Progress 12/23/14 to 09/30/17
Outputs Target Audience:The target audience were fresh produce and animal prodcerswhich included cattle and poutry Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?The PI went for training on metagenomics: Whole Genomic Sequencing Symposium Whole Genome Sequencing for Food Safety Symposium SEPTEMBER 28-30, 2016 Chicago Marriott Southwest at Burr Ridge Burr Ridge, Illinois The program has improved the communication with our stakeholders and were in position to share their challenges in both produce and animal production systems. Three students were also engaged to increase their participation in food safety research activities. PI attended and presented at International Association of Food Protection (IAFP) Annual Meeting, 2017, Tampa, Florida How have the results been disseminated to communities of interest?The results of this project have been disseminated to communitesthrough workshops, booklets, and fact sheets. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts What was accomplished under these goals?
Monitoring antimicrobial resistant bacteria and identifying sources of resistance genes in farmland and local foods will contribute to the efforts of mitigating antimicrobial resistance in foods systems. Objective 1: Determine the extent of antibiotic use in Tennessee agriculture A questionnaire based survey was developed and conducted among 25 animal producers in different counties in Tennessee. The farmers were asked key questions including type of livestock and number on the farm, how/when were the antibiotics used, treated sick animals only, disease control plans, did the farm maintain written records for antibiotic treatments including medicated feeds, written plans for treating sick animals with antibiotics, was the veterinarian's advice sought before administering antibiotics, who determined when and which antibiotics to be used, any collaboration with a veterinarian and following administration of an antibiotic, was the course of treatment completed. The animal on the farms ranged from 3 to 200 per farm. The data collected was analyzed and 72% of animals kept were cattle, followed by chicken (16%), pigs (4%), goats (4%), and sheep (4%). The most surprising practice was that 52 % of the farmers indicated not to seek veterinarian's advice before administering antibiotics; only 40% indicated to collaborate with a veterinarian. Two farmers (8%) indicated not to have a veterinarian involved on their farms. Ten farms (56%) indicated that animals had at least developed some infections including mastitis, diarrhea, respiratory infections, and skin/foot and as a result the farmers administered antibiotics to the animals. A large number of animal producers dis not judiciously use antibiotics on their sick animals and indicated poor record keeping. The lack of knowledge on what pathogens cause specific diseases and education guidelines for antimicrobial use has led to overuse of antibiotics on farms. Objective 2: Determine zoonotic and indicator bacteria in growing crops, soil, animal manure and feed, water sources, and locally produced foods retailed in farmers markets Produce farms, animal farms and farmers' markets were contacted through the Tennessee State University extension program. At each visit, 3 samples per food type (produce, chicken, and eggs), environmental samples (water, soil, animal manure) were collected, transported in coolers with ice and analyzed within 24 h. Escherichia coli, Salmonella spp., Shigella spp. and other bacteria including, Enterobacter spp, and Klebsiella in the Enterobacteriaceae were isolated from local foods (produce, eggs, water from farms, animal manure, and soil. Objective 3: Determine the resistance of Salmonella, Shigella, Escherichia coli, Enterococcus, and Listeria to cephalosporin, carbapenems, aminoglycosides, fluoroquinolones, macrolides, and tetracyclines. Data on antibiotic resistant profiles of both pathogenic and commensal bacteria Escherichia coli, Salmonella spp., Shigella spp. and other bacteria including, Enterobacter spp, and Klebsiella were isolated from local foods, water from farms, animal manure, and soil. Pathogenic bacteria such as Shigella and Salmonella; and indicator bacteria including Escherichia coli, Klebsiella , and other showed resistance to ampicillin, cefpodoxime, tetracycline, erythromycin, cefotaxime, gentamicin, and other antibiotics used in human medicine. Escherichia coli which is an indicator bacteria showed the following antimicrobial resistance patterns. Salmonella which is also an indicator bacteria indicated several resistance patterns.. MAR index values indicate the usage of antibiotics in the environment. Cumulatively, the antibiotic resistance patterns showed MAR index values ranging from 0.08 to 0.70. The highest MAR index among the bacteria was for Escherichia coli (MAR