Combined Effects Of Antibiotics Research Articles

Combined toxicity of microplastics and antibiotics towards aquatic ecosystems: A case study in Beiluo River, China

Microplastics (MPs) can adsorb antibiotics to form complex pollutants that seriously threaten the health of freshwater ecosystems. However, few studies have examined the combined ecotoxicity of MPs and multiple antibiotics in natural aquatic environments. In this study, we examined the effects of combined exposure to antibiotics and MPs on aquatic community structure in natural settings in the Beiluo River using eDNA analysis. The results revealed that antibiotic pollution in the Beiluo River mainly originates from animal husbandry and agriculture, whereas MPs mainly originate from agriculture and urban sewage. The community structure of aquatic organisms was significantly correlated with the concentrations of antibiotics and MPs and that the combined effects of antibiotics and MPs were highly interpretable for cyanobacteria and phytoplankton. The effect of MPs on antibiotic toxicity is highly dependent on the antibiotic type and MP particle size. These insights highlight that co-exposure to MPs and antibiotics in the natural environment is a common phenomenon and that co-exposure to MPs can alter the toxicological profile of antibiotics. The results enhance our comprehensive understanding of the complex combined effects of MPs and antibiotics on aquatic organisms, emphasising the necessity of informed scientific management of these emerging contaminants.

Synergistic impacts of antibiotics and heavy metals on Hermetia illucens: Unveiling dynamics in larval gut bacterial communities and microbial metabolites

Hermetia illucens larvae showcases remarkable bioremediation capabilities for both antibiotics and heavy metal contaminants. However, the distinctions in larval intestinal microbiota arising from the single and combined effects of antibiotics and heavy metals remain poorly elucidated. In this study, we delved into the details of larval intestinal bacterial communities and microbial metabolites when exposed to single and combined contaminants of oxytetracycline (OTC) and hexavalent chromium (Cr(VI)). After conversion, single contaminant-spiked substrate showed 75.5% of OTC degradation and 95.2% of Cr(VI) reductiuon, while combined contaminant-spiked substrate exhibited 71.3% of OTC degradation and 93.4% of Cr(VI) reductiuon. Single and combined effects led to differences in intestinal bacterial communities, mainly reflected in the genera of Enterococcus, Pseudogracilibacillus, Gracilibacillus, Wohlfahrtiimonas, Sporosarcina, Lysinibacillus, and Myroide. Moreover, these effects also induced differences across various categories of microbial metabolites, which categorized into amino acid and its metabolites, benzene and substituted derivatives, carbohydrates and its metabolites, heterocyclic compounds, hormones and hormone-related compounds, nucleotide and its metabolites, and organic acid and its derivatives. In particular, the differences induced OTC was greater than that of Cr(VI), and combined effects increased the complexity of microbial metabolism compared to that of single contaminant. Correlation analysis indicated that the bacterial genera, Preudogracilibacillus, Enterococcus, Sporosarcina, Lysinibacillus, Wohlfahrtiimonas, Ignatzschineria, and Fusobacterium exhibited significant correlation with significant differential metabolites, these might be used as indicators for the resistance and bioremediation of OTC and Cr(VI) contaminants. These findings are conducive to further understanding that the metabolism of intestinal microbiota determines the resistance of Hermetia illucens to antibiotics and heavy metals.

Effects of environmentally relevant concentrations of oxytetracycline and sulfadiazine on the bacterial communities, antibiotic resistance genes, and functional genes are different between maize rhizosphere and bulk soil.

