Nanopore-Based Surveillance of Zoonotic Bacterial Pathogens in Farm-Dwelling Peridomestic Rodents

The effective control of rodent populations on farms is crucial for food safety, as rodents are reservoirs and vectors for several zoonotic pathogens. Clear links have been identified between rodents and farm-level outbreaks of pathogens throughout Europe and Asia; however, comparatively little research has been devoted to studying the rodent–agricultural interface in the USA. Here, we address this knowledge gap by metabarcoding bacterial communities of rodent pests collected from Minnesota and Wisconsin food animal farms. We leveraged the Oxford Nanopore MinION sequencer to provide a rapid real-time survey of putative zoonotic foodborne pathogens, among others. Rodents were live trapped (n = 90) from three dairy and mixed animal farms. DNA extraction was performed on 63 rodent colons along with 2 shrew colons included as outgroups in the study. Full-length 16S amplicon sequencing was performed. Our farm-level rodent-metabarcoding data indicate the presence of multiple foodborne pathogens, including Salmonella spp., Campylobacter spp., Staphylococcus aureus, and Clostridium spp., along with many mastitis pathogens circulating within five rodent species (Microtus pennsylvanicus, Mus musculus, Peromyscus leucopus, Peromyscus maniculatus, and Rattus norvegicus) and a shrew (Blarina brevicauda). Interestingly, we observed a higher abundance of enteric pathogens (e.g., Salmonella) in shrew feces compared to the rodents analyzed in our study. Knowledge gained from our research efforts will directly inform and improve farm-level biosecurity efforts and public health interventions to reduce future outbreaks of foodborne and zoonotic disease.

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Molecular surveillance of zoonotic bacterial pathogens in farm dwelling peridomestic rodents across the upper Midwest, USA

10.1101/2021.07.28.454187 ◽

2021 ◽

Author(s):

Nusrat A. Jahan ◽

Laramie L. Lindsey ◽

Evan J. Kipp ◽

Bradley J Heins ◽

Amy M. Runck ◽

...

Keyword(s):

Foodborne Pathogens ◽

Amplicon Sequencing ◽

Effective Control ◽

Human Pathogens ◽

Blarina Brevicauda ◽

Food Borne ◽

16S Amplicon Sequencing ◽

Oxford Nanopore ◽

Animal Farms ◽

The Usa

The effective control of rodent populations on farms is a critical component of food-safety, as rodents are reservoirs and vectors for many foodborne pathogens in addition to several zoonotic pathogens. The functional role of rodents in the amplification and transmission of pathogens is likely underappreciated. Clear links have been identified between rodents and outbreaks of pathogens throughout Europe and Asia, however, comparatively little research has been devoted to studying this rodent-agricultural interface in the USA, particularly across the Midwest. Here, we address this existing knowledge gap by characterizing the metagenomic communities of rodent pests collected from Minnesota and Wisconsin food animal farms. We leveraged the Oxford Nanopore MinION sequencer to provide a rapid real-time survey of the putative zoonotic food-borne and other human pathogens. Rodents (mice and rats) were live trapped from three dairy and mixed animal farms. Tissues and fecal samples were collected from all rodents. DNA extraction was performed on 90 rodent colons along with 2 shrew colons included as outgroups in the study. Full-length 16S amplicon sequencing was performed with the MinION. Our data suggests the presence of putative foodborne pathogens including Salmonella spp., Campylobacter spp., Staphylococcus aureus, and Clostridium spp., along with many important mastitis pathogens. A critically important observation is that we discovered these pathogens within all five species of rodents (Microtus pennsylvanicus, Mus musculus, Peromyscus leucopus, Peromyscus maniculatus, and Rattus norvegicus) and shrew (Blarina brevicauda) in varying abundances. Interestingly, we observed a higher abundance of enteric pathogens (e.g. Salmonella) in shrew feces compared to the rodents analyzed in our study, however more data is required to establish that connection. Knowledge gained from our research efforts will directly inform and improve upon farm-level biosecurity efforts and public health interventions to reduce future outbreaks of foodborne and zoonotic disease.

