scholarly journals Metagenomic characterization of creek sediment microbial communities from a major agricultural region in Salinas, California

2019 ◽  
Author(s):  
Brittany J. Suttner ◽  
Eric R. Johnston ◽  
Luis H. Orellana ◽  
Luis M. Rodriguez-R ◽  
Janet K. Hatt ◽  
...  

ABSTRACTLittle is known about the public health risks associated with natural creek sediments that are affected by runoff and fecal pollution from agricultural and livestock practices. For instance, the persistence of foodborne pathogens originating from agricultural activities such as Shiga Toxin-producing E. coli (STEC) in such sediments remains poorly quantified. Towards closing these knowledge gaps, the water-sediment interface of two creeks in the Salinas River Valley was sampled over a nine-month period using metagenomics and traditional culture-based tests for STEC. Our results revealed that these sediment communities are extremely diverse and comparable to the functional and taxonomic diversity observed in soils. With our sequencing effort (~4 Gbp per library), we were unable to detect any pathogenic Escherichia coli in the metagenomes of 11 samples that had tested positive using culture-based methods, apparently due to relatively low pathogen abundance. Further, no significant differences were detected in the abundance of human- or cow-specific gut microbiome sequences compared to upstream, more pristine (control) sites, indicating natural dilution of anthropogenic inputs. Notably, a high baseline level of metagenomic reads encoding antibiotic resistance genes (ARGs) was found in all samples and was significantly higher compared to ARG reads in metagenomes from other environments, suggesting that these communities may be natural reservoirs of ARGs. Overall, our metagenomic results revealed that creek sediments are not a major sink for anthropogenic runoff and the public health risk associated with these sediment microbial communities may be low.IMPORTANCECurrent agricultural and livestock practices contribute to fecal contamination in the environment and the spread of food and water-borne disease and antibiotic resistance genes (ARGs). Traditionally, the level of pollution and risk to public health is assessed by culture-based tests for the intestinal bacterium, E. coli. However, the accuracy of these traditional methods (e.g., low quantification, and false positive signal when PCR-based) and their suitability for sediments remains unclear. We collected sediments for a time series metagenomics study from one of the most highly productive agricultural regions in the U.S. in order to assess how agricultural runoff affects the native microbial communities and if the presence of STEC in sediment samples can be detected directly by sequencing. Our study provided important information on the potential for using metagenomics as a tool for assessment of public health risk in natural environments.

2020 ◽  
Vol 86 (6) ◽  
Author(s):  
Brittany Suttner ◽  
Eric R. Johnston ◽  
Luis H. Orellana ◽  
Luis M. Rodriguez-R ◽  
Janet K. Hatt ◽  
...  

ABSTRACT Little is known about the public health risks associated with natural creek sediments that are affected by runoff and fecal pollution from agricultural and livestock practices. For instance, the persistence of foodborne pathogens such as Shiga toxin-producing Escherichia coli (STEC) originating from these practices remains poorly quantified. Towards closing these knowledge gaps, the water-sediment interface of two creeks in the Salinas River Valley of California was sampled over a 9-month period using metagenomics and traditional culture-based tests for STEC. Our results revealed that these sediment communities are extremely diverse and have functional and taxonomic diversity comparable to that observed in soils. With our sequencing effort (∼4 Gbp per library), we were unable to detect any pathogenic E. coli in the metagenomes of 11 samples that had tested positive using culture-based methods, apparently due to relatively low abundance. Furthermore, there were no significant differences in the abundance of human- or cow-specific gut microbiome sequences in the downstream impacted sites compared to that in upstream more pristine (control) sites, indicating natural dilution of anthropogenic inputs. Notably, the high number of metagenomic reads carrying antibiotic resistance genes (ARGs) found in all samples was significantly higher than ARG reads in other available freshwater and soil metagenomes, suggesting that these communities may be natural reservoirs of ARGs. The work presented here should serve as a guide for sampling volumes, amount of sequencing to apply, and what bioinformatics analyses to perform when using metagenomics for public health risk studies of environmental samples such as sediments. IMPORTANCE Current agricultural and livestock practices contribute to fecal contamination in the environment and the spread of food- and waterborne disease and antibiotic resistance genes (ARGs). Traditionally, the level of pollution and risk to public health are assessed by culture-based tests for the intestinal bacterium Escherichia coli. However, the accuracy of these traditional methods (e.g., low accuracy in quantification, and false-positive signal when PCR based) and their suitability for sediments remain unclear. We collected sediments for a time series metagenomics study from one of the most highly productive agricultural regions in the United States in order to assess how agricultural runoff affects the native microbial communities and if the presence of Shiga toxin-producing Escherichia coli (STEC) in sediment samples can be detected directly by sequencing. Our study provided important information on the potential for using metagenomics as a tool for assessment of public health risk in natural environments.


2019 ◽  
Vol 12 (7) ◽  
pp. 984-993 ◽  
Author(s):  
Md. Abdus Sobur ◽  
Abdullah Al Momen Sabuj ◽  
Ripon Sarker ◽  
A. M. M. Taufiqur Rahman ◽  
S. M. Lutful Kabir ◽  
...  