Antibiotic contamination in soil has become a major concern worldwide. At present, it is not clear how two co-existed antibiotics with environmentally relevant concentrations would affect soil bacterial community structure, the abundances of antibiotic resistance genes (ARGs) and functional genes, and whether the effects of antibiotics would differ between rhizosphere and bulk soil. We conducted a greenhouse pot experiment to grow maize in a loess soil treated with oxytetracycline (OTC) or sulfadiazine (SDZ) or both at an environmentally relevant concentration (1 mg kg-1) to investigate the effects of OTC and SDZ on the rhizosphere and bulk soil bacterial communities, abundances of ARGs and carbon (C)-, nitrogen (N)-, and phosphorus (P)-cycling functional genes, and on plant growth and plant N and P nutrition. The results show that the effects of environmentally relevant concentrations of OTC and SDZ on bacterial communities and abundances of ARGs and functional genes differ between maize rhizosphere and bulk soil. The effects of two antibiotics resulted in a higher absolute abundances of accA, tet(34), tnpA-04, and sul2 in the rhizosphere soil than in the bulk soil and different bacterial community compositions and biomarkers in the rhizosphere soil and the bulk soil. However, OTC had a stronger inhibitory effect on the abundances of a few functional genes in the bulk soil than SDZ did, and their combination had no synergistic effect on plant growth, ARGs, and functional genes. The role of co-existed OTC and SDZ decreased shoot height and increased root N concentration. The results demonstrate that environmentally relevant concentrations of antibiotics shift soil microbial community structure, increase the abundances of ARGs, and reduce the abundances of functional genes. Furthermore, soil contamination with antibiotics can diminish agricultural production via phytotoxic effects on crops, and combined effects of antibiotics on plant growth and nutrient uptake should be considered.

Mechanism and synergistic effect of sulfadiazine (SDZ) and cadmium toxicity in spinach (Spinacia oleracea L.) and its alleviation through zinc fortification

Cadmium (Cd) and antibiotic’s tendency to accumulate in edible plant parts and fertile land is a worldwide issue. The combined effect of antibiotics and heavy metals on crops was analyzed, but not mitigation of their toxicity. This study investigated the potential of zinc oxide nanoparticles (ZnO NPs) to alleviate the SDZ and Cd toxicity (alone/combined) to promote spinach growth. Results revealed that the ZnO 200 mg L−1 spray decreased the malondialdehyde (MDA) 14%, hydrogen peroxide (H2O2) 13%, and electrolyte leakage (EL) 7%, and increased the superoxide dismutase (SOD) 8%, peroxidase (POD) 25%, catalase (CAT) 39% and ascorbate peroxidase (APX) 12% in spinach leaves under combined SDZ+Cd (25 mg Kg−1 +50 mg Kg−1) stress compared to ZnO 100 mg L−1 spray. Likewise, ZnO NPs 200 mg L−1 spray enhanced the zinc (Zn) 97%, iron (Fe) 86%, magnesium (Mg) 35%, manganese (Mn) 8%, and potassium (K) 23% in shoots under combined SDZ+Cd (25 mg Kg−1 +50 mg Kg−1) stress compared to ZnO 100 mg L−1 spray. Further, ZnO 200 mg L−1 spray reduced Cd uptake in roots by 9% and shoots 15% under combined SDZ+Cd (25 mg Kg−1 +50 mg Kg−1) stress compared to ZnO 100 mg L−1. Overall, ZnO NPs alleviated the SDZ and Cd toxicity and enhanced spinach growth in all treatments.

Combined effects of antibiotics and acidified nitrite on biofilm formation by beta-lactamase-producing uropathogenic bacteria

Background and objective: The increased prevalence of extended- spectrum beta-lactamase producing Enterobacteriaceae has increased the use of last-resort antimicrobial drugs like Carbapenems. An alternative idea is to use new combinations of common antibiotics. The aim of the present study is to examine whether acidified nitrite has the ability to enhance the activity of beta-lactam antibiotics against the biofilm formation and bacterial growth of beta-lactamase- producing uropathogenic isolates. Methods: In this cross-sectional study, a total of 37 beta-lactamase- producing uropathogens were collected from patients at Urology Department at Rizgary Teaching Hospital in Erbil/Iraq. Biofilm formation was determined using a 96-well tissue culture plate assay. The ability to produce beta-lactamase production was detected by a phenotypic confirmatory combination disk diffusion test. The sub-minimal inhibitory concentration of antibiotics alone and in combination with each of acidified nitrite, ascorbic acid and sodium nitrite towards biofilm formation were observed. Results: The combination of cefotaxime with each of acidified nitrite (P <0.001), ascorbic acid (P <0.001) and sodium nitrite (P = 0.003) significantly enhanced the effect of cefotaxime against the biofilm producing activity of beta-lactamase producing uropathogens. Furthermore, with ceftazidime identical synergistic results were obtained with ascorbic acid (P = 0.001) and acidified nitrite (P = 0.007). Conclusion: Acidified nitrite significantly improved the activity of beta-lactam antibiotics against the biofilm mass of beta-lactamase producing uropathogens.

