. Results Out of the 204 children aged below 5 years with diarrhoeal disease 19/204 (9.3%, [95%CI, 5.7–13.7%]) of them were positive for bacterial growth, of which 17/204(8.3%) were Shigella species and 2/204(1%) were Salmonella species. Both Shigella and Salmonella isolates were 100% susceptible to norfloxacin, nalidixic acid and kanamycin. However, isolates of Shigella showed 100, 76.5 and 64.7% resistance to ampicillin, gentamicin and cotrimoxazole respectively while Salmonella species were highly resistant to ampicillin and gentamicin (100% each). Conclusions Salmonella and Shigella species is prevalent in the current study area. Among the tested antibiotics, norfloxacin, nalidixic acid and kanamycin were found to be most effective for both isolates. Both species are developing resistance to the commonly prescribed antibiotic. Therefore, culture based bacterial species identification and antimicrobial susceptibility testing services are strongly recommended to avoid empirical treatment in the study area.

Nalidixic acid is an antibiotic drug used for treatment of Salmonellosis, a gastrointestinal infection. DNA gyrase subunit A (GyrA) of Salmonella typhimurium is the drug target for nalidixic acid. Resistance of GyrA to nalidixic acid, because of a point mutation in S. typhimurium, was recently reported. Substitution of Phe in place of Ser at locus 83 in GyrA of S. typhimurium has been experimentally associated with nalidixic acid resistance. Despite recent efforts, the mechanism of this resistance is not well understood. In this investigation we used computational techniques to address this shortcoming. Our results showed that contact with residue Arg 91 is certainly important for efficient binding of nalidixic acid to the target protein, and that mutation of this residue results in 180° rotation of the antibiotic in its binding pocket, around its own long axis. It is hoped these findings may enable development of new antibiotics against resistant forms of Salmonella.

The role of DNA gyrase in F’lac plasmid conjugation was studied using Escherichia coli gyrA43 (Ts), gyrB41(Ts), and dnaA46(Ts) thermosensitive mutants as donor or recipient organisms, and a rifampicin or nalidixic acid-resistant J-53 strain in the presence or absence of nalidixic acid. Mating experiments were also performed employing Hfr derivatives of the thermosensitive strains. Conjugation was carried out in broth for 60 min using a standard method at permissive and non-permissive (32 and 43 °C) temperatures, with or without drugs. At 32 °C, nalidixic acid reduced the number of transconjugants by about 97 % in comparison to the control, while at 43 °C, the drug inhibited F’lac transfer by about 98 % from dnaA46(Ts) mutant and by about 6.5 % from gyrA43(Ts) and 15 % from gyrB41(Ts) hosts. Using the temperature-sensitive mutants as recipient strains, the transconjugants found were approximately the same under all conditions. The number of transconjugants did not change significantly when nalidixic acid-resistant strains were used as donor or recipient strains. Lastly, nalidixic acid reduced the number of transconjugants from Hfr selected in the above mutants under all experimental conditions. These findings suggest that F’lac transfer does not involve DNA gyrase activity.

We evaluated the antibiogram profile of Escherichia coli (n = 300) isolated from selected rivers in Osun State, Nigeria. The identities of the E. coli isolates were confirmed by polymerase chain reaction (PCR) technique. Susceptibility of the isolates to 20 antibiotics conventionally used in clinical cases was assessed in vitro by the standardized agar disc-diffusion method. All the isolates were susceptible to imipenem, meropenem, amikacin and gatilofloxacin. The isolates were variously susceptible to the other antibiotics as follows: ciprofloxacin (96 %), kanamycin (95 %), neomycin (92 %), streptomycin (84 %), chloramphenicol (73 %), nalidixic acid (66 %), nitrofurantoin (64 %), gentamycin (63 %), doxycycline (58 %), cefepime (57 %), tetracycline (49 %) and cephalothin (42 %). The multiple antibiotic resistance indexing ranged from 0.50 to 0.80 for all the sampling locations and exceeded the threshold value of 0.2, suggesting the origin of the isolates to be of high antimicrobial usage. Our findings signify an increase in the incidence of antimicrobial resistance of E. coli towards conventionally used antibiotics necessitating proper surveillance programmes towards the monitoring of antimicrobial resistance determinants in water bodies.

A novel binuclear squire planar complex of nalidixic acid with Ag(I) metal ion with the formula [Ag(Nal)2] has been synthesized. The synthesized metal complex was characterized using CHN analysis, Fourier-transformed infra-red (FT-IR), thermo gravimetric analysis (TGA), differential scanning calorimetry (DSC), ultra violet-visible (Uv-vis) and single-crystal X-ray diffraction (XRD). The newly synthesized complex shows more advanced antifungal activity compared to the parent quinolone against four fungi, namely Pythium aphanidermatum, Sclerotinia rolfsii, Rhizoctonia solani and Rhizoctonia bataticola.

