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The prevalence of ampicillin-resistant opportunistic pathogenic bacteria undergoing selective stress of heavy metal pollutants in the Xiangjiang River, China
2021
Wang, Qing | Xu, Yan | Liu, Lei | Li, Lin-Yun | Lin, Huai | Wu, Xin-Yan | Bi, Wen-Jing | Wang, Lidao | Mao, Da-Qing | Luo, Yi
The emergence of clinically relevant β-lactam-resistant bacteria poses a serious threat to human health and presents a major challenge for medical treatment. How opportunistic pathogenic bacteria acquire antibiotic resistance and the prevalence of antibiotic-resistant opportunistic pathogenic bacteria in the environment are still unclear. In this study, we further confirmed that the selective pressure of heavy metals contributes to the increase in ampicillin-resistant opportunistic pathogens in the Xiangjiang River. Four ampicillin-resistant opportunistic pathogenic bacteria (Pseudomonas monteilii, Aeromonas hydrophila, Acinetobacter baumannii, and Staphylococcus epidermidis) were isolated on Luria-Bertani (LB) agar plates and identified by 16S rRNA sequencing. The abundance of these opportunistic pathogenic bacteria significantly increased in the sites downstream of the Xiangjiang River that were heavily influenced by metal mining activities. A microcosm experiment showed that the abundance of β-lactam resistance genes carried by opportunistic pathogenic bacteria in the heavy metal (Cu²⁺ and Zn²⁺) treatment group was 2–10 times higher than that in the control. Moreover, heavy metals (Cu²⁺ and Zn²⁺) significantly increased the horizontal transfer of plasmids in pathogenic bacteria. Of particular interest is that heavy metals facilitated the horizontal transfer of conjugative plasmids, which may lead to the prevalence of multidrug-resistant pathogenic bacteria in the Xiangjiang River.
Afficher plus [+] Moins [-]Titanium dioxide nanoparticles enhance mortality of fish exposed to bacterial pathogens
2015
Jovanović, Boris | Whitley, Elizabeth M. | Kimura, Kayoko | Crumpton, Adam | Palić, Dušan
Nano-TiO2 is immunotoxic to fish and reduces the bactericidal function of fish neutrophils. Here, fathead minnows (Pimephales promelas) were exposed to low and high environmentally relevant concentration of nano-TiO2 (2 ng g−1 and 10 μg g−1 body weight, respectively), and were challenged with common fish bacterial pathogens, Aeromonas hydrophila or Edwardsiella ictaluri. Pre-exposure to nano-TiO2 significantly increased fish mortality during bacterial challenge. Nano-TiO2 concentrated in the kidney and spleen. Phagocytosis assay demonstrated that nano-TiO2 has the ability to diminish neutrophil phagocytosis of A. hydrophila. Fish injected with TiO2 nanoparticles displayed significant histopathology when compared to control fish. The interplay between nanoparticle exposure, immune system, histopathology, and infectious disease pathogenesis in any animal model has not been described before. By modulating fish immune responses and interfering with resistance to bacterial pathogens, manufactured nano-TiO2 has the potential to affect fish survival in a disease outbreak.
Afficher plus [+] Moins [-]Grass carps co-exposed to environmentally relevant concentrations of cypermethrin and sulfamethoxazole bear immunodeficiency and are vulnerable to subsequent Aeromonas hydrophila infection
2020
Zhao, Hongjing | Wang, Yu | Guo, Menghao | Mu, Mengyao | Yu, Hongxian | Xing, Mingwei
The aquatic ecosystem is seriously damaged because of the heavy use of pesticides and antibiotics. Fish is the indispensable link between environmental pollution and human health. However, the toxic effects of environment-related concentrations of pesticides and antibiotics in fish have not been thoroughly studied. In this study, grass carps exposed to cypermethrin (CMN, 0.651 μg/L) or/and sulfamethoxazole (SMZ, 0.3 μg/L) for 42 days caused oxidative stress, apoptosis and immunodeficiency in the spleen of grass carps. CMN or/and SMZ exposure led to oxidative damage (consumption of antioxidant enzymes (superoxide dismutase and catalase)) and lipid peroxidation (accumulation of malondialdehyde), induced apoptosis (increases in TUNEL index, Bax/bcl-2, p53, puma and Caspase family expression). In addition, the levels of immunoglobulin M (IgM), complement 3 (C3) were significantly decreased in all treatment groups, which trend was also found in C-reactive protein in CMN and MIX group, and lysozyme in MIX group. Transcription of almost all genes involved in the Toll-like receptors (TLR) signaling pathway was up-regulated under CMN or/and SMZ exposure. However, when subsequently attacked by Aeromonas hydrophila for 2 days, the TLR pathway was inhibited in spleens of all treatment groups accompanied by higher mortality. Overall, the environmentally relevant concentration of CMN and SMZ damages the immune system, triggering oxidative stress and apoptosis in carps. And by affecting the conduction of TLR signaling pathway, CMN or/and SMZ exposure inhibits the innate immune response of fish and reducing their disease resistance. This study highlights the importance of rational and regulated use of these pesticides and antibiotics.