New insights into the effects of antibiotics and copper on microbial community diversity and carbon source utilization.

Residual antibiotics (ABs) and heavy metals (HMs) are continuously released from soil, reflecting their intensive use and contamination of water and soil, posing an environmental problem of great concern. Relatively few studies exist of the functional diversity of soil microorganisms under the combined action of ABs and HMs. To address this deficiency, BIOLOG ECO microplates and the Integrated Biological Responses version 2 (IBRv2) method were used to comprehensively explore the effects of single and combined actions of copper (Cu) and enrofloxacin (ENR), oxytetracycline (OTC), and sulfadimidine (SM2) on the soil microbial community. The results showed that the high concentration (0.80mmol/kg) compound group had a significant effect on average well color development (AWCD) and OTC showed a dose-response relationship. The results of IBRv2 analysis showed that the single treatment group of ENR or SM2 had a significant effect on soil microbial communities, and the IBRv2 of E1 was 5.432. Microbes under ENR, SM2, and Cu stress had more types of available carbon sources, and all treatment groups were significantly more enriched with microorganisms having D-mannitol and L-asparagine as carbon sources. This study confirms that the combined effects of ABs and HMs can inhibit or promote the function of soil microbial communities. In addition, this paper will provide new insights into IBRv2 as an effective method to evaluate the impacts of contaminants on soil health.

Effects of Sulfamethoxazole and Florfenicol on Growth, Antioxidant Capacity, Immune Responses and Intestinal Microbiota in Pacific White Shrimp Litopenaeus vannamei at Low Salinity.

Antibiotic residue may pose a serious risk to aquaculture, and the culture of Litopenaeus vannamei in a low-salinity environment is a growing trend over the world. Here, we aimed to understand the combined effect of low salinity and sulfamethoxazole (SMZ) and florfenicol (FLO) antibiotics on L. vannamei. The growth performance, immune functions, antioxidant capacity and intestinal microbiota were investigated. Compared with the control group, the weight gain and survival rate significantly decreased (p < 0.05) in shrimp after they were exposed to low-salinity (salinity 3) water and the mixture of antibiotics and low-salt conditions for 28 days. The antioxidant activities of SOD and T-AOC, shown at low salinity and in the higher concentration of the SMZ treatment group (SMZH), were significantly decreased, while the GST activity was significantly increased in each treatment group in comparison with the control group. The expression of immune-related genes, including TOLL, LvIMD, PPO and HSP, in the low concentration of the SMZ treatment group (SMZL) was higher than that in the other groups. The diversity of intestine microbiota was disturbed with a lower Shannon index in the low-salinity and SMZH groups, and a higher Simpson index in the SMZH group. Proteobacteria, Actinobacteria and Bacteroidetes were the dominant phyla in the gut of L. vannamei. At the genus level, Microbacterium, Shewanella, Aeromonas, Acinetobacter, Gemmobacter, Paracoccus and Lysobacter were significantly decreased in the low-salinity group. However, the abundance of opportunistic pathogens belonging to the genus Aeromonas in the FLO group was increased. The predicted microbe-mediated functions showed that the pathway for "amino acid metabolism" and "replication and repair" was significantly inhibited in both the low-salinity and antibiotic-exposed groups. All the findings in this study indicate that the combined effect of antibiotics and low salinity on L. vannamei negatively impacted the physiological and intestinal microbiota functions.