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Integrated genomic view of SARS-CoV-2 in India

Wellcome Open Research ◽

10.12688/wellcomeopenres.16119.1 ◽

2020 ◽

Vol 5 ◽

pp. 184 ◽

Cited By ~ 1

Author(s):

Pramod Kumar ◽

Rajesh Pandey ◽

Pooja Sharma ◽

Mahesh S. Dhar ◽

Vivekanand A. ◽

...

Keyword(s):

Host Response ◽

Genomic Data ◽

Amplicon Sequencing ◽

Climatic Conditions ◽

Molecular Surveillance ◽

Multiple Introductions ◽

Oxford Nanopore ◽

The World ◽

The Usa ◽

Local Transmission

Background: India first detected SARS-CoV-2, causal agent of COVID-19 in late January 2020, imported from Wuhan, China. From March 2020 onwards, the importation of cases from countries in the rest of the world followed by seeding of local transmission triggered further outbreaks in India. Methods: We used ARTIC protocol-based tiling amplicon sequencing of SARS-CoV-2 (n=104) from different states of India using a combination of MinION and MinIT sequencing from Oxford Nanopore Technology to understand how introduction and local transmission occurred. Results: The analyses revealed multiple introductions of SARS-CoV-2 genomes, including the A2a cluster from Europe and the USA, A3 cluster from Middle East and A4 cluster (haplotype redefined) from Southeast Asia (Indonesia, Thailand and Malaysia) and Central Asia (Kyrgyzstan). The local transmission and persistence of genomes A4, A2a and A3 was also observed in the studied locations. The most prevalent genomes with patterns of variance (confined in a cluster) remain unclassified, and are here proposed as A4-clade based on its divergence within the A cluster. Conclusions: The viral haplotypes may link their persistence to geo-climatic conditions and host response. Multipronged strategies including molecular surveillance based on real-time viral genomic data is of paramount importance for a timely management of the pandemic.

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Gut microbiome composition in the Hispanic Community Health Study/Study of Latinos is shaped by geographic relocation, environmental factors, and obesity

Genome Biology ◽

10.1186/s13059-019-1831-z ◽

2019 ◽

Vol 20 (1) ◽

Cited By ~ 14

Author(s):

Robert C. Kaplan ◽

Zheng Wang ◽

Mykhaylo Usyk ◽

Daniela Sotres-Alvarez ◽

Martha L. Daviglus ◽

...

Keyword(s):

Community Health ◽

Gut Microbiome ◽

Alpha Diversity ◽

Amplicon Sequencing ◽

Shannon Index ◽

Health Study ◽

Rrna Gene ◽

Cross Sectional ◽

Hispanic Community ◽

The Usa

Abstract Background Hispanics living in the USA may have unrecognized potential birthplace and lifestyle influences on the gut microbiome. We report a cross-sectional analysis of 1674 participants from four centers of the Hispanic Community Health Study/Study of Latinos (HCHS/SOL), aged 18 to 74 years old at recruitment. Results Amplicon sequencing of 16S rRNA gene V4 and fungal ITS1 fragments from self-collected stool samples indicate that the host microbiome is determined by sociodemographic and migration-related variables. Those who relocate from Latin America to the USA at an early age have reductions in Prevotella to Bacteroides ratios that persist across the life course. Shannon index of alpha diversity in fungi and bacteria is low in those who relocate to the USA in early life. In contrast, those who relocate to the USA during adulthood, over 45 years old, have high bacterial and fungal diversity and high Prevotella to Bacteroides ratios, compared to USA-born and childhood arrivals. Low bacterial diversity is associated in turn with obesity. Contrasting with prior studies, our study of the Latino population shows increasing Prevotella to Bacteroides ratio with greater obesity. Taxa within Acidaminococcus, Megasphaera, Ruminococcaceae, Coriobacteriaceae, Clostridiales, Christensenellaceae, YS2 (Cyanobacteria), and Victivallaceae are significantly associated with both obesity and earlier exposure to the USA, while Oscillospira and Anaerotruncus show paradoxical associations with both obesity and late-life introduction to the USA. Conclusions Our analysis of the gut microbiome of Latinos demonstrates unique features that might be responsible for health disparities affecting Hispanics living in the USA.