Afficher plus [+] Moins [-]Environmental concentrations of antibiotics impair zebrafish gut health
2018
Zhou, Li | Limbu, Samwel Mchele | Shen, Meilin | Zhai, Wanying | Qiao, Fang | He, Anyuan | Du, Zhen-Yu | Zhang, Meiling
Antibiotics have been widely used in human and veterinary medicine to both treat and prevent disease. Due to their high water solubility and low bioavailability, many antibiotic residues have been found in aquatic environments. Fish are an indispensable link between the environmental pollution and human health. However, the chronic effects of environmental concentrations of antibiotics in fish have not been thoroughly investigated. Sulfamethoxazole (SMX) and oxytetracycline (OTC) are frequently detected in aquatic environments. In this study, zebrafish were exposed to SMX (260 ng/L) and OTC (420 ng/L) for a six-week period. Results indicated that exposure to antibiotics did not influence weight gain of fish but increased the metabolic rate and caused higher mortality when treated fish were challenged with Aeromonas hydrophila. Furthermore, exposure to antibiotics in water resulted in a significant decrease in intestinal goblet cell numbers, alkaline phosphatase (AKP), acid phosphatase (ACP) activities, and the anti-oxidant response while there was a significant increase in expression of inflammatory factors. Antibiotic exposure also disturbed the intestinal microbiota in the OTC-exposed group. Our results indicated that environmental antibiotic concentrations can impair the gut health of zebrafish. The potential health risk of antibiotic residues in water should be evaluated in the future.
Afficher plus [+] Moins [-]Application of docking and active site analysis for enzyme linked biodegradation of textile dyes
2019
Srinivasan, Shantkriti | Sadasivam, Senthil Kumar | Gunalan, Seshan | Shanmugam, Gnanendra | Kothandan, Gugan
Growth of textile industries led to production of enormous dye varieties. These textile dyes are largely used, chemically stable and easy to synthesize. But they are recalcitrant and persist as less biodegradable pollutants when discharged into waterbodies. Potential use of enzyme-linked bioremediation of textile dyes will control their toxicity in waterbodies. Bioinformatics and Molecular docking tool provides an insight into remediation mechanism by predicting susceptibility of dye degradation using oxidoreductive enzymes. In this study, six dyes, Reactive Red F3B, Remazol Red RGB, Joyfix Red RB, Joyfix Yellow MR, Remazol Blue RGB and Turquoise CL-5B of azo, anthraquinone and phthalocyanine molecular class were identified as potential targets for degradation by laccase and azoreductase of Aeromonas hydrophila in addition to Lysinibacillus sphaericus through in silico docking tool BioSolveIT-FlexX. Azoreductase breaks azo bonds by ping-pong mechanism whereas laccase decolorizes dyes by free radical mechanism which is not specific in nature. Results were analyzed based on parameters like stability, catalytic action and selectivity for enzyme-dye interactions. Amino acids of enzymes interacted with several dyes substantiating variations in active site for enzyme-ligand binding affinity. This suggests the role of enzymes in decolorizing an extensive variety of textile dyes, thereby, aiding in understanding the enzyme mechanisms in Bioremediation.
Afficher plus [+] Moins [-]Influence of anthropogenic activities on the spatial distribution of heterotrophic, indicator and pathogenic bacteria around Andaman and Nicobar Islands
2021
Srinivas, T.N.R. | Sudharani, P. | Behera, Swarnaprava | Dutta, S. | Sampath Kumar, G. | Sathibabu, Y. | Geethika, G. | Swagata, C. | Pratim Kandar, P.