A264 ORAL IRON SUPPLEMENTATION SHAPES THE RECOVERY OF GUT THE MICROBIOTA AND PROMOTES CARCINOGENESIS IN THE APCMIN/+ MOUSE MODEL

Abstract Background Colorectal cancer (CRC) induces anemia in a large proportion of patients and is usually treated with oral iron supplementation. Surgery, the main treatment for CRC, is routinely accompanied by prophylactic antibiotics to avoid infection. Moreover, the concomitance of oral iron supplementation and antibiotics treatment is highly prevalent in general population. However, the combined effect of antibiotics and high luminal iron concentration in the gut on the microbiota and intestinal homeostasis remains unknown. Purpose This study sought to identify possible deleterious effects of the recovery from antibiotics exposure of the gut microbiota under oral iron supplementation. Method In order to assess the effects of luminal iron on the recovery of the gut microbiota, WT mice were subjected to antibiotic treatment with different concentrations of dietary iron. Then, to explore potential deleterious effects of this setup on CRC promotion and progression mediated by the human CRC gut microbiota, CRC model ApcMin/+ mice were subjected to antibiotics prior to gavage with fecal samples from anemic CRC patients, then fed different concentrations of dietary iron. The composition of the gut microbiota and its recovery after these interventions were assessed in stool samples by 16S rRNA. Gut microbiota functions were inferred by using the PICRUSt2, and short chain fatty acid (SCFA) concentration in feces were assessed by liquid-chromatography mass spectrometry. CRC progression and promotion were assessed by tumour counts and immunohistochemistry. Result(s) Recovery from antibiotics under high luminal iron concentration shifted the gut microbiota toward a Bacteroidetes phylum-dominant composition. Three bacterial species characterized as CRC initiators were significantly more abundant. Nitrogen, pentose phosphate, cyanoamino acid metabolisms were higher after recovery under oral iron supplementation. Antibiotic exposure induced a long-term increase in SCFAs linked to gut inflammation, propionate and succinate. For mice under high luminal iron concentration, they showed a lack of recovery in baseline levels of butyrate, a SCFA that inhibits cancer cell proliferation in the gut. In addition, an increase in CRC burden and progression were found in ApcMin/+ mice receiving fecal sample from patients and oral iron supplementation. Conclusion(s) Gut microbiota recovery from antibiotic exposure under oral iron supplementation is frequent in CRC patients, but is also common in the general population. This study identifies possible deleterious effects of the concomitance of these two disruptive agents of the gut microbiota on the CRC progression. Please acknowledge all funding agencies by checking the applicable boxes below Other Please indicate your source of funding; NSERC, ICM Disclosure of Interest None Declared

Synergistic effect of copper oxide nanoparticles and chloramphenicol antibiotic on MexA gene expression of pump efflux system in drug-resistant Pseudomonas aeruginosa isolates

Background. Pseudomonas aeruginosa is a causative agent of nosocomial infections. Chloramphenicol is a broad-spectrum, inexpensive, and common antibiotic. MexAB-OprM is a secretory pump that causes the innate resistance of this bacterium. This study aimed to evaluate the expression of MexA gene under the treatment of copper nanoparticles and chloramphenicol. Methods. In this study, 49 samples were collected from 11 laboratories in Mashhad, Iran.Abundance of Me gene MexA was done by polymerase chain reaction (PCR) method, and antibiotic susceptibility assessment was performed by disk diffusion method. To calculate minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC), copper nanoparticles, and a combination of both, the broth dilution method was performed. Microdilution method and real time PCR technique were used to determine the effective dilution of copper oxide nanoparticles and chloramphenicol antibiotics and expression of MexA gene, respectively. Result. The identity of 49 strains was confirmed. All strains had the MexA gene and were resistant to more than two antibiotics. The MIC of bacterial strains was 250 μg/ml for copper oxide nanoparticles and 62.50 μg/ml for chloramphenicol antibiotic. Compared to antibiotics, copper nanoparticles had a greater effect on MIC, though it was lower than the combined effect of antibiotics and copper oxide nanoparticles. Conclusion. Copper oxide nanoparticles or the synergistic effect of copper oxide nanoparticles and chloramphenicol antibiotics are effective in reducing MexA gene expression. Practical Implications. Nanoparticles are an important option for use as an alternative to or supplement to antibiotics. Also, copper oxide nanoparticles are more effective to inhibit bacterial growth when used in combination with antibiotics.