A total of 1,187 Vibrio cholerae isolates were received during 2011 cholera outbreaks from 16 provinces in different geographical location to Iranian reference Health laboratory. A random selection was performed, and 61 isolates were subjected to further investigations. Cholera cases were come up from May with nine cases and reached to its maximum rate at August (57 cases) and continued to October after which a fall occurred in September. All of the isolates were susceptible to three antimicrobial agents including ciprofloxacin, cefixime, and ampicillin. The highest rate of resistance was seen to nalidixic acid (96.7 %) and co-trimoxazole (91.8 %). Clonality of isolates was investigated through genotyping by PFGE method. A total of seven pulsotypes were obtained from 61 isolates under study. The pulsotypes were highly related with only 1-3 bands differences. Three pulsotypes (PT5, PT6, and PT7) constituted 93.4 % of total isolates. One environmentally isolated strain showed distinct pattern from clinical specimens. This strain although had no any evidence in identified cholera infections, highlighted selecting more environmental specimens in any future outbreaks as long as human samples. In conclusion, emergence and dominance of Ogawa serotypes after about 7 years in Iran are alarming due to fear of import of new V. cholerae clones from out of the country. Approximately, one third of patients in 2011 cholera outbreak in Iran were of Afghan or Pakistani nationality which makes the hypothesis of import of Ogawa serotype strains from neighboring countries more documented and signifies the need to monitor and protect the boundaries.

Background The Campylobacter species usually causes infection between humans and livestock interaction via livestock breeding. The studies of the Campylobacter species thus far in all clinical isolates were to show the many kinds of antibiotic phenomenon that were produced. Their integrons cause the induction of antibiotic resistance between bacterial species in the Campylobacter species. Results The bacterial strains from the diarrhea of pediatric patient which isolated by China Medical University Hospital storage bank. These isolates were identified by MALDI-TOF mass spectrometry. The anti-microbial susceptibility test showed that Campylobacter species resistant to cefepime, streptomycin, tobramycin and trimethoprim/sulfamethoxazole (all C. jejuni and C. coli isolates), ampicillin (89% of C. jejuni; 75% of C. coli), cefotaxime (78% of C. jejuni; 100% of C. coli), nalidixic acid (78% of C. jejuni; 100% of C. coli), tetracycline (89% of C. jejuni; 25% C. coli), ciprofloxacin (67% of C. jejuni; 50% C. coli), kanamycin (33% of C. jejuni; 75% C. coli) and the C. fetus isolate resisted to ampicillin, cefotaxime, nalidixic acid, tetracycline, ciprofloxacin, kanamycin by disc-diffusion method. The effect for ciprofloxacin and tetracycline of the Campylobacter species was tested using an E-test. The tet, erm, and integron genes were detected by PCR assay. According to the sequencing analysis (type I: dfr12-gcuF-aadA2 genes and type II: dfrA7 gene), the cassette type was identified. The most common gene cassette type (type I: 9 C. jejuni and 2 C. coli isolates; type II: 1 C. coli isolates) was found in 12 class I integrase-positive isolates. Conclusions Our results suggested an important information in the latency of Campylobacter species with resistance genes, and irrational antimicrobial use should be concerned.

The spread of Extended Spectrum β-lactamases (ESBLs) among Enterobacteriaceae and other Gram-Negative pathogens in the community and hospitals represents a major challenge to combat infections. We conducted a study to assess the prevalence and genetic makeup of ESBL-type resistance in bacterial isolates causing community- and hospital-acquired urinary tract infections. A total of 172 isolates of Enterobacteriaceae were collected in Dar es Salaam, Tanzania, from patients who met criteria of community and hospital-acquired urinary tract infections. We used E-test ESBL strips to test for ESBL-phenotype and PCR and sequencing for detection of ESBL genes. Overall 23.8% (41/172) of all isolates were ESBL-producers. ESBL-producers were more frequently isolated from hospital-acquired infections (32%, 27/84 than from community-acquired infections (16%, 14/88, p

Wastewater use for crop irrigation and aquaculture is commonly practiced by communities situated close to wastewater treatment ponds. The objective of this study was to characterize Salmonella spp. and their antimicrobial susceptibility patterns among isolates from wastewater and Tilapia fish. A total of 123 Salmonella spp. isolates were isolated from 52 water and 21 fish intestinal samples. Genotyping of Salmonella spp. isolates was done by Pulsed-field Gel Electrophoresis (PFGE). Antimicrobial susceptibility testing was done by the minimal inhibitory concentration (MIC) technique. A total of 123 Salmonella spp. isolates represented 13 different serovars and 22 PFGE groups. Salmonella serovars showed resistance to 8 out of 14 antimicrobials; sulfamethaxazole (94%), streptomycin (61%), tetracycline (22%), ciprofloxacin and nalidixic acid (17%), trimethoprim (11%); gentamycin and chloramphenicol (6%). Salmonella Kentucky, S. Chandans, S. Durban and S. Kiambu showed multiple antimicrobial resistance to 7, 6 and 3 antimicrobials, respectively. This study has demonstrated that wastewater at the study sites is contaminated with Salmonella spp. which are resistant to common antimicrobials used for treatment of diseases in humans. Wastewater may, therefore, contaminate pristine surface water bodies and foodstuffs including fish and irrigated crops as well as food handlers.