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SARS-CoV-2 does not have a strong effect on the nasopharyngeal microbial composition

Scientific Reports ◽

10.1038/s41598-021-88536-6 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Tzipi Braun ◽

Shiraz Halevi ◽

Rotem Hadar ◽

Gilate Efroni ◽

Efrat Glick Saar ◽

...

Keyword(s):

Clinical Microbiology ◽

Amplicon Sequencing ◽

Viral Disease ◽

Nasopharyngeal Swab ◽

Microbial Composition ◽

16S Amplicon Sequencing ◽

The World ◽

Health And Disease ◽

Longitudinal Sampling ◽

Potential Interactions

AbstractThe coronavirus disease 2019 (COVID-19) has rapidly spread around the world, impacting the lives of many individuals. Growing evidence suggests that the nasopharyngeal and respiratory tract microbiome are influenced by various health and disease conditions, including the presence and the severity of different viral disease. To evaluate the potential interactions between Severe Acute Respiratory Syndrome Corona 2 (SARS-CoV-2) and the nasopharyngeal microbiome. Microbial composition of nasopharyngeal swab samples submitted to the clinical microbiology lab for suspected SARS-CoV-2 infections was assessed using 16S amplicon sequencing. The study included a total of 55 nasopharyngeal samples from 33 subjects, with longitudinal sampling available for 12 out of the 33 subjects. 21 of the 33 subjects had at least one positive COVID-19 PCR results as determined by the clinical microbiology lab. Inter-personal variation was the strongest factor explaining > 75% of the microbial variation, irrespective of the SARS-CoV-2 status. No significant effect of SARS-CoV-2 on the nasopharyngeal microbial community was observed using multiple analysis methods. These results indicate that unlike some other viruses, for which an effect on the microbial composition was noted, SARS-CoV-2 does not have a strong effect on the nasopharynx microbial habitants.

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Meta-Apo improves accuracy of 16S-amplicon-based prediction of microbiome function

BMC Genomics ◽

10.1186/s12864-020-07307-1 ◽

2021 ◽

Vol 22 (1) ◽

Author(s):

Gongchao Jing ◽

Yufeng Zhang ◽

Wenzhi Cui ◽

Lu Liu ◽

Jian Xu ◽

...

Keyword(s):

16S Rrna ◽

Large Scale ◽

Low Cost ◽

Human Microbiome ◽

Amplicon Sequencing ◽

Training Sample ◽

Rrna Gene ◽

16S Amplicon Sequencing ◽

Cross Platform ◽

Functional Profiles

Abstract Background Due to their much lower costs in experiment and computation than metagenomic whole-genome sequencing (WGS), 16S rRNA gene amplicons have been widely used for predicting the functional profiles of microbiome, via software tools such as PICRUSt 2. However, due to the potential PCR bias and gene profile variation among phylogenetically related genomes, functional profiles predicted from 16S amplicons may deviate from WGS-derived ones, resulting in misleading results. Results Here we present Meta-Apo, which greatly reduces or even eliminates such deviation, thus deduces much more consistent diversity patterns between the two approaches. Tests of Meta-Apo on > 5000 16S-rRNA amplicon human microbiome samples from 4 body sites showed the deviation between the two strategies is significantly reduced by using only 15 WGS-amplicon training sample pairs. Moreover, Meta-Apo enables cross-platform functional comparison between WGS and amplicon samples, thus greatly improve 16S-based microbiome diagnosis, e.g. accuracy of gingivitis diagnosis via 16S-derived functional profiles was elevated from 65 to 95% by WGS-based classification. Therefore, with the low cost of 16S-amplicon sequencing, Meta-Apo can produce a reliable, high-resolution view of microbiome function equivalent to that offered by shotgun WGS. Conclusions This suggests that large-scale, function-oriented microbiome sequencing projects can probably benefit from the lower cost of 16S-amplicon strategy, without sacrificing the precision in functional reconstruction that otherwise requires WGS. An optimized C++ implementation of Meta-Apo is available on GitHub (https://github.com/qibebt-bioinfo/meta-apo) under a GNU GPL license. It takes the functional profiles of a few paired WGS:16S-amplicon samples as training, and outputs the calibrated functional profiles for the much larger number of 16S-amplicon samples.