Single and combined effects of antibiotics and nanoplastics from surgical masks and plastic bottles on pathogens

Over the last decade, pollution of plastics and antibiotics has increased in its threat to the environment and human health. However, very limited information is available concerning impact of co-presence of plastics and antibiotics on environment and human health. Moreover, the potential ingestion and inhalation of nano(micro)plastics due to the disposable materials has dramatically increased. With the outbreak and spread of the COVID-19 in the world, disposable surgical masks and plastic bottles have been widely used by the public, and their rapid use and improper dispensing can cause to increase plastic pollution risk on human. However, impacts of co-presence of nano(micro)plastics and antibiotics on pathogens have yet been demonstrated. Therefore, this study aims to investigate the impact the individual and combined influences of nano-sized plastics (surgical mask and plastic bottles) and antibiotics (amoxicillin and spiramycin) towards the main susceptible bacterium (Staphylococcus epidermidis, Staphylococcus aureus, Bacillus subtilis, Escherichia coli, and Pseudomonas aeruginosa) by microbial activity, biofilm formation and their biochemical characteristics. The results showed that antimicrobial efficiencies of the tested antibiotics were reduced (approximately 10–98%) with the plastics. Moreover, the biochemical pathways of the microbial activity changed by the plastics entrance. Polymer structure and sorption play the role on the reduction in the inhibition of pathogens. In the meantime, the biofilm formation changed and characteristic of the extracellular polymeric substance with the co-presence of plastics and antibiotics mostly depended on the polymer structure, exposure time and sorption.

A34 ANTIBIOTIC TREATMENT AND ORAL IRON SUPPLEMENTATION SHAPE THE COMPOSITION, RECOVERY AND FUNCTION OF THE GUT MICROBIOTA

Abstract Background Colorectal cancer (CRC) induces anemia in a large proportion of patients and is usually treated with oral iron supplementation. Surgery, the main treatment for CRC, is routinely accompanied by prophylactic antibiotics to avoid infection. However, the combined effect of antibiotics and luminal iron in the gut on the microbiota and intestinal homeostasis remains unknown. Aims We aim to characterize the dynamics of gut microbiota composition and recovery from antibiotic exposure under iron-sufficient and iron-supplemented diets in mice. We will investigate how microbial shifts induced by antibiotics and iron influence the function of the gut microbiota and metabolites in the gut. Methods Mice were subjected to antibiotic treatment with different concentrations of dietary iron. The composition of the gut microbiota and its recovery after these interventions were assessed in stool samples by 16S rRNA sequencing before and after antibiotic exposure and during the recovery period. Gut microbiota functions were inferred by using the PICRUSt2 prediction tool, and short chain fatty acid (SCFA) concentration in feces were assessed by liquid-chromatography mass spectrometry. Results Recovery from antibiotics under high luminal iron concentration shifted the gut microbiota toward a Bacteroidetes phylum-dominant composition. Four bacterial species characterized as CRC markers and/or CRC initiators were significantly more abundant, and nitrogen and pentose phosphate metabolism were higher after recovery under oral iron supplementation. Antibiotic exposure induced a long-term increase in SCFAs linked to gut inflammation, propionate and succinate, and was independent of luminal iron concentration. For mice recovering from antibiotics under high luminal iron concentration, they showed a lack of recovery in baseline levels of butyrate, a SCFA that inhibits cancer cell proliferation in the gut. Conclusions Gut microbiota recovery from antibiotic exposure under oral iron supplementation is frequent in CRC patients, but is also common in the general population. This study identifies possible deleterious effects of the concomitance of these two disruptive agents of the gut microbiota and may lead to modifications in the management of anemia in patients with CRC. Funding Agencies CIHR Bourse de Mérite Rougier-Armandie en recherche médicale de la Faculté de médecine

Toxicity and combined effects of antibiotics and nano ZnO on a phosphorus-removing Shewanella strain in wastewater treatment