Aim: The present study was carried out to determine load of total bacteria, Escherichia coli and Salmonella spp. in dairy farm and its environmental components. In addition, the antibiogram profile of the isolated bacteria having public health impact was also determined along with identification of virulence and resistance genes by polymerase chain reaction (PCR) under a one-health approach. Materials and Methods: A total of 240 samples of six types (cow dung - 15, milk - 10, milkers' hand wash - 10, soil - 10 water - 5, and vegetables - 10) were collected from four dairy farms. For enumeration, the samples were cultured onto plate count agar, eosin methylene blue, and xylose-lysine deoxycholate agar and the isolation and identification of the E. coli and Salmonella spp. were performed based on morphology, cultural, staining, and biochemical properties followed by PCR. The pathogenic strains of E. coli stx1, stx2, and rfbO157 were also identified through PCR. The isolates were subjected to antimicrobial susceptibility test against 12 commonly used antibiotics by disk diffusion method. Detection of antibiotic resistance genes ereA, tetA, tetB, and SHV were performed by PCR. Results: The mean total bacterial count, E. coli and Salmonella spp. count in the samples ranged from 4.54±0.05 to 8.65±0.06, 3.62±0.07 to 7.04±0.48, and 2.52±0.08 to 5.87±0.05 log colony-forming unit/g or ml, respectively. Out of 240 samples, 180 (75%) isolates of E. coli and 136 (56.67%) isolates of Salmonella spp. were recovered through cultural and molecular tests. Among the 180 E. coli isolates, 47 (26.11%) were found positive for the presence of all the three virulent genes, of which stx1 was the most prevalent (13.33%). Only three isolates were identified as enterohemorrhagic E. coli. Antibiotic sensitivity test revealed that both E. coli and Salmonella spp. were found highly resistant to azithromycin, tetracycline, erythromycin, oxytetracycline, and ertapenem and susceptible to gentamycin, ciprofloxacin, and imipenem. Among the four antibiotic resistance genes, the most observable was tetA (80.51-84.74%) in E. coli and Salmonella spp. and SHV genes were the lowest one (22.06-25%). Conclusion: Dairy farm and their environmental components carry antibiotic-resistant pathogenic E. coli and Salmonella spp. that are potential threat for human health which requires a one-health approach to combat the threat.


2019 ◽  
Author(s):  
Chengzhen L. Dai ◽  
Claire Duvallet ◽  
An Ni Zhang ◽  
Mariana G. Matus ◽  
Newsha Ghaeli ◽  
...  

AbstractThe spread of bacterial antibiotic resistance across human and environmental habitats is a global public health challenge. Wastewater has been implicated as a major source of antibiotic resistance in the environment, as it carries resistant bacteria and resistance genes from humans into natural ecosystems. However, different wastewater environments and antibiotic resistance genes in wastewater do not all present the same level of risk to human health. In this study, we investigate the public health relevance of antibiotic resistance found in wastewater by combining metagenomic sequencing with risk prioritization of resistance genes, analyzing samples across urban sewage system environments in multiple countries. We find that many of the resistance genes commonly found in wastewater are not readily present in humans. Ranking antibiotic resistance genes based on their potential pathogenicity and mobility reveals that most of the resistance genes in wastewater are not clinically relevant. Additionally, we show that residential wastewater resistomes pose greater risk to human health than those in wastewater treatment plant samples, and that residential wastewater can be as risky as hospital effluent. Across countries, differences in antibiotic resistance in residential wastewater can, in some cases, reflect differences in antibiotic drug consumption. Finally, we find that the flow of antibiotic resistance genes is influenced by geographical distance and environmental selection. Taken together, we demonstrate how different analytical approaches can provide greater insights into the public health relevance of antibiotic resistance in wastewater.


2019 ◽  
Vol 16 (5) ◽  
pp. 347 ◽  
Author(s):  
Xiaolin Lai ◽  
Pingxiao Wu ◽  
Bo Ruan ◽  
Juan Liu ◽  
Zehua Liu ◽  
...  