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A Review of Alternative Management Tactics Employed for the Control of Various Cockroach Species (Order: Blattodea) in the USA

Insects ◽

10.3390/insects12060550 ◽

2021 ◽

Vol 12 (6) ◽

pp. 550

Author(s):

Ameya D. Gondhalekar ◽

Arthur G. Appel ◽

Gretchen M. Thomas ◽

Alvaro Romero

Keyword(s):

Applied Research ◽

Research Effort ◽

Effective Control ◽

Control Methods ◽

Alternative Tactics ◽

Alternative Control ◽

The Public ◽

Concurrent Use ◽

The Usa ◽

Alternative Control Methods

Effective control of domestic and peridomestic cockroaches requires integrated approaches that emphasize concurrent use of chemicals with alternative control tactics. An integrated pest management (IPM) approach is particularly justified in environments where satisfactory cockroach control cannot be achieved due to multiple factors including development of insecticide aversion and resistance in some cockroach species, and poor sanitation or structural issues that foster infestations. While a flurry of research effort has been devoted to study alternative tactics for cockroach control, only a few of them have been evaluated in the context of IPM programs. This review focuses on examining studies on alternative tactics that are proven efficacious, economical, and logistically feasible for their inclusion in IPM programs for important domestic and peridomestic cockroaches in the USA. Management programs that educate the public on cockroach biology, behavior, and the importance of sanitation; use of traps to monitor infestation levels; apply targeted low impact insecticides such as baits, have demonstrated a greater success for effective and sustainable control of cockroaches when compared to an insecticide-only approach. Incorporation of other alternative control methods to IPM programs will require more applied research that validates their use in real-world scenarios and demonstrates their cost-effectiveness.

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The Use of Bacteriophages in the Poultry Industry

Animals ◽

10.3390/ani10050872 ◽

2020 ◽

Vol 10 (5) ◽

pp. 872 ◽

Cited By ~ 6

Author(s):

Katarzyna Żbikowska ◽

Monika Michalczuk ◽

Beata Dolka

Keyword(s):

Foodborne Pathogens ◽

Pathogenic Bacteria ◽

Bacterial Resistance ◽

Health And Safety ◽

Legal Framework ◽

Multidrug Resistant ◽

Poultry Production ◽

Salmonella Spp ◽

Food Contact Surfaces ◽

Public Health And Safety

The emergence of multidrug-resistant infections and antibiotic failures have raised concerns over human and veterinary medicine worldwide. Poultry production has had to confront the problems of an alarming increase in bacterial resistance, including zoonotic pathogens. According to the European Food Safety Authority (EFSA), campylobacteriosis and salmonellosis have been the most frequently reported human foodborne diseases linked to poultry. This situation has strongly stimulated a renewal of scientists’ interest in bacteriophages (phages) since the beginning of the 21st century. Bacteriophages are the viruses of bacteria. They are abundant in nature, and accompany bacteria in each environment they colonize, including human microbiota. In this review, we focused on the use of bacteriophages as therapeutic agents to treat infections and reduce counts of pathogenic bacteria in poultry, as biocontrol agents to eliminate foodborne pathogens on/in food, and also as disinfectants to reduce contamination on food-contact surfaces or poultry carcasses in industrial conditions. Most of the phage-based products are targeted against the main foodborne pathogens, such as Campylobacter jejuni, Salmonella spp., Escherichia coli, Listeria monocytogenes, Staphylococcus aureus, and Clostridium perfringens. Phages are currently addressed at all stages of the poultry production "from farm to fork", however, their implementation into live birds and food products still provokes discussions especially in the context of the current legal framework, limitations, as well as public health and safety.