Antibiotics and nanoparticles, which are emerging contaminants, can occur simultaneously in biological wastewater treatment systems, potentially resulting in complex interactive effects. This study investigated the effects of individual and complex zinc oxide nanoparticles (nZnO) and antibiotics (quinolone and sulfonamide), on the Shewanella strain used to remove phosphorus (PO43-), metabolic processes, as well as its complexing and toxicity mechanisms. The inhibition of PO43- removal increased from 30.7% to 100.0% with increased nZnO concentrations (half maximal effective concentration (EC50) = 1.1 mg Zn/L) by affecting poly-p and glycogen metabolites. The combined exposure to nZnO and ciprofloxacin/norfloxacin (CIP/NOR) had a significant antagonistic effect on the removal of PO43- and on the metabolism of poly-p and glycogen in phosphate-accumulating organisms (PAOs), whereas the complexing of sulfonamide and nZnO had no significant additional effect. Thus, the complexing of nanoparticles and antibiotics exhibited different toxicity effects from the antibiotic structure-based complex reactions. These results can be used to improve wastewater treatment processes and reduce risks associated with wastewater discharge.

Hormetic dose-dependent response about typical antibiotics and their mixtures on plasmid conjugative transfer of Escherichia coli and its relationship with toxic effects on growth

Bacterial resistance caused by the abuse of antibiotics has attracted worldwide attention. However, there are few studies exploring bacterial resistance under the environmental exposure condition of antibiotics that is featured by low-dose and mixture. In this study, sulfonamides (SAs), sulfonamide potentiators (SAPs) and tetracyclines (TCs) were used to determine the effects of antibiotics on plasmid RP4 conjugative transfer of Escherichia coli (E. coli) under single or combined exposure, and the relationship between the effects of antibiotics on conjugative transfer and growth was investigated. The results show that the effects of single or binary antibiotics on plasmid RP4 conjugative transfer all exhibit a hormetic phenomenon. The linear regression reveals that the concentrations of the three antibiotics promoting conjugative transfer are correlated with the concentrations promoting growth and the physicochemical properties of the compounds. The combined effects of SAs-SAPs and SAs-TCs on plasmid conjugative transfer are mainly synergistic and antagonistic. While SAPs provide more effective concentrations for the promotion of conjugative transfer in SAs-SAPs mixtures, SAs play a more important role in promoting conjugative transfer in SAs-TCs mixtures. Mechanism explanation shows that SAs, SAPs and TCs inhibit bacterial growth by acting on their target proteins DHPS, DHFR and 30S ribosomal subunit, respectively. This study indicates that toxic stress stimulates the occurrence of conjugative transfer and promotes the development of bacterial resistance, which will provide a reference for resistance risk assessment of antibiotic exposure.

Bioactive compounds from the African medicinal plant Cleistochlamys kirkii as resistance modifiers in bacteria.

Cleistochlamys kirkii (Benth) Oliv. (Annonaceae) is a medicinal plant traditionally used in Mozambique to treat infectious diseases. The aim of this study was to find resistance modifiers in C.kirkii for Gram-positive and Gram-negative model bacterial strains. One of the most important resistance mechanisms in bacteria is the efflux pump-related multidrug resistance. Therefore, polycarpol (1), three C-benzylated flavanones (2-4), and acetylmelodorinol (5) were evaluated for their multidrug resistance-reverting activity on methicillin-susceptible and methicillin-resistant Staphylococcus aureus and Escherichia coli AG100 and AG100 A strains overexpressing and lacking the AcrAB-TolC efflux pump system. The combined effects of antibiotics and compounds (2 and 4) were also assessed by using the checkerboard microdilution method in both S.aureus strains. The relative gene expression of the efflux pump genes was determined by real-time reverse transcriptase quantitative polymerase chain reaction. The inhibition of quorum sensing was also investigated. The combined effect of the antibiotics and compound 2 or 4 on the methicillin-sensitive S.aureus resulted in synergism. The most active compounds 2 and 4 increased the expression of the efflux pump genes. These results suggested that C.kirkii constituents could be effective adjuvants in the antibiotic treatment of infections.