Environmental contextAntibiotic resistance by microorganisms in the natural environment poses a threat to ecosystems and public health. We report findings suggesting kaolinite can effectively inhibit the development of antibiotic resistance genes in microorganisms, and present a new understanding of the molecular mechanisms that promote the development of antibiotic resistance. These results are critical to mitigating environmental and public health risks resulting from the abuse of antibiotics. AbstractAntibiotic resistance and antibiotic resistance genes (ARGs) in the natural environment pose a threat to ecosystems and public health; therefore, better strategies are needed to mitigate the emergence of resistance. This study examined the expression of ARGs in Escherichia coli (E. coli) after exposure to sub-MIC (minimum inhibitory concentration) antibiotics for 15 days in the presence and absence of kaolinite. The results of the real-time polymerase chain reaction (PCR) showed that the expression levels of the eight target genes of E. coli adhering to kaolinite were relatively decreased, and the MIC results also indicated that the final resistance was lower than that of the strains without kaolinite. A close relationship between E. coli and kaolinite was also revealed, as well as a unique interfacial interaction. In addition, the differential protein expression was further analysed to detect proteins and genes associated with ARGs mutations, and then the underlying mechanisms of cell growth and metabolism were identified under low dose ampicillin stress to elucidate the role of kaolinite in the process. Molecular mechanisms analysis determined that when cells adhering to kaolinite were stressed, transport of ampicillin to the periplasmic space was reduced, and the redox metabolism of bacteria was promoted to combat the harsh environment. Moreover, cells synthesised related peptides or proteins under the action of ribosomal proteins to prevent toxic damage. Therefore, this work not only provides new insights into the cellular response to antibiotic stress, but also provides a topic for more research on methods to delay the emergence of ARGs.


2014 ◽  
Vol 77 (3) ◽  
pp. 388-394 ◽  
Author(s):  
LUCAS M. WIJNANDS ◽  
ELLEN H. M. DELFGOU-van ASCH1 ◽  
MARIEKE E. BEEREPOOT-MENSINK ◽  
ALICE van der MEIJ-FLORIJN ◽  
IFE FITZ-JAMES ◽  
...  

Recent outbreaks with vegetable or fruits as vehicles have raised interest in the characterization of the public health risk due to microbial contamination of these commodities. Because qualitative and quantitative data regarding prevalence and concentration of various microbes are lacking, we conducted a survey to estimate the prevalence and contamination level of raw produce and the resulting minimally processed packaged salads as sold in The Netherlands. A dedicated sampling plan accounted for the amount of processed produce in relation to the amount of products, laboratory capacity, and seasonal influences. Over 1,800 samples of produce and over 1,900 samples of ready-to-eat mixed salads were investigated for Salmonella enterica serovars, Campylobacter spp., Escherichia coli O157, and Listeria monocytogenes. The overall prevalence in raw produce varied between 0.11% for E. coli O157 and L. monocytogenes and 0.38% for Salmonella. Prevalence point estimates for specific produce/pathogen combinations ranged for Salmonella from 0.53% in iceberg lettuce to 5.1% in cucumber. For Campylobacter, this ranged from 0.83% in endive to 2.7% in oak tree lettuce. These data will be used to determine the public health risk posed by the consumption of ready-to-eat mixed salads in The Netherlands.


2020 ◽  
Vol 18 (3) ◽  
pp. 292-305
Author(s):  
M. M. Majedul Islam ◽  
Md. Atikul Islam

Abstract A Quantitative Microbial Risk Assessment (QMRA) technique was applied to assess the public health risk from exposure to infectious microorganisms at bathing areas of three rivers in Bangladesh. The QMRA assessed the probability of illness due to the accidental ingestion of river water impacted by untreated sewage. The simplified QMRA was based on average concentrations of four reference pathogens Escherichia coli (E. coli) O157:H7, Cryptosporidium spp, norovirus and rotavirus relative to indicator bacterium E. coli. Public health risk was estimated as the probability of infection and illness from a single exposure of bathers. The risks of illness were ranged from 7 to 10% for E. coli O157:H7, 13 to 19% for Cryptosporidium, 7 to 10% for norovirus and 12 to 17% for rotavirus. The overall risk of illness at the rivers was slightly higher in children (9–19%) compared to adults (7–16%). The risks of illness in individuals exposed to the river bathing were unacceptably high, exceeding the USEPA acceptable risk of 3–6 illnesses per hundred bathing events. This study gives a basis for reducing the burden of disease in the population by applying appropriate risk management. Findings and methods of this study will be helpful for other countries with similar socio-economic and geographic settings.


Diversity ◽  
2021 ◽  
Vol 13 (6) ◽  
pp. 230
Author(s):  
Shan Wan ◽  
Min Xia ◽  
Jie Tao ◽  
Yanjun Pang ◽  
Fugen Yu ◽  
...  

In this study, we used a metagenomic approach to analyze microbial communities, antibiotic resistance gene diversity, and human pathogenic bacterium composition in two typical landfills in China. Results showed that the phyla Proteobacteria, Bacteroidetes, and Actinobacteria were predominant in the two landfills, and archaea and fungi were also detected. The genera Methanoculleus, Lysobacter, and Pseudomonas were predominantly present in all samples. sul2, sul1, tetX, and adeF were the four most abundant antibiotic resistance genes. Sixty-nine bacterial pathogens were identified from the two landfills, with Klebsiella pneumoniae, Bordetella pertussis, Pseudomonas aeruginosa, and Bacillus cereus as the major pathogenic microorganisms, indicating the existence of potential environmental risk in landfills. In addition, KEGG pathway analysis indicated the presence of antibiotic resistance genes typically associated with human antibiotic resistance bacterial strains. These results provide insights into the risk of pathogens in landfills, which is important for controlling the potential secondary transmission of pathogens and reducing workers’ health risk during landfill excavation.


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