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Insects as vectors of foodborne pathogenic bacteria

Terrestrial Arthropod Reviews ◽

10.1163/187498311x543989 ◽

2011 ◽

Vol 4 (1) ◽

pp. 5-16 ◽

Cited By ~ 21

Author(s):

Jeffrey Blazar ◽

Marc Allard ◽

E. Kurt Lienau

Keyword(s):

Food Safety ◽

Foodborne Pathogens ◽

Pathogenic Bacteria ◽

Current Knowledge ◽

Disease Outbreaks ◽

House Fly ◽

American Cockroach ◽

Health Concern ◽

Foodborne Disease ◽

Salmonella Spp

AbstractFood safety is an important consideration worldwide. To maintain and improve our current knowledge of foodborne disease outbreaks, we must understand some of the more imminent issues related to food safety. A variety of agents are responsible for transmitting the estimated 76 million cases of illnesses caused by foodborne pathogens every year. This review explores why insects pose a serious health concern, in terms of worldwide food safety initiatives, by looking at evidence in published <abs>Food safety is an important consideration worldwide. To maintain and improve our current knowledge of foodborne disease outbreaks, we must understand some of the more imminent issues related to food safety. A variety of agents are responsible for transmitting the estimated 76 million cases of illnesses caused by foodborne pathogens every year. This review explores why insects pose a serious health concern, in terms of worldwide food safety initiatives, by looking at evidence in published literature. We highlight at least eleven different species of insects, including the lesser mealworm, Alphitobius diaperinus (Panzer); secondary screwworm, Cochliomyia macellaria (Fabricius); synanthropic flies [flesh fly, Sarcophaga carnaria (L.); house fly, Musca domestica (L.); fruit fly, Drosophila melanogaster (Meigen); and stable fly, Stomoxys calcitrans (L.)], American cockroach, Periplaneta americana (L.); German cockroach, Blatella germanica (L.); Oriental cockroach, Blatta orientalis (L.); Pacific beetle cockroach, Diploptera punctata (Eschscholtz); and Speckled feeder cockroach, Nauphoeta cinerea (Olivier), which act as vectors for Salmonella spp. or Escherichia coli and illustrate how these insects are successful vectors of foodborne disease outbreaks. We propose that insects be considered as one of the latest issues in food safety initiatives. Not only are some insects extremely important contributors to diseases, but now we suggest that more research into insects as potential carriers of E. coli and Salmonella spp., and therefore as contributing to foodborne disease outbreaks, is granted.

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Utilizing the Public GenomeTrakr Database for Foodborne Pathogen Traceback v1

10.17504/protocols.io.bxw3ppgn ◽

2021 ◽

Author(s):

Ruth E Timme ◽

Maria Sanchez ◽

Marc Allard

Keyword(s):

Real Time ◽

Foodborne Pathogens ◽

Pathogen Detection ◽

Foodborne Pathogen ◽

Reference Database ◽

Surveillance Program ◽

Environmental Isolates ◽

The Public ◽

The World ◽

The Usa

This protocol outlines the all the steps necessary to become a GenomeTrakr data contributor. GenomeTrakr is an international genomic reference database of mostly food and environmental isolates from foodborne pathogens. The data and analyses are housed at the National Center for Biotechnology Information (NCBI), which is a database freely available to anyone in the world. The Pathogen Detection browser at NCBI computes daily cluster results adding the newly submitted data to the existing phylogenetic clusters of closely related genomes. Contributors to this database can see how their new isolates are related to the real-time foodborne pathogen surveillance program established in the USA and a few other countries, and at the same time adding valuable new data to the reference database. ------ Although originally published as a Chapter in Methods and Protocols, Foodborne Bacterial Pathogens, the protocol has since been adapted and split into four separate protocols all of which are contained in this collection.

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