Synthesis, characterization and evaluation of antimicrobial and cytotoxic activities of biogenic silver nanoparticles synthesized from Streptomyces xinghaiensis OF1 strain

We report synthesis of silver nanoparticles (AgNPs) from Streptomyces xinghaiensis OF1 strain, which were characterised by UV–Vis and Fourier transform infrared spectroscopy, Zeta sizer, Nano tracking analyser, and Transmission electron microscopy. The antimicrobial activity of AgNPs alone, and in combination with antibiotics was evaluated against bacteria, namely Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and Bacillus subtilis, and yeasts viz., Candida albicans and Malassezia furfur by using micro-dilution method. The minimum inhibitory concentration (MIC) and minimum biocidal concentration of AgNPs against bacterial and yeast strains were determined. Synergistic effect of AgNPs in combination with antibacterial and antifungal antibiotics was determined by FIC index. In addition, MTT assay was performed to study cytotoxicity of AgNPs alone and in combination with antibiotics against mouse fibroblasts and HeLa cell line. Biogenic AgNPs were stable, spherical, small, polydispersed and capped with organic compounds. The variable antimicrobial activity of AgNPs was observed against tested bacteria and yeasts. The lowest MIC (16 µg ml−1) of AgNPs was found against P. aeruginosa, followed by C. albicans and M. furfur (both 32 µg ml−1), B. subtilis and E. coli (both 64 µg ml−1), and then S. aureus and Klebsiella pneumoniae (256 µg ml−1). The high synergistic effect of antibiotics in combination with AgNPs against tested strains was found. The in vitro cytotoxicity of AgNPs against mouse fibroblasts and cancer HeLa cell lines revealed a dose dependent potential. The IC50 value of AgNPs was found in concentrations of 4 and 3.8 µg ml−1, respectively. Combination of AgNPs and antibiotics significantly decreased concentrations of both antimicrobials used and retained their high antibacterial and antifungal activity. The synthesis of AgNPs using S. xinghaiensis OF1 strain is an eco-friendly, cheap and nontoxic method. The antimicrobial activity of AgNPs could result from their small size. Remarkable synergistic effect of antibiotics and AgNPs offer their valuable potential in nanomedicine for clinical application as a combined therapy in the future.

Effects of multiple antibiotics exposure on denitrification process in the Yangtze Estuary sediments

Denitrification is a dominant reactive nitrogen removal pathway in most estuarine and coastal ecosystems, and plays a significant role in regulating N2O release. Although multiple antibiotics residues are widely detected in aquatic environment, combined effects of antibiotics on denitrification remain indistinct. In this work, 5 classes of antibiotics (sulfonamides, chloramphenicols, tetracyclines, macrolides, and fluoroquinolones) were selected to conduct orthogonal experiments in order to explore their combined effects on denitrification. 15N-based denitrification and N2O release rates were determined in the orthogonal experiments, while denitrifying functional genes were examined to illustrate the microbial mechanism of the combined antibiotics effect. Denitrification rates were inhibited by antibiotics treatments, and synergistic inhibition effect was observed for multiple antibiotics exposure. Different classes of antibiotics had different influence on N2O release rates, but multiple antibiotics exposure mostly led to stimulatory effect. Abundances of denitrifying functional genes were inhibited by multiple antibiotics exposure due to the antimicrobial properties, and different inhibition on denitrifiers may be the major mechanism for the variations of N2O release rates. Combined effects of antibiotics on denitrification may lead to nitrate retention and N2O release in estuarine and coastal ecosystems, and consequently cause cascading environmental problems, such as greenhouse effects and hyper-eutrophication.

Enhanced antibacterial effect of antibiotics in combination with silver nanoparticles against animal pathogens