To examine the influence of anthropogenic activities on the marine ecosystem around the Andaman and Nicobar Islands, a study was conducted to investigate the abundance of heterotrophic, indicator and pathogenic bacteria during the monsoon season. We noticed the higher abundance of heterotrophic, indicator (Escherichia coli and Enterococcus faecalis) and pathogenic bacterial counts (Aeromonas hydrophila, Enterobacter aerogens, Escherichia coli strain O157:H7, Shigella dysenteriae and Vibrio parahaemolyticus) near the 10ᵒ channel, which is the principal route to reach Andaman mainland. Most of the stations are offshore (chosen to cover shipping tracks) at a distance ranging from 41 to 266 km from the coast. The nearest station to the coast was at 21 km away. Apart from those mentioned above, several other pathogenic bacteria were also detected such as Klebsiella pneumoniae, Salmonella paratyphi, Vibrio cholera and Vibrio vulnificus but they are sparsely detected at few stations only.
Afficher plus [+] Moins [-]Biomarkers (glutathione S-transferase and catalase) and microorganisms in soft tissues of Crassostrea rhizophorae to assess contamination of seafood in Brazil
2020
Ribeiro, Eliane Braga | Noleto, Katherine Saldanha | de Oliveira, Suelen Rosana Sampaio | Batista de Jesus, Wanda | de Sousa Serra, Ilka Márcia Ribeiro | Almeida, Zafira da Silva de | de Sousa de Oliveira Mota Andrade, Ticianne | de Araújo Soares, Rômulo | Antonio, Ícaro Gomes | Santos, Débora Martins Silva | Jorge, Marianna Basso | Fortes Carvalho Neta, Raimunda Nonata
The goal of this study was to evaluate biomarkers (glutathione S-transferase and catalase) and microorganisms in soft tissues of Crassostrea rhizophorae to assess possible contamination of seafood in Brazil. The oysters were sampled from a reference area (Ports 1 and 2) and an impacted area (Ports 3 and 4) in Brazil (São Luís Island, Maranhão). Six attributes were examined in sampled oysters: glutathione S-transferase activity, catalase activity, concentrations of total coliforms and thermotolerant coliforms, and levels of Escherichia coli and Aeromonas hydrophila. Water samples were analysed for aluminium, cadmium, iron, manganese, lead, mercury, phenolics, and polychlorinated biphenyls. We found that Ports 3 and 4 are impacted by several contaminants (mercury, phenolics, and polychlorinated biphenyls), while Ports 1 and 2 are still relatively free of these contaminants. Changes in enzymes activity as well as the highest tissue bacterial concentrations were recorded in oysters from Ports 3 and 4 during the rainy season.
Afficher plus [+] Moins [-]A novel biomarker for marine environmental pollution of CAT from Mytilus coruscus
2018
Bao, Miaomiao | Huo, Liping | Wu, Jiong | Ge, Delong | Lv, Zhenming | Chi, Changfeng | Liao, Zhi | Liu, Huihui
Bivalves use anti-oxidative enzyme systems to defend themselves against excessive reactive oxygen species, which are often catalyzed by environmental pollution. As a key member of anti-oxidative enzyme family, catalase plays a crucial role in scavenging the high level of reactive oxygen species to protect organisms against various oxidative stresses. In this study, a catalase homologue was identified from Mytilus coruscus (named McCAT, KX957929). The open reading frame of McCAT was 1844bp with a 5′ untranslated region of 341bp and a 3′ untranslated region of 927bp. The deduced amino acid sequence was 512 residues in length with theoretical pI/MW 8.02/57.91kDa. BLASTn and phylogenetic analyses strongly suggested that it was a member of catalase, also known as CAT family for its conserved catalytic site motif and proximal heme-ligand signature motif. Real-time fluorescence quantitative PCR showed that constitutive expression of McCAT was occurred, with increasing order in mantle, adductor, gill, hemocyte, gonad and hepatopancreas. It was observed that bacterial infection and heavy metals stimulation up-regulated McCAT mRNA expression in hepatopancreas with time-dependent manners. The maximum expression appeared at 8h after pathogenic bacteria injecting, with 15-fold in Vibrio parahemolyticus and 60-fold in Aeromonas hydrophila than that of 0h. The highest point of McCAT mRNA appeared at different times for exposure to heavy metals with copper at day 5 (0.1mg/L 30-fold, 0.5mg/L 15-fold, 1.5mg/L 6-fold) and plumbum at day 3 (3.0mg/L 20-fold). The enzymatic activity analysis found that McCAT activity in the gill of M. coruscus was affected by heavy metals concentration. The results suggested that McCAT plays a significant role in antioxidation and the expression of McCAT can be used as a biomarker for detection of marine environmental pollution.
