Hydrothermal carbonization of livestock mortality for the reduction of pathogens and microbially-derived DNA

doi:10.1007/s11783-017-0930-x

Front. Environ. Sci. Eng.

2017, Vol. 11

Issue (3) : 9 https://doi.org/10.1007/s11783-017-0930-x

RESEARCH ARTICLE

Hydrothermal carbonization of livestock mortality for the reduction of pathogens and microbially-derived DNA

Thomas F. Ducey¹(

), Jessica C. Collins², Kyoung S. Ro¹, Bryan L. Woodbury³, D. Dee Griffin⁴

¹. Coastal Plains Soil, Water, and Plant Research Center, Agricultural Research Service, United States Department of Agriculture, Florence, SC?29501, USA
². South Carolina Governor’s School for Science and Mathematics, Hartsville, SC 29550, USA
³. U.S. Meat Animal Research Center, Agricultural Research Service, United States Department of Agriculture, Clay Center, NE 68933, USA
⁴. University of Nebraska-Lincoln, Great Plains Veterinary Education Center, Lincoln, NE 68588, USA

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Abstract

Hydrothermal carbonization treatment eliminates pathogens and microbial DNA.

Hydrothermal carbonization treatment worked at both 150°C and 200°C.

Hydrothermal carbonization treatment worked in both bovine bone and tissue.

30 minute treatment was sufficient for pathogen kill and complete DNA degradation.

Hydrothermal carbonization (HTC), utilizing high temperature and pressure, has the potential to treat agricultural waste via inactivating pathogens, antibiotic resistance genes (ARG), and contaminants of emerging concern (CEC) in a environmental and economical manner. Livestock mortality is one facet of agricultural waste that can pose a threat to the surrounding environment. While several methods are utilized to treat livestock mortality, there remains a paucity of data on the elimination of microbially-derived DNA in these treatment practices. This DNA, most notably ARGs, if it survives treatment can be reintroduced in agricultural environments where it could potentially be passed to pathogens, posing a risk to animal and human populations. HTC treatments have been successfully utilized for the treatment of CECs, however very little is understood on how ARGs survive HTC treatment. This study aims to fill this knowledge gap by examining the survivability of microbially-derived DNA in the HTC treatment of livestock mortality. We examined three treatment temperatures (100°C, 150°C, and 200°C) at autogenic pressures at three treatment times (30, 60, and 240 min). We examined the amplification of a plasmid-borne reporter gene carried byEscherichia coli DH10B introduced to both beef bone and tissue. Results indicate that while all three temperatures, at all treatment times, were suitable for complete pathogen kill, only temperatures of 150°C and 200°C were sufficient for eliminating microbial DNA. These results serve as the basis for future potential HTC treatment recommendations for livestock mortality when considering the elimination of pathogens and ARGs.

Keywords High-temperature carbonization Microbial DNA Livestock mortality

Corresponding Author(s): Thomas F. Ducey

Issue Date: 13 April 2017

Cite this article:

Thomas F. Ducey,Jessica C. Collins,Kyoung S. Ro, et al. Hydrothermal carbonization of livestock mortality for the reduction of pathogens and microbially-derived DNA[J]. Front. Environ. Sci. Eng., 2017, 11(3): 9.

URL:

https://academic.hep.com.cn/fese/EN/10.1007/s11783-017-0930-x
https://academic.hep.com.cn/fese/EN/Y2017/V11/I3/9

Fig.1 Example of bone sample for testing pathogen and microbially-derived DNA reduction. Panel A represents pre-treatment, Panel B represents post-treatment.

Fig.2 Representative temperature and pressure profiles from Parr reactor for 150℃ (Panel A) and 200℃ (Panel B) with four hour treatment times.

Fig.3 Escherichia coli DH10B CFU after HTC treatment. Inoculant and control samples were grown at 37℃.

Fig.4 Amplification results from DNA extractions after HTC treatment of bovine bone samples at 150℃ (Panel A) and 200℃ (Panel B). Lanes are as follows: 1) inoculant; 2) negative control; 3) 30 min; 4) 60 min; 5) 240 min; 6) New England Biolabs 2-log DNA ladder; 7) positive control.

Fig.5 Amplification results from DNA extractions after HTC treatment of bovine tissue samples at 150℃ (Panel A) and 200℃ (Panel B). Lanes are as follows: 1) inoculant; 2) negative control; 3) 30 min; 4) 60 min; 5) 240 min; 6) New England Biolabs 2-log DNA ladder; 7) positive control.

Fig.6 Representative Bioanalyzer results from DNA extractions after HTC treatment for 30 min at 150℃ and 200℃, along with untreated plasmid. The lower marker is 35 bp while the upper marker is 10,380 bp in length. Ladder lengths (in base pairs) are on left hand side.

Tab.1 Conditions (sample temperatures, reactor pressures, and treatment times) during HTC treatment of livestock mortality bone and tissue samples

Fig.7 Amplification results from DNA extractions after treatment of bovine bone (Panel A) and tissue (Panel B) samples at 100℃. Lanes are as follows: 1) inoculant; 2) negative control; 3) 30 min; 4) 60 min; 5) 240 min; 6) New England Biolabs 2-log DNA ladder; 7) positive control.

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