Antibiotic resistant bacteria are a serious health risk in both human and veterinary medicine. Several studies have shown that silver nanoparticles (AgNPs) exert a high level of antibacterial activity against antibiotic resistant strains in humans. The aim of this study was to evaluate the antibacterial effects of a combined therapy of AgNPs and antibiotics against veterinary bacteria that show resistance to antibiotics. A microdilution checkerboard method was used to determine the minimal inhibitory concentrations of both types of antimicrobials, alone and in combination. The fractional inhibitory concentration index was calculated and used to classify observed collective antibacterial activity as synergistic, additive (only the sum of separate effects of drugs), indifferent (no effect) or antagonistic.From the 40 performed tests, seven were synergistic, 17 additive and 16 indifferent. None of the tested combinations showed an antagonistic effect. The majority of synergistic effects were observed for combinations of AgNPs given together with gentamicin, but the highest enhancement of antibacterial activity was found with combined therapy together with penicillin G against Actinobacillus pleuropneumoniae. A. pleuropneumoniae and Pasteurella multocida originally resistant to amoxycillin, gentamicin and colistin were sensitive to these antibiotics when combined with AgNPs. The study shows that AgNPs have potential as adjuvants for the treatment of animal bacterial diseases.

In vitro Synergism between Chloroquine and Antibiotics against Orientia tsutsugamushi.

BackgroundTo investigate whether chloroquine enhances the effect of antibiotics against Orientia tsutsugamushi, the causative organism of scrub typhus, we compared the effect of antibiotics in combination with chloroquine with the effect of antibiotics alone in vitro.Materials and MethodsThe Boryong or AFSC-4 strain was inoculated into ECV304 cells, and incubated in medium containing doxycycline (4 µg/mL), rifampin (4 µg/mL), azithromycin (0.5 µg/mL), chloroquine (1 µg/mL), and each of these antibiotics in combination with chloroquine for 7 d. Immunofluorescence (IF) staining for O. tsutsugamushi was performed 4 hr and 7 d after inoculation of the bacteria, and IF-positive foci were enumerated.ResultsChloroquine inhibited the growth of O. tsutsugamushi by 15.5%. In combination with chloroquine, the antimicrobial effects increased by 4.4% for doxycycline (a 92.9% reduction of bacterial numbers for doxycycline versus a 97.3% reduction for doxycycline plus chloroquine), 4.6% for rifampin (90.0% versus 94.6%), and 8.3% for azithromycin (86.9% versus 95.2%). The antimicrobial effect of the antibiotics alone was significantly different compared to the combined effect of antibiotics and chloroquine (Wilcoxon signed-rank test, P = 0.001).ConclusionsThe combined use of chloroquine with an antibiotic for the treatment of O. tsutsugamushi infections may be useful for increasing the efficacy of the antibiotics.

Stress of Five Heavy Metals on the Resistance of Isolates from Swine Wastewater to Four Antibiotics

Co-existence of heavy metals and antibiotics becomes increasingly common in environmental pollution. To investigate the stress of heavy metals on microbial resistance to antibiotics, fifty-six strains of bacteria were initially isolated from some swine water in Guangzhou city, based on their resistance to four antibiotics (cefradine, norfloxacin, amoxicillin, tetracycline) and five heavy metals (Pb2+, Cr(VI), Hg2+, Cu2+, Zn2+), a gram-negative isolate, Pseudomonas putida XX6, was selected to study the detail stress rules of heavy metals on its resistance to antibiotics. The antibiotics incidences of these isolates were in the order of norfloxacin>amoxicillin>cefradine>tetracycline, and that of P. putida XX6 was cefradine>amoxicillin≈tetracycline>norfloxacin. The addition of heavy metals made all isolates’ resistance to antibiotics decrease, and Cr(VI) impacted their resistance to norfloxacin most obviously. If the concentration of heavy metals was the most important factor affecting the resistance of P. putida XX6 to the antibiotics? There was a positive correlation between the bacterial resistances to antibiotics and heavy metals of low concentrations, and the correlation turned to negative with the concentrations of heavy metals increasing. But the bacterial resistance to amoxicillin or cefradine remained irrelevant to the concentrations of Cr (VI) or Pb2+. Results showed that the combined effect of antibiotics and heavy metals could alter their individual effect on bio-toxicity as well as on the biological removal capability of pollutants.

β-lactam antibiotic-induced vancomycin resistant メチシリン耐性黄色ブドウ球菌株の検出方法と抗菌薬の併用効果

β-lactam antibiotic-induced vancomycin resistant メチシリン耐性黄色ブドウ球菌株の検出方法と抗菌薬の併用効果