Afficher plus [+] Moins [-]Polyethylene microplastic exposure and concurrent effect with Aeromonas hydrophila infection on zebrafish
2022
Ding, Ning | Jiang, Lin | Wang, Xiao | Wang, Chun | Geng, Yue | Zhang, Jianxin | Sun, Yingxue | Zhang, Yanping | Yuan, Quan | Liu, Hong
Microplastics are widely distributed in the environment, raising significant concerns owing to their potential negative effects on humans. Zebrafish were used in this study to assess the toxicity of microplastic exposure. Adult zebrafish were exposed to polyethylene (PE) microplastics with smooth clustered sphere shapes and diameters of 75–100 µm for 35 days. Survival rates of the zebrafish were not significantly affected, whereas growth rates were. Analyses on oxidative stress-related enzyme activities showed that glutathione (GSH), glutathione peroxidase (GSH-PX), and glutathione s-transferase (GST) production in the intestines was stimulated when exposed to low concentrations of microplastics (0.1 and 1 mg/L), while superoxide dismutase (SOD), catalase (CAT), GSH, and GSH-PX production was suppressed when exposed to 10 mg/L microplastics. Enzyme activities in the muscles were much less affected. Intestinal injuries and changes in colony structure in the intestines were observed in zebrafish following exposure to microplastics. After 35 days of exposure, concurrent exposure to microplastics and Aeromonas hydrophila did not increase zebrafish mortality compared with those challenged by bacteria alone. This study confirms that intestinal enzyme activities of zebrafish are altered by exposure to PE microplastics but mortality and bacterial infection were not significantly affected under the tested conditions.
Afficher plus [+] Moins [-]Biofabrication of ecofriendly copper oxide nanoparticles using Ocimum americanum aqueous leaf extract: analysis of in vitro antibacterial, anticancer, and photocatalytic activities
2021
Manikandan, Dinesh Babu | Arumugam, Manikandan | Veeran, Srinivasan | Sridhar, Arun | Krishnasamy Sekar, Rajkumar | Perumalsamy, Balaji | Ramasamy, Thirumurugan
Nanotechnology tends to be a swiftly growing field of research that actively influences and inhibits the growth of bacteria/cancer. Noble metal nanoparticles (NPs) such as silver, copper, and gold have been used to damage bacterial and cancer growth over recent years; however, the toxicity of higher NPs concentrations remains a major issue. The copper oxide nanoparticles (CuONPs) were therefore fabricated using a simple green chemistry approach. Biofabricated CuONPs were characterized using UV-visible, FE-SEM with EDS, HR-TEM, FT-IR, XRD, Raman spectroscopy, and XPS analysis. Formations of CuONPs have been observed by UV-visible absorbance peak at 360.74 nm. The surface morphology of the CuONPs showed the spherical structure and size (~ 68 nm). The EDS spectrum of CuONPs has proved to be the key signals of copper (Cu) and oxygen (O) components. FT-IR analysis, to validate the important functional biomolecules (O–H, C=C, C–H, C–O) are responsible for reduction and stabilization of CuONPs. The monoclinic end-centered crystalline structures of CuONPs were confirmed with XRD planes. The electrochemical oxygen states of the CuONPs have been studied using spectroscopy of the Raman and X-ray photoelectron. After successful preparation, CuONPs examined their antibacterial, anticancer, and photocatalytic activities. Green-fabricated CuONPs were promising antibacterial candidate against human pathogenic gram-negative bacteria Escherichia coli, Vibrio cholerae, Salmonella typhimurium, Klebsiella pneumoniae, Aeromonas hydrophila, and Pseudomonas aeruginosa. CuONPs were demonstrated the excellent anticancer activity against A549 human lung adenocarcinoma cell line. Furthermore, CuONPs exhibited photocatalytic degradation of azo dyes such as eosin yellow (EY), rhodamine 123 (Rh 123), and methylene blue (MB). Biofabricated CuONPs may therefore be an important biomedical research for the aid of bacterial/cancer diseases and photocatalytic degradation of azo dyes.
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