Anthrax, a fatal zoonotic disease caused by spore forming bacteria Bacillus anthracis is predominantly witnessed among herbivorous animals that are highly sus- ceptible to the pathogen. Typically B. anthracis spores existing in soil, initiate the infection during grazing and animal to animal spread has been rarely reported. A sus- pected outbreak of anthrax was observed among cattle in Gundlupet taluk neighboring Bandipur national park and tiger reserve during January 2013. Blood samples collected from domestic cattle in Gundlupet region were examined by EA1 sandwich ELISA and immunocapture PCR. Of the 64 samples examined, 12 recovered during the course of outbreak were found positive by the diagnostic methods and further resulted in recovery of B. anthracis isolates that were confirmed by standard methods indicating prevalence of the pathogen among diseased cattle. The outbreak could be contained by the administration of oxytetracycline and live spore vaccine. The isolates recovered from the cattle were susceptible to the major antibiotics penicillin, ampi- cillin, ciprofloxacin, nalidixic acid, tetracycline, erythro- mycin and chloramphenicol whereas resistance could be seen for the antibiotic neomycin (25 %). The ambiguity for initial symptomatic identification due to the absence of bleeding from natural orifices could be overcome by the EA1 immunoassay kits employed in this study.

The level of antibiotic multiresistance of Campylobacter strains from chicken was evaluated and responses to the bactericidal effects of organic acids were analyzed. Strains (76) isolated from chicken were analyzed for resistance to antibiotics and acid shock. A high strain resistance to fluoroquinolones (ciprofloxacin and nalidixic acid) was observed, with cross resistance to both drugs in 41% of strains. A low resistance was observed for amoxicillin, erythromycin, and gentamicin. Campylobacter jejuni was the most resistant species. Antibacterial activities against multiresistant Campylobacter strains were observed using acetic, citric, and ascorbic acids at minimum bactericidal concentrations (MBC) ranging from 0.3 to 3 mg/mL. Acetic acid was the most efficient acid with the lowest MBC value. However, a contact time of 4 h was required for an efficient effect against Campylobacter contaminated chicken skin. Using organic acids in the poultry production chain can reduce propagation of antibiotic multiresistant strains of Campylobacter.

Streptococcus pneumoniae is a leading cause of respiratory tract infections, including pneumonia. In recent years, due to the incidence of resistance to commonly used antibiotics, quinolones have been taken into consideration in treatment of S. pneumoniae. The present study aimed to investigate the quinolone resistance rate among pneumococcus isolates from Shiraz. This cross-sectional-descriptive study was performed on 45 strains of S. pneumoniae isolated from clinical samples. Initially, using biochemical tests and PCR technique with specific primers of lytA gene, S. pneumoniae strains were identified. Subsequently, antibiotic resistance to quinolones was evaluated by CLSI criteria. Among 45 strains isolated from clinical specimen, resistance to nalidixic acid, ciprofloxacin, ofloxacin, norfloxacin and levofloxacin were estimated 88.22, 73.43, 53.33, 48.88 and 42.22 %, respectively. The highest frequency of resistance corresponded to the people with pneumonia. Besides, the greatest resistance to all antibiotics related to the age group of 31–40. Statistical analysis indicated no significant association between antibiotic resistance and disease type or age groups. In concurrence with other investigations, our results showed that the level of resistance to quinolones is increasing in Shiraz. Hence, the quinolone resistance in this region is a serious issue; therefore, the regional pattern of antimicrobial resistance on therapeutic regimes must be taken into consideration.

A study was conducted to evaluate the risk of Salmonella contamination and its survival at different stages in chill ready-to-eat poultry meat products chain. Samples (n = 181) were collected and examined for the presence of Salmonella species. The bacteria were initially identified against polyvalent antisera "O" and "H", followed by confirmation with 16 s rDNA. The single Salmonella, isolate from the tested food samples showed 99.8 % phylogenetic similarity with Salmonella enterica. It was further evaluated for antibiotic sensitivity pattern and found resistant to four antibiotics including ampicillin, chloramphenicol, tetracycline and nalidixic acid. Salmonella associated with ready-to-eat poultry meat products were found active at storage temperature ≥4 °C in a challenge study. It was revealed that shape and weight of the meat pieces have direct influence on the reduction of pathogens during microwave heating. The 30 and 60 s microwaving (with radiation power fixed at 900 W) was found ineffective for the elimination of target bacteria (10(6)-10(7) CFU/g) in meat pieces with weight ≥90 g. Salmonella enterica was able to survive in simulated gastric fluid. The storage temperature and microwaving were found critical point for the transfer of pathogens through ready-to-eat poultry meat products to consumer in chill ready-to-eat poultry meat chain.