Index=0.7). lation, identification, and classification of bacteria in local foods and environmental samples (soil, water, animal manure). We found a varying degree of antimicrobial resistance and ESBL- producing EC in poultry farms and retail meats. The prevalence of ESBL-producing E. coli in retail chicken (73.6%) The occurrence of ESBL-producingE. coli(82%) in chicken feces was significantly higher as compared to retail chicken, soil, and feed. We found ESBL-EC (42.3%) in soil within the vicinity of the poultry farms. The prevalence of ESBL-EC in feed (24%) was significantly lower as compared to other environmental samples. ESBL-EC isolates from poultry farms and retail chicken meat. The dominant resistance pattern was ERY-STR (36.5%; 42/115), followed by ERY-STR-KAN-NAL (9.6%, 11/115), ERY (7.0%, 8/115) and ERY-STR-KAN-TET-VAN-AMC (6.1%, 7/ 115). The least (0.9%; 1/115) resistance patterns were AMK-CIP-ERY-STR-CHL-KAN-NAL-TET-VAN-COL, ERY-CHL-STR-NOV-NAL-TET-VAN-COL-AMC, and among others (Table 2). Highly multi-resistant ESBL-EC (> 3 resistances) is displayed in our study. Among ESBL-producing E. coli isolates from feces, highest resistance against erythromycin (100 %), followed by streptomycin (100 %), tetracycline (95.1%), kanamycin (92.7%), and nalidixic acid (73.2%); moderate resistance rates were observed for chloramphenicol (68.3%), ampicillin (43.9%); and relatively low resistance rates were observed for , novobioxn (34.1%), vancomycin (31.7%), colistin and ciprofloxacin (12.2%), and amikacin (4.9%). Among ESBL-producing E. coli isolates from retail chicken, high resistance rates were observed for erythromycin (100.0%) streptomycin (, 92.3%), tetracycline 84.6% and nalidixic acid, (74.4%); moderate resistance rates were observed for kanamycin (51.3%), and relatively low resistance rates were observed for chloramphenicol, ampicillin, colistin, ciprofloxacin, and amikacin. Objective 4: Education and outreach on judicious use of antibiotics. Data from the project indicated farms were contaminated with antibiotic resistant bacteria. From these results, there is need to educate farmers on prudent use of antibiotics and record keeping. A workshop was conducted to educate the stakeholders on judicious use of antibiotics. During the workshop, there was discussion on management practices, dosing and treatment regiments for antibiotic and how antibiotics select for antibiotic resistance. During this workshop, produce growers and animal farmers were engaged in the learning process by asking them to comment on their own experiences. After the workshop, farmers understood the importance of judicious use of antibiotics. Booklets on judicious use of antibiotic were developed and delivered to farmers. Products, Results and Measurable Outcomes: (1) A questionnaire survey instrument on management practices, record keeping, dosing and treatment regiments on animal farms was developed and disseminated; (2) workshop on judicious use of antimicrobials in agriculture, aimed at delivering science based information on judicious use of antibiotics to agricultural commodity producers; (3) antimicrobial resistant data on both pathogenic and commensal bacteria from farms was collected to provide logical corridors to mitigate antibiotic-resistance in Tennessee food system; (4) 25 farmers, now part of educated community ready to accept policy changes aimed at mitigating antimicrobial resistance; (5) producers implementing 'Best Management Practices', or BMPs on their promises; (6) two journal articles are published; (7) three presentations made at the International Association of Food Protection, and Association of 1890 Research Directors (ARD), (8) three undergraduate students were engaged and trained on microbial analysis of environmental samples from animal and produce farms (9) one graduate thesis was produced. An educated agricultural community will allow informed practical decisions thereby forming a solid basis for broadly accepting policy changes aimed at limiting antibiotic resistant bacteria in the environment and ultimately in the food chain.
Publications
- Type:
Journal Articles
Status:
Submitted
Year Published:
2017
Citation:
A. Kilonzo-Nthenge, et al,. 2017. Extended-spectrum lactamase producing Escherichia coli in Small-scaled Poultry Farms and Retail Chicken. Submitted to Journal of Food Science and Engineering, September 2017.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2017
Citation:
A. Kilonzo-Nthenge, et al. 2017. Extended-spectrum ?-Lactamase Producing Escherichia coli in Feed, Manure, and Soil from the Poultry Farm Environment: P1-221, IAFP 2017
- Type:
Theses/Dissertations
Status:
Awaiting Publication
Year Published:
2017
Citation:
Joy Oshiorenua Igbafe, Agnes Kilonzo, Samuel Nahashon. 2017. Evaluating the efficacy of probiotic to reduce the colonization of Salmonella in chicken.
ETD Collection for Tennessee State University. http://digitalscholarship.tnstate.edu/dissertations
- Type:
Books
Status:
Other
Year Published:
2017
Citation:
Booklets: Prudent use of Antimicrobial Agent on Small-scaled Poultry and Cattle Farms(in the preparation)
- Type:
Journal Articles
Status:
Published
Year Published:
2016
Citation:
A. Kilonzo-Nthenge, S. N. Nahashon, S. Godwin, Siqin liu, and D. Long. 2016. Prevalence and Antimicrobial Resistance of
Enterobacteriaceae in Shell Eggs from Small-Scale Poultry Farms and Farmers' Markets. Food Prot. 79: 2031-2037
- Type:
Other
Status:
Published
Year Published:
2016
Citation:
Booklets on Egg Safety from Farm to Fork. 2016. TSU-16-0253 (A)-17090 were developed and distributed to farmers in Tennessee.
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Progress 10/01/15 to 09/30/16
Outputs Target Audience:The target audience were Tennessee farmers with 10 to 200 farm animals which included cattle, chicken, pigs, goats, and sheep. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?The PI went for training on metagenomics: Whole Genomic Sequencing Symposium Whole Genome Sequencing for Food Safety Symposium SEPTEMBER 28-30, 2016 Chicago Marriott Southwest at Burr Ridge Burr Ridge, Illinois How have the results been disseminated to communities of interest?
Nothing Reported
What do you plan to do during the next reporting period to accomplish the goals? Determine antmicrobial resistance of foodborne pathogens isolated from local foods and environmental samples (soil, water, soil, and animal manure). The following antibiotics will be used in the resistance study: cephalosporin, carbapenems, aminoglycosides, fluoroquinolones, macrolides, and tetracycline. Conduct training workshop on judicious use of antibiotics to animal producers across Tennessee
Impacts What was accomplished under these goals?
Use of antibiotic in farm animals A questionnaire based survey was developed and conducted among 25 animal producers in different counties in Tennesee. The farmers were asked key questions including type of livestock and number on the farm, how/when were the antibiotics used, treated sick animals only, disease control plans, did the farm maintain written records for antibiotic treatments including medicated feeds, written plans for treating sick animals with antibiotics, was the veterinarian's advice sought before administering antibiotics, who determinedwhen and which antibiotics to be used, any collaboration with a veterinarianand following administration of an antibiotic, was the course of treatment completed. The animal on the farms ranged from 3 to 200 per farm. The data collected was analyzed and 72% of animals kept were cattle, followed by chicken (16%), pigs (4%), goats (4%), and sheep (4%). The most surprising practice was that 52 % of the farmers indicated not to seek veterinarian's advice before administering antibiotics; only 40% indicated to collaborate with a veterinarian. Two farmers (8%) indicated not to have a veterinarian involved on their farms. Tenfarms (56%) indicated that animals had at least developed some infections includingmastitis, diarrhea, respiratory infections, and skin/foot and as a result thye farmers administered antibiotics to the animals. Zoonotic and Indicator Bacteria in local foods and agricultural lands During the growing season of 2015 and 2016, produce farms and farmers' markets were visited 2 times per week, subject on the availability of the products. The farms were contacted through the Tennessee State University extension program. At each visit, 3 samples per food type (produce, chicken, eggs), environmental samples (water, soil, animal manure) were collected, transported in coolers with ice and analyzed within 24 h. Each sample was placed in a sterile sampling bag and labeled with identification letter, origin, and date of collection. The microbial quality of food, water, soil, manure was determined by standard quantitative, biochemical, and PCR techniques. Antimicrobial resistance of isolated bacteria was also determined. Amikacin (AMI); Amoxicillin/ clavulanic acid (AMO); Ampicillin (AMP); Azithromycin (AZI); Cefoxitin (FOX); Cefpodoxime (CPD); Chloramphenicol (CHL); Ciprofloxacin (CIP); Kanamycin (KAN); Nalidixic Acid (NAL); Streptomycin (STR); Tetracycline (TET). Erythromycin (ERY), Cefotaxime (CTX), Gentamicin (GEN) were some of the antibiotics used in this study. The antimicrobial susceptibilitywas determined using the Bauer and Kirby disk diffusion technique on Mueller-Hinton Agar Escherichia coli, Salmonella spp., Shigella spp. and other bacteria including, Enterobacter spp, and Klebsiella in the Enterobacteriaceae were isolated from local foods, water from farms, animal manure, and soil. Escherichia coli which is an indicator bacteria showed the following antimicrobial resistance patterns AMP-CPD-CIP- CHL ERY- KAN- NAL-TET, AMP-CPD-ERY-STR-TE, AMP- CTX- CHO-ERY- KAM- STY- VAN, AMP-CTX-CHO-ER-STY-VAN, AMP-CTX-CHO-ERY-KAM-VAN, AMP-ERY-KAM-STY-VAN, CTX-CHO- ERY-KAM-VAN, AMP-CTX- ERY-VAN, ERY-KAM-VAN, ERY-VAN. Salmonella which is also anindicator bacteria had the following resistance patterns AMP-CTX-CHO-ERY-KAM-STY-VAN, AMP- CHO-ERY-KAM-STY-VAN, AMP- ERY- KAM-VAN-AMP- ERY-VA, ERY-STY- VAN. Some of the other antimicrobial resistance patterns determined were AMP-CTX-CHO-ERY-KAM-STY-VAN, AMP- CHO-ERY-KAM-STY-VAN, AMP- ERY- KAM-VAN-AMP- ERY-VA, ERY-STY- VAN, , ERY-VAN, AMP-CTX-CHO-ERY-KAM-STY-VAN, AMP-CTX-CHO-ERY-STY-VAN , AMP-ERY-VAN, AMP-ERY-STY-VAN, AMP-ERY-STY-VAN, AMP-ERY-VAN, AMP- CTX-CHO-ERY-STY-VAN, and AMP-CTX-CHO-ERY- KAM-VAN. The MAR index values of isolated bacteria wasalso evaluated in this study. MAR index values indicate the usage of antibiotics in the environment. Cumulatively, the antibiotic resistance patterns showed MAR index values ranging from 0.08 to 0.70. The highest MARindex among thebacteria was for Escherichia coli (MAR Index=0.7) Students were also engaged to increase their participation in food safety research activities. Through this program two graduate and one undergraduate students were trained on isolation, identification, and classification of bacteria in local foods and environmental samples (soil, water, animal manure). Students were also trained onAntibiotic Sensitivity Testing.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2016
Citation:
A. Kilonzo-Nthenge, S. N. Nahashon, S. Godwin, Siqin liu, and D. Long. 2016. Prevalence and Antimicrobial Resistance of Enterobacteriaceae in Shell Eggs from Small-Scale Poultry Farms and Farmers' Markets. Food Prot. 79: 2031-2037
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Progress 12/23/14 to 09/30/15
Outputs Target Audience:The target audience reached were small to medium-sized fresh produce growers, small scale produce growers, farmers' markets, and graduate students Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?
Nothing Reported
How have the results been disseminated to communities of interest?
Nothing Reported
What do you plan to do during the next reporting period to accomplish the goals?Data on antibiotic use on food animals will be collected from anima producers and analyzed for antimicrobial resistance. More samples from farms and water in agricultural lands will be analyzed for bacterial contamination and resistance.
Impacts What was accomplished under these goals?
As emerging food contaminant in the food chain, antibiotic resistant bacteria has become a public health concern. The potential threat to human health as a result of inappropriate antibiotic use in food animals is significant, as pathogenic-resistant organisms in these animals are poised to enter the food supply and could be broadly distributed in food products. Antibiotic use in food animals may represent a risk to human health and therefore, it's important to investigate the use of antimicrobials on farms. It is important to determine the level of antimicrobial use in food animal production and identify the environmental reservoirs of antimicrobial resistant bacteria in Tennessee agro-ecosystems. Objective 1: Determine the extent of antibiotic use in Tennessee agriculture: To acquire feedback on the usage of antimicrobial usage on anima farms, a survey was developed and distributed to animal reproducers. This objective will help examine the scope and nature of antibiotics used in food animals. The data obtained will be used for public discussions and give guidance to future directions for agricultural use of antibiotics. Currently, the suvveys on antimicrobial use on food animals are being collected from animal producers. Objective 2: Determine zoonotic and indicator bacteria in growing crops, soil, animal manure, feed, water sources, and locally produced foods retailed in farmers markets: Microbiological analyses of eggs from locals farms and farmers markets, water form agricultural lands, fresh produce was conducted to determine, coliforms, aerobic microbe contamination levels, generic E. coli, E. coli O157:H7, and Salmonella. Antimicrobial susceptibility testing was carried out on the isolated bacteria and the Kirby-Bauer disk diffusion method was used to determine the resistance. Microbial analyses indicated that shell eggs can harbor resistant foodborne and commensal bacteria and failure to properly handle raw eggs, positions a potential health hazard to consumers. Bacterial isolates from water in agricultural lands and fresh produce are being analyzed for antimicrobial resistance